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The high burden of malaria and HIV/AIDS prevents economic and social progress in developing countries. A continuing need exists for development of novel drugs and treatment regimens for both diseases in order to address the tolerability and long-term safety concerns associated with current treatment options and the emergence of drug resistance. We describe new spiro-β-lactam derivatives with potent (nM) activity against HIV and Plasmodium and no activity against bacteria and yeast. The best performing molecule of the series, BSS-730A, inhibited both HIV-1 and HIV-2 replication with an IC50 of 13 ± 9.59 nM and P. berghei hepatic infection with an IC50 of 0.55 ± 0.14 μM with a clear impact on parasite development. BSS-730A was also active against the erythrocytic stages of P. falciparum, with an estimated IC50 of 0.43 ± 0.04 μM. Time-of-addition studies showed that BSS-730A potentially affects all stages of the HIV replicative cycle, suggesting a complex mechanism of action. BSS-730A was active against multidrug-resistant HIV isolates, with a median 2.4-fold higher IC50 relative to control isolates. BSS-730A was equally active against R5 and X4 HIV isolates and displayed strong synergism with the entry inhibitor AMD3100. BSS-730A is a promising candidate for development as a potential therapeutic and/or prophylactic agent against HIV and Plasmodium.

Objective: Renal artery aneurysms are a rare condition; however, the rate of diagnosis has been increasing, because of the increasing use of complementary diagnostic methods. The best treatment strategy for RAAs remains controversial. Data on ex-vivo surgery associated with kidney autotransplantation are scarce. As a result, the goal of this study was to describe this technique and to report our results. Methods: A retrospective monocentric study was undertaken using the clinical records and images of 35 patients diagnosed with renal artery aneurysm from 01/01/2010 to 31/12/2018. Indications for ex vivo surgery and autotransplantation were complex aneurysms with diameter >20 mm or rapid growth or symptomatic aneurysms or women wishing to become pregnant. Complex aneurysms were defined by anatomical criteria (bifurcation of the renal artery and its primary branches or hilar aneurysms) and/or physiological criteria (when time of warm ischemia in in-situ reconstruction is expected to last more than 45 minutes). The technique of ex-vivo surgery and autotransplantation consists of performing a nephrectomy, renal cooling, treatment of aneurysm in banking and implantation of the kidney in the homolateral iliac fossa. Results: A total of 35 patients with 56 renal artery aneurysms (26 women, mean age 52.4 years-minimum and maximum 16 and 74 years) were included. Of these, 27 were treated by surgery and 8 were followed clinically. Among those treated surgically, 24 performed ex vivo surgery associated with autotransplantation. Regarding ex vivo surgery, nephrectomy was performed by laparoscopic surgery in 24 of the 27 surgeries, the mean surgical time was 5.3 hours, the median warm ischemia time was 4 minutes and the length of hospital stay was 12.2 days. Mortality was 0% and the kidney patency rate was 93% with a follow up of 47.2 months. Of the 17 patients with hypertension, 6 cured it, 4 improved and 7 maintained hypertension. Conclusion: Kidney autotransplantation appears to be efficient for most complex RAA with the possibility to minimize surgical aggression by performing laparoscopic nephrectomy.

Congenital deafness or early acquired deafness affects 1 to 3 out of 1000 newborns without risk factors and 20 to 40 out of 1000 newborns with risk factors. The universal newborn hearing screening enables its early identification. Children with congenital deafness/early acquired deafness have a higher prevalence of other conditions, especially ophthalmologic and neurodevelopmental ones, and at least 30% to 40% have at least one associated comorbidity.We carried out a cross-sectional, multicenter study in which 83% (n = 30) of the hospitals/maternity hospitals of the National Health Service participated.All surveyed hospitals/maternity hospitals routinely performed universal newborn hearing screening to all newborns before discharge; 63% referred children with risk factors for hearing loss to Otorhinolaryngology. All children with congenital deafness/early acquired deafness are referred to: Pediatrics in 23% hospitals/maternity hospitals. In 23 hospitals/maternity hospitals, all children with congenital deafness/early acquired deafness are referred to: Speech Therapy in 44% hospitals/ maternity hospitals; Ophthalmology in 17% hospitals/maternity hospitals; National System of Early Intervention in Childhood in 30% hospitals/maternity hospitals; 22% of hospitals/maternity hospitals refer all children with congenital deafness/early acquired deafness, with no identified cause, to Clinical Genetics clinics. The number of diagnoses of deafness in the years 2014 and 2015 was 2.5 and 1.5 per 1000 newborns, respectively, in 15 hospitals/maternity hospitals.Awareness of universal newborn hearing screening seems to be widely spread in the National Health Service. The number of children with SC / SPA, as well as the percentage of different types of deafness diagnosed, were identical to those found in other studies and shows its importance. The assessment / follow-up of these children by specialties other than the otolaryngology was heterogeneous in different health entities and revealed that not all children with risk factors for deafness follow up advised by existing standards.Results show that Portugal made an important path in the screening and follow-up of children with SC / SPA. It is important, with the ultimate aim of continually improving the care of these children, to reflect on the involvement of specialties other than otolaryngology, such as the National Early Childhood Intervention System in the follow-up of these children.Introdução: A surdez congénita ou precocemente adquirida afeta 1 a 3 por cada 1000 recém-nascidos sem fatores de risco e 20 a 40/1000 com fatores de risco. O rastreio auditivo neonatal universal permite a sua identificação precoce. As crianças com surdez congénita/precocemente adquirida têm uma maior prevalência de outras patologias, especialmente oftalmológicas e do neurodesenvolvimento, tendo pelo menos 30% a 40% uma comorbilidade associada.Material e Métodos: Realizámos um estudo transversal, multicêntrico onde participaram 83% (n = 30) dos hospitais/maternidades do Serviço Nacional de Saúde.Resultados: Todos os hospitais/maternidades inquiridos realizam, por rotina, o rastreio auditivo neonatal universal a todos os recém-nascidos antes da alta; 63% encaminham para Otorrinolaringologia crianças com fatores de risco de surdez. Todas as crianças com surdez congénita/precocemente adquirida são encaminhadas para Pediatria em 23% hospitais/maternidades. Em 23 hospitais/maternidades todas as crianças com surdez congénita/precocemente adquirida são encaminhadas para: Terapia da Fala em 44% hospitais/maternidades; Oftalmologia em 17% hospitais/maternidades; Sistema Nacional de Intervenção Precoce na Infância (SNIPI) em 30% hospitais/maternidades; referenciação para Genética de todas as crianças com surdez congénita/ precocemente adquirida, sem causa identificada, em 22% hospitais/maternidades. O número de diagnósticos de surdez nos anos de 2014 e 2015 foi de 2,5 e 1,5 por cada1000 recém-nascidos, respetivamente, em 15 dos hospitais/maternidades.Discussão: O rastreio auditivo neonatal universal parece estar amplamente difundido no Serviço Nacional de Saúde. O número de crianças com SC/SPA tal como a percentagem dos diferentes tipos de surdez diagnosticados, foram idênticos aos encontrados noutros estudos e mostra a indiscutível importância do rastreio. A avaliação/acompanhamento destas crianças por outras especialidades, além da Otorrinolaringologia, mostrou-se heterogéneo nas diferentes entidades de saúde e revelou que nem todas as crianças com fatores de risco de surdez realizam o seguimento aconselhado pelas normas existentes.Conclusão: Os resultados mostram que Portugal realizou um percurso importante no âmbito do rastreio e seguimento das crianças com SC/SPA. Importa, com o fim último da melhoria continua da prestação de cuidados a estas crianças, refletir sobre o envolvimento de outras especialidades, além da Otorrinolaringologia, tal como do Sistema Nacional de Intervenção Precoce na Infância no seguimento destas crianças.

Hybrid compounds may play a critical role in the context of the malaria eradication agenda, which will benefit from therapeutic tools active against the symptomatic erythrocytic stage of Plasmodium infection, and also capable of eliminating liver stage parasites. To address the need for efficient multistage antiplasmodial compounds, a small library of 1,2,4,5-tetraoxane-8- aminoquinoline hybrids, with the metabolically labile C-5 position of the 8-aminoquinoline moiety blocked with aryl groups, was synthesized and screened for antiplasmodial activity and metabolic stability. The hybrid compounds inhibited development of intra-erythrocytic forms of the multidrug-resistant Plasmodium falciparum W2 strain, with EC50 values in the nM range, and with low cytotoxicity against mammalian cells. The compounds also inhibited the development of P. berghei liver stage parasites, with the most potent compounds displaying EC50 values in the low μM range. SAR analysis revealed that unbranched linkers between the endoperoxide and 8-aminoquinoline pharmacophores are most beneficial for dual antiplasmodial activity. Importantly, hybrids were significantly more potent than a 1:1 mixture of 8-aminoquinoline-tetraoxane, highlighting the superiority of the hybrid approach over the combination therapy. Furthermore, aryl substituents at C-5 of the 8-aminoquinoline moiety improve the compounds' metabolic stability when compared with their primaquine (i.e. C-5 unsubstituted) counterparts. Overall, this study reveals that blocking the quinoline C-5 position does not result in loss of dual-stage antimalarial activity, and that tetraoxane-8- aminoquinoline hybrids are an attractive approach to achieve elimination of exo- and intraerythrocytic parasites, thus with the potential to be used in malaria eradication campaigns.

1. Catarina S. Oliveira, MD, MMed* 2. Ines Pessanha, MD, MMed† 3. Liliana Santos, MD, MMed† 4. Marta Machado, MD, MMed* 5. Raquel Zenha, MD* 1. *Department of Pediatrics, Baixo Vouga Medical Center, Aveiro, Portugal 2. †Department of Pediatric Surgery, Coimbra Hospital and Universitary Centre, Coimbra, Portugal A 21-day-old boy presents to the emergency department for the third time in 24 hours due to a 2-day history of an umbilical mass noted to be draining fluid. The mass is growing in size and increasing its drainage. The patient has an unremarkable medical history, and his umbilical stump fell off when he was 15 days old. During the patient’s first 2 trips to the emergency department, it was assumed that he had a bleeding granuloma. He was first treated with silver nitrate and then with a porcine gelatin absorbable sponge. The physical examination during this third visit reveals a comfortable boy with an umbilical mass that is draining a blood-tinged greenish-yellow liquid. Abdominal palpation seems to cause airflow from a small stoma in the umbilical mass (Fig 1). A catheter is inserted into the stoma, and airflow through the catheter is noted. Additional imaging confirms the diagnosis. Figure 1. Photograph of the umbilical mass where a small stoma is seen (white arrow). The physical examination suggests a patent omphalomesenteric duct (POMD). Ultrasonography is performed, revealing a hypoechogenic area between the umbilical mass and the small bowel. No other abdominal anomalies are observed. To better characterize the hypoechogenic mass, fistulography is performed and shows a patent duct between the stoma of the umbilical mass …

Fifteen Amaryllidaceae alkaloids (1–15) of various structural types were isolated by standard chromatographic methods from fresh bulbs of Narcissus poeticus cv. Pink Parasol. The chemical structures were elucidated by MS, and 1D and 2D NMR spectroscopic analyses, and by comparison with literature data. Narcipavline (5) and narcikachnine (6) are reported here for the first time. In their structure are combined two basic structural types of Amaryllidaceae alkaloids (galanthamine- and galanthindole-structural types), which represent a new structural type of these compounds. Alkaloids isolated in sufficient amounts were evaluated for their human erythrocytic acetylcholinesterase, and human serum butyrylcholinesterase (HuBuChE) inhibition activity using Ellman’s method. Z-Gly-Pro-p-nitroanilide was used as substrate in the prolyl oligopeptidase (POP) assay. Untested alkaloids were also screened for their cytotoxic activity against a small panel of human cancer cells, which spanned cell lines from different tissue types. In parallel, MRC-5 human fibroblasts were employed to determine overall toxicity against noncancerous cells. Some compounds were evaluated for their antiprotozoal activity. The newly isolated alkaloid narcipavline (5) showed interesting HuBuChE inhibition activity (IC50 = 24.4 ± 1.2 µM), and norlycoramine (11) demonstrated promising POP inhibition (IC50 = 0.21 ± 0.01 mM).

The incidence of multiple gestations is increasing worldwide and many studies have shown higher perinatal morbidity and mortality rates in monochorionic twins compared to dichorionic. The aim of this study was to assess the twin population born at a tertiary center and to evaluate the impact of chorionicity on perinatal outcomes of twin pregnancies.Retrospective study of all twins born in a tertiary center from January 2004 to December 2013.In this period, 1051 twins were born, related to 540 gestations (26.7% monochorionic; 73.3% dichorionic). There was no statistical significant difference between the groups concerning obstetric complications. The monochorionic group had a higher incidence of intrauterine growth restriction (20.5 vs 11.3%, p0.001), lower mean maternal age (29.9 vs 31.9 years, p0.001), lower mean gestational age (33.4 vs 34.3 weeks, p0.05) and lower mean birth weight (1943 vs 2147 g, p0.001). Monochorionic twins had a higher incidence of hyaline membrane disease (7 vs 4%, p0.05), sepsis (10.3 vs 5.8%, p0.05) and anemia (9.5 vs 5.4%, p0.05). There were no statistical significant differences concerning necrotizing enterocolitis, intraperiventricular hemorrhage or retinopathy of prematurity. Perinatal mortality was higher in the monochorionic group (5.2 vs 2.9%, p0.05).Monochorionic twins represent considerable challenges to both obstetricians and neonatologists and should be monitored and delivered at tertiary centers.Currently gemelarity has a major impact on total births. It would be interesting to develop protocols to standardize clinical approach to twins.Introdução: A incidência da gestação múltipla tem vindo a aumentar em todo o mundo e vários estudos têm demonstrado taxas de morbilidade e mortalidade mais elevadas nos gémeos monocoriónicos comparativamente com os bicoriónicos. Os objetivos deste trabalho foram: caracterizar a população de gémeos fruto de gravidez bifetal nascidos num hospital nível três e avaliar o impacto da corionicidade na morbimortalidade perinatal.Material e Métodos: Estudo retrospetivo de todos os gémeos fruto de gravidez bifetal nascidos num hospital nível três entre janeiro de 2004 e dezembro de 2013.Resultados: Neste período nasceram 1051 gémeos, fruto de 540 gestações (26,7% monocoriónicos; 73,3% bicoriónicos). Não houve diferença estatisticamente significativa entre os dois grupos no respeitante às complicações obstétricas. No grupo monocoriónico verificou-se uma incidência mais elevada de restrição do crescimento intra-uterino (20,5 vs 11,3%, p0,001), idade materna mais baixa (29,9 vs 31,9 anos, p0,001), idade gestacional mais baixa (33,4 vs 34,3 semanas, p0,05) e peso de nascimento mais baixo (1943 vs 2147 g, p0,001). Os gémeos monocoriónicos tiveram uma incidência mais elevada de doença de membrana hialina (7 vs 4%, p0,05), sépsis (10,3 vs 5,8%, p0,05) e anemia (9,5 vs 5,4%, p0,05). Não se encontrou diferença estatisticamente significativa relativamente à ocorrência de enterocolite necrotizante, hemorragia intraperiventricular ou retinopatia da prematuridade. A mortalidade perinatal foi mais elevada no grupo monocoriónico (5,2 vs 2,9%, p0,05).Discussão: Os gémeos monocoriónicos representam um desafio para obstetras e neonatologistas, devendo a vigilância pré-natal e o parto ser realizados em centros de referência.Conclusão: A gemelaridade tem atualmente um importante impacto nos nascimentos, pelo que seria interessante desenvolver protocolos que uniformizassem a prática clínica na abordagem a estes recém-nascidos.

The restricted pipeline of drugs targeting the liver stage of

Thrombospondin type I repeat (TSR) domains are commonly O-fucosylated by protein O-fucosyltransferase 2 (PoFUT2), and this modification is required for optimal folding and secretion of TSR-containing proteins. The human malaria parasite Plasmodium falciparum expresses proteins containing TSR domains, such as the thrombospondin-related anonymous protein (TRAP) and circumsporozoite surface protein (CSP), which are O-fucosylated. TRAP and CSP are present on the surface of sporozoites and play essential roles in mosquito and human host invasion processes during the transmission stages. Here, we have generated PoFUT2 null-mutant P. falciparum and Plasmodium berghei (rodent) malaria parasites and, by phenotyping them throughout their complete life cycle, we show that PoFUT2 disruption does not affect the growth through the mosquito stages for both species. However, contrary to what has been described previously by others, P. berghei PoFUT2 null mutant sporozoites showed no deleterious motility phenotypes and successfully established blood stage infection in mice. This unexpected result indicates that the importance of O-fucosylation of TSR domains may differ between human and RODENT malaria parasites; complicating our understanding of glycosylation modifications in malaria biology.

Perinatal stroke is often difficult to diagnose and an established treatment has not yet been validated, except for symptomatic measures. Herein, we propose to test the neuroprotective potential of the intravenous injection of retinoic acid-loaded nanoparticles (RA-NP) upon ischemic injury to the immature brain. The role of RA-NP on endothelial cells and organotypic slice cultures exposed to oxygen and glucose deprivation was assessed by evaluating markers pertaining to survival, proliferation, oxidative stress (NO, ROS), neuronal damage (enolase), vascular oxidation (p47phox) and microglia activation (CD68). Data showed that RA-NP (3 μg/ml) increased endothelial proliferation and survival, and normalized NO and ROS levels. The intravenous administration of RA-NP (10 μg/g) prevented ischemic injury in the hippocampus of 2-day-old mice by inhibiting cell death and normalizing markers of neurovascular function and inflammation. In sum, systemic administration of RA-NP protected neurovascular integrity and the inflammatory milieu from ischemia in the immature brain, highlighting its therapeutic value for perinatal stroke.

Recent WHO guidelines on control of human immunodeficiency virus (HIV) call for the widespread use of antiretroviral (AR) therapy (ART) for people living with HIV. Given the considerable overlap between infections by HIV and Plasmodium, the causative agent of malaria, it is important to understand the impact of AR compounds and ART regimens on infections by malaria parasites. We undertook a systematic approach to identify AR drugs and ART drug combinations with inhibitory activity against the obligatory hepatic stage of Plasmodium infection. Our in vitro screen of a wide array of AR drugs identified the non-nucleoside reverse transcriptase inhibitors efavirenz and etravirine (ETV), and the protease inhibitor nelfinavir, as compounds that significantly impair the development of the rodent malaria parasite P. berghei in an hepatoma cell line. Furthermore, we show that WHO-recommended ART drug combinations currently employed in the field strongly inhibit Plasmodium liver infection in mice, an effect that may be significantly enhanced by the inclusion of ETV in the treatment. Our observations are the first report of ETV as an anti-Plasmodial drug, paving the way for further evaluation and potential use of ETV-containing ARTs in regions of geographical overlap between HIV and Plasmodium infections.

Angiogenesis and inflammation are clearly interconnected and interdependent processes that are dysregulated in a series of systemic and brain pathologies. Herein, key aspects regarding endothelial cell function and tissue remodelling that are particularly affected or aggravated by inflammation are presented. Most importantly, the cellular and molecular mechanisms involved in the vascular regulation of the inflammatory processes occurring in several brain disorders and how they impact on disease/injury progression are detailed, highlighting potential targets for therapy. Finally, nanomedicine-based approaches designed to overcome limitations pertaining to low systemic bioavailability, light, pH and temperature sensitivity and/or rapid degradation of these targets and to optimise their mode of action are discussed. Ultimately, we expect this review to provide new insight and to suggest novel approaches for the treatment of blood-brain barrier dysfunction per se or as a means to treat the injured or diseased central nervous system. This article is protected by copyright. All rights reserved.

Stroke is a leading cause of death in adult life, closely behind ischemic heart disease, and causes a significant and abiding socioeconomic burden. However, current therapies are not able to ensure full neurologic and/or sequelae-free recovery to all stroke survivors. We believe treatment efficacy and patient rehabilitation could be enhanced significantly by targeting blood-brain barrier (BBB) deregulation and inflammation-induced barrier loss that occurs after stroke. In this pathological context, bone marrow-derived endothelial progenitor cells (EPC) enter the bloodstream towards the lesion site, but their insufficient numbers and impaired angiogenic ability compromise neurovascular regeneration. In this context, cell-based therapies have become increasingly appealing since treating patients with large numbers of mesenchymal or hematopoietic stem/progenitor cells alone may boost repair. However, this approach could be met with several challenges in terms of logistics and cost; hence, the development of a drug delivery system suitable for intravenous administration and functionalized for selective uptake by circulating EPC could enhance their restorative potential without perceived complications. The ability to encapsulate proangiogenic and anti-inflammatory agents, such as retinoic acid, and to safely and easily deliver them systemically may open new therapeutic perspectives for the treatment of cerebrovascular disorders. Copyright © 2017 John WileySons, Ltd.

A structure-activity relationship study was performed with ten 8-aminoquinoline-squaramides compounds active against liver stage malaria parasites, using human hepatoma cells (Huh7) infected by Plasmodium berghei parasites. In addition, their blood-schizontocidal activity was assessed against chloroquine-resistant W2 strain Plasmodium falciparum. Compound 3 was 7.3-fold more potent than the positive control primaquine against liver-stage parasites, illustrating the importance of the squarate moiety to activity.

© 2015 American Chemical Society, The syntheses and antiplasmodial activities of various substituted aminoquinolines coupled to an adamantane carrier are described. The compounds exhibited pronounced in vitro and in vivo activity against Plasmodium berghei in the Thompson test. Tethering a fluorine atom to the aminoquinoline C(3) position afforded fluoroaminoquinolines that act as intrahepatocytic parasite inhibitors, with compound 25 having an IC50 = 0.31 μM and reducing the liver load in mice by up to 92% at 80 mg/kg dose. Screening our peroxides as inhibitors of liver stage infection revealed that the tetraoxane pharmacophore itself is also an excellent liver stage P. berghei inhibitor (78: IC50 = 0.33 μM). Up to 91% reduction of the parasite liver load in mice was achieved at 100 mg/kg. Examination of tetraoxane 78 against the transgenic 3D7 strain expressing luciferase under a gametocyte-specific promoter revealed its activity against stage IV-V Plasmodium falciparum gametocytes (IC50 = 1.16 ± 0.37 μM). To the best of our knowledge, compounds 25 and 78 are the first examples of either an 4-aminoquinoline or a tetraoxane liver stage inhibitors., This research was supported by The Ministry of Science and Technological Development of Serbia (grant nos. 172008 and 41019), the Serbian Academy of Sciences and Arts, Fundação para a Ciência e Tecnologia, Portugal (grant PTDC/SAU-MIC/117060/2010), the Bill & Melinda Gates Foundation (grant OPP1040394).

The potential of azaaurones as dual-stage antimalarial agents was investigated by assessing the effect of a small library of azaaurones on the inhibition of liver and intraerythrocytic lifecycle stages of the malaria parasite. The whole series was screened against the blood stage of a chloroquine-resistant Plasmodium falciparum strain and the liver stage of P. berghei, yielding compounds with dual-stage activity and sub-micromolar potency against erythrocytic parasites. Studies with genetically modified parasites, using a phenotypic assay based on the P. falciparum Dd2-ScDHODH line, which expresses yeast dihydroorotate dehydrogenase (DHODH), showed that one of the azaaurone derivatives has the potential to inhibit the parasite mitochondrial electron-transport chain. The global urgency in finding new therapies for malaria, especially against the underexplored liver stage, associated with chemical tractability of azaaurones, warrants further development of this chemotype. Overall, these results emphasize the azaaurone chemotype as a promising scaffold for dual-stage antimalarials.

© 2015 Elsevier Masson SAS. All rights reserved., A series of novel pyrimidine-primaquine hybrids were synthesized and their effectiveness against the blood and liver stages of malaria parasites was evaluated. The hybrids displayed enhanced liver stage in vitro activity against P. berghei liver stage infection., We gratefully acknowledge financial assistance from CSIR, New Delhi and Fundação para a Ciência e Tecnologia, Portugal (grant PTDC/SAU-MIC/117060/2010 to MP). H.K. thanks CSIR, New Delhi for senior research fellowship (F. No. 09/254 (0195)/2009-EMR-I).

© 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim, Malaria continues to be a major cause of morbidity and mortality to this day, and resistance to drugs like chloroquine has led to an urgent need to discover novel chemical entities aimed at new targets. Here, we report the discovery of a novel class of potential antimalarial compounds containing an indolizinoindolone scaffold. These novel enantiopure indolizinoindolones were synthesized, in good-to-excellent yields and excellent diastereoselectivities, by cyclocondensation reaction of (S)- or (R)-tryptophanol and 2-acyl benzoic acids, followed by intramolecular α-amidoalkylation. Interestingly, we were able to synthesize for the first time 7,13b-cis indolizinoindolones in a two-step route. The novel compounds showed promising activity against erythrocytic stages of the human malaria parasite, Plasmodium falciparum, and liver stages of the rodent parasite Plasmodium berghei. In particular, an (S)-tryptophanol-derived isoindolinone was identified as a promising starting scaffold to search for novel antimalarials, combining excellent activity against both stages of the parasite's life cycle with low cytotoxicity and excellent metabolic and chemical stability in vitro., This work was supported by Fundação para a Ciência e a Tecnologia (FCT) through iMed.ULisboa (UID/DTP/04138/2013) and research projects PTDC/QUI-QUI/111664/2009 and PTDC/SAU-MIC/117060/2010. M. M. M. Santos would like to acknowledge FCT, “Programa Operacional Potencial Humano” and the European Social Fund for the IF Program (IF/00732/2013).

In a follow-up study to our reports of N-cinnamoylated chloroquine and quinacrine analogues as promising dual-stage antimalarial leads with high in vitro potency against both blood-stage Plasmodium falciparum and liver-stage Plasmodium berghei, we decided to investigate the effect of replacing the cinnamoyl moiety with other acyl groups. Thus, a series of N-acylated analogues were synthesized, and their activities against blood- and liver-stage Plasmodium spp. were assessed along with their in vitro cytotoxicities. Although the new N-acylated analogues were found to be somewhat less active and more cytotoxic than their N-cinnamoylated counterparts, they equally displayed nanomolar activities in vitro against blood-stage drug-sensitive and drug-resistant P. falciparum, and significant in vitro liver-stage activity against P. berghei. Therefore, it is demonstrated that simple N-acylated surrogates of classical antimalarial drugs are promising dual-stage antimalarial leads.

Background Anti-malarial resistance in Plasmodium falciparum remains an obstacle for malaria control. Resistance-associated genes were analysed in Brazilian samples over four decades to evaluate the impact of different treatment regimens on the parasite genetic profile. Methods Samples were collected on filter paper from patients infected in the Amazon region from 1984 to 2011. DNA was extracted with Chelex® 100 and monoinfection confirmed by PCR. SNPs in the pfcrt, pfmdr1, pfdhfr and pfdhps genes were assessed by PCR-RFLP. The pfmdr1 copy number was estimated using real time quantitative PCR with SYBR® Green. Parasite response was assessed ex vivo with seven concentrations of each anti-malarial. Patients were treated according to Brazilian guidelines: quinine plus tetracycline or mefloquine in period 1 and ACT in period 2. Results All 96 samples presented the pfcrt 76T mutant throughout the assessed periods. In addition, all isolates showed ex vivo chloroquine resistance. The pfmdr1 86Y was detected in 1.5% of samples in period 1, and in 25% in period 2. All samples presented the pfmdr1 1246Y. The analysis of pfmdr1 copy number showed amplification in 37.3% in period 1 and in 42% in period 2. Mutations in pfdhfr were shown as follows: 51I in all samples in period 1 and in 81.2% in period 2; 59R in 6.4% in period 2. The pfdhfr 108N and the pfdhps 437G were seen in all samples along time; the pfdhps 540E in 93.7% in period 1 and in 75% in period 2. Conclusions The 76T mutation associated to chloroquine resistance is still present in the parasite population, although this anti-malarial was withdrawn from the chemotherapy of P. falciparum in Brazil in the mid-1980s. All isolates assayed ex vivo for chloroquine showed resistant phenotype and 76T. No association was observed between pfmdr1 mutations and resistance to quinine, mefloquine and artemisinin derivatives. Additionally, the pfdhfr 108N mutation was detected in all samples throughout the evaluated periods, demonstrating fixation of the mutant allele in the parasite population. Changes in Brazilian national guidelines for the malaria chemotherapy in the last 27 years yielded a discreet genetic impact in the parasite population.

To get insight into the relevance of targeting hemozoin (Hz) crystals, two isomeric series, N5,N10-bis-alkylamine (2a-k) and N10,O11-bis-alkylamine (3a-k) indolo[3,2-b]quinolines, were evaluated for their in vitro activity against chloroquine (CQ)-resistant and sensitive strains of Plasmodium falciparum. In general, compounds of series 3 were more active than isomers 2, with IC50/LD50 ranging from 25/233 nM (3i) to 1.3 (3a)/10.7 (3b) μM. SAR analyses showed that lipophilicity and chlorine substitution at C3 increased both cytostatic and cytocidal activities. Both series bound to hematin monomer, inhibited β-hematin formation in vitro, delayed intraerythrocytic parasite development with apparent inhibition of Hz biocrystallization, and showed higher cytocidal activity against schizonts. In addition, cytostatic and cytocidal activities of series 3, but not those of isomers 2, correlated with calculated vacuole accumulation ratios, suggesting different capacities of 2 and 3 to bind to the Hz crystal face {001} exposed on the vacuole aqueous medium and different mechanisms of cytocidal potency.

Plasmodium falciparum, the causative agent of the most lethal form of malaria, is becoming increasingly resistant to most available drugs. A convenient approach to combat parasite resistance is the development of analogues of classical antimalarial agents, appropriately modified in order to restore their relevance in antimalarial chemotherapy. Following this line of thought, the design, synthesis and in vitro evaluation of N-cinnamoylated quinacrine surrogates, 9-(N-cinnamoylaminobutyl)-amino-6-chloro-2-methoxyacridines, is reported. The compounds were found to be highly potent against both blood-stage P.falciparum, chloroquine-sensitive 3D7 (IC50=17.0-39.0nM) and chloroquine-resistant W2 and Dd2 strains (IC50=3.2-41.2 and 27.1-131.0nM, respectively), and liver-stage P.berghei (IC50=1.6-4.9M) parasites. These findings bring new hope for the possible future "rise of a fallen angel" in antimalarial chemotherapy, with a potential resurgence of quinacrine-related compounds as dual-stage antimalarial leads. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

© 2014 Elsevier Masson SAS. All rights reserved., A library comprising 44 diversely substituted aurones derivatives was synthesized by straightforward aldol condensation reactions of benzofuranones and the appropriately substituted benzaldehydes. Microwave enhanced synthesis using palladium catalyzed protocols was introduced as a powerful strategy for extending the chemical space around the aurone scaffold. Additionally, Mannich-base derivatives, containing a 7-aminomethyl-6-hydroxy substitution pattern at ring A, were also prepared. Screening against the chloroquine resistant Plasmodium falciparum W2 strain identified novel aurones with IC50 values in the low micromolar range. The most potent compounds contained a basic moiety, with the ability to accumulate in acidic digestive vacuole of the malaria parasite. However, none of those aurones revealed significant activity against hemozoin formation and falcipain-2, two validated targets expressed during the blood stage of P. falciparum infection and functional in digestive vacuole of the parasite. Overall, this study highlight (i) the usefulness of aurones as platforms for synthetic procedures using palladium catalyzed protocols to rapidly deliver lead compounds for further optimization and (ii) the potential of novel aurone derivatives as promising antimalarial compounds., This work was supported by Fundação para a Ciência e Tecnologia (FCT, Portugal) through projects PTDC/SAU-FAR/118459/2010 and Pest-OE/SAL/UI4013/2011. A.S.N. and M.P.C. acknowledge FCT for grants SFRH/BD/41276/2007 and SFRH/BD/61611/2009, respectively. P.J.R. is a Doris Duke Charitable Foundation Distinguished Clinical Scientist.

A 3-year-old boy presented with pigmented (brownish) macules on his hands (figure 1), left arm (figure 2) and face (figure 3). The lesions appeared days after the boy played with common rue and were best noticed after sun exposure. Topical treatment with methylprednisolone 0.1% cream led to complete resolution within 2 weeks. Figure 1 Irregular pigmented lesions on both hands. Figure 2 Irregular pigmented lesion on left arm. Figure 3 Irregular facial pigmented lesion. Rue ( Ruta …

The isolation of bioactive compounds from medicinal plants, based on traditional use or ethnomedical data, is a highly promising potential approach for identifying new and effective antimalarial drug candidates. The purpose of this review was to create a compilation of the phytochemical studies on medicinal plants used to treat malaria in traditional medicine from the Community of Portuguese-Speaking Countries (CPSC): Angola, Brazil, Cape Verde, Guinea-Bissau, Mozambique and São Tomé and Príncipe. In addition, this review aimed to show that there are several medicinal plants popularly used in these countries for which few scientific studies are available. The primary approach compared the antimalarial activity of native species used in each country with its extracts, fractions and isolated substances. In this context, data shown here could be a tool to help researchers from these regions establish a scientific and technical network on the subject for the CPSC where malaria is a public health problem.

Transepidermal water loss (TEWL) is a measure of the steady-state water vapour flux crossing the skin to the external environment and it has been used extensively to characterise skin barrier function. We have previously hypothesised that in vivo TEWL is directly related to the reciprocal of the diffusional permeation pathlength through the stratum corneum (SC). The aim of the present paper is to validate experimentally this hypothesis. Ninety volunteers were recruited and TEWL and corneocyte surface areas were measured for six anatomic sites. The number of cell layers in the SC was calculated for each anatomic site in order to estimate the geometric pathlength for water efflux. Significant anatomic site variability was found for both TEWL and corneocyte surface area which were inversely correlated. A direct reciprocal relationship between TEWL and pathlength was determined, with TEWL values tending to zero when corneocytes are infinitely large. In general, skin sites with smaller corneocytes have fewer cell layers, with shorter permeation pathlengths and higher TEWL values. The results confirm our previous hypothesis and suggest that TEWL may be used to characterise the permeation routes for different anatomic sites.

Recently, latex type I hypersensibility reactions were frequently described, mainly in children with myelomeningocele (MMC), which maintain earlier intimate and frequent contact with latex products. Allergic food cross-reactions are well known for many years. Nowadays, latex allergy is frequently associated with food allergies to avocado, banana, kiwi, grapefruit, papaya, chestnut and peach.The objective of this study consisted in identifying the number of patients with myelomeningocele (MMC) and banana-latex clinico-laboratorial sensitization.Questionnaires were applied to 33 children with MMC and blood was collected from 30 to perform latex IgE (RAST Pharmacia), and from 29 to banana IgE. Specific IgE equal or above class I was considered positive.Four children related histories of latex contact urticaria, and one child related a history of urticaria and diarrhea with banana. 14/30 (46.6%) were latex IgE positive, and 4/29 (13.7%) to banana. There was no statistically significant association between variables.We identified high prevalence of latex and banana allergies in patients with MMC and we need more studies to analyze the prevalence of food-latex allergic cross-reactions in children with MMC.

Chloroquine is an antimalarial agent with strong activity against the blood stage of Plasmodium infection, but with low activity against the parasite's liver stage. In addition, the resistance to chloroquine limits its clinical use. The discovery of new molecules possessing multistage activity and overcoming drug resistance is needed. One possible strategy to achieve this lies in combining antimalarial quinolones with the pharmacological effects of transition metals. We investigated the antimalarial activity of four platinum(ii) complexes composed of chloroquine and phosphine ligands, denoted as WV-90, WV-92, WV-93 and WV-94. In comparison with chloroquine, the complexes were less potent against the chloroquine-sensitive 3D7 strain but they were as active as chloroquine in inhibiting the chloroquine-resistant W2 strain of P. falciparum. Regarding selectivity, the complexes WV-90 and WV-93 displayed higher indexes. Unlike chloroquine, the complexes act as irreversible parasiticidal agents against trophozoites and the WV-93 complex displayed activity against the hepatic stage of P. berghei. The in vivo suppression activity against P. berghei in the Peters 4 day test displayed by the complexes was similar to that of chloroquine. However, the efficacy in an established P. berghei infection in the Thompson test was superior for the WV-93 complex compared to chloroquine. The complexes' antimalarial mechanism of action is initiated by inhibiting the hemozoin formation. While chloroquine efficiently inhibits hemozoin, parasites treated with the platinum complexes display residual hemozoin crystals. This is explained since the interaction of the platinum complexes with ferriprotoporphyrin is weaker than that of chloroquine. However, the complexes caused a loss of mitochondrial integrity and subsequent reduction in mitochondrial activity, and their effects on mitochondria were more pronounced than those in the chloroquine-treated parasites. The dual effect of the platinum complexes may explain their activity against the hemozoin-lacking parasites (hepatic stage), where chloroquine has no activity. Our findings indicate that the platinum(ii)-chloroquine complexes are multifunctional antimalarial compounds and reinforce the importance of metal complexes in antimalarial drug discovery.

Copyright © 2017 Mendes et al. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license., Avermectins are powerful endectocides with an established potential to reduce the incidence of vector-borne diseases. Here, we show that several avermectins inhibit the hepatic stage of Plasmodium infection in vitro Notably, ivermectin potently inhibits liver infection in vivo by impairing parasite development inside hepatocytes. This impairment has a clear impact on the ensuing blood stage parasitemia, reducing disease severity and enhancing host survival. Ivermectin has been proposed as a tool to control malaria transmission because of its effects on the mosquito vector. Our study extends the effect of ivermectin to the early stages of mammalian host infection and supports the inclusion of this multipurpose drug in malaria control strategies., A.M.M., P.M., and M.P. acknowledge the Fundação para a Ciência e Tecnologia, Portugal, for grants SFRH/BPD/80693/2011, SFRH/BD/71098/2010, and Investigador FCT (2013), respectively. This work was supported by Fundação para a Ciência e Tecnologia (FCT, Portugal) grant PTDC/SAU-MIC/117060/2010.

Background: Drug resistance in Plasmodium falciparum has posed an obstacle to effective treatment and challenges many malaria control programmes in endemic areas. In Angola, until 2003, chloroquine (CQ) was used as first-line therapy for uncomplicated malaria. It was replaced initially by amodiaquine and, in 2006, by artemisinin-based combination therapy (ACT) with artemether-lumefantrine (AL, Coartem®). Efficacy study of ACT, conducted in Angola between 2004 and 2005, showed a baseline efficacy of ≈99 %. Methods: 103 malaria patients were enrolled according to WHO proceedings. Patients were followed up with clinical and parasitological evaluations for 28 days, parasite density and identification was evaluated by microscopy, the pfmsp2 were genotyped by nested-PCR, to distinguish parasite recrudescence from new infections; the polymorphisms at codons 86 and 1246 of pfmdr1 gene, and 769 of pfatp6 gene were assessed by PCR-RFLP and sequencing for pfk13-propeller genotype. Results: The cure rate was 91.3 %. The obtained results showed that from 103 patients, 12.6 % (n = 13) still had parasitaemia 1 day after the treatment was finished. On day 0, of the 94 evaluated samples, wild-type alleles were identified in 73.4 % (n = 69) for pfmdr1 N86Y position and only one sample carried the mutant allele (Y) for pfmdr1 1246; 14 % of these samples showed increased pfmdr1 copy number; 100 % (n = 21) had wild-type allele of k13 gene in all the studied positions. Discussion: These results showed changes in parasite profile susceptibility to AL in comparison to the baseline data from 2002 to 2004 and on the genotyping characteristics; the clinical outcome after treatment with AL did not link a particular genotype with treatment failure; observed changes do not provide sufficient evidence for a treatment policy change, but they suggest that a carefully monitoring is needed in this area. publishersversion published