Therapeutic Potential of Marine-Derived Cyclic Peptides as Antiparasitic Agents.

Parasitic diseases still compromise human health. Some of the currently available therapeutic drugs have limitations considering their adverse effects, questionable efficacy, and long treatment, which have encouraged drug resistance. There is an urgent need to find new, safe, effective, and affordable antiparasitic drugs. Marine-derived cyclic peptides have been increasingly screened as candidates for developing new drugs. Therefore, in this review, a systematic analysis of the scientific literature was performed and 25 marine-derived cyclic peptides with antiparasitic activity (1-25) were found. Antimalarial activity is the most reported (51%), followed by antileishmanial (27%) and antitrypanosomal (20%) activities. Some compounds showed promising antiparasitic activity at the nM scale, being active against various parasites. The mechanisms of action and targets for some of the compounds have been investigated, revealing different strategies against parasites.

Cyclic Peptides in Pipeline: What Future for These Great Molecules?

Cyclic peptides are molecules that are already used as drugs in therapies approved for various pharmacological activities, for example, as antibiotics, antifungals, anticancer, and immunosuppressants. Interest in these molecules has been growing due to the improved pharmacokinetic and pharmacodynamic properties of the cyclic structure over linear peptides and by the evolution of chemical synthesis, computational, and in vitro methods. To date, 53 cyclic peptides have been approved by different regulatory authorities, and many others are in clinical trials for a wide diversity of conditions. In this review, the potential of cyclic peptides is presented, and general aspects of their synthesis and development are discussed. Furthermore, an overview of already approved cyclic peptides is also given, and the cyclic peptides in clinical trials are summarized.

Increased expression of NLRP3 inflammasome components in granulosa cells and follicular fluid interleukin(IL)-1beta and IL-18 levels in fresh IVF/ICSI cycles in women with endometriosis.

The inflammasomes are a family of recently described multi-protein cytoplasmic sensors that orchestrate the inflammatory response and participate in a variety of inflammatory conditions. We hypothesized that the activation of pyrin domain­containing protein 3 (NLRP3) inflammasome by granulosa cells (hGCs) may be activated in women with endometriosis and influence oocyte maturation and IVF outcomes. We performed a cross-sectional study to investigate the NLRP3 inflammasome status in follicular fluid (FF) and in hGCs from 44 women undergoing controlled ovarian stimulation for IVF/ICSI. Study subjects were divided into two groups according to the infertility etiology: group with tubal or male factor (control, n = 22) vs. group with endometriosis (n = 22). The FF IL-1beta and IL-18 levels in the endometriosis group were significantly higher than those in the non-endometriosis group, i.e., 5010 pg/mL and 2738 pg/mL, respectively (p < 0.05). No correlation was found between clinical pregnancy and live birth rate and analyzed inflammasome component levels (p > 0.05). In addition, the hGCs from endometriosis women demonstrated high expression of NLRP3 inflammasome at both protein and mRNA levels. Higher expression of inflammasome components within the ovary compartment may result from the exaggerated inflammatory state associated with endometriosis and thus impact the fertility of these women.

Retinoic acid (all-trans) presents antioxidant properties within human ovary and reduces progesterone production by human granulosa cells.

Both vitamin A and E support female reproduction and embryonic development. These vitamins have been associated with decreased fertility or failure to end the pregnancy in animals. An observational study was conducted on follicular fluid (FF) samples to determine the concentrations of fat-soluble vitamins of women undergoing in vitro fertilization and its correlation with assisted reproductive technology characteristics and pregnancy outcomes. Moreover, the effects of all-trans-retinoic acid (atRA) and alpha-tocopherol on granulosa cell viability, apoptosis, autophagy and hormonal production were evaluated. No association was identified between fat-soluble vitamin concentrations in FF and infertility aetiology, body mass index or woman's age. There were differences in follicular antioxidant profiles and ovarian response stimulation. In vitro evaluation of atRA and alpha-tocopherol reveals that, at physiological concentrations, both compounds may affect the viability of granulosa cells. In addition, these compounds are able to protect granulosa cells from oxidative stress, as well as to affect estradiol and progesterone production. Our data suggest that atRA and alpha-tocopherol levels should be well controlled as they may have implications in the function and viability of granulosa cells and highlights retinol as a marker of the oxidative defenses within ovary environment.

Low Doses of Resveratrol Protect Human Granulosa Cells from Induced-Oxidative Stress.

Resveratrol is a phytoalexin present in plant-derived foods, including grape's skin, cocoa, and peanuts. Evidence suggests that it has beneficial effects on human health because of its antioxidant properties. However, there is limited knowledge about the part played by resveratrol in ovarian function. In this paper, the influence of resveratrol on granulosa cells (GC) was evaluated. In addition to being the main estradiol producers, GC are in direct contact with the oocyte, playing a fundamental role in its growth and development. The cell line COV434 and human granulosa cells (hGC), obtained from women undergoing assisted reproductive technology (ART), were used. GC were treated with resveratrol (0.001-20 µM) at different times (24-72 h). Low concentrations of this compound suggest a protective role, as they tend to reduce ROS/RNS formation after inducement of stress. On the contrary, high concentrations of resveratrol affect GC viability and steroidogenic function. As it may act as a direct modulator of GC oxidative balance, this work may help to clarify the impact of resveratrol on GC and the usefulness of this antioxidant as adjunct to infertility treatments.

Concentrations of the endocannabinoid N-arachidonoylethanolamine in the follicular fluid of women with endometriosis: the role of M1 polarised macrophages.

Although N-arachidonoylethanolamine (AEA; also known as anandamide) is present in human follicular fluid (FF), its regulation remains unknown. Therefore, the aims of the present study were to: (1) investigate the relationships between FF AEA concentrations in women undergoing assisted reproductive technology and their age, body mass index, ART characteristics and fertility treatment outcomes; and (2) assess how different inflammatory patterns may trigger AEA production by human granulosa cells (hGCs). FF AEA concentrations were higher in women undergoing IVF than in those undergoing intracytoplasmic sperm injection group. FF AEA median concentrations were lower in women undergoing ART because of male factor infertility than in women with endometriosis (1.6 vs 2.5nM respectively), but not women with tubal, hormonal or unexplained infertility (1.6, 2.4 and 1.9nM respectively). To evaluate the effects of macrophages on AEA production by hGCs, hGCs were cocultured with monocyte-derived macrophages. The conditioned medium from M1 polarised macrophages increased AEA production by hGCs. This was accompanied by an increase in AEA-metabolising enzymes, particularly N-acyl phosphatidylethanolamine-specific phospholipase D. The results of the present study show that high FF AEA concentrations in patients with endometriosis may be associated with the recruitment of inflammatory chemokines within the ovary, which together may contribute to the decreased reproductive potential of women with endometriosis. Collectively, these findings add a new player to the hormone and cytokine networks that regulate fertility in women.

Orally administrated chitosan microspheres bind Helicobacter pylori and decrease gastric infection in mice.

Persistent Helicobacter pylori (H. pylori) infection is related to 90% of gastric cancers. With bacterial resistance rising and treatment inefficiency affecting 15% of the patients, alternative treatments urge. Chitosan microspheres (ChMics) have been proposed as an H. pylori-binding system. This work evaluates ChMics biocompatibility, mucopenetration and capacity to treat H. pylori infection in mice after oral administration. ChMics of different size (XL, ∼120 µm and XS, ∼40 µm) and degree of acetylation (6% and 16%) were developed and revealed to be able to adhere both human and mouse-adapted H. pylori strains without cytotoxicity towards human gastric cells. Ex vivo studies showed that smaller (XS) microspheres penetrate further within the gastric foveolae, suggesting their ability to reach deeply adherent bacteria. In vivo assays showed 88% reduction of infection when H. pylori-infected mice (C57BL/6) were treated with more mucoadhesive XL6 and XS6 ChMics. Overall, ChMics clearly demonstrate ability to reduce H. pylori gastric infection in mice, with chitosan degree of acetylation being a dominant factor over microspheres' size on H. pylori removal efficiency. These results evidence the strong potential of this strategy as an antibiotic-free approach to fight H. pylori infection, where microspheres are orally administered, bind H. pylori in the stomach, and remove them through the gastrointestinal tract. STATEMENT OF SIGNIFICANCE: Approximately 90% of gastric cancers are caused by the carcinogenic agent Helicobacter pylori, which infects >50% of the world population. Bacterial resistance, reduced antibiotic bioavailability, and the intricate distribution of bacteria in mucus and within gastric foveolae hamper the success of most strategies to fight H. pylori. We demonstrate that an antibiotic-free therapy based on bare chitosan microspheres that bind and remove H. pylori from stomach can achieve 88% reduction of infection from H. pylori-infected mice. Changing size and mucoadhesive properties, microspheres can reach different areas of gastric mucosa: smaller and less mucoadhesive can penetrate deeper into the foveolae. This promising, simple and inexpensive strategy paves the way for a faster bench-to-bedside transition, therefore holding great potential for clinical application.

The endocannabinoid 2-arachidonoylglycerol promotes endoplasmic reticulum stress in placental cells.

Proliferation, differentiation and apoptosis of trophoblast cells are required for normal placental development. Impairment of those processes may lead to pregnancy-related diseases. Disruption of endoplasmic reticulum (ER) homeostasis has been associated with several reproductive pathologies including recurrent pregnancy loss and preeclampsia. In the unfolded protein response (UPR), specific ER-stress signalling pathways are activated to restore ER homeostasis, but if the adaptive response fails, apoptosis is triggered. Protein kinase RNA-like endoplasmic reticulum kinase (PERK), inositol-requiring enzyme 1 (IRE1) and Activating transcription factor 6 (ATF6) are central players in UPR and in ER-stress-induced apoptosis, as well as downstream transcription factors, as C/EBP homologous protein (CHOP). Our previous studies have shown that the endocannabinoid 2-arachidonoylglycerol (2-AG) modulates trophoblast cell turnover. Nevertheless, the role of ER-stress on 2-AG induced apoptosis and cannabinoid signalling in trophoblast has never been addressed. In this work, we used BeWo cells and human primary cytotrophoblasts isolated from term-placenta. The expression of ER-stress markers was analysed by qRT-PCR and Western blotting. ROS generation was assessed by fluorometric methods, while apoptosis was detected by the evaluation of caspase -3/-7 activities and Poly (ADP-ribose) polymerase (PARP) cleavage. Our findings indicate that 2-AG is able to induce ER-stress and apoptosis. Moreover, the eukaryotic initiation factor 2 (eIF2α)/CHOP pathway involved in ER-stress-induced apoptosis is triggered through a mechanism dependent on cannabinoid receptor CB2 activation. The results bring novel insights on the importance of ER-stress and cannabinoid signalling on 2-AG mechanisms of action in placenta.

Dissimilar effects of curcumin on human granulosa cells: Beyond its anti-oxidative role.

Global infertility prevalence has been increasing in recent decades, mainly due to advanced reproductive age. Concerned women look for dietary supplements with antioxidant properties advertised as a natural way to increase fertility. Curcumin (CUR) is a polyphenol with antioxidant and anti-inflammatory properties. CUR elicits apoptotic cell death as evidenced in some tumor cells. In this work, the effect of CUR on granulosa cells (GC) was studied. GC surround the oocyte, providing nutrient exchange and hormone production, necessary for its development. COV434 cell line and primary human granulosa cells (hGC) cultures from patients undergoing Assisted Reproductive Technology (ART) were used. GC were treated with CUR (0.001-50 µM) at different times (24-72 h). Low concentrations of CUR showed an increase on cell viability. Likewise, it leads to a decrease in ROS/RNS formation after stress induction, suggesting a protective role. Changes in hormonal levels were not observed. In contrast, high concentrations of CUR triggered a reduction on cell viability and a programmed cell death mechanism. Allied to the above results, high doses of CUR affected hormonal function of GCs. Our work reinforces the benefits of dietary supplements, namely CUR, on the main functions of GC and, consequently, on reproductive success.

The synthetic cannabinoid WIN-55,212 induced-apoptosis in cytotrophoblasts cells by a mechanism dependent on CB1 receptor.

The endocannabinoid system has evolved as a key regulator in several pathological and physiological processes, including placentation, decidualization and implantation. In addition, it is known that Cannabis and cannabinoids negatively affect female reproduction. Although, the biological action of synthetic cannabinoids, such as WIN-55,212, in human fertility and pregnancy outcome remain to be unveiled. A tight balance between proliferation, differentiation and apoptosis of trophoblast cells is required for placental development and pregnancy outcome. Therefore, in this work, the effects of the synthetic cannabinoid WIN-55,212 in placental cytotrophoblast cells were explored. For that, it was used a human choriocarcinoma cell line, BeWo cells, and primary cultures of human cytotrophoblasts isolated from term placentas. Results demonstrate that this synthetic cannabinoid induces cell cycle arrest. We also observed that cell viability loss was associated with a disruption of mitochondrial membrane potential and activation of caspases -9 and -3/-7 independently of reactive oxygen species (ROS) production or recruitment of the endoplasmic reticulum stress marker CHOP. Moreover, these effects were prevented by pre-incubation with a selective cannabinoid receptor 1 (CBR1) antagonist (AM281). Thus, our results provide strong evidences of the apoptotic process induced by WIN-55,212 through the activation of the CBR1, which may reveal the impact of cannabinoids consumption during placental development.

Cannabinoid-induced autophagy: Protective or death role?

Autophagy, the "self-digestion" mechanism of the cells, is an evolutionary conserved catabolic process that targets portions of cytoplasm, damaged organelles and proteins for lysosomal degradation, which plays a crucial role in development and disease. Cannabinoids are active compounds of Cannabis sativa and the most prevalent psychoactive substance is Δ(9)-tetrahydrocannabinol (THC). Cannabinoid compounds can be divided in three types: the plant-derived natural products (phytocannabinoids), the cannabinoids produced endogenously (endocannabinoids) and the synthesized compounds (synthetic cannabinoids). Various studies reported a cannabinoid-induced autophagy mechanism in cancer and non-cancer cells. In this review we focus on the recent advances in the cannabinoid-induced autophagy and highlight the molecular mechanisms involved in these processes.

Trauma inguinal penetrante com formação de fístula arteriovenosa e pseudoaneurisma: relato de caso / Acquired arteriovenous fistula and pseudoaneurysm secondary to penetrating inguinal trauma: a case report

Os autores apresentam um relato de caso de vítima de acidente de trabalho com ferimento penetrante em região inguinal direita com peça metálica em espiral, que evoluiu com fístula arteriovenosa da artéria femoral profunda com a veia femoral profunda associado a pseudoaneurisma envolvendo essas estruturas e a veia femoral comum. As fístulas arteriovenosas ocorrem frequentemente após traumas e a associação com pseudoaneurisma é fato raro, devendo ser tratadas precocemente após seu diagnóstico. O ultrassom duplex é atualmente o exame mais utilizado para a avaliação inicial e a arteriografia, o padrão ouro para diagnóstico. No paciente em questão foi realizado tratamento convencional com abordagem cirúrgica direta, sutura arterial e ligaduras venosas. Entretanto, nos dias atuais a cirurgia endovascular e a compressão guiada por ultrassom são métodos terapêuticos que têm sido utilizados com sucesso. O paciente evoluiu sem intercorrência, recebendo alta para acompanhamento ambulatorial com preservação do membro.

This article describes the case of a work accident victim with a penetrating wound to the right inguinal region caused by a metal spiral. The patient developed an arteriovenous fistula between the deep femoral artery and deep femoral vein, combined with a pseudoaneurysm surrounding these structures and the common femoral vein. Arteriovenous fistulas frequently occur after traumas, but the combination of fistula and pseudoaneurysm is rare. It is recommended that they be treated immediately after diagnosis. Duplex ultrasound is the most widely used method for initial assessment and arteriography is the gold standard for diagnosis of arteriovenous fistulas. Endovascular surgery has recently been used successfully in such cases. However, this patient was treated conventionally using a direct surgical approach, arterial suture and venous ligatures, and the limb was saved. The patient developed no complications and was discharged to outpatients follow-up.

Influence of LH and high-density lipoprotein cholesterol (HDL-C) on metformin response in women with polycystic ovary syndrome.

OBJECTIVE: To search for predictors of metformin response in women with polycystic ovary syndrome (PCOS) through a detailed analysis of clinical and laboratory parameters. STUDY DESIGN: We designed a prospective study to investigate clinical and laboratory parameters to search for predictors of metformin response in women with PCOS. A total of 53 PCOS patients were given metformin 850 mg twice a day for 6 months, after which patients were classified as responders or non-responders. Parameters analyzed for comparison between the two groups were: plasma fasting insulin glucose/insulin ratio; oral glucose tolerance test (OGTT) with insulin (120 min); HOMA and QUICKI tests; total cholesterol and fractions, triglycerides; LH, FSH, estradiol, progesterone, testosterone, androstenedione, 17-OH progesterone, and DHEAS. RESULTS: From all patients, 30 (56.6%) were responders and 23 (43.3%) were non-responders. Multinomial analysis showed that the positive response to metformin was associated with higher levels of basal LH (p=0.038) and lower levels of high-density lipoprotein cholesterol (HDL-C) (p=0.015). CONCLUSION: In weight-matched PCOS subjects, laboratory markers might predict the metformin response. Higher levels of basal LH and lower levels of HDL-C are correlated with a positive response to metformin treatment in PCOS subjects.

Cesárea prévia como fator de risco para o descolamento prematuro da placenta / Previous cesarean section as a risk factor for abruptio placentae

Investigar a relação entre o antecedente de cesárea e a ocorrência do descolamento prematuro da placenta (DPP). MÉTODOS: estudo retrospectivo em que foram avaliados os dados referentes a 6495 partos realizados no período entre abril de 2001 e janeiro de 2004. Foram adotados como critérios de inclusão: diagnóstico de DPP confirmado por exame da placenta após o parto, gestação única, peso do recém-nascido superior a 500 g e idade gestacional acima de 22 semanas e ausência de história de trauma abdominal na gestação atual. Para cada caso de DPP incluído no estudo foram selecionados cinco controles, obedecendo ao seguinte pareamento: paridade, idade gestacional (< ou >30 semanas), diagnóstico materno de síndrome hipertensiva na gestação índice, antecedente de cicatriz uterina prévia não relacionada à operação cesariana, diagnóstico de rotura prematura de membranas ou diagnóstico de polidrâmnio. A análise univariada das variáveis contínuas foi realizada utilizando-se o teste t de Student e as variáveis categóricas foram avaliadas por meio de teste exato de Fisher ou teste de chi2, com níveis descritivos (p) menores que 0,05 considerados significantes. RESULTADOS: 34 casos de pacientes com diagnóstico de DPP preencheram os critérios de inclusão (incidência de 0,52 por cento). Para o grupo controle foram selecionadas 170 pacientes que obedeceram aos critérios de pareamento propostos. No grupo de pacientes com DPP, 26,5 por cento apresentavam antecedente de parto cesárea (9 casos), ao passo que, no grupo controle, esse antecedente foi observado em 21,2 por cento das pacientes (36 casos). Não houve diferença estatisticamente significativa na incidência de cesárea prévia entre os dois grupos estudados (p=0,65, OR=1,34, IC 95 por cento=0,53-3,34). CONCLUSAO: o aspecto abordado neste estudo, isto é, a associação do DPP em pacientes com cicatriz uterina de cesárea, não pôde ser confirmado com a presente casuística