RESUMO
Fungal pollution of indoor environments contributes to several allergic symptoms and represents a public health problem. It is well-established that 1-octen-3-ol, also known as mushroom alcohol, is a fungal volatile organic compound (VOC) commonly found in damp indoor spaces and responsible for the typical musty odor. Previously it was reported that exposure to 1-octen-3-ol induced inflammations and disrupted mitochondrial morphology and bioenergetic rate in Drosophila melanogaster. The aim of this study was to examine the influence of 1-octen-3-ol on dehydrogenase activity, apoptotic biomarkers, levels of nitric oxide (NO) and reactive oxygen species (ROS), as well as antioxidant enzymes activities. D. melanogaster flies were exposed to an atmosphere containing 1-octen-3-ol (2.5 or ∞l/L) for 24 hr. Data demonstrated that 1-octen-3-ol decreased dehydrogenases activity and NO levels but increased ROS levels accompanied by stimulation of glutathione-S-transferase (GST) and superoxide dismutase (SOD) activities without altering caspase 3/7 activation. These findings indicate that adverse mitochondrial activity effects following exposure of D. melanogaster to 1-octen-3-ol, a fungal VOC, may be attributed to oxidant stress. The underlying mechanisms involved in adverse consequences of indoor fungal exposure appear to be related to necrotic but not apoptotic mechanisms. The adverse consequences were sex-dependent with males displaying higher sensitivity to 1-octen-3-ol. Based upon on the fact that the fly genome shares nearly 75% of disease-related genes to human exposure to this fungus may explain the adverse human responses to mold especially for males.
Assuntos
Poluentes Atmosféricos , Compostos Orgânicos Voláteis , Animais , Antioxidantes/farmacologia , Drosophila melanogaster , Masculino , Óxido Nítrico , Octanóis , Oxirredutases , Espécies Reativas de Oxigênio , Compostos Orgânicos Voláteis/análise , Compostos Orgânicos Voláteis/toxicidadeRESUMO
Urine is one of the biological matrices most used for detecting human contamination, as it is representative and easily obtained via noninvasive sampling. This study proposes a fast, accurate, and ecological method based on liquid-liquid microextraction with low-temperature partition (µLLE/LTP). It was validated to determine nine pesticides (lindane, alachlor, aldrin, chlorpyrifos, dieldrin, endrin, DDT, bifenthrin, and permethrin) in human urine, in association with gas chromatography coupled with mass spectrometry (GC-MS). The technique was optimized through a factorial design. The best conditions for the simultaneous extraction of the analytes comprised the addition of 600 µL of water and 600 µL of acetonitrile (extracting solvent) to a 500-µL urine sample, followed by vortexing for 60 s. By freezing the samples for 4 h, it was possible to extract the pesticides and perform the extract clean-up simultaneously. The parameters selectivity, linearity, limit of detection (LOD), limit of quantification (LOQ), precision, and accuracy were used to appraise the performance of the method. Good values of selectivity and linearity (R2 > 0.990), LOQ (0.39-1.02 µg L-1), accuracy (88-119% recovery), and precision (%CV ≤ 15%) were obtained. The µLLE/LTP-GC-MS method was applied to authentic urine samples collected from volunteers in Southeast Brazil.
Assuntos
Clorpirifos , Microextração em Fase Líquida , Resíduos de Praguicidas , Praguicidas , Clorpirifos/análise , Cromatografia Gasosa-Espectrometria de Massas/métodos , Humanos , Resíduos de Praguicidas/análise , Praguicidas/análiseRESUMO
Fungal volatile organic compounds (VOCs) comprise a group of compounds commonly found in damp or water-damaged indoor places affecting air quality. Indoor fungal pollution is a severe threat to human health, contributing to the onset of allergic diseases. The compound 1-octen-3-ol, known as "mushroom alcohol", is the most abundant VOC and confers the characteristic mold odor. Exposure to 1-octen-3-ol induces inflammatory markers and episodes of allergic rhinitis and conjunctivitis; however, the effects of this compound towards mitochondria are fairly known. The present study aimed to evaluate the effects of 1-octen-3-ol on inflammatory targets and on mitochondrial morphology and bioenergetic rate in D. melanogaster. Drosophilas were exposed by inhalation to 2.5 µL/L and 5 µL/L of 1-octen-3-ol for 24 h. Observation showed a decreasing in the survival and locomotor ability of flies. Superoxide dismutase (SOD) activity was induced whereas Catalase (CAT) activity was inhibited. Analysis of the mitochondria respiration, detected inhibition of complex I and II in the electron transport chain and a decreased bioenergetic rate. Electronic microscopy provided morphological insights of the mitochondrial status in which a disarrangement in mitochondrial cristae profile was observed. 1-Octen-3-ol induced increased activity of caspase 3/7 and ERK phosphorylation. The mRNA relative steady-state levels of p38MAPK and JNK were down-regulated, whereas NF-κB and p53 were up-regulated. In parallel, nitrite levels were induced in relation to the non-exposed group. These findings point to the mitochondria as a crucial target for the toxicity of 1-octen-3-ol in parallel with activation of pro-inflammatory factors and apoptotic signaling pathway cascade.
Assuntos
Drosophila melanogaster/efeitos dos fármacos , Mitocôndrias/efeitos dos fármacos , Octanóis/toxicidade , Compostos Orgânicos Voláteis/toxicidade , Poluição do Ar , Poluição do Ar em Ambientes Fechados/efeitos adversos , Animais , Antioxidantes/metabolismo , Apoptose/efeitos dos fármacos , Drosophila melanogaster/enzimologia , Drosophila melanogaster/genética , Feminino , Fungos/metabolismo , Expressão Gênica/efeitos dos fármacos , Humanos , Masculino , Mitocôndrias/metabolismo , Mitocôndrias/ultraestrutura , Atividade Motora/efeitos dos fármacos , Octanóis/análise , Compostos Orgânicos Voláteis/análiseRESUMO
The entomotoxic potential of Manilkara rufula crude extract (CEMR) and its aqueous (AFMR) and methanolic (MFMR) fractions were evaluated against Nauphoeta cinerea cockroaches. The results point out to a direct modulation of octopaminergic and cholinergic pathways in insect nervous system. CEMR induced an anti-acetylcholinesterase (AChE) effect in cockroach brain homogenates. CEMR significantly decreased the cockroach heart rate in semi-isolated heart preparations. CEMR also caused a broad disturbance in the insect behavior by reducing the exploratory activity. The decreased antennae and leg grooming activities, by different doses of CEMR, mimicked those of phentolamine activity, a selective octopaminergic receptor antagonist. The lethargy induced by CEMR was accompanied by neuromuscular failure and by a decrease of sensilla spontaneous neural compound action potentials (SNCAP) firing in in vivo and ex vivo cockroach muscle-nerve preparations, respectively. AFMR was more effective in promoting neuromuscular paralysis than its methanolic counterpart, in the same dose. These data validate the entomotoxic activity of M. rufula. The phentolamine-like modulation induced in cockroaches is the result of a potential direct inhibition of octopaminergic receptors, combined to an anti-AChE activity. In addition, the modulation of CEMR on octopaminergic and cholinergic pathways is probably the result of a synergism between AFMR and MFMR chemical compounds. Further phytochemical investigation followed by a bio-guiding protocol will improve the molecular aspects of M. rufula pharmacology and toxicology to insects.
Assuntos
Baratas , Manilkara , Acetilcolinesterase , Animais , Colinérgicos , ÁrvoresRESUMO
Prasiola crispa is a macroscopic green algae found in abundance in Antarctica ice free areas. Prasiola crispan-hexaneextract (HPC) induced insecticidal activity in Nauphoeta cinerea cockroaches after 24 h of exposure. The chemical analysis of HPC revealed the presence of the followingphytosterols: ß-sitosterol, campesterol and stigmasterol. The incubation of cockroach semi-isolated heart preparations with HPC caused a significant negative chronotropic activity in the heartbeats. HPC affected the insect neuromuscular function by inducing a complete inhibition of the cockroach leg-muscle twitch tension. When the isolated phytosterols were injected at in vivo cockroach neuromuscular preparations, there was a progressive inhibition of muscle twitches on the following order of potency: ß-sitosterol > campesterol > stigmasterol. HPC also provoked significant behavioral alterations, characterized by the increase or decrease of cockroach grooming activity, depending on the dose assayed. Altogether, the results presented here corroborate the insecticide potential of Prasiola crispa Antarctic algae. They also revealed the presence of phytosterols and the involvement of these steroidal compounds in the entomotoxic activity of the algae, potentially by modulating octopaminergic-cholinergic pathways. Further phytochemical-combined bioguided analysis of the HPC will unveil novel bioactive compounds that might be an accessory to the insecticide activity of the algae.
Assuntos
Clorófitas/química , Baratas , Inseticidas/química , Fitosteróis/análise , Extratos Vegetais/química , Animais , Regiões Antárticas , Hexanos/química , Inseticidas/isolamento & purificação , Dose Letal Mediana , Extratos Vegetais/isolamento & purificaçãoRESUMO
Trichomonas vaginalis is an extracellular parasite that binds to the epithelium of the human urogenital tract and causes the sexually transmitted infection, trichomoniasis. In view of increased resistance to drugs belonging to the 5-nitroimidazole class, new treatment alternatives are urgently needed. In this study, eight semisynthetized triterpene derivatives were evaluated for in vitro anti-T. vaginalis activity. Ursolic acid and its derivative, 3-oxime-urs-12-en-28-oic-ursolic acid (9), presented the best anti-T. vaginalis activity when compared to other derivatives, with minimum inhibitory concentration (MIC) at 25 µM. Moreover, 9 was active against several T. vaginalis fresh clinical isolates. Hemolysis assay demonstrated that 9 presented a low hemolytic effect. Importantly, 25 µM 9 was not cytotoxic against the Vero cell lineage. Finally, we demonstrated that compound 9 acts synergistically with metronidazole against a T. vaginalis metronidazole-resistant isolate. This report reveals the high potential of the triterpenoid derivative 9 as trichomonicidal agent.
Assuntos
Antitricômonas/farmacologia , Sinergismo Farmacológico , Metronidazol/farmacologia , Tricomoníase/tratamento farmacológico , Vaginite por Trichomonas/tratamento farmacológico , Trichomonas vaginalis/efeitos dos fármacos , Triterpenos/farmacologia , Animais , Morte Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Chlorocebus aethiops , Resistência a Medicamentos , Quimioterapia Combinada , Feminino , Células HeLa , Hemólise/efeitos dos fármacos , Humanos , Testes de Sensibilidade Microbiana , Tricomoníase/parasitologia , Vaginite por Trichomonas/parasitologia , Triterpenos/química , Células Vero , Ácido UrsólicoRESUMO
Rhinella icterica is a poisonous toad whose toxic secretion has never been studied against entomotoxic potential. Sublethal doses of Rhinella icterica toxic secretion (RITS) were assayed in Nauphoeta cinerea cockroaches, in order to understand the physiological and behavioral parameters, over the insect central and peripheral nervous system. RITS (10⯵g/g) injections, induced behavioral impairment as evidenced by a significant decrease (38⯱â¯14%) in the distance traveled (pâ¯<â¯.05), followed by an increase (90⯱â¯6%) of immobile episodes (pâ¯<â¯.001, nâ¯=â¯28, respectively). In cockroaches semi-isolated heart preparations, RITS (16⯵g/200⯵l) induced a significant irreversible dose-dependent negative chronotropism, reaching ~40% decrease in heart rate in 20â¯min incubation. In in vivo cockroach neuromuscular preparations, RITS (20, 50 and 100⯵g/g of animal weight) induced a time-dependent inhibition of twitch tension that was complete for 20⯵g/g, in 120â¯min recordings. RITS (10⯵g/g) also induced a significant increase in the insect leg grooming activity (128⯱â¯10%, nâ¯=â¯29, pâ¯<â¯.01), but not in the antennae counterparts. The RITS increase in leg grooming activity was prevented in 90% by the pretreatment of cockroaches with phentolamine (0.1⯵g/g). The electrophysiological recordings of spontaneous neural compound action potentials showed that RITS (20⯵g/g) induced a significant increase in the number of events, as well as in the rise time and duration of the potentials. In conclusion, RITS showed to be entomotoxic, being the neuromuscular failure and cardiotoxic activity considered the main deleterious effects. The disturbance of the cockroaches' behavior together with the electrophysiological alterations, may unveil the presence of some toxic components present in the poison with inherent biotechnological potentials.
Assuntos
Bufonidae/fisiologia , Baratas/efeitos dos fármacos , Octopamina/farmacologia , Pele/metabolismo , Toxinas Biológicas/toxicidade , Potenciais de Ação/efeitos dos fármacos , Animais , Comportamento Animal/efeitos dos fármacos , Baratas/metabolismo , Relação Dose-Resposta a Droga , Asseio Animal/efeitos dos fármacos , Frequência Cardíaca/efeitos dos fármacos , Técnicas In Vitro , Junção Neuromuscular/efeitos dos fármacos , Octopamina/metabolismo , Fentolamina/farmacologia , Toxinas Biológicas/metabolismoRESUMO
Tritrichomonas foetus infects the bovine urogenital tract, causing bovine trichomoniasis. Significant economic losses may occur due to infertility and abortion among cattle. Trichomonas vaginalis is the causative agent of trichomoniasis; the most common but overlooked non-viral sexually transmitted disease worldwide. Human and bovine trichomoniasis present treatment restrictions and efforts to identify new alternatives are essential. The present study evaluated the anti-trichomonads activities of seven fractions from northwest endemic plant Manilkara rufula. Flavonoids and condensed tannins were identified from these fractions by LC-DAD-MS/MS and MALDI-MS/MS. Altogether, the results demonstrated for the first time the structural description of tannins from leaves of M. rufula and the relation of these compounds with anti-T. vaginalis and anti-T. foetus activities. Overall, this report reveals the potential of M. rufula fractions against both parasites and shows new alternatives to treat the infection caused by trichomonads.
Assuntos
Antitricômonas/farmacologia , Flavonoides/farmacologia , Manilkara/química , Extratos Vegetais/farmacologia , Taninos/farmacologia , Trichomonas vaginalis/efeitos dos fármacos , Tritrichomonas foetus/efeitos dos fármacos , Antitricômonas/química , Antitricômonas/isolamento & purificação , Brasil , Linhagem Celular , Sobrevivência Celular , Cromatografia Líquida , Células Epiteliais/efeitos dos fármacos , Flavonoides/química , Flavonoides/isolamento & purificação , Células HeLa , Humanos , Estrutura Molecular , Extratos Vegetais/química , Extratos Vegetais/isolamento & purificação , Folhas de Planta/química , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Taninos/química , Taninos/isolamento & purificação , Trichomonas vaginalis/fisiologia , Tritrichomonas foetus/fisiologiaRESUMO
Trichomonas vaginalis causes trichomoniasis; the most common but overlooked non-viral sexually transmitted disease worldwide. The treatment is based at 5'-nitroimidazoles, however, failure are related to resistance of T. vaginalis to chemotherapy. Caatinga is a uniquely Brazilian region representing a biome with type desert vegetation and plants present diverse biological activity, however, with few studies. The aim of this study was to investigate the activity against T. vaginalis of different plants from Caatinga and identify the compounds responsible by the activity. A bioguided fractionation of Manilkara rufula was performed and four major compounds were identified: caproate of α-amyrin (1b), acetate of ß-amyrin (2a), caproate of ß-amyrin (2b), and acetate of lupeol (3a). In addition, six derivatives of α-amyrin (1), ß-amyrin (2) and lupeol (3) were synthesized and tested against the parasite. Ursolic acid (5) reduced about 98% of parasite viability after 2h of incubation and drastic ultrastructural alterations were observed by scanning electron microscopy. Moreover, 5 presented high cytotoxicity to HMVII and HeLa cell line and low cytotoxicity against Vero line at 50 µM (MIC against the parasite). Metronidazole effect against T. vaginalis resistant isolate was improved when in association with 5.
Assuntos
Extratos Vegetais/farmacologia , Plantas Medicinais/química , Trichomonas vaginalis/efeitos dos fármacos , Avaliação Pré-Clínica de Medicamentos , Plantas Medicinais/classificaçãoRESUMO
Extracellular ATP may act as a danger signalling molecule, inducing inflammation and immune responses in infection sites. The ectonucleotidases NTPDase and ecto-5'-nucleotidase are enzymes that modulate extracellular nucleotide levels; these enzymes have been previously characterised in Trichomonas vaginalis. Iron plays an important role in the complex trichomonal pathogenesis. Herein, the effects of iron on growth, nucleotide hydrolysis and NTPDase gene expression in T. vaginalis isolates from female and male patients were evaluated. Iron from different sources sustained T. vaginalis growth. Importantly, iron from haemoglobin (HB) and haemin (HM) enhanced NTPDase activity in isolates from female patients and conversely reduced the enzyme activity in isolates from male patients. Iron treatments could not alter the NTPDase transcript levels in T. vaginalis. Furthermore, our results reveal a distinct ATP, ADP and AMP hydrolysis profile between isolates from female and male patients influenced by iron from HB and HM. Our data indicate the participation of NTPDase and ecto-5'-nucleotidase in the establishment of trichomonas infection through ATP degradation and adenosine production influenced by iron.
Assuntos
Hemina/química , Hemoglobinas/química , Ferro/fisiologia , Nucleotídeos/metabolismo , Trichomonas vaginalis/enzimologia , 5'-Nucleotidase/metabolismo , Difosfato de Adenosina/metabolismo , Monofosfato de Adenosina/metabolismo , Adenosina Trifosfatases/metabolismo , Trifosfato de Adenosina/metabolismo , Técnicas de Cultura de Células , Feminino , Expressão Gênica/efeitos dos fármacos , Humanos , Hidrólise , Ferro/administração & dosagem , Masculino , RNA de Protozoário , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Tricomoníase/enzimologia , Trichomonas vaginalis/efeitos dos fármacos , Trichomonas vaginalis/crescimento & desenvolvimentoRESUMO
Trichomonas vaginalis is a flagellate protozoan that causes trichomonosis, a sexually transmitted disease of worldwide importance. However, the infection has long received much less attention than other parasitic and sexually transmitted diseases. This negligence leads to poor diagnosis and underestimated prevalence values, and consequently, it has been associated to increasing acquisition and transmission of HIV, pregnancy outcomes, infertility, pelvic inflammatory disease, and cervical and prostate cancer. In view of increased resistance to drugs belonging to the nitroimidazole class, new treatment alternatives are urgently needed. Natural products provide an immeasurable wealth of active molecules, and a great number of new drugs have been originated from these compounds. In addition, new synthetic products or derivatives from old drugs also provide an alternative to treat trichomonosis. Albeit many studies have been performed with natural products against T. vaginalis, none of them progressed to clinical trials. Overall, inadequate financial investments are made, and no alternative treatment for trichomonosis has been discovered; meanwhile, hundreds of thousands of people will remain infected and suffering the serious consequences of this nonviral STD. Thus, it is highlighted that clinical trials for better understanding the potential in vitro are necessary and urgent in order to furnish a new therapeutic alternative for trichomonosis treatment. The current review attempts to give an overview on the potential of natural and synthetic products as antitrichomonal.
Assuntos
Antitricômonas/farmacologia , Azóis/farmacologia , Produtos Biológicos/farmacologia , Tricomoníase/tratamento farmacológico , Trichomonas vaginalis/efeitos dos fármacos , Antitricômonas/síntese química , Antitricômonas/isolamento & purificação , Organismos Aquáticos/química , Azóis/química , Produtos Biológicos/isolamento & purificação , Resistência a Medicamentos , Humanos , Plantas/química , Tricomoníase/parasitologiaRESUMO
To investigate the transmission of species of Cryptosporidium Tyzzer, 1907 in Timis County, Romania, 48 isolates of Cryptosporidium coccidia from 11 children, 29 calves and eight pigs were characterised by molecular analysis of two loci (SSU rRNA and 60-kDa glycoprotein gene). Overall, 22 isolates were amplified and sequence analyses revealed that all isolates were Cryptosporidium parvum Tyzzer, 1912. Two subtype families were identified, IIa and IId. Subtype IIdA22G1 (n = 4) was the single C. parvum subtype found in children. Subtypes found in calves included IIdA27G1 (n = 8), a novel subtype, IIdA25G1 (n = 5), IIdA22G1 (n = 2), IIdA21G1a (n = 1), and IIaA16G1R1 (n = 1). Subtype IIdA26G1 was found in a pig. These results were significantly different from previous Romanian reports, as the five subtypes of family IId identified in this study were never identified previously in this country. Thus, cattle may be a source of Cryptosporidium infections for humans and the transmission dynamics of C. parvum in Romania is more complex than previously believed.
RESUMO
Trichomonas vaginalis is a flagellated parasite that causes trichomonosis, the most common non-viral sexually transmitted disease (STD) in the world. Worryingly, trichomonosis is associated to increased transmission of HIV. Due to high frequency of the infection during pregnancy and the development of metronidazole-resistant isolates, therapeutic alternatives to 5-nitroimidazole are being searched. Triterpenes are natural products presenting several biological activities such as anti-protozoal activity. The aim of this study was to evaluate the in vitro anti-T. vaginalis activity from betulinic and ursolic acids, as well as semisynthetic derivatives obtained. Compounds obtained from betulinic acid presented better activity than those from ursolic acid. Piperazine derivatived from betulinic acid presented minimum inhibitory concentration (MIC) value of 91.2 µM, and the kinetic growth curve performed with parasites treated with this most active compound revealed complete inhibition of trophozoite proliferation at 2 h of incubation and total abolition of trophozoite growth in 24 h, revealing that the piperazine derivative is an efficient trichomonacidal molecule. The same compound promoted total erythrocyte lysis and lactate dehydrogenase (LDH) liberation of 83 and 100% (at 45.6 and 91.2 µM, respectively), indicating parasite membrane damage. The piperazine derivative demonstrated cytotoxic effect against the HMVII and HeLa cell lineages at the MIC value. This is the first report of semisynthetic triterpenoid derivatives with anti-T. vaginalis activity, revealing the high potential of these compounds as trichomonacidal agents.
Assuntos
Antitricômonas/farmacologia , Trichomonas vaginalis/efeitos dos fármacos , Triterpenos/farmacologia , Resistência a Medicamentos , Eritrócitos/efeitos dos fármacos , Feminino , Células HeLa , Hemólise , Humanos , Masculino , Metronidazol/farmacologia , Testes de Sensibilidade Microbiana , Triterpenos Pentacíclicos , Piperazina , Piperazinas/farmacologia , Trichomonas vaginalis/crescimento & desenvolvimento , Ácido Betulínico , Ácido UrsólicoRESUMO
AIMS: The aim of this study was to assess the quality of life, psychiatric morbidity, and the psychosocial adjustment of adolescents and young adults with CHD, and determine which variables play a role in buffering stress and promoting resilience and which ones have a detrimental effect; and to investigate the situation on school performance and failures, social and family support, physical limitations, and body image of these patients. METHODS: The study enrolled 137 CHD patients (79 male), with age ranging from 12 to 26 years old (M=17.60±3.450 years). The patients were interviewed regarding social support, family educational style, self-image, demographic information, and physical limitations. They responded to questions in a standardised psychiatric interview (SADS-L) and completed self-reported questionnaires for the assessment of quality of life (WHOQOL-BREF) and psychosocial adjustment (YSR/ASR). RESULTS: We found a 19.7% lifetime prevalence of psychopathology in our patients (27.6% in female and 13.9% in male). Of them, 48% had retentions in school (M=1.61 year±0.82). The perception of quality of life in CHD patients is better compared with the Portuguese population in the social relationships and environmental dimensions. However, it is worse in complex forms of CHD than in moderate-to-mild ones, in cyanotic versus acyanotic patients, in moderate-to-severe versus mild residual lesions, in patients submitted versus those not submitted to surgery, in patients with versus without physical limitations, and patients who have need for medication versus those who do not. Social support is very important in improving quality of life of patients in all dimensions as well as academic performance. CONCLUSIONS: Female patients and patients with poor academic performance and poor social support have worse psychosocial adjustment and perception of quality of life.
Assuntos
Cardiopatias Congênitas/psicologia , Transtornos Mentais/epidemiologia , Transtornos Mentais/psicologia , Qualidade de Vida/psicologia , Adolescente , Adulto , Criança , Comorbidade , Avaliação Educacional , Feminino , Cardiopatias Congênitas/epidemiologia , Humanos , Entrevista Psicológica , Masculino , Portugal/epidemiologia , Prevalência , Distribuição por Sexo , Apoio Social , Adulto JovemRESUMO
BACKGROUND: Carbapenemase-producing Enterobacterales (CPE) is an increasing problem in healthcare settings. This study aimed to identify the source of a CPE outbreak that occurred in 2022, in a tertiary hospital in the North of Portugal, to identify exposed patients, and to assess the risk of becoming CPE-positive following hospital admission. METHODS: A multi-disciplinary investigation was conducted including descriptive, analytical, and molecular epidemiology, environmental screening, and assessment of infection control measures. Clinical and environmental isolates were analyzed using whole-genome sequencing and phylogenetic analysis. Additionally, a prospective observational cohort study was conducted to further investigate the risk factors associated with the emergence of new cases in cohorts of CPE-negative admitted patients. RESULTS: We observed the presence of multispecies KPC-, IMP-, and/or NDM-producing isolates. Genetically indistinguishable clinical and environmental isolates were found on the same room/ward. The ST45 KPC-3-producing Klebsiella pneumoniae clone was the responsible for the outbreak. During patients' treatment, we detected the emergence of resistance to ceftazidime-avibactam, associated with mutations in the blaKPC-3 gene (blaKPC-46, blaKPC-66 and blaKPC-124, the last variant never previously reported), suggesting a vertical evolutionary trajectory. Patients aged ≥ 75 years, hygiene/feeding-care dependent, and/or subjected to secretion aspiration were risk factors for CPE colonization after hospital admission. Additionally, cases with previous admission to the emergency department suggest that CPE dissemination may occur not only during hospitalization but also in the emergency department. CONCLUSION: Overall, the study highlights that selection pressure with antibiotics, like ceftazidime-avibactam, is a contributing factor to the emergence of new ß-lactamase variants and antibiotic resistance. It also shows that the hospital environment can be a significant source of CPE transmission, and that routine use of infection control measures and real-time molecular epidemiology investigations are essential to ensure the long-term termination of CPE outbreaks and prevent future resurgences.
Assuntos
Infecções por Klebsiella , Humanos , Portugal/epidemiologia , Infecções por Klebsiella/epidemiologia , Infecções por Klebsiella/tratamento farmacológico , Filogenia , Estudos Prospectivos , beta-Lactamases/genética , beta-Lactamases/uso terapêutico , Proteínas de Bactérias/genética , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Surtos de Doenças , Genômica , Hospitais , Klebsiella pneumoniae , Testes de Sensibilidade MicrobianaRESUMO
Pancreatic ductal adenocarcinoma (PDAC), a lethal disease, requires a grasp of its biology for effective therapies. Exosomes, implicated in cancer, are poorly understood in living systems. Here we use the genetically engineered mouse model (ExoBow) to map the spatiotemporal distribution of exosomes from healthy and PDAC pancreas in vivo to determine their biological significance. We show that, within the PDAC microenvironment, cancer cells establish preferential communication routes through exosomes with cancer associated fibroblasts and endothelial cells. The latter being a conserved event in the healthy pancreas. Inhibiting exosomes secretion in both scenarios enhances angiogenesis, underscoring their contribution to vascularization and to cancer. Inter-organ communication is significantly increased in PDAC with specific organs as most frequent targets of exosomes communication occurring in health with the thymus, bone-marrow, brain, and intestines, and in PDAC with the kidneys, lungs and thymus. In sum, we find that exosomes mediate an organized intra- and inter- pancreas communication network with modulatory effects in vivo.
Assuntos
Carcinoma Ductal Pancreático , Exossomos , Neoplasias Pancreáticas , Camundongos , Animais , Exossomos/patologia , Células Endoteliais/patologia , Linhagem Celular Tumoral , Movimento Celular , Neoplasias Pancreáticas/patologia , Carcinoma Ductal Pancreático/genética , Carcinoma Ductal Pancreático/patologia , Pâncreas/patologia , Microambiente TumoralRESUMO
OBJECTIVE: Congenital heart disease (CHD) is a chronic illness with a high frequency in the worldwide population, and is normally diagnosed at birth or in uterus. Because of better conditions in diagnosis and early medical and surgical treatment, patients have survival rates of 90% and go further and further in life, facing different challenges in life cycle. In this study, we tested the effects of different demographic, clinical and psychosocial variables on the perception of quality of life (QOL), on psychosocial adjustment (PSA) and psychiatric morbidity (PM) of adolescents and young adults with CHD. We aimed to evaluate QOL, PM and PSA of adolescents and young adults with CHD and to determine which variables (demographic, clinical, and psychosocial) play a role in buffering stress and promoting resilience and which ones have a detrimental effect. METHODS: The study enrolled 150 CHD patients (87 males and 63 females), 12 to 26 years (17.45±3.373 years). The participants were interviewed regarding social support, family educational style, self-image, demographic information and physical limitations. They responded to questions in a standardized psychiatric interview (SADS-L) and completed self-reports questionnaires for assessment of QOL (WHOQOL-BREF) and PSA (YSR/ASR). RESULTS: We found a 18.7% lifetime prevalence of psychopathology in our participants (25.4% in females and 13.8% in males). 57.1% had retentions in school (1.53±0.804 year). The perception of QOL of CHD patients is better compared to the Portuguese population in the social relationships, environmental, physical and general dimensions. However, it is worse in female CHD patients and patients with poor academic performance and social support as well as in patients with complex or cyanotic CHD, moderate-to-severe residual lesions and physical limitations, and undergoing surgery. All of these variables, except presence of cyanosis, are also associated to a worse PSA. CONCLUSIONS: Female patients and patients with poor academic performance and poor social support refer worse PSA and QOL.
Assuntos
Cardiopatias Congênitas/psicologia , Adaptação Psicológica , Adolescente , Adulto , Criança , Feminino , Humanos , Masculino , Morbidade , Qualidade de Vida , Apoio Social , Adulto JovemRESUMO
Over the past few decades, research has suggested that cognitive variables play a key role in sleep disorders, particularly, in insomnia. The SLOC (Sleep Locus of Control Scale) evaluates the sleep locus of control, which is associated with the degree to which an individual attributes her/his experiences of sleep to chance or internal causes. The aim of this study was to develop the first translation and adaptation of the SLOC into the European Portuguese, as well as to analyze its psychometric properties. In this study, it was recruited a sample of 2029 Portuguese Higher Education students, aged ≥ 18 years, where approximately 75% of the sample were women and 25% men. The results showed that the SLOC had acceptable internal consistency value (α = .64), considering that it is a measure with a reduced number of items. As in the original study, a principal component analysis with varimax rotation identified two components. A parallel analysis was also conducted, identifying two factors. The correlation between the two subscales "internal sleep locus of control" and "chance sleep locus of control" was positive albeit of low magnitude (r = .15). Through the analysis carried out, it was also observed that individuals with "insomnia", relative to those without, had a more chance sleep locus of control. Overall, these findings show similarities with the original study. The SLOC seems to be a useful psychological assessment measure to be used in clinical and research settings.
RESUMO
Sleep disorders are catching attention worldwide as they can induce dyshomeostasis and health issues in all animals, including humans. Circadian rhythms are biological 24-hour cycles that influence physiology and behavior in all living organisms. Sleep is a crucial resting state for survival and is under the control of circadian rhythms. Studies have shown the influence of sleep on various pathological conditions, including metabolic diseases; however, the biological mechanisms involving the circadian clock, sleep, and metabolism regulation are not well understood. In previous work, we standardized a sleep disturbance protocol and, observed that short-time sleep deprivation and sleep-pattern alteration induce homeostatic sleep regulation, locomotor deficits, and increase oxidative stress. Now, we investigated the relationship between these alterations with the circadian clock and energetic metabolism. In this study, we evaluated the expression of the circadian clock and drosophila insulin-like peptides (DILPs) genes and metabolic markers glucose, triglycerides, and glycogen in fruit flies subjected to short-term sleep disruption protocols. The sleep disturbance altered the expression of clock genes and DILPs genes expression, and modulated glucose, triglycerides, and glycogen levels. Moreover, we demonstrated changes in mTor/dFoxo genes, AKT phosphorylation, and dopamine levels in nocturnal light-exposed flies. Thus, our results suggest a connection between clock genes and metabolism disruption as a consequence of sleep disruption, demonstrating the importance of sleep quality in health maintenance.
Assuntos
Relógios Circadianos , Drosophila , Animais , Humanos , Sono/fisiologia , Ritmo Circadiano/fisiologia , Privação do Sono/metabolismo , Glucose , Glicogênio/metabolismo , Expressão Gênica , Triglicerídeos , Regulação da Expressão Gênica , Relógios Circadianos/genéticaRESUMO
Yoghurt fermented under sub-lethal high pressure (10, 20, 30 and 40 MPa at 43 °C), and afterward placed under refrigeration (4 °C for 23 days) was studied and compared with yoghurt fermented at atmospheric pressure (0.1 MPa). For a deeper analysis, metabolite fingerprinting by nuclear magnetic resonance (NMR), sugars and organic acids assessment by high performance liquid chromatography (HPLC), total fatty acids (TFA) determination and quantification by gas chromatography with a flame ionization detector (GC-FID) were performed. Metabolomic analyses revealed that only 2,3-butanediol, acetoin, diacetyl and formate vary with the increase of pressure and probable relation with pressure influenced diacetyl reductase, acetoin reductase and acetolactate decarboxylase. Yoghurts fermented at 40 MPa had the lowest content in lactose (39.7 % of total sugar reduction) and the less content in TFA (56.1 %). Further research is of interest to understand more about fermentation processes under sub-lethal high pressure.