Essential oil of oregano (Origanum vulgare L.) reduces infection and proliferation of Toxoplasma gondii in BeWo cells with induction of autophagy and death of tachyzoites through a mechanism similar to necrosis.

Toxoplasmosis poses a global health threat, ranging from asymptomatic cases to severe, potentially fatal manifestations, especially in immunocompromised individuals and congenital transmission. Prior research suggests that oregano essential oil (OEO) exhibits diverse biological effects, including antiparasitic activity against Toxoplasma gondii. Given concerns about current treatments, exploring new compounds is important. This study was to assess the toxicity of OEO on BeWo cells and T. gondii tachyzoites, as well as to evaluate its effectiveness in in vitro infection models and determine its direct action on free tachyzoites. OEO toxicity on BeWo cells and T. gondii tachyzoites was assessed by MTT and trypan blue methods, determining cytotoxic concentration (CC50), inhibitory concentration (IC50), and selectivity index (SI). Infection and proliferation indices were analyzed. Direct assessments of the parasite included reactive oxygen species (ROS) levels, mitochondrial membrane potential, necrosis, and apoptosis, as well as electron microscopy. Oregano oil exhibited low cytotoxicity on BeWo cells (CC50: 114.8 µg/mL ± 0.01) and reduced parasite viability (IC50 12.5 ± 0.06 µg/mL), demonstrating 9.18 times greater selectivity for parasites than BeWo cells. OEO treatment significantly decreased intracellular proliferation in infected cells by 84% after 24 h with 50 µg/mL. Mechanistic investigations revealed increased ROS levels, mitochondrial depolarization, and lipid droplet formation, linked to autophagy induction and plasma membrane permeabilization. These alterations, observed through electron microscopy, suggested a necrotic process confirmed by propidium iodide labeling. OEO treatment demonstrated anti-T. gondii action through cellular and metabolic change while maintaining low toxicity to trophoblastic cells.

Trilobolide-6-O-isobutyrate from Sphagneticola trilobata acts by inducing oxidative stress, metabolic changes and apoptosis-like processes by caspase 3/7 activation of human lung cancer cell lines.

BACKGROUND: Lung cancer, a chronic and heterogeneous disease, is the leading cause of cancer-related death on a global scale. Presently, despite a variety of available treatments, their effectiveness is limited, often resulting in considerable toxicity and adverse effects. Additionally, the development of chemoresistance in cancer cells poses a challenge. Trilobolide-6-O-isobutyrate (TBB), a natural sesquiterpene lactone extracted from Sphagneticola trilobata, has exhibited antitumor effects. Its pharmacological properties in NSCLC lung cancer, however, have not been explored. PURPOSE: This study evaluated the impact of TBB on the A549 and NCI-H460 tumor cell lines in vitro, examining its antiproliferative properties and initial mechanisms of cell death. METHODS: TBB, obtained at 98 % purity from S. trilobata leaves, was characterized using chromatographic techniques. Subsequently, its impact on inhibiting tumor cell proliferation in vitro, TBB-induced cytotoxicity in LLC-MK2, THP-1, AMJ2-C11 cells, as well as its effects on sheep erythrocytes, and the underlying mechanisms of cell death, were assessed. RESULTS: In silico predictions have shown promising drug-likeness potential for TBB, indicating high oral bioavailability and intestinal absorption. Treatment of A549 and NCI-H460 human tumor cells with TBB demonstrated a direct impact, inducing significant morphological and structural alterations. TBB also reduced migratory capacity without causing toxicity at lower concentrations to LLC-MK2, THP-1 and AMJ2-C11 cell lines. This antiproliferative effect correlated with elevated oxidative stress, characterized by increased levels of ROS, superoxide anion radicals and NO, accompanied by a decrease in antioxidant markers: SOD and GSH. TBB-stress-induced led to changes in cell metabolism, fostering the accumulation of lipid droplets and autophagic vacuoles. Stress also resulted in compromised mitochondrial integrity, a crucial aspect of cellular function. Additionally, TBB prompted apoptosis-like cell death through activation of caspase 3/7 stressors. CONCLUSION: These findings underscore the potential of TBB as a promising candidate for future studies and suggest its viability as an additional component in the development of novel anticancer drugs prototypes.

Leishmania amazonensis infection regulates oxidate stress in hyperglycemia and diabetes impairing macrophage's function and immune response.

Leishmaniasis is a group of infectious diseases caused by protozoa of the Leishmania genus and its immunopathogenesis results from an unbalanced immune response during the infection. Diabetes is a chronic disease resulting from dysfunction of the body's production of insulin or the ability to use it properly, leading to hyperglycemia causing tissue damage and impairing the immune system. AIMS: The objective of this work was to evaluate the effects of hyperglycemia and diabetes during Leishmania amazonensis infection and how these conditions alter the immune response to the parasite. METHODS: An in vitro hyperglycemic stimulus model using THP-1-derived macrophages and an in vivo experimental diabetes with streptozotocin (STZ) in C57BL/6 mice was employed to investigate the impact of diabetes and hyperglicemia in Leishmania amazonensis infection. RESULTS: We observed that hyperglycemia impair the leishmanicidal capacity of macrophages derived from THP-1 cells and reverse the resistance profile that C57BL/6 mice have against infection by L. amazonensis, inducing more exacerbated lesions compared to non-diabetic animals. In addition, the hyperglycemic stimulus favored the increase of markers related to the phenotype of M2 macrophages. The induction of experimental diabetes in C57BL/6 mice resulted in a failure in the production of nitric oxide (NO) in the face of infection and macrophages from diabetic animals failed to process and present Leishmania antigens, being unable to activate and induce proliferation of antigen-specific lymphocytes. CONCLUSION: Together, these data demonstrate that diabetes and hyperglycemia can impair the cellular immune response, mainly of macrophages, against infection by parasites of the genus Leishmania.

3,3',5,5'-Tetramethoxybiphenyl-4,4'diol triggers oxidative stress, metabolic changes, and apoptosis-like process by reducing the PI3K/AKT/NF-κB pathway in the NCI-H460 lung cancer cell line.

Lung cancer is one of the leading causes of cancer-related deaths in men and women worldwide. Current treatments have limited efficacy, cause significant side effects, and cells can develop drug resistance. New therapeutic strategies are needed to discover alternative anticancer agents with high efficacy and low-toxicity. TMBP, a biphenyl obtained by laccase-biotransformation of 2,6-dimethoxyphenol, possesses antitumor activity against A549 adenocarcinoma cells. Without causing damage to sheep erythrocytes and mouse peritoneal macrophages of BALB/c mice. In addition to being classified as a good oral drug according to in-silico studies. This study evaluated the in-vitro cytotoxic effect of TMBP on lung-cancer cell-line NCI-H460 and reports mechanisms on immunomodulation and cell death. TMBP treatment (12.5-200 µM) inhibited cell proliferation at 24, 48, and 72 h. After 24-h treatment, TMBP at IC50 (154 µM) induced various morphological and ultrastructural changes in NCI-H460, reduced migration and immunofluorescence staining of N-cadherin and ß-catenin, induced increased reactive oxygen species and nitric oxide with reduced superoxide radical-anion, increased superoxide dismutase activity and reduced glutathione reductase. Treatment also caused metabolic stress, reduced glucose-uptake, intracellular lactate dehydrogenase and lactate levels, mitochondrial depolarization, increased lipid droplets, and autophagic vacuoles. TMBP induced cell-cycle arrest in the G2/M phase, death by apoptosis, increased caspase-3/7, and reduced STAT-3 immunofluorescence staining. The anticancer effect was accompanied by decreasing PI3K, AKT, ARG-1, and NF-κB levels, and increasing iNOS. These results suggest its potential as a candidate for use in future lung anticancer drug design studies.

Case Report: Alternaria alternata keratitis.

Background: Alternaria spp are ubiquitous pigmented filamentous fungi that can cause opportunistic human infections. These molds can be found in healthy individuals and the most frequently observed clinical manifestations are skin infections, ocular mycosis, rhinosinusitis, among others. Immunosuppression (both local and systemic) as well as contact or trauma with contaminated matter are important risk and predisposing factors for ocular mycosis. Accurate diagnosis with microscopy and culture is crucial since infections by filamentous fungi are sight-threatening, and clinically indistinguishable from bacterial disease. In general terms, conventional antifungal drugs are effective. Case Presentation: We report a case of Alternaria alternata keratitis after ocular trauma with biological material in a 44-year-old man's functional single eye, which had a good clinical and functional evolution after topical therapy with voriconazole. Conclusion: This case accounts the importance of thinking about other etiological diagnoses in infectious keratitis refractory to the established therapy and with a history of trauma with biological material.

Pleural empyema due to Rhodotorula mucilaginosa: A rare yet severe complication of a previously undiagnosed cancer patient.

Rhodotorula mucilaginosa (R. mucilaginosa) has been increasingly recognized as an emerging opportunistic pathogen causing invasive fungal infection, mainly in immunosuppressed patients. We report the case of a previously undiagnosed lung cancer patient with a pleural empyema due to R. mucilaginosa.

Painless rash in a transplant patient.

Dermatophytes are common keratinophilic fungi responsible for superficial skin infections. Deep dermatophytosis is a rare form of invasive skin infection described in immunocompromised patients. We report the case of a 65-year-old man with a history of an orthotopic liver transplant for hepatocarcinoma 6 months earlier, who presented with small painless erythematous papules in lower limbs, some of which were umbilicated. Skin biopsy showed an intense non-necrotizing granulomatous reaction in the dermis around fungal structures. Trichophyton rubrum was identified as the causal agent through culture and internal transcribed spacer sequencing.

Exophiala Keratitis following Descemet Stripping Automated Endothelial Keratoplasty.

PURPOSE: To report a case with Exophiala spp. keratitis in a Portuguese patient. METHODS: A case report with deep corneal brown-pigmented infiltrates that developed 2 months after a Descemet Stripping Automated Endothelial Keratoplasty (DSAEK) for pseudophakic bullous keratopathy. RESULTS: Diagnosis was established by positive direct examination and cultures from the surgically obtained corneal button. Slit-lamp images and anterior segment optical coherence tomography (AS-OCT) scans were obtained. CONCLUSION: This is the first described case of fungal keratitis caused by Exophiala spp. in Portugal and, to our knowledge, the first case following DSAEK in the literature.

Revisão do arranjo das compras públicas a partir de um contexto de crise / Nota Técnica n. 68 (Diset): Revisão do arranjo das compras públicas a partir de um contexto de crise / Português

A pandemia da Covid-19 expôs ao mundo a fragilidade dos arranjos institucionais de suprimentos para os sistemas de saúde mundiais. Insumos básicos para o atendimento de saúde, como equipamentos de proteção individuais (EPIs), respiradores artificiais e higienizantes/saneantes, passaram a ser disputados por diversos países e a ensejar comportamentos fraticidas entre sistemas de saúde privado e público de diferentes esferas no Brasil: pelo menos seis estados e diversas prefeituras editaram atos administrativos para requisitar insumos e produtos como respiradores durante a pandemia, levando a União a reagir na Justiça e a defender uma centralização das compras desses produtos. Entes privados como a Associação Nacional de Hospitais Privados (ANAHP), a Confederação das Santas Casas de Misericórdia, Hospitais e Entidades Filantrópicas (CMB) e a Confederação Nacional de Saúde (CNS) questionaram judicialmente o privilégio dos hospitais públicos na distribuição de EPIs. Nesse contexto de ruptura da cadeia de suprimentos e de concorrência predatória entre compradores, a Organização Mundial da Saúde (OMS) recomenda, além da racionalização da demanda e do uso dos insumos hospitalares, a gestão centralizada dessas aquisições e a coordenação do abastecimento das unidades de saúde para evitar excessos e quebras de estoques (WHO, 2020). A transposição dessa reflexão para o cenário brasileiro requer uma apreciação dos desafios e das oportunidades de aperfeiçoar os mecanismos de agregação de demandas e as sistemáticas de compras conjuntas disponíveis para os gestores públicos envolvidos no enfrentamento da emergência de saúde pública da Covid-19, com maior colaboração entre os entes públicos e o possível envolvimento de entes privados que atendam majoritariamente pacientes do Sistema Único de Saúde (SUS), ao mesmo tempo que se preserve a transparência e se facilite a fiscalização. Algumas alterações legais e procedimentais são necessárias, parte delas no nível mais geral das compras e contratações, e sua implementação em caráter experimental, se alcançar resultados satisfatórios, pode se tornar definitiva e inclusive ser expandida para outros contextos (objetos de contratação, modalidades, poderes e entes). A nota tem mais cinco seções além desta introdução. Na próxima, fazemos uma motivação para um arcabouço de compras centralizadas em saúde. Em seguida, varremos algumas experiências internacionais de compras centralizadas e o que tem sido feito no Brasil a respeito. As seções seguintes detalham, então, o arcabouço legal para se implementar um Sistema de Registro de Preços de Emergência, que chamamos de Acordo-Marco, e um leque de propostas de implementação para esse sistema. A última seção consolida as considerações finais.

Donor-derived fatal hyperinfection strongyloidiasis in renal transplant recipient.

Strongyloides stercoralis is a nematode, endemic in tropical and subtropical areas. Strongyloidiasis has been reported in recipients of hematopoietic stem cells, kidney, liver, heart, intestine, and pancreas, eventually presenting as disseminated strongyloidiasis and hyperinfection syndrome (SHS) which is associated with high mortality. We report one case of a recent renal transplant recipient, who presented with gastrointestinal and respiratory symptoms, evolving into shock. The identification of Strongyloides stercoralis in the bronchoalveolar lavage (BAL) lead us to the diagnosis of SHS. Treatment with subcutaneous ivermectin was started, however the patient did not survive. Retrospective serum donor analysis allowed us to identify the donor as the source of infection.

Compras públicas centralizadas em situações de emergência e calamidade pública / Texto para Discussão (TD) 2575: Compras públicas centralizadas em situações de emergência e calamidade pública

A pandemia da Covid-19 expôs ao mundo a fragilidade dos arranjos institucionais de suprimentos e suscitou a concorrência predatória entre compradores. Neste contexto, a racionalização da demanda e do uso dos insumos hospitalares e a gestão centralizada dessas aquisições são estratégias recomendadas, e transpô-las para o cenário brasileiro requer uma apreciação dos desafios e das oportunidades disponíveis para os gestores públicos. As compras colaborativas ou conjuntas são um dos instrumentos mais efetivos para reduzir custos em sistemas de saúde, mas nem a redução de custos deve ser o único objetivo, nem a mera agregação de compras é condição necessária para tal redução. O estado de calamidade pública instituído em fevereiro de 2020 propiciou o rápido surgimento de um direito provisório com vistas a suprir mais rápida e eficazmente as urgentes necessidades de aquisições e contratações, entre as quais destacamos a previsão de utilização da dispensa de para seleção de fornecedores para atas de registro de preços, a ampliação do uso do regime diferenciado de contratações, o pagamento antecipado de despesas contratadas, e o aumento temporário do limite de valor de dispensa de licitação. Contudo, os avanços podem não ser ainda suficientes e aparenta ser necessário estruturar uma sistemática inovadora para as contratações públicas, como a adoção dos acordos-quadro multifornecedores. Neste trabalho, traçamos três estratégias para viabilizá-los, além de oferecermos três cenários operacionais de implantação, de acordo com a plataforma a ser utilizada (Siasg/Comprasnet ou marketplace ou ambos). A pandemia pode ser uma ótima oportunidade de acelerarmos a convergência para essas compras coordenadas.

The Covid-19 pandemic exposed the fragility of institutional supply arrangements to the world and sparked predatory competition among buyers. In this context, the rationalization of demand and use of medical inputs and the centralized management of these acquisitions are recommended strategies, and transposing them to the Brazilian scenario requires an appreciation of the challenges and opportunities available to public managers. Collaborative or joint purchases are one of the most effective instruments to reduce costs in healthcare systems, but neither cost reduction should be the sole objective, nor is the mere aggregation of purchases a necessary condition for such reduction. The state of public calamity instituted in February 2020 led to the rapid emergence of a provisional Law with a view to meeting more quickly and effectively the urgent needs for acquisitions and contracts, among which we highlight the provision for the use of bidding waivers in awarding frame contracts (Brazilian Price Registration System), the expansion of the use of the (more flexible) Differentiated Contracting Regime, introduction of advanced payments, and the temporary increase of the upper value threshold of the bidding . However, progress may not be sufficient yet and it appears necessary to structure an innovative system for public procurement, such as the adoption of multi-supplier framework agreements. In this article, we outline three strategies to render them feasible, in addition to offering three operational deployment scenarios, according to the platform to be used (Siasg/Comprasnet or marketplace or both). The pandemic can be a great opportunity to accelerate convergence for these coordinated purchases.

Aspergillus Species and Antifungals Susceptibility in Clinical Setting in the North of Portugal: Cryptic Species and Emerging Azoles Resistance in A. fumigatus.

Aspergillus spp. are agents of a broad-spectrum of diseases among humans. Their growing resistance to azoles, the cornerstone in the management of human aspergillosis, is a worrisome problem around the world. Considering lack of data from Portugal on this topic, particularly from the northern region, a retrospective surveillance study was planned to assess frequency of cryptic Aspergillus species and azoles resistance. A total of 227 clinical isolates, mainly from the respiratory tract (92.1%), collected from three hospitals serving a population of about three million people, were studied for their epidemiology and antifungal susceptibility patterns determined by the E.DEF.9.3 protocol of EUCAST. Employing molecular methods, seven Aspergillus complexes were identified; Aspergillus fumigatus sensu stricto was the most frequent isolate (86.7%). A 7.5% prevalence of cryptic species was found; A. welwitschiae (A. niger complex-3.1%) and A. lentulus (A. fumigatus complex-2.2%) were the most frequent. Amongst cryptic species, it was found a percentage of resistance to voriconazole, posaconazole and isavuconazole of 47.1, 82.4, and 100%, respectively. Five A. fumigatus sensu stricto showed pan-azole resistance. Sequencing their cyp51A gene revealed the presence of one isolate with TR46/Y121F/T289A mutation and two isolates with TR34/L98H mutation. This study emphasizes the need to identify strains to the species level and to evaluate their antifungal susceptibility in all human originated Aspergillus spp. isolates, particularly those from invasive aspergillosis.

Cerebral tuberculomas in a patient with hairy cell leukaemia treated with cladribine.

A 67-year-old man was treated with cladribine for hairy cell leukaemia. A few weeks later, he presented with persistent headaches, intermittent hypoesthesia of the right upper limb and language impairment. Brain CT scan showed 3 contrast-enhancing lesions. MRI revealed infracentimetric nodular lesions with restricted diffusion. One of the lesions was surgically removed and tested positive for acid-fast bacilli. Moreover, Mycobacterium tuberculosis was confirmed by PCR. Antituberculous drugs were prescribed for 12 months, with complete resolution of neurological deficits. This case highlights the risk of mycobacterial infections associated with both hairy cell leukaemia and cladribine use, and the importance of screening and treatment of latent forms of tuberculosis in patients undergoing treatment with immunosuppressive drugs.

Rhodotorula mucilaginosa Fungemia and Pleural Tuberculosis in an Immunocompetent Patient: An Uncommon Association.

We report an atypical case of Rhodotorula mucilaginosa fungemia coexisting with pleural tuberculosis, in an immunocompetent host. The patient was an inhaled drug abuser and worked in a fruit market. The diagnosis of Rhodotorula mucilaginosa infection was established by the isolation of the yeast in two blood cultures followed by a good response to amphotericin B treatment. Persistent evening fever and pleural effusion led to the second diagnosis-pleural tuberculosis. In the last 5 years, this was the only case of Rhodotorula mucilaginosa fungemia in our hospital and the first case in the literature that documents Rhodotorula mucilaginosa fungemia associated with pleural tuberculosis.

Ultraviolet-B radiation effects on phenolic profile and flavonoid content of Kalanchoe pinnata.

Ultraviolet-B radiation is an important abiotic factor that can stimulate the production of secondary metabolites, including polyphenolic compounds. Kalanchoe pinnata (Crassulaceae) is a medicinal plant popularly used in Brazil for treating wounds and inflammation. This species is rich in phenolic compounds, which could account for some of its biological activities, including antileishmanial, antihypertensive and antibacterial properties. We investigated the effects of supplemental UV-B radiation on the phenolic profile, antioxidant activity and total flavonoid content of leaves of K. pinnata. Plants were grown under white light (W - control) and supplemental UV-B radiation (W+UVB). Supplemental UV-B radiation enhanced the total flavonoid content of the leaf extracts, without affecting the antioxidant activity or yield of extracts. Analysis by TLC and HPLC of W and W+UVB leaf extracts revealed quantitative and qualitative differences in their phenolic profiles. W+UVB extracts contained a higher diversity of phenolic compounds and a larger amount of quercitrin, an important bioactive flavonoid of this species. This is the first report of the use of ImageJ® program to analyze a TLC visualized by spraying with NP-PEG reagent. UV-B radiation is proposed as a supplemental light source in K. pinnata cultivation in order to improve its flavonoid composition.

Cryptococcal infection in non-HIV immunosuppressed patients - Three case reports in a nephrology setting.

Cryptococcal infection has been increasing among immunosuppressed population. We report three cases of Cryptococcus neoformans infection in immunosuppressed patients - two renal transplanted and one with lupus nephritis. Early infection (<3months) was diagnosed in two - an allograft Cryptococcus infection and a central nervous system involvement. The third, a 10-year transplant vintage patient, presented with cryptococcal meningitis. Amphotericin B provided good clinical outcomes. We outline the importance of suspicion for cryptococcal infection in immunosuppressed patients.