Antimicrobial and antibiofilm activities of Inga cylindrica trypsin inhibitor.

Inga cylindrica, a tropical fruit tree of the Fabaceae family (subfamily Mimosoideae), is native to South America. The seeds from this family are an essential source of trypsin inhibitors, which display promising bioactivity for increasing host defense against pathogens. The aim of the present study was to characterize the antimicrobial and antibiofilm activities of the trypsin inhibitor extracted from I. cylindrica seeds, ICTI. ICTI demonstrated antifungal activity with a minimum inhibitory concentration (MIC) of 32.11 µmol.L-1, and a minimum fungicidal concentration (MFC) of 32.1 µmol.L-1, against Cryptococcus gattii, Candida albicans, Candida glabrata and Candida guilliermondii. Combining ICTI with Amphotericin B had a significant synergistic effect, reducing the concentration of the antibiotic by 75% for C. albicans and 94% for C. gatti. The significant increase (16 x) in activity observed with ergosterol (1.01 mol.L-1) for C. albicans and C. gatti, and the lack of activity against bacterial strains, suggests that ICTI interferes with the integrity of the fungal cell membrane. Treatment with ICTI at 10 x MIC resulted in a 51% reduction in biofilm formation and a 56% decrease in mature biofilm colonies for C. albicans. Finally, ICTI displayed no toxicity in the in vivo Galleria mellonella model. Given its antifungal and antibiofilm properties, ICTI could be a promising candidate for the development of new antimicrobial drug prototypes.

Guarana (Paullinia cupana) as a potential tool for mesenchymal stromal cells priming in regenerative medicine.

Mesenchymal stromal cells (MSCs) have therapeutic potential due to their abilities of differentiation, immunomodulation, and migration to injured tissues, potentiating such effects when cells are activated. Guarana (Paullinia cupana) is a tropical plant species found in South America that is known for its antioxidant, stimulant, and cicatricial effects. The guarana extract is composed of many substances and caffeine is the main component. The objective was to evaluate the effects of guarana and caffeine on MSCs. After the initial characterization, MSCs were treated with Paullinia cupana (10, 100, and 1000 µg/mL) or caffeine (0.4, 4, and 40 µg/mL) for 24 h. MSCs treatment with 1000 µg/mL guarana increased cell polarity, viability, cell migration to chemoattractant, antioxidant potential, and liberation of extracellular vesicles (EVs), while it reduced the levels of autophagy. MSCs treated with 100 and 1000 µg/mL guarana or 40 µg/mL caffeine showed a decrease of cell proliferation. No treatment affected the cellular area and cell cycle of MSCs. The study shows in vitro evidence that guarana could be a promising alternative for activating MSCs to promote better cellular products for future clinical therapies.

Ontogenetic and sexual differences in the venom of Bothrops moojeni: insights from a litter and its mother.

Variability in snake venom composition is well-documented and crucial for understanding snake ecology and predicting snakebites. In this study, we characterize the venom composition and biological activities of newborn female and male Bothrops moojeni and their mother. Our results reveal significant differences between the venom of newborn females and males, demonstrating a broad and diverse range of proteins. The venoms of newborn females showed higher serine protease effects, increased hemorrhagic activity, and greater lethality compared to the venom of newborn males. However, no differences were observed in phospholipase A2 and coagulant activity. The differences in protein composition and toxic activities between maternal and neonatal venom, as well as between the venoms of newborn females and males, contribute to understanding the diverse outcomes of snakebites. These results underscore the importance of considering sex and ontogeny in understanding venom composition in snakes.

Guarana (Paullinia cupana) as a potential tool for mesenchymal stromal cells priming in regenerative medicine

Mesenchymal stromal cells (MSCs) have therapeutic potential due to their abilities of differentiation, immunomodulation, and migration to injured tissues, potentiating such effects when cells are activated. Guarana (Paullinia cupana) is a tropical plant species found in South America that is known for its antioxidant, stimulant, and cicatricial effects. The guarana extract is composed of many substances and caffeine is the main component. The objective was to evaluate the effects of guarana and caffeine on MSCs. After the initial characterization, MSCs were treated with Paullinia cupana (10, 100, and 1000 μg/mL) or caffeine (0.4, 4, and 40 μg/mL) for 24 h. MSCs treatment with 1000 μg/mL guarana increased cell polarity, viability, cell migration to chemoattractant, antioxidant potential, and liberation of extracellular vesicles (EVs), while it reduced the levels of autophagy. MSCs treated with 100 and 1000 μg/mL guarana or 40 μg/mL caffeine showed a decrease of cell proliferation. No treatment affected the cellular area and cell cycle of MSCs. The study shows in vitro evidence that guarana could be a promising alternative for activating MSCs to promote better cellular products for future clinical therapies.

Extubação paliativa em uma unidade de terapia intensiva cardiológica ­ um relato de caso / Palliative extubation in a cardiac intensive care unit ­ a case report

INTRODUÇÃO: A extubação paliativa é descrita como a retirada do tubo traqueal e da ventilação mecânica, quando todas as possibilidades de desmame ventilatório falharam ou o prognóstico do paciente é desfavorável. É um procedimento que evita prolongar a vida de maneira artificial e deve ser acordado juntamente com os familiares e a equipe. OBJETIVO: Relatar um caso de extubação paliativa numa unidade de terapia intensiva cardiológica. MÉTODO: Relato de caso RESULTADOS: Paciente do sexo feminino, portadora de Miocardiopatia Dilatada deu entrada no pronto-socorro em Insuficiência Cardíaca perfil C. Após a realização das medidas farmacológicas e o uso de ventilação não invasiva, não houve resposta satisfatória, evoluindo para intubação orotraqueal. A extubação ocorreu após 6 dias, porém em 24 horas houve falha de extubação, seguida de parada cardiorrespiratória (PCR) de 15 minutos. Foram realizadas todas as medidas pós PCR e mesmo a compensação clínica, não houve resposta neurológica favorável para progredir com uma nova extubação, desse modo, a equipe definiu os Cuidados Paliativos. O filho da paciente trouxe o relato de que ela não gostaria de viver de forma artificial e mesmo estando apreensivo com a possibilidade de óbito iminente após a extubação, visto que a mãe não tinha resposta favorável, ele não gostaria que mantivéssemos as medidas invasivas. Após uma conferência familiar com consenso entre a equipe e os familiares optou-se por realizar a extubação paliativa, para tal houve a suspensão de medicamentos não essenciais e foi otimizado as medicações para prevenir o estridor laríngeo. Para a extubação, foi realizado aspiração de alívio e o teste de respiração espontânea com resposta satisfatória e após 19 dias de intubação a paciente foi extubada, o procedimento ocorreu de maneira tranquila, não houve necessidade de aspiração ou suplementação com oxigênio, a paciente permaneceu com um Glasgow 6, eupneica em ar ambiente com uma fisionomia confortável, sem sinais de desconforto respiratório ou de dor. O atendimento fisioterapêutico foi mantido visando manter o conforto, a paciente teve o acompanhamento dos familiares com visita estendida e veio a falecer após 4 dias da extubação. CONCLUSÃO: A extubação paliativa ocorreu com o suporte da equipe e dos familiares, livre de dor ou desconforto e a paciente veio a falecer 4 dias após o procedimento. Esse relato de caso reforça a importância do alinhamento de condutas, da presença da família e a possibilidade de sobrevida após uma extubação paliativa.

High levels of NRF2 sensitize temozolomide-resistant glioblastoma cells to ferroptosis via ABCC1/MRP1 upregulation.

Glioblastoma patients have a poor prognosis mainly due to temozolomide (TMZ) resistance. NRF2 is an important transcript factor involved in chemotherapy resistance due to its protective role in the transcription of genes involved in cellular detoxification and prevention of cell death processes, such as ferroptosis. However, the relation between NRF2 and iron-dependent cell death in glioma is still poorly understood. Therefore, in this study, we analyzed the role of NRF2 in ferroptosis modulation in glioblastoma cells. Two human glioblastoma cell lines (U251MG and T98G) were examined after treatment with TMZ, ferroptosis inducers (Erastin, RSL3), and ferroptosis inhibitor (Ferrostatin-1). Our results demonstrated that T98G was more resistant to chemotherapy compared to U251MG and showed elevated levels of NRF2 expression. Interestingly, T98G revealed higher sensitivity to ferroptosis, and significant GSH depletion upon system xc- blockage. NRF2 silencing in T98G cells (T98G-shNRF2) significantly reduced the viability upon TMZ treatment. On the other hand, T98G-shNRF2 was resistant to ferroptosis and reverted intracellular GSH levels, indicating that NRF2 plays a key role in ferroptosis induction through GSH modulation. Moreover, silencing of ABCC1, a well-known NRF2 target that diminishes GSH levels, has demonstrated a similar collateral sensitivity. T98G-siABCC1 cells were more sensitive to TMZ and resistant to Erastin. Furthermore, we found that NRF2 positively correlates with ABCC1 expression in tumor tissues of glioma patients, which can be associated with tumor aggressiveness, drug resistance, and poor overall survival. Altogether, our data indicate that high levels of NRF2 result in collateral sensitivity on glioblastoma via the expression of its pro-ferroptotic target ABCC1, which contributes to GSH depletion when the system xc- is blocked by Erastin. Thus, ferroptosis induction could be an important therapeutic strategy to reverse drug resistance in gliomas with high NRF2 and ABCC1 expression.

White LED phototherapy as an improved treatment for neonatal jaundice.

With the aim of improving phototherapy for neonatal jaundice (hyperbilirubinemia), this study investigates the degradation of unconjugated bilirubin under irradiance by conventional light and by white, red, green, and blue LED sources in vitro. The absorption spectra of bilirubin under these different light sources are compared. The results demonstrate that white LED phototherapy promotes more efficient bilirubin degradation than conventional blue-light therapy. This study provides a basis for the design of novel phototherapy devices for the treatment of hyperbilirubinemia.

Detection of Mycoplasma Synoviae and Other Pathogens in Laying Hens with Respiratory Signs in the Rearing and Production Phases

The present study aimed to investigate the occurrence of Mycoplasma synoviae (MS), M. gallisepticum (MG), Ornitobacterium rhinotracheale (OR), Avibacterium paragallinarum (AP), Pasteurella multocida (PM) and Infectious Bronchitis Virus (IBV) in laying hens with respiratory clinical signs in two phases of production. 140 tracheal swabs and 140 blood samples were collected from laying hens in the rearing and production phases, the chickens belonged to six farms (A-F) located in the state of São Paulo, Brazil. The samples were analyzed by PCR for MG, MS, OR, AP, PM and IBV and by ELISA for MG and MS. The highest frequencies observed by PCR were for MS at farms B and C with 95 and 100% positivity, followed by MG at farms D and E with 35% and 65%, IBV with 35% at farm F and ORT with 15% at farm A. All flocks were positive for MG and MS in serology. Although MG and IBV have been detected, this can be explained by the vaccination protocols, since live attenuated vaccines are widely used for immunization against these pathogens. It was also possible to detect OR and AP thorugh PCR in some flocks. The occurrence of several etiological agents that cause respiratory diseases in laying hens was confirmed by PCR and serology, with MS being the most prevalent and being present in all farms studied.(AU)

Detection of Mycoplasma Synoviae and Other Pathogens in Laying Hens with Respiratory Signs in the Rearing and Production Phases

The present study aimed to investigate the occurrence of Mycoplasma synoviae (MS), M. gallisepticum (MG), Ornitobacterium rhinotracheale (OR), Avibacterium paragallinarum (AP), Pasteurella multocida (PM) and Infectious Bronchitis Virus (IBV) in laying hens with respiratory clinical signs in two phases of production. 140 tracheal swabs and 140 blood samples were collected from laying hens in the rearing and production phases, the chickens belonged to six farms (A-F) located in the state of São Paulo, Brazil. The samples were analyzed by PCR for MG, MS, OR, AP, PM and IBV and by ELISA for MG and MS. The highest frequencies observed by PCR were for MS at farms B and C with 95 and 100% positivity, followed by MG at farms D and E with 35% and 65%, IBV with 35% at farm F and ORT with 15% at farm A. All flocks were positive for MG and MS in serology. Although MG and IBV have been detected, this can be explained by the vaccination protocols, since live attenuated vaccines are widely used for immunization against these pathogens. It was also possible to detect OR and AP thorugh PCR in some flocks. The occurrence of several etiological agents that cause respiratory diseases in laying hens was confirmed by PCR and serology, with MS being the most prevalent and being present in all farms studied.

Differential responses of genes and enzymes associated with ROS protective responses in the sugarcane smut fungus.

The fungal pathogen Sporisorium scitamineum causes sugarcane smut disease. We have previously shown that resistant sugarcane plants induce ROS, coinciding with a delay in fungal colonization. Here, we investigated whether the fungus modifies the enzymatic antioxidant system in vitro and when colonizing sugarcane tissues in response to ROS. In vitro, the exposure to ROS did not affect cell integrity, and a combination of superoxide dismutases (SOD) and catalases (CAT) were active. In vitro, the fungus did not alter the expression of the transcriptional regulator Yap1 and the effector Pep1. The fungus activated distinct enzymes when colonizing plant tissues. Instead of CAT, S. scitamineum induced glutathione peroxidase (Gpx) expression only when colonizing smut-resistant plants. Yap1 had an earlier expression in both smut-susceptible and -resistant plants, with no apparent correlation with the expression of antioxidant genes sod, cat, gpx, or external redox imbalance. The expression of the effector pep1 was induced only in smut-resistant plants, potentially in response to ROS. These results collectively suggest that S. scitamineum copes with oxidative stress by inducing different mechanisms depending on the conditions (in vitro/in planta) and intensity of ROS. Moreover, the effector Pep1 is responsive to the stress imposed only by the sugarcane resistant genotype.

Pontine Myelinolysis Caused by Hypovolemic Hypernatremia.

Introduction. Central pontine myelinolysis is characterized by the occurrence of acute demyelinating lesions of cells in the pons secondary to abrupt oscillations of serum osmolarity. Its exact incidence is not well defined, but studies show a prevalence of 0.25 to 0.5% in the general population, 2.5% in the intensive care unit, and up to 10% in patients with risk factors, such as chronic liver disease and hepatic transplantation, alcoholism, malnutrition, diuretic therapy, electrolyte imbalance, hypoglycemia, and hyperglycemia. Case Report. A 70-year-old white female with extranodal diffuse large B-cell non-Hodgkin's lymphoma (extensive mass on the left anterior chest wall), stage IVA, developed pontine myelinolysis secondary to hypovolemic acute hypernatremia, which occurred due to diarrhea caused by chemotherapy (rituximab, cyclophosphamide, doxorubicin, and vincristine). Discussion. Pontine myelinolysis occurs most often due to the rapid correction of chronic hyponatremia. But here, we describe a case of the disease secondary to the occurrence of hypovolemic acute hypernatremia in a patient with a hematological malignancy under treatment, who was on chronic treatment with thiazide diuretics and who presented with other electrolyte disturbances as risk factors for the development of pontine myelinolysis.

Phlebotomines (Diptera: Psychodidae) and Mosquitoes (Diptera: Culicidae) Surrounding an Environmental Protection Zone in the Metropolitan Region of Natal: Use of Light-Emitting Diode (LED) Bulbs in Entomological Surveillance.

Entomological surveillance is very important for parasite and arbovirus vector control programs. Light traps with incandescent bulbs are used to attract insects and analyze the factors that contribute to the occurrence of species surrounding an environmental protection zone. Phlebotomine and mosquito abundance and their diversity were analyzed. Captures occurred monthly using six CDC light traps with two incandescent bulbs, two blue and two red LED lights. A total of 2211 phlebotomines of seven species and 4486 mosquitoes belonging to 20 taxa were captured. Different phlebotomine and mosquito species were found in the forest and peridomestic environments, with a predominance of the sand fly Evandromyia walkeri (Newstead) and the mosquito Coquillettidia venezuelensis (Theobald). There was a significant difference in the abundance of sand flies captured with the three bulbs tested, the blue bulb being the most efficient. The Shannon-Wiener diversity index showed that the trap equipped with a red LED light obtained a higher value than that of the blue LED and incandescent bulb. Analyses showed that the potential vectors and non-vectors of the two groups circulate between the forest and the peridomestic environment, suggesting an adaptation process of species to the altered environment. An incandescent light bulb can be substituted by an LED bulb, without compromising the sensitivity of the method. A blue LED is indicated for the abundant capture of mosquitoes and sand flies, while a red LED obtains a better result in terms of species diversity per capture.

Prevalência e estudo genético de Mycoplasma gallisepticum e M. synoviae em poedeiras comerciais, na região centro-oeste do estado de São Paulo, Brasil / [Prevalence and genetic study of Mycoplasma gallisepticum and M. synoviae strains from commercial laying hens in the Midwest region of Sao Paulo state, Brazil]

O objetivo deste trabalho foi estudar a prevalência de MG e MS e a filogenia das cepas circulantes, comparando-as com outras já descritas em poedeiras comerciais no Brasil. Foram coletados 140 suabes traqueais de poedeiras comerciais com sinais respiratórios em seis granjas da região centro-oeste de São Paulo. As amostras foram avaliadas por PCR, com posterior sequenciamento e análise filogenética das cepas identificadas. Das 140 amostras, 16,4% foram positivas para MG e 68,6% para MS. Houve diferença significativa nas frequências de MG e MS por granja, segundo o teste G de independência (P<0,05). Todas as cepas identificadas de MG e MS de granjas distintas apresentaram similaridade tanto pela lipoproteína para MG quanto pela região 16s rRNA para MS. Neste estudo, foi possível observar altas prevalências dos agentes estudados, sendo a de MS maior que a de MG. Foi detectada infecção mista por MG e MS em 11,4% das amostras e sabe-se que esses micoplasmas podem agir de forma sinérgica, agravando o quadro respiratório. As cepas circulantes identificadas, pela análise das regiões gênicas da lipoproteína para MG e 16S rRNA para MS, são similares em todas as granjas estudadas.(AU)

The aim of this study was to evaluate the prevalence of MG and MS and the phylogeny of the circulating strains, comparing them with others already described in commercial laying hens from Brazil. A total of 140 tracheal swabs were collected from commercial laying hens with respiratory signs in six farms from the western region of São Paulo state. The samples were analyzed by PCR with subsequent sequencing and phylogenetic analysis of the identified strains. From the 140 samples, 68.6% were positive for MS and 16.4% for MG. There was a significant difference in the frequencies of MG and MS per farm according to G Test of independence (P<0.05). All strains identified as MG and MS from distinct farms presented similarity both by lipoprotein to MG and by 16s rRNA region to MS. In this study, it was possible to observe a high prevalence of MS compared to MG. Mixed MG and MS infection was detected in 11.4% of the samples. These mycoplasmas may act synergistically, worsening the respiratory signs. The circulating strains identified by analysis of the lipoprotein for MG and 16S rRNA for MS are similar on all poultry farms studied.(AU)

Prevalência e estudo genético de Mycoplasma gallisepticum e M. synoviae em poedeiras comerciais, na região centro-oeste do estado de São Paulo, Brasil / [Prevalence and genetic study of Mycoplasma gallisepticum and M. synoviae strains from commercial laying hens in the Midwest region of Sao Paulo state, Brazil]

O objetivo deste trabalho foi estudar a prevalência de MG e MS e a filogenia das cepas circulantes, comparando-as com outras já descritas em poedeiras comerciais no Brasil. Foram coletados 140 suabes traqueais de poedeiras comerciais com sinais respiratórios em seis granjas da região centro-oeste de São Paulo. As amostras foram avaliadas por PCR, com posterior sequenciamento e análise filogenética das cepas identificadas. Das 140 amostras, 16,4% foram positivas para MG e 68,6% para MS. Houve diferença significativa nas frequências de MG e MS por granja, segundo o teste G de independência (P<0,05). Todas as cepas identificadas de MG e MS de granjas distintas apresentaram similaridade tanto pela lipoproteína para MG quanto pela região 16s rRNA para MS. Neste estudo, foi possível observar altas prevalências dos agentes estudados, sendo a de MS maior que a de MG. Foi detectada infecção mista por MG e MS em 11,4% das amostras e sabe-se que esses micoplasmas podem agir de forma sinérgica, agravando o quadro respiratório. As cepas circulantes identificadas, pela análise das regiões gênicas da lipoproteína para MG e 16S rRNA para MS, são similares em todas as granjas estudadas.(AU)

The aim of this study was to evaluate the prevalence of MG and MS and the phylogeny of the circulating strains, comparing them with others already described in commercial laying hens from Brazil. A total of 140 tracheal swabs were collected from commercial laying hens with respiratory signs in six farms from the western region of São Paulo state. The samples were analyzed by PCR with subsequent sequencing and phylogenetic analysis of the identified strains. From the 140 samples, 68.6% were positive for MS and 16.4% for MG. There was a significant difference in the frequencies of MG and MS per farm according to G Test of independence (P<0.05). All strains identified as MG and MS from distinct farms presented similarity both by lipoprotein to MG and by 16s rRNA region to MS. In this study, it was possible to observe a high prevalence of MS compared to MG. Mixed MG and MS infection was detected in 11.4% of the samples. These mycoplasmas may act synergistically, worsening the respiratory signs. The circulating strains identified by analysis of the lipoprotein for MG and 16S rRNA for MS are similar on all poultry farms studied.(AU)

Socio-epidemiological characterisation of blood donors with asymptomatic Leishmania infantum infection from three Brazilian endemic regions and analysis of the transfusional transmission risk of visceral leishmaniasis.

OBJECTIVES: This study assessed the prevalence of visceral leishmaniasis in blood donors from three endemic regions in Brazil and evaluated the risk of transmission by transfusion. BACKGROUND: Despite strong evidence of the transmission of visceral leishmaniasis through blood transfusion, the real risk, an essential condition for taking effective measures to control this serious disease, has not been determined. METHODS: A multicentre study was performed in highly endemic areas. Candidates eligible for their first blood donation underwent a socio-epidemiological interview, and blood samples were collected for enzyme-linked immunosorbent assay (ELISA) analysis, Western blot and polymerase chain reaction (PCR). Patients transfused with red blood cells or random platelet concentrates collected from these donors were also studied. The results were analysed using descriptive statistics and prevalence estimates, with significance defined as p-values <0·05. RESULTS: Of the 608 eligible donors, 37 (6·1%) were positive for visceral leishmaniasis as per ELISA. The socio-epidemiological analysis showed a significantly higher prevalence in non-Caucasians (p = 0·008). Among 296 patients who received blood components from these donors, the pre-transfusion seropositivity was 7·7%, and 13 patients received blood positive for Leishmania infantum. Six patients were followed up for 90 days, of which two (33·3%) had serological conversion at 60 days. CONCLUSIONS: The results confirm the high prevalence of L. infantum seropositivity among donors in the three regions; the seroconversion in a short period of time in two of six patients suggests the possibility of transmission of the infection by transfusion.