Understanding the mechanism of action of protease inhibitors in controlling the growth of the Candida Genus: potential candidates for development of new antifungal molecules.

There is a growing imperative for research into alternative compounds for the treatment of the fungal infections. Thus, many studies have focused on the analysis of antifungal proteins and peptides from different plant sources. Among these molecules are protease inhibitors (PIs). Previously, PIs present in the peptide-rich fractions called PEF1, PEF2 and PEF3 were identified from Capsicum chinense seeds, which have strong activity against phytopathogenic fungi. The aim of this study was to evaluate the mechanism of action and antimicrobial activity of PIs from PEF2 and PEF3 on the growth of yeasts of the genus Candida. In this work, analyses of their antimicrobial activity and cell viability were carried out. Subsequently, the mechanism of action by which the PIs cause the death of the yeasts was evaluated. Cytotoxicity was assessed in vitro by erythrocytes lysis and in vivo in Galleria mellonella larvae. PEF2 and PEF3 caused 100% of the growth inhibition of C. tropicalis and C. buinensis. For C. albicans inhibition was approximately 60% for both fractions. The PEF2 and PEF3 caused a reduction in mitochondrial functionality of 54% and 46% for C. albicans, 26% and 30% for C. tropicalis, and 71% and 68% for C. buinensis, respectively. These fractions induced morphological alterations, led to membrane permeabilization, elevated ROS levels, and resulted in necrotic cell death in C. tropicalis, whilst demonstrating low toxicity toward host cells. From the results obtained here, we intend to contribute to the understanding of the action of PIs in the control of fungal diseases of medical importance.

Transcriptome-based Phylogeny of the Semi-aquatic Bugs (Hemiptera: Heteroptera: Gerromorpha) Reveals Patterns of Lineage Expansion in a Series of New Adaptive Zones.

Key innovations enable access to new adaptive zones and are often linked to increased species diversification. As such, innovations have attracted much attention, yet their concrete consequences on the subsequent evolutionary trajectory and diversification of the bearing lineages remain unclear. Water striders and relatives (Hemiptera: Heteroptera: Gerromorpha) represent a monophyletic lineage of insects that transitioned to live on the water-air interface and that diversified to occupy ponds, puddles, streams, mangroves and even oceans. This lineage offers an excellent model to study the patterns and processes underlying species diversification following the conquest of new adaptive zones. However, such studies require a reliable and comprehensive phylogeny of the infraorder. Based on whole transcriptomic datasets of 97 species and fossil records, we reconstructed a new phylogeny of the Gerromorpha that resolved inconsistencies and uncovered strong support for previously unknown relationships between some important taxa. We then used this phylogeny to reconstruct the ancestral state of a set of adaptations associated with water surface invasion (fluid locomotion, dispersal and transition to saline waters) and sexual dimorphism. Our results uncovered important patterns and dynamics of phenotypic evolution, revealing how the initial event of water surface invasion enabled multiple subsequent transitions to new adaptive zones on the water surfaces. This phylogeny and the associated transcriptomic datasets constitute highly valuable resources, making Gerromorpha an attractive model lineage to study phenotypic evolution.

Lipid droplet accumulation occurs early following Salmonella infection and contributes to intracellular bacterial survival and replication.

Salmonellosis is a public health problem caused by Salmonella sp., a highly adapted facultative intracellular pathogen. After internalization, Salmonella sp. Manipulates several host processes, mainly through the activation of the type III secretion system (T3SS), including modification of host lipid metabolism and lipid droplet (LD) accumulation. LDs are dynamic and complex lipid-rich organelles involved in several cellular processes. The present study investigated the mechanism involved in LD biogenesis in Salmonella-infected macrophages and its role in bacterial pathogenicity. Here, we reported that S. Typhimurium induced a rapid time-dependent increase of LD formation in macrophages. The LD biogenesis was demonstrated to depend on Salmonella's viability and SPI1-related T3SS activity, with the participation of Toll-Like Receptor (TLR) signaling. We also observed that LD accumulation occurs through TLR2-dependent signaling and is counter-regulated by TLR4. Last, the pharmacologic modulation of LD formation by inhibiting diacylglycerol O-acyltransferase 1 (DGAT1) and cytosolic phospholipase A2 (cPLA2) significantly reduced the intracellular bacterial proliferation and impaired the prostaglandin E2 (PGE2 ) synthesis. Collectively, our data suggest the role of LDs on S. typhimurium intracellular survival and replication in macrophages. This data set provides new perspectives for future investigations about LDs in host-pathogen interaction.

300 Million years of coral treaders (Insecta: Heteroptera: Hermatobatidae) back to the ocean in the phylogenetic context of Arthropoda.

Among hundreds of insect families, Hermatobatidae (commonly known as coral treaders) is one of the most unique. They are small, wingless predaceous bugs in the suborder Heteroptera. Adults are almost black in colour, measuring about 5 mm in body length and 3 mm in width. Thirteen species are known from tropical coral reefs or rocky shores, but their origin and evolutionary adaptation to their unusual marine habitat were unexplored. We report here the genome and metagenome of Hermatobates lingyangjiaoensis, hitherto known only from its type locality in the South China Sea. We further reconstructed the evolutionary history and origin of these marine bugs in the broader context of Arthropoda. The dated phylogeny indicates that Hexapoda diverged from their marine sister groups approximately 498 Ma and that Hermatobatidae originated 192 Ma, indicating that they returned to an oceanic life some 300 Myr after their ancestors became terrestrial. Their origin is consistent with the recovery of tropical reef ecosystems after the end-Triassic mass extinction, which might have provided new and open niches for them to occupy and thrive. Our analyses also revealed that both the genome changes and the symbiotic bacteria might have contributed to adaptations necessary for life in the sea.

Environmental vulnerability evolution in the Brazilian Amazon.

Decision making and environmental policies are mainly based on propensity level to impact in the area. The propensity level can be determined through artificial intelligence techniques included in geotechnological universe. Thus, this study aimed to determine the areas of greatest vulnerability to human activities, in Amazon biome, through MODIS images of Land use and land cover (LULC) from the 2001 and 2013. Remote sensing, Euclidean distance, Fuzzy logic, AHP method and analysis of net variations were applied to specialize the classes of vulnerability in the states belonging to the Amazon Biome. From the results, it can be seen that the class that most evolved in a positive net gain during the evaluated period was "very high" and the one that most reduced was "high", showing that there was a transition from "high" to "very high" risk areas. The states with the largest areas under "very high" risk class were Mato Grosso (101,100.10 km2) and Pará (81,010.30 km2). It is concluded that the application of remote sensing techniques allows the determination and assessment of the environmental vulnerability evolution. Mitigation measures urgently need to be implemented in the Amazon biome. The methodology can be extended to any other area of the planet.

Cooption of the pteridine biosynthesis pathway underlies the diversification of embryonic colors in water striders.

Naturalists have been fascinated for centuries by animal colors and color patterns. While widely studied at the adult stage, we know little about color patterns in the embryo. Here, we study a trait consisting of coloration that is specific to the embryo and absent from postembryonic stages in water striders (Gerromorpha). By combining developmental genetics with chemical and phylogenetic analyses across a broad sample of species, we uncovered the mechanisms underlying the emergence and diversification of embryonic colors in this group of insects. We show that the pteridine biosynthesis pathway, which ancestrally produces red pigment in the eyes, has been recruited during embryogenesis in various extraocular tissues including antennae and legs. In addition, we discovered that this cooption is common to all water striders and initially resulted in the production of yellow extraocular color. Subsequently, 6 lineages evolved bright red color and 2 lineages lost the color independently. Despite the high diversity in colors and color patterns, we show that the underlying biosynthesis pathway remained stable throughout the 200 million years of Gerromorpha evolutionary time. Finally, we identified erythropterin and xanthopterin as the pigments responsible for these colors in the embryo of various species. These findings demonstrate how traits can emerge through the activation of a biosynthesis pathway in new developmental contexts.

An analysis of the quality indicators of the organ donation process in São Paulo, Brazil.

BACKGROUND: Few studies have investigated whether quality initiatives in the process of organ donation yield better results of the organ donation process. OBJECTIVE: To analyze whether the indicators of the organ donation process in Brazilian hospitals meet the standards established by the Organ Donation European Quality System (ODEQUS). DESIGN: We evaluated the quality of the organ donation in a selected group of Brazilian hospitals served by the Nucleus of Organ Procurement (NOP) using standards of the ODEQUS. RESULTS: Structural and process indicators had 100% conformity. Indicators of results showed a family consent rate of 61% (29% lower than ODEQUS goal); a conversion rate of potential donors to effective donors of 47% (28% below the goal); and a 12% rate of sudden cardiac arrest (higher than the quality limit). CONCLUSIONS: Our findings highlight the importance for the development of quality initiatives in identifying gaps and weaknesses in the process that should be corrected or even restructured, therefore maximizing the number of donors and organs transplanted. Hospitals that participate in the NOP process met 61% of the quality indicators proposed by ODEQUS. Identification of potential donors, family consent, conversion, and sudden cardiac arrest rates are areas that did not conform to ODEQUS standard and revealed a great opportunity for improvement.

Cytogenetics analysis and testis morphology of aquatic species of the families Belostomatidae, Gelastocoridae, Gerridae, Notonectidae, and Veliidae (Heteroptera).

The Heteroptera have holocentric chromosomes with kinetic activity restricted to the end of chromosomes. The first meiotic division is reductional for the autosomes and equational for the sexual. Only a few species of this suborder have been analyzed. In this study, we observed the morphologies of the testes of the Heteroptera species Belostoma anurum (Herrich-Schäffer, 1948), Belostoma micantulum (Stal, 1858), Gelastocoris angulatus (Melin, 1929), Gelastocoris flavus flavus (Guérin-Méneville, 1844), Rheumatobates crassifemur crassifemur (Esaki, 1926), Buenoa amnigenus (White, 1879), Buenoa unguis (Truxal, 1953), Martarega brasiliensis (Truxal, 1949), Martarega membranácea (White, 1879), Martarega uruguayensis (Berg, 1883), Rhagovelia tenuipes (Champion, 1898) and Rhagovelia zela (Drake, 1959). We found that the testes of these species can be round, round/spiral, or elongated/spiral. The size of the prophase I cells was found to vary, with the smallest ones being detected in B. micantulum and Rha. zela, the largest in G. f. flavus, and ones of intermediate size in R. c. crassifemur and M. brasiliensis. With respect to the chromosome complement, we verified the presence of 2n = 16: (14A+XY, B. micantulum and G. angulatus), 21: (20A+X0, R. c. crassifemur), 23: (22A+X0, Rha. zela and Rha. tenuipes), 25: (24A+X0, Bu. amnigenus and Bu. unguis; 22A+2m+X0, M. membranacea), 27: (24A+2m+X0, M. brasiliensis and M. uruguayensis), 29: (26A+X1X2Y, B. anurum), and 35: (30A+X1X2X3X4Y, G. f. flavus). We found that the features of spermatogenesis in these species are similar to those of other previously described Heteroptera species, differing only in testicular morphology, chromosome number, and sex chromosome system.

Two new species of Rhagovelia (Hemiptera: Heteroptera: Veliidae) from Costa Rica, with a key and new records from the country.

Rhagovelia danpolhemi sp. nov., from the Caribbean Slope of Costa Rica and R. springerae sp. nov., from the northern portion of the country, are described, illustrated, compared with similar species, and included in a key to species recorded from the country. Rhagovelia calopa Drake & Harris, R. spinosa Gould, and R. tantilla Drake & Harris are recorded for the first time from Costa Rica. New provincial records are provided for R. acuminata Bacon, R. angustipes Uhler, R. armata (Burmeister), R. bisignata Bacon, R. crassipes Champion, R. elegans Uhler, R. femoralis Champion, R. solida Bacon, R. spinigera Champion, R. tenuipes Champion, and R. uncinata Champion. Additional distributional information is given for these species and for R. formosa Bacon and R. scabra Bacon.

New species and notes on Microveliinae from northern South America (Hemiptera: Heteroptera: Veliidae).

Euvelia mazzucconiae sp. nov., from northern Colombia is described, illustrated, and included in an updated identification key to the species of the genus. A map with the known geographical distribution of the species of Euvelia is presented. Additionally, two species from Colombia previously placed in the genus Paravelia are transferred to Microvelia and a species of the latter genus is synonymized, such that Paravelia acantha Padilla-Gil, 2013 = Microvelia acantha (Padilla-Gil, 2013) comb. nov., Paravelia fanera Padilla-Gil, 2013 = Microvelia fanera (Padilla-Gil, 2013) comb. nov., and Microvelia amrishi Makhan 2014 = Microvelia mimula White 1879 syn. nov.

A new Rhagovelia (Hemiptera: Heteroptera: Veliidae) from the Brazilian Amazon, with a key to species of the robusta group known from the country.

Rhagovelia zecai sp. nov., from the Brazilian Amazon, is described, illustrated, compared with similar species, and included in a key to species of the robusta group recorded from the country.

The genus Paravelia Breddin, 1898 (Hemiptera: Heteroptera: Veliidae) in Brazil, with descriptions of eight new species.

Eight new species of Paravelia Breddin, 1898 from Brazil are described and illustrated: P. amapaensis sp. nov. from Amapá State, P. bipunctata sp. nov. from Minas Gerais, Mato Grosso and Mato Grosso do Sul States, P. bilobata sp. nov. and P. polhemusi sp. nov. from Mato Grosso State, P. bahiana sp. nov. from Bahia State, P. lacrymosa sp. nov. from Minas Gerais State, P. micromaculata sp. nov. from Maranhão State, and P. ornata sp. nov. from Amazonas State. The genus is redescribed, with photos of the dorsal view for nineteen species: P. basalis (Spinola), P. biae Spangler, P. boliviana Breddin, P. bullialata Polhemus & Polhemus, P. capillata (Drake& Harris), P. capixaba Moreira, Nessimian & Rúdio, P. conata (Hungerford), P. dilatata Polhemus & Polhemus, P. foveata Polhemus & Polhemus, P. itatiayana (Drake), P. lanemeloi Moreira & Barbosa, P. manausana Polhemus & Polhemus, P. nieseri Moreira & Barbosa, P. platensis (Berg), P. recens (Drake & Harris), P. rotundanotata (Hungerford), P. spinifera Polhemus & Polhemus, P. splendoris (Drake & Harris) and P. williamsi (Hungerford). Three of these species are recorded for the first time from Brazil: P. platensis, P. spinifera and P. williamsi. Also, the macropterous forms of P. capixaba and P. dilatata are described. Lastly, an identification key to the 36 species of Paravelia recorded from Brazil and a checklist of described species are presented.

Foveavelia, a new South American genus of Veliinae (Hemiptera: Heteroptera: Veliidae).

Background: Semiaquatic bugs (Hemiptera: Heteroptera: Gerromorpha) are distributed worldwide and play fundamental roles in limnic ecosystems. They are the most successful group of organisms to occupy the air-water interface, are important models to study ecology and evolution, and can be relevant tools in biomonitoring. Veliidae is the second most speciose family of semiaquatic bugs, but its internal classification, including subfamilies and genera, is artificial and based on symplesiomorphies. One of these non-monophyletic entities is Paravelia Breddin, 1898, the largest genus in the subfamily Veliinae. Results: In an effort to better classify the Veliinae, we describe Foveavelia to hold five South American species previously placed in Paravelia. The new genus is characterized by the following combination of features: unusual coarse cuticular punctures throughout the thorax and abdomen; a pair of small, frosty, pubescent areas formed by a very dense layer of short setae on the anterior lobe of the pronotum; fore tibial grasping comb present only in males; middle tibia with a row of elongate dark-brown trichobothria-like setae on the distal third, decreasing in size distally; macropterous specimens with the apical macula of the forewings elongate and constricted at mid-length, reaching the wing apex; and the male proctiger with a pair of anterodorsal projections. Besides the description, a key to the species of Foveavelia is provided, accompanied by illustrations and a species distribution map.

A synopsis of the genus Cylindrostethus Fieber 1861 (Insecta: Hemiptera: Gerridae).

Cylindrostethus Fieber, 1861 is one of the most striking genera of water striders (Insecta: Hemiptera: Gerridae) and has Pantropical distribution. Members of this group can be recognized by the very long, cylindrical body; the short antennomere IV; the short labium not reaching the mesosternum; and by characteristics of the abdomen of males and females. Although Neotropical representatives of the genus have been revised, there are pending taxonomic issues related to this fauna, and that of the Eastern Hemisphere has been barely studied in recent years. Here, we present a short note about the authorship of Cylindrostethus, an updated key to all species of the genus, a new synonymy, and the description of a previously unknown macropterous male of C. hungerfordi Drake and Harris.

Exploring the physicochemical properties of the integration of Tristearoyl uridine in Langmuir monolayers: An approach to cell membrane modeling for prodrugs.

Understanding the mechanisms by which drugs interact with cell membranes is crucial for unraveling the underlying biochemical and biophysical processes that occur on the surface of these membranes. Our research focused on studying the interaction between an ester-type derivative of tristearoyl uridine and model cell membranes composed of lipid monolayers at the air-water interface. For that, we selected a specific lipid to simulate nontumorigenic cell membranes, namely 1,2-dihexadecanoyl-sn-glycero-3-phospho-l-serine. We noted significant changes in the surface pressure-area isotherms, with a noticeable shift towards larger areas, which was lower than expected for ideal mixtures, indicating monolayer condensation. Furthermore, the viscoelastic properties of the interfacial film demonstrated an increase in both the elastic and viscous parameters for the mixed film. We also observed structural alterations using vibrational spectroscopy, which revealed an increase in the all-trans to gauche conformers ratio. This confirmed the stiffening effect of the prodrug on the lipid monolayer. In summary, this study indicates that this lipophilic prodrug significantly impacts the lipid monolayer's thermodynamic, rheological, electrical, and molecular characteristics. This information is crucial for understanding how the drug interacts with specific sites on the cellular membrane. It also has implications for drug delivery, as the drug's passage into the cytosol may involve traversing the lipid bilayer.

Description of a New Species and New Records of Naucoridae (Hemiptera: Heteroptera: Nepomorpha) from Eastern Brazil.

The aquatic bug family Naucoridae (Hemiptera: Heteroptera: Nepomorpha) is currently represented in Brazil by 68 species. Although the diversity of the family has been the target of several recent studies, large areas of the country are still unexplored and several species that have been deposited in entomological collections are waiting for a formal description. Aiming to fill these knowledge gaps, a series of expeditions were carried out in six states of eastern Brazil between 2018 and 2023: Alagoas, Bahia, Ceará, Espírito Santo, Pernambuco, and Sergipe. The fieldwork targeted protected areas, but surrounding regions were also explored. The material obtained, in addition to specimens previously deposited in a national entomological collection, revealed the existence of Australambrysus margaritifer Jordão, Santos and Moreira, a new species herein described, and new records for other 11 species and two subspecies belonging to the genera Carvalhoiella De Carlo, 1963, Limnocoris Stål, 1876, Maculambrysus Reynoso-Velasco and Sites 2021, and Pelocoris Stål, 1876.

Synergistic action of synthetic peptides and amphotericin B causes disruption of the plasma membrane and cell wall in Candida albicans.

The objective of this work was to evaluate the combination of synthetic peptides based on the γ-core motif of defensin PvD1 with amphotericin B (AmB) at different concentrations against Candida albicans. We applied the checkerboard assay using different concentrations of the commercial drug AmB and the synthetic peptides γ31-45PvD1++ and γ33-41PvD1++ against C. albicans, aiming to find combinations with synergistic interactions. Between these two interactions involving γ31-45PvD1++ and AmB, an additive effect was observed. One such interaction occurred at concentrations of 0.009 µM of peptide γ31-45PvD1++ and 13.23 µM of AmB and another condition of 0.019 µM of peptide γ31-45PvD1++ and 6.61 µM of AmB. The other two concentrations of the interaction showed a synergistic effect in the combination of synthetic peptide γ31-45PvD1++ and AmB, where the concentrations were 1.40 µM peptide γ31-45PvD1++ and 0.004 µM AmB and 0.70 µM γ31-45PvD1++ peptide and 0.002 µM AmB. We proceeded with analysis of the mechanism of action involving synergistic effects. This examination unveiled a range of impactful outcomes, including the impairment of mitochondrial functionality, compromise of cell wall integrity, DNA degradation, and a consequential decline in cell viability. We also observed that both synergistic combinations were capable of causing damage to the plasma membrane and cell wall, causing leakage of intracellular components. This discovery demonstrates for the first time that the synergistic combinations found between the synthetic peptide γ31-45PvD1++ and AmB have an antifungal effect against C. albicans, acting on the integrity of the plasma membrane and cell wall.

Defensin-like peptides from Capsicum chinense induce increased ROS, loss of mitochondrial functionality, and reduced growth of the fungus Colletotrichum scovillei.

In the present study, we identified and characterized two defensin-like peptides in an antifungal fraction obtained from Capsicum chinense pepper fruits and inhibited the growth of Colletotrichum scovillei, which causes anthracnose. AMPs were extracted from the pericarp of C. chinense peppers and subjected to ion exchange, molecular exclusion, and reversed-phase in a high-performance liquid chromatography system. We investigated the endogenous increase in reactive oxygen species (ROS), the loss of mitochondrial functioning, and the ultrastructure of hyphae. The peptides obtained from the G3 fraction through molecular exclusion chromatography were subsequently fractionated using reverse-phase chromatography, resulting in the isolation of fractions F1, F2, F3, F4, and F5. The F1-Fraction suppressed C. scovillei growth by 90, 70.4, and 44% at 100, 50, and 25 µg mL-1, respectively. At 24 h, the IC50 and minimum inhibitory concentration were 21.5 µg mL-1 and 200 µg mL-1, respectively. We found an increase in ROS, which may have resulted in an oxidative burst, loss of mitochondrial functioning, and cytoplasm retraction, as well as an increase in autophagic vacuoles. MS/MS analysis of the F1-Fraction indicated the presence of two defensin-like proteins, and we were able to identify the expression of three defensin sequences in our C. chinense fruit extract. The F1-Fraction was also found to inhibit the activity of insect α-amylases. In summary, the F1-Fraction of C. chinense exhibits antifungal activity against a major pepper pathogen that causes anthracnose. These defensin-like compounds are promising prospects for further research into antifungal and insecticide biotechnology applications. © 2024 Society of Chemical Industry.

Accumulation of Lipid Droplets Induced by Listeria monocytogenes in Macrophages: Implications for Survival and Evasion of Innate Immunity.

Listeriosis, caused by Listeria monocytogenes (L.m.), poses a significant public health concern as one of the most severe foodborne diseases. The pathogenesis of L.m. involves critical steps such as phagosome rupture and escape upon internalization. Throughout infection, L.m. influences various host processes, including lipid metabolism pathways, yet the role of lipid droplets (LDs) remains unclear. Here, we reported a rapid, time-dependent increase in LD formation in macrophages induced by L.m. LD biogenesis was found to be dependent on L.m. viability and virulence genes, particularly on the activity of the pore-forming protein listeriolysin O (LLO). The prevention of LD formation by inhibiting diacylglycerol O-acyltransferase 1 (DGAT1) and cytosolic phospholipase A2 (cPLA2) significantly reduced intracellular bacterial survival, impaired prostaglandin E2 (PGE2) synthesis, and decreased IL-10 production. Additionally, inhibiting LD formation led to increased levels of TNF-α and IFN-ß. Collectively, our data suggest a role for LDs in promoting L.m. cell survival and evasion within macrophages.

Contamination in a brazilian river: a risk of exposure to untreated effluents.

Due to natural or anthropogenic activities, various chemical contaminants, such as toxic metals and organic compounds, enter water systems and can have multiple deleterious effects on many living organisms. The Todos os Santos River (TSR) in Brazil receives a high discharge of untreated effluents from various sources. The purpose of this study was to determine trace element concentrations in water samples from the TSR, evaluate the toxicological effects of these elements by using the (onion) test, and verify the physical-chemical parameters to establish the risk of exposure related to the contaminated river. The samples were taken in 2011 at six selected points (two upstream of an urban area, two in the urban area, and two downstream of the urban area) with physical-chemical parameters evaluated. The concentrations of Al (21.63-1688.84 µg L), P (38.59-1760.87 µg L), and Fe (478.9-8296.3 µg L) were above the maximum levels permitted under Brazilian law (and the World Health Organization); little dissolved O was observed. Based on the test, genotoxic and mutagenic effects may occur in the river and could be due to anthropogenic increases in metal content. These important results have serious implications for the ecosystem as well as the health of the people who use the water.