Vaccine Strategies for Cryptococcus neoformans.

Cryptococcus neoformans infections are a major worldwide concern as current treatment strategies are becoming less effective in alleviating the infection. The most extreme and fatal cases are those of immunocompromised individuals. Clinical treatments for cryptococcosis are limited to a few classes of approved drugs, and due to a rise in drug resistance, these drugs are becoming less effective. Therefore, it is essential to develop innovative ways to control this infection. Vaccinations have emerged as a safe, viable, and cost-effective solution to treat a number of diseases over the years. Currently, there are no clinically available vaccines to treat cryptococcal infections, but a number of studies have shown promising results in animal models. Here, we present step-by-step experimental protocols using live-attenuated or heat-killed C. neoformans cells as a vaccination strategy in a preventive or in a therapeutic murine model of cryptococcosis.

Combination therapy of itraconazole and an acylhydrazone derivative (D13) for the treatment of sporotrichosis in cats.

Acylhydrazone (AH) derivatives represent a novel category of anti-fungal medications that exhibit potent activity against Sporothrix sp., both in vitro and in a murine model of sporotrichosis. In this study, we demonstrated the anti-fungal efficacy of the AH derivative D13 [4-bromo-N'-(3,5-dibromo-2-hydroxybenzylidene)-benzohydrazide] against both planktonic cells and biofilms formed by Sporothrix brasiliensis. In a clinical study, the effect of D13 was then tested in combination with itraconazole (ITC), with or without potassium iodide, in 10 cats with sporotrichosis refractory to the treatment of standard of care with ITC. Improvement or total clinical cure was achieved in five cases after 12 weeks of treatment. Minimal abnormal laboratory findings, e.g., elevation of alanine aminotransferase, were observed in four cats during the combination treatment and returned to normal level within a week after the treatment was ended. Although highly encouraging, a larger and randomized controlled study is required to evaluate the effectiveness and the safety of this new and exciting drug combination using ITC and D13 for the treatment of feline sporotrichosis. IMPORTANCE: This paper reports the first veterinary clinical study of an acylhydrazone anti-fungal (D13) combined with itraconazole against a dimorphic fungal infection, sporotrichosis, which is highly endemic in South America in animals and humans. Overall, the results show that the combination treatment was efficacious in ~50% of the infected animals. In addition, D13 was well tolerated during the course of the study. Thus, these results warrant the continuation of the research and development of this new class of anti-fungals.

SAR study of N'-(Salicylidene)heteroarenecarbohydrazides as promising antifungal agents.

Clinically available antifungal drugs have therapeutic limitations due to toxicity, narrow spectrum of activity, and intrinsic or acquired drug resistance. Thus, there is an urgent need for new broad-spectrum antifungal agents with low toxicity and a novel mechanism of action. In this context, we have successfully identified several highly promising lead compounds, i.e., aromatic N'-(salicylidene)carbohydrazides, exhibiting excellent antifungal activities against Cryptococcus neoformans, Candida albicans, Aspergillus fumigatus and several other fungi both in vitro and in vivo. Building upon these highly promising results, 71 novel N'-(salicylidene)heteroarenecarbohydrazides 5 were designed, synthesized and their antifungal activities examined against fungi. Based on the SAR study, four highly promising lead compounds, i.e., 5.6a, 5.6b, 5.7b and 5.13a were identified, which exhibited excellent potency against C. neoformans, C. albicans and A. fumigatus, and displayed impressive time-kill profiles against C. neoformans with exceptionally high selectivity indices (SI ≥ 500). These four lead compounds also showed synergy with clinical antifungal drugs, fluconazole, caspofungin (CS) and amphotericin B against C. neoformans. For the SAR study, we also employed quantitative structure-activity relationship (QSAR) analysis by taking advantage of the accumulated data on a large number of aromatic and heteroaromatic N'-(salicylidene)carbohydrazides, which successfully led to rational design and selection of promising compounds for chemical synthesis and biological evaluation.

Lipidome remodeling in response to nutrient replenishment requires the tRNA modifier Deg1/Pus3 in yeast.

In the yeast Saccharomyces cerevisiae, the absence of the pseudouridine synthase Pus3/Deg1, which modifies tRNA positions 38 and 39, results in increased lipid droplet (LD) content and translational defects. In addition, starvation-like transcriptome alterations and induced protein aggregation were observed. In this study, we show that the deg1 mutant increases specific misreading errors. This could lead to altered expression of the main regulators of neutral lipid synthesis which are the acetyl-CoA carboxylase (Acc1), an enzyme that catalyzes a key step in fatty acid synthesis, and its regulator, the Snf1/AMPK kinase. We demonstrate that upregulation of the neutral lipid content of LD in the deg1 mutant is achieved by a mechanism operating in parallel to the known Snf1/AMPK kinase-dependent phosphoregulation of Acc1. While in wild-type cells removal of the regulatory phosphorylation site (Ser-1157) in Acc1 results in strong upregulation of triacylglycerol (TG), but not steryl esters (SE), the deg1 mutation more specifically upregulates SE levels. In order to elucidate if other lipid species are affected, we compared the lipidomes of wild type and deg1 mutants, revealing multiple altered lipid species. In particular, in the exponential phase of growth, the deg1 mutant shows a reduction in the pool of phospholipids, indicating a compromised capacity to mobilize acyl-CoA from storage lipids. We conclude that Deg1 plays a key role in the coordination of lipid storage and mobilization, which in turn influences lipid homeostasis. The lipidomic effects in the deg1 mutant may be indirect outcomes of the activation of various stress responses resulting from protein aggregation.

Role of sphingolipids in the host-pathogen interaction.

Fungal pathogens have been under the spotlight as their expanding geographic range combined with their potential harm to vulnerable populations turns them into increasingly threats to public health. Therefore, it is ultimately important to unveil the mechanisms associated with their infection process for further new treatment discovery. With this purpose, sphingolipid-based research has gained attention over the last years as these molecules have key properties that can regulate fungal pathogenicity. Here we discuss some of these properties as well as their role in fungal diseases, focusing on the subgroup of glycosphingolipids, as they represent promising molecules for drug discovery and for the development of fungal vaccines.

Antimicrobial Spectrum of Activity and Mechanism of Action of Linear Alpha-Helical Peptides Inspired by Shrimp Anti-Lipopolysaccharide Factors.

Shrimp antilipopolysaccharide factors (ALFs) form a multifunctional and diverse family of antimicrobial host defense peptides (AMPs) composed of seven members (groups A to G), which differ in terms of their primary structure and biochemical properties. They are amphipathic peptides with two conserved cysteine residues stabilizing a central ß-hairpin that is understood to be the core region for their biological activities. In this study, we synthetized three linear (cysteine-free) peptides based on the amino acid sequence of the central ß-hairpin of the newly identified shrimp (Litopenaeus vannamei) ALFs from groups E to G. Unlike whole mature ALFs, the ALF-derived peptides exhibited an α-helix secondary structure. In vitro assays revealed that the synthetic peptides display a broad spectrum of activity against both Gram-positive and Gram-negative bacteria and fungi but not against the protozoan parasites Trypanosoma cruzi and Leishmania (L.) infantum. Remarkably, they displayed synergistic effects and showed the ability to permeabilize bacterial membranes, a mechanism of action of classical AMPs. Having shown low cytotoxicity to THP-1 human cells and being active against clinical multiresistant bacterial isolates, these nature-inspired peptides represent an interesting class of bioactive molecules with biotechnological potential for the development of novel therapeutics in medical sciences.

On the wave of the crustin antimicrobial peptide family: From sequence diversity to function.

Crustins represent the largest and most diverse family of antimicrobial peptides (AMPs) found in crustaceans. They are classically defined as disulfide-rich peptides/polypeptides holding a typical Whey Acidic Protein (WAP) domain at the C-terminal end. This WAP domain has eight cysteine residues forming a tightly packed structure, the four-disulfide core (4DSC) motif, that is also found in other proteins displaying protease inhibitory properties. Crustins are highly diverse in terms of primary structure, size and biochemical features, thus exhibiting a series of biological functions beyond their antimicrobial properties. In order to better categorize the distinct crustin members, different classification systems have been proposed. In this review, we discuss the current classification systems and explore the biological implication of the impressive molecular diversity of this unique AMP family. We also summarize the recent findings on the role of these effectors in crustacean immunity and homeostasis as well as in host-microbe interactions.

Expression of Transient Receptor Potential Channel Genes and Their Isoforms in Alpha-Cells and Beta-Cells of Human Islets of Langerhans.

Expression of the transient receptor potential (TRP) channel genes and their isoforms in the alpha-cells and the beta-cells of the human islets of Langerhans has not been studied in detail. In this study, we have analyzed the RNA sequencing data obtained from purified human alpha-cells and beta-cells to identify the genes and their isoforms that are expressed differentially in these two cell types. We found that TRPC1, TRPC4, TRPC7, TRPM3, and TRPML1 were differentially expressed in these two cell types. TRPC1, TRPM3, and TRPML1 were expressed at a higher level in the beta-cells than in the alpha-cells. TRPC4 and TRPC7 were expressed at a higher level in the alpha-cells than in the beta-cells. The TRPC4-206 isoform was expressed at a 45-fold higher level in the alpha-cells compared to the beta-cells. Expression of TRPM3-202 was 200-fold and TRPM3-209 was 25-fold higher in the beta-cells than in the alpha-cells. Our study has demonstrated the relative abundance of expression of the TRP channel genes and their isoforms in the human alpha-cells and the beta-cells.

Microevolution of Trypanosoma cruzi reveals hybridization and clonal mechanisms driving rapid genome diversification.

Protozoa and fungi are known to have extraordinarily diverse mechanisms of genetic exchange. However, the presence and epidemiological relevance of genetic exchange in Trypanosoma cruzi, the agent of Chagas disease, has been controversial and debated for many years. Field studies have identified both predominantly clonal and sexually recombining natural populations. Two of six natural T. cruzi lineages (TcV and TcVI) show hybrid mosaicism, using analysis of single-gene locus markers. The formation of hybrid strains in vitro has been achieved and this provides a framework to study the mechanisms and adaptive significance of genetic exchange. Using whole genome sequencing of a set of experimental hybrids strains, we have confirmed that hybrid formation initially results in tetraploid parasites. The hybrid progeny showed novel mutations that were not attributable to either (diploid) parent showing an increase in amino acid changes. In long-term culture, up to 800 generations, there was a variable but gradual erosion of progeny genomes towards triploidy, yet retention of elevated copy number was observed at several core housekeeping loci. Our findings indicate hybrid formation by fusion of diploid T. cruzi, followed by sporadic genome erosion, but with substantial potential for adaptive evolution, as has been described as a genetic feature of other organisms, such as some fungi.

Madelung Deformity - Esthetic and Functional Outcomes from the Surgical Treatment with Distal Radial Dome Osteotomy and Vickers Ligament Section.

Objective The present study aimed to evaluate esthetic and functional outcomes from the surgical treatment of Madelung deformity in children. Method This is a retrospective study of pediatric patients with Madelung deformity who were surgically treated with dome osteotomy of the distal radius and Vickers ligament section from 2015 to 2018. Patients with a minimum postoperative follow-up period of 12 months were included. Demographic data, surgical technique, clinical and radiographic outcomes were analyzed. Pre and postoperative radiographic evaluation consisted of ulnar tilt, lunate subsidence lunate fossa angle, and palmar carpal displacement measurements. The postoperative clinical evaluation consisted of ranges of motion of the wrist, visual analog scale (VAS) and Disabilities of the Arm, Shoulder and Hand (DASH) score. Results Four patients were included, two with idiopathic Madelung deformity and two with bone dysplasia. All patients were females and presented bilateral disease. Six wrists were operated on. The median age at surgery was 15.5 years old, and the median postoperative follow-up time was of 37.5 months. The postoperative radiographic analysis revealed an average correction of 8.8 ± 7.5° for ulnar tilt, 3.0 ± 3.9 mm for lunate subsidence 8.2 ± 6.6° for lunate fossa angle, and 4.7 ± 2.6 mm for palmar carpal displacement. Average postoperative ranges of motion of the wrist joint were 75.8 ± 3.4° for flexion, 62.5 ± 14.1° for extension, 25.7 ± 2.9° for radial deviation, 40.0 ± 2.9° for ulnar deviation, 88.3 ± 2.4° for pronation, and 82.5 ± 2.5° for supination. The median VAS was 1 for residual pain, 0 for functional deficit, 0 for esthetic impairment, and 10 for recommending the surgical procedure. The median DASH score was 0. Conclusion Madelung deformity treatment using dome osteotomy of the distal radius and Vickers ligament section results in excellent esthetic and functional outcomes.

Madelung Deformity - Esthetic and Functional Outcomes from the Surgical Treatment with Distal Radial Dome Osteotomy and Vickers Ligament Section / Deformidade de Madelung - Resultado estético e funcional do tratamento cirúrgico com osteotomia em cúpula do rádio distal e secção do ligamento de Vickers

Abstract Objective The present study aimed to evaluate esthetic and functional outcomes from the surgical treatment of Madelung deformity in children. MethodThis is a retrospective study of pediatric patients with Madelung deformity who were surgically treated with dome osteotomy of the distal radius and Vickers ligament section from 2015 to 2018. Patients with a minimum postoperative follow-up period of 12 months were included. Demographic data, surgical technique, clinical and radiographic outcomes were analyzed. Pre and postoperative radiographic evaluation consisted of ulnar tilt, lunate subsidence lunate fossa angle, and palmar carpal displacement measurements. The postoperative clinical evaluation consisted of ranges of motion of the wrist, visual analog scale (VAS) and Disabilities of the Arm, Shoulder and Hand (DASH) score. Results Four patients were included, two with idiopathic Madelung deformity and two with bone dysplasia. All patients were females and presented bilateral disease. Six wrists were operated on. The median age at surgery was 15.5 years old, and the median postoperative follow-up time was of 37.5 months. The postoperative radiographic analysis revealed an average correction of 8.8 ± 7.5° for ulnar tilt, 3.0 ± 3.9 mm for lunate subsidence 8.2 ± 6.6° for lunate fossa angle, and 4.7 ± 2.6 mm for palmar carpal displacement. Average postoperative ranges of motion of the wrist joint were 75.8 ± 3.4° for flexion, 62.5 ± 14.1° for extension, 25.7 ± 2.9° for radial deviation, 40.0 ± 2.9° for ulnar deviation, 88.3 ± 2.4° for pronation, and 82.5 ± 2.5° for supination. The median VAS was 1 for residual pain, 0 for functional deficit, 0 for esthetic impairment, and 10 for recommending the surgical procedure. The median DASH score was 0. Conclusion Madelung deformity treatment using dome osteotomy of the distal radius and Vickers ligament section results in excellent esthetic and functional outcomes.

Resumo Objetivo Avaliar o resultado estético e funcional do tratamento cirúrgico da deformidade de Madelung em idade pediátrica. MétodoEstudo retrospectivo dos pacientes com deformidade de Madelung em idade pediátrica tratados cirurgicamente através de osteotomia em cúpula do rádio distal e secção do ligamento de Vickers entre 2015 e 2018. Foram incluídos doentes com tempo de seguimento pós-operatório mínimo de 12 meses. Foram analisados dados demográficos, técnica cirúrgica, resultados clínicos e radiográficos. A avaliação radiográfica pré e pós-operatória consistiu na medição da inclinação ulnar, do afundamento semilunar, do ângulo da fossa semilunar e do desvio palmar do carpo. A avaliação clínica pós-operatória consistiu na medição das amplitudes articulares do punho, escala visual analógica (EVA) e score Disabilities of the Arm, Shoulder and Hand (DASH). Resultados Foram incluídos quatro pacientes, dois com Madelung idiopática e dois com displasia óssea, todos do sexo feminino e com doença bilateral. Foram operados 6 punhos, a idade mediana à data de cirurgia foi 15,5 anos, e o tempo mediano de seguimento pós-operatório foi de 37,5 meses. Na análise radiográfica pós-operatória, verificou-se uma correção média de 8,8 ± 7,5° da inclinação ulnar, de 3 ± 3,9 mm do afundamento semilunar, de 8,2 ± 6,6° do ângulo da fossa semilunar e de 4,7 ± 2,6 mm do desvio palmar do carpo. Na avaliação da amplitude articular média pós-operatória, registrou-se uma flexão de 75,8 ± 3,4°; extensão de 62,5 ± 14,1°; desvio radial de 25,7 ± 2,9°; desvio cubital de 40,0 ± 2,9; pronação de 88,3 ± 2,4°; supinação de 82,5 ± 2,5°. Registou-se EVA mediana para dor residual = 1, défice funcional = 0, prejuízo estético = 0, e recomendação de procedimento cirúrgico = 10. A mediana do score DASH foi 0. Conclusão O tratamento da deformidade de Madelung através da osteotomia em cúpula do rádio distal e secção do ligamento de Vickers permite obter um excelente resultado estético e funcional.

Influence of the Inclination of the Incisal Edge of Planas Direct Tracks on Deciduous Dentition with Anterior Crossbite: Finite-Elements Study.

OBJECTIVE: This study aimed to evaluate the influence of three different inclinations of the incisal edge of Planas direct tracks (PDTs) on the upper deciduous incisors (15, 30, and 45 degrees) through simulations using the finite-elements method. MATERIALS AND METHODS: A three-dimensional virtual model of tooth 51 was elaborated by using the Rhinoceros computer-aided design (CAD) software. A mesh was constructed by using the Patran software, and the evaluations were processed by using the ANSYS 15.0 software. The geometry of the elements used consisted of a triangular-base tetrahedron composed of 2,167,386 elements and 3,012,995 nodal points. RESULTS: Progressive alterations of proportional intensity and distribution were seen in the areas of tension on the dentoalveolar structures, with increasing inclination of the edge of the PDT. The tractive contact tensions were seen to be concentrated in the vestibular apical thirds and palatine cervical thirds, while the compressive contact tensions were in the palatine apical thirds and vestibular cervical thirds. CONCLUSION: It was concluded that a 30-degree inclination for the PDT was most appropriate for the cases of anterior crossbite in the deciduous dentition. Nevertheless, because this was a laboratory evaluation, clinical criteria and complementary examinations for each case need to be taken into consideration in making therapeutic decisions.

Big defensin from the scallop Argopecten purpuratus ApBD1 is an antimicrobial peptide which entraps bacteria through nanonets formation.

Big defensins is a large family of antimicrobial peptides found in restricted groups of invertebrates, in particular mollusks where they have highly diversified. Big defensins are composed of a highly hydrophobic N-terminal region and a C-terminal region containing six cysteine residues whose arrangement is identical to that of vertebrate ß-defensins. They have been shown to be active against both Gram-positive and Gram-negative bacteria and fungi. Antimicrobial aggregates called nanonets entrapping and killing bacteria have been recently described for the hydrophobic N-terminal region of the Cg-BigDef1 from the oyster Crassostrea gigas. To determine whether nanonets formation is a conserved trait of mollusk big defensins, we assessed the potential entrapping of bacteria through nanonets of the big defensin from the scallop Argopecten purpuratus, ApBD1. Recombinant ApBD1 was produced with a thrombin-cleavable N-terminal His6 tag, followed by the mature peptide carrying a mutation of the last cysteine residue of the C-terminal region by and arginine, named rApBD1(C87R). This mutation did not apparently affect the three-dimensional structure and the biological properties of rApBD1(C87R), as evidenced by in silico modeling and in vitro antimicrobial assays. Strong immune staining of rApBD1(C87R) in numerous areas surrounding bacteria was observed by confocal microscopy, suggesting that rApBD1(C87R) entraps bacteria in peptide aggregates similar to those reported to the oyster big defensin. This study suggests the conservation of bactericidal activity and nanonet formation across big defensins from bivalve mollusks.

Electrodiagnostic Testing Characteristics of Diabetic People with Carpal Tunnel Syndrome.

Objective The present study aimed to correlate electroneuromyography (ENMG) findings in diabetic and nondiabetic subjects with carpal tunnel syndrome (CTS). Methods In total, 154 patients were evaluated in a hand surgery outpatient clinic. All ENMG tests were bilaterally performed by a single neurologist. Qualitative variables were described for all patients with CTS according to their diabetic status, and the chi-squared test was used to reveal any association. A joint model was adjusted to determine the influence of diabetes on ENMG severity in CTS patients. Results The sample consisted of 117 women and 37 men, with an average age of 56.9 years old. Electroneuromyography demonstrated bilateral CTS in 82.5% of the patients. Diabetes was identified in 21.4% of the cases. Severe ENMG was prevalent. Conclusion There was no association between diabetes and ENMG severity in patients with CTS. Level of evidence IV, case series.

Regulation of sphingolipid synthesis by the G1/S transcription factor Swi4.

SBF (Swi4/Swi6 Binding Factor) complex is a crucial regulator of G1/S transition in Saccharomyces cerevisiae. Here, we show that SBF complex is required for myriocin resistance, an inhibitor of sphingolipid synthesis. This phenotype was not shared with MBF complex mutants nor with deletion of the Swi4p downstream targets, CLN1/CLN2. Based on data mining results, we selected putative Swi4p targets related to sphingolipid metabolism and studied their gene transcription as well as metabolite levels during progression of the cell cycle. Genes which encode key enzymes for the synthesis of long chain bases (LCBs) and ceramides were periodically transcribed during the mitotic cell cycle, having a peak at G1/S, and required SWI4 for full transcription at this stage. In addition, HPLC-MS/MS data indicated that swi4Δ cells have decreased levels of sphingolipids during progression of the cell cycle, particularly, dihydrosphingosine (DHS), C24-phytoceramides and C24-inositolphosphoryl ceramide (IPC) while it had increased levels of mannosylinositol phosphorylceramide (MIPC). Furthermore, we demonstrated that both inhibition of de novo sphingolipid synthesis by myriocin or SWI4 deletion caused partial arrest at the G2/M phase. Importantly, our lipidomic data demonstrated that the sphingolipid profile of WT cells treated with myriocin resembled that of swi4Δ cells, with lower levels of DHS, IPC and higher levels of MIPC. Taken together, these results show that SBF complex plays an essential role in the regulation of sphingolipid homeostasis, which reflects in the correct progression through the G2/M phase of the cell cycle.

Electrodiagnostic Testing Characteristics of Diabetic People with Carpal Tunnel Syndrome / Características eletrofisiológicas das pessoas diabéticas com síndrome do túnel do carpo

Abstract Objective The present study aimed to correlate electroneuromyography (ENMG) findings in diabetic and nondiabetic subjects with carpal tunnel syndrome (CTS). Methods In total, 154 patients were evaluated in a hand surgery outpatient clinic. All ENMG tests were bilaterally performed by a single neurologist. Qualitative variables were described for all patients with CTS according to their diabetic status, and the chi-squared test was used to reveal any association. A joint model was adjusted to determine the influence of diabetes on ENMG severity in CTS patients. Results The sample consisted of 117 women and 37 men, with an average age of 56.9 years old. Electroneuromyography demonstrated bilateral CTS in 82.5% of the patients. Diabetes was identified in 21.4% of the cases. Severe ENMG was prevalent. Conclusion There was no association between diabetes and ENMG severity in patients with CTS. Level of evidence IV, case series.

Resumo Objetivo O presente trabalho teve por objetivo verificar se existe correlação entre a síndrome do túnel do carpo (STC) e eletroneuromiografia (ENMG) de pacientes diabéticos e não diabéticos. Métodos Foram avaliados 154 pacientes em um ambulatório de cirurgia da mão. Todas as ENMGs avaliadas foram realizadas por um único neurologista, bilateralmente. As variáveis qualitativas foram descritas para todas as pessoas em acompanhamento devido à STC segundo a presença de diabetes e foi verificada a associação com uso do teste qui-quadrado. Foi ajustado o modelo conjunto para verificar a influência da diabetes na gravidade da ENMG em pacientes com STC. Resultados Foram incluídos no presente estudo 117 mulheres e 37 homens, com média de idade de 56,9 anos. Eletroneuromiografia demonstrando STC bilateral foi observada em 82,5% das pessoas. Pessoas diabéticas foram identificadas em 21,4% dos casos. Eletroneuromiografia com padrão grave foi prevalente. Conclusão Não houve associação entre a presença de diabetes e a gravidade da ENMG em pessoas com STC. Nível de evidência IV, série de casos.

Repeat-Driven Generation of Antigenic Diversity in a Major Human Pathogen, Trypanosoma cruzi.

Trypanosoma cruzi, a zoonotic kinetoplastid protozoan parasite, is the causative agent of American trypanosomiasis (Chagas disease). Having a very plastic, repetitive and complex genome, the parasite displays a highly diverse repertoire of surface molecules, with pivotal roles in cell invasion, immune evasion and pathogenesis. Before 2016, the complexity of the genomic regions containing these genes impaired the assembly of a genome at chromosomal level, making it impossible to study the structure and function of the several thousand repetitive genes encoding the surface molecules of the parasite. We here describe the genome assembly of the Sylvio X10/1 genome sequence, which since 2016 has been used as a reference genome sequence for T. cruzi clade I (TcI), produced using high coverage PacBio single-molecule sequencing. It was used to analyze deep Illumina sequence data from 34 T. cruzi TcI isolates and clones from different geographic locations, sample sources and clinical outcomes. Resolution of the surface molecule gene distribution showed the unusual duality in the organization of the parasite genome, a synteny of the core genomic region with related protozoa flanked by unique and highly plastic multigene family clusters encoding surface antigens. The presence of abundant interspersed retrotransposons in these multigene family clusters suggests that these elements are involved in a recombination mechanism for the generation of antigenic variation and evasion of the host immune response on these TcI strains. The comparative genomic analysis of the cohort of TcI strains revealed multiple cases of such recombination events involving surface molecule genes and has provided new insights into T. cruzi population structure.

Perfil de docentes humanistas, fenomenológicos e existenciais: análise de editais de concursos / The profile of humanistic, phenomenological and existential educators: an analysis of public selection notice / Perfil de docentes humanistas, fenomenológicos y existenciales: análisis de convocatorias de concursos

Resumo Com base no argumento de que existe um mercado de trabalho acadêmico público que indica um perfil formativo para contratar profissionais para atuar no ensino superior, objetiva-se analisar quais conhecimentos circulam em editais de concurso para a área humanista, fenomenológica e existencial. Inicialmente, percebe-se que no Brasil é comum vincular as perspectivas humanistas, fenomenológicas e existenciais de Psicologia com as perspectivas fenomenológicas e existenciais da Filosofia. Em seguida, apresenta-se o plano de coleta e a análise dos dados conforme o método da análise de conteúdo. Foram compilados 24 editais, dos quais se extraíram 241 pontos de conhecimento cujas unidades de registro temáticas foram organizadas segundo seis eixos temáticos que remetem ao perfil formativo estudado. Este aponta para uma composição de conhecimentos relacionados: aos fundamentos históricos, epistemológicos e filosóficos; aos entendimentos teórico-conceituais; às práticas clínicas; às atuações no campo da saúde; à formação para a pesquisa e intervenção; e à reflexão acerca de fenômenos atuais. Conclui-se que esse perfil formativo é vasto, plural, complexo e difícil de ser atingido/dominado em sua totalidade, considerando as explicações, delimitações, articulações, aplicações e implicações das mencionadas perspectivas. Por fim, questiona-se o motivo disso e se lança outra possibilidade de pesquisa em instituições privadas.

Abstract Following the argument that there is a public academic paper market that indicates the profile of professionals to work in higher education, this text analyzes the content of the knowledge required in public tenders for the phenomenological and existential humanist area. Initially, it can be observed that in Brazil it is common for the humanist, phenomenological and existential perspectives of psychology and philosophy to be associated. The data collection and analysis followed the content analysis method. Twenty-four public notices were selected, of which 241 points of knowledge were extracted whose recording thematic units were organized according to six thematic axes that refer to the formative profile studied. The knowledge detected was related to: historical, epistemological and philosophical foundations; theoretical-conceptual understandings; clinical practices; activities in the health field; training for research and intervention; and the understanding of current phenomena. This formative profile is shown to be broad, plural, complex and difficult to be reached/mastered in full, considering the explanations, determinations, articulations, applications and implications of the aforementioned mentioned perspectives. Lastly, the reason for such is questioned and another possibility of research in private institutions is proposed.

Resumen Con base en el argumento de que existe un mercado de trabajo académico público que indica un perfil formativo en la contratación de profesionales para actuar en la enseñanza superior, se pretende analizar qué conocimientos circulan en convocatorias de concurso público para el área humanista, fenomenológica y existencial. Inicialmente, se sitúa que en Brasil es común vincular las perspectivas humanistas, fenomenológicas y existenciales de psicología y con las perspectivas fenomenológicas y existenciales de filosofía. A continuación, se presenta el plan de recolección y el análisis de los datos según el método del análisis de contenido. Se han compilado 24 convocatorias, de las cuales se extrajeron 241 puntos de conocimiento cuyas unidades de registro temáticas fueron organizadas según seis ejes temáticos que remiten al perfil formativo estudiado. Este apunta a una composición de conocimientos relacionados a: los fundamentos históricos, epistemológicos y filosóficos; los entendimientos teórico-conceptuales; las prácticas clínicas; las actuaciones en el campo de la salud; la formación para la investigación e intervención; y la reflexión sobre fenómenos actuales. Se concluye que ese perfil formativo es vasto, plural, complejo y difícil de ser alcanzado/dominado en su totalidad, considerando las explicaciones, delimitaciones, articulaciones, aplicaciones e implicaciones de las mencionadas perspectivas. Se cuestiona el motivo de ello y se lanza otra posibilidad de investigación en instituciones privadas.

Subungual exostosis - treatment results with preservation of the nail bed.

Subungual exostosis is a rare benign osteocartilaginous tumor, of unknown etiology, that affects the subungual part of the distal phalanx and predominantly affects adolescent females. The objective of this study is to evaluate the results of surgical treatment of subungual exostosis in pediatric patients by means of surgical excision and nail preservation. We present a retrospective study, including pediatric patients with subungual exostosis who underwent surgical treatment in a single institution, over a 6-year period, We describe the surgical technique used, the characteristics of the patients and lesions, and evaluate the results obtained in terms of function, satisfaction and complications. Thirteen patients were included in this study, seven female (54%) and six male (46%), median age 11 years old. The most common location was the hallux, in eight patients (73%). Patients had good functional results with a median American Orthopedic Foot & Ankle Society score of 100 [80; 100] and a median personal satisfaction score of 9. The median follow-up was 30 [3;68] months. In our patients, subungual excision with nail bed preservation, allows a good balance between the radical excision of the exostosis and the preservation of the natural protection granted by the native nail, yielding good functional results and patient satisfaction.

A Type IIa crustin from the pink shrimp Farfantepenaeus paulensis (crusFpau) is constitutively synthesized and stored by specific granule-containing hemocyte subpopulations.

Crustins are cysteine-rich antimicrobial peptides (AMPs) widely distributed across crustaceans. From the four described crustin Types (I to IV), crustins from the subtype IIa are the most abundant and diverse members found in penaeid shrimp. Despite the critical role of Type IIa crustins in shrimp antimicrobial defenses, there is still limited information about their synthesis and antimicrobial properties. Here, we report the subcellular localization and the antibacterial spectrum of crusFpau, a Type IIa crustin from the pink shrimp Farfantepenaeus paulensis. The recombinantly expressed crusFpau showed antimicrobial activity against both Gram-positive and Gram-negative bacteria at low concentrations. Results from immunofluorescence using anti-rcrusFpau antiserum revealed that crusFpau is synthetized and stored by both granular and semigranular hemocytes, but not by hyaline cells. Interestingly, not all granular and semigranular hemocytes stained for crusFpau, revealing that this crustin is produced by specific granule-containing hemocyte subpopulations. Finally, we showed that the granule-stored peptides are not constitutively secreted into the plasma of healthy animals.