Metagenomic binning of PacBio HiFi data prior to assembly reveals a complete genome of Cosmopolites sordidus (Germar) (Coleopterea: Curculionidae, Dryophthorinae) the most damaging arthropod pest of bananas and plantains.

PacBio HiFi sequencing was employed in combination with metagenomic binning to produce a high-quality reference genome of Cosmopolites sordidus. We compared k-mer and alignment reference based pre-binning and post-binning approaches to remove contamination. We were also interested to know if the post-binning approach had interspersed bacterial contamination within intragenic regions of Arthropoda binned contigs. Our analyses identified 3,433 genes that were composed with reads identified as of putative bacterial origins. The pre-binning approach yielded a C. sordidus genome of 1.07 Gb genome composed of 3,089 contigs with 98.6% and 97.1% complete and single copy genome and protein BUSCO scores respectively. In this article we demonstrate that in this case the pre-binning approach does not sacrifice assembly quality for more stringent metagenomic filtering. We also determine post-binning allows for increased intragenic contamination increased with increasing coverage, but the frequency of gene contamination increased with lower coverage. Future work should focus on developing reference free pre-binning approaches for HiFi reads produced from eukaryotic based metagenomic samples.

The impact of frailty on death, discharge destination and modelling accuracy in patients receiving organ support on the intensive care unit.

Background: This study aims to identify any effect of frailty in altering the risk of death or poor outcome already associated with receipt of organ support on ICU. It also aims to assess the performance of mortality prediction models in frail patients. Methods: All admissions to a single ICU over 1-year were prospectively allocated a Clinical Frailty Score (CFS). Logistic regression analysis was used to investigate the effect of frailty on death or poor outcome (death/discharge to a medical facility). Logistic regression analysis, area under the Receiver Operator Curve (AUROC) and Brier scores were used to investigate the ability of two mortality prediction models, ICNARC and APACHE II, to predict mortality in frail patients. Results: Of 849 patients, 700 (82%) patients were not frail, and 149 (18%) were frail. Frailty was associated with a stepwise increase in the odds of death or poor outcome (OR for each point rise of CFS = 1.23 ([1.03-1.47]; p = .024) and 1.32 ([1.17-1.48]; p = <.001) respectively). Renal support conferred the greatest odds of death and poor outcome, followed by respiratory support, then cardiovascular support (which increased the odds of death but not poor outcome). Frailty did not modify the odds already associated with organ support. The mortality prediction models were not modified by frailty (AUROC p = .220 and .437 respectively). Inclusion of frailty into both models improved their accuracy. Conclusions: Frailty was associated with increased odds of death and poor outcome, but did not modify the risk already associated with organ support. Inclusion of frailty improved mortality prediction models.

Evaluation of Characteristics and Building Applications of Multi-Recycled Concrete Aggregates from Precast Concrete Rejects.

The construction industry must meet current environmental requirements, mostly those pertaining to the reduction in construction and demolition waste and the consumption of raw materials. The use of recycled concrete aggregates can be part of the solution, but one question that arises is how many times recyclables can be recycled. This unknown involves other related queries regarding the properties and possible uses of repeated recycled concrete aggregates. This research is derived from the precast concrete industry, where multi-recycling is a pressing need. From good-quality parent concretes, three cycles of recycled concrete aggregates were produced and analysed. The final results are promising due to the good quality of the recycled and multi-recycled concrete aggregates obtained. Not only can they be used in low-level applications (backfilling) as usual, but they can also be used for more demanding purposes, such as graded aggregates, cement-treated road bases and concrete pavements. Their use in structural concrete is feasible, but it will be dependent on the water absorption level and the amount of recycled aggregate substitution. This research proves the viability of multi-recycled concrete aggregates with all of the associated environmental benefits.

Caracterización y manejo del nódulo tiroideo en niños, en el Servicio de Cirugía de Cabeza, Cuello y Máxilo Facial del Instituto Nacional de Salud del Niño - Breña, 2000 - 2020 / Characterization and management of the thyroid nodule in children, in the Head, Neck and Maxillo facial Surgery Service of the Instituto Nacional de Salud del Niño - Breña, 2000 - 2020

RESUMEN Introducción. El nódulo tiroideo en niños generalmente es asintomático, de presentación múltiple y aislada, con dificultad para instaurar el riesgo de malignidad, sobre todo si no es palpable. Objetivo. Determinar las características epidemiológicas, clínicas, ecográficas y manejo quirúrgico del nódulo tiroideo en pacientes del Servicio de Cirugía de Cabeza, Cuello y Máxilo Facial del Instituto Nacional de Salud del Niño (INSN) - Breña, 2000 -2020. Métodos. Estudio observacional, descriptivo y retrospectivo. Se usó la clasificación Bethesda para los estudios citológicos. Los datos fueron analizados en el programa SPSS versión 22. Las variables cualitativas fueron expresadas en frecuencias absolutas y relativas; y las cuantitativas en medidas de tendencia central y dispersión. Resultados. Fueron 66 casos, el promedio de edad fue 10,94 ± 0,88 años, el 48,5% entre 11 a 15 años, el 78,8% fueron del sexo femenino, un 89,4% no referían antecedente familiar de cáncer. Predominó la forma asintomática (74,2%). Ecográficamente el 43,9% de nódulos se localizaron en lóbulo derecho, presentación única (68,2%) y menor a dos centímetros (43,9%). El 82,6% de los 23 casos Bethesda II fueron hemitiroidectomizados y el 86,4% de 22 casos Bethesda VI fueron tiroidectomizados. Conclusiones. En pacientes pediátricos, los nódulos tiroideos se presentaron mayormente en mujeres de 11 a 15 años, sin antecedente familiar de cáncer, asintomáticos, localizados en lóbulo derecho, únicos y menor a dos centímetros. El grado II de la clasificación Bethesda fue la más frecuente. La hemitiroidectomía fue la intervención quirúrgica más utilizada.

ABSTRACT Introduction. The thyroid nodule in children is usually asymptomatic, multiple and isolated presentation, with difficulty in establishing the risk of malignancy, especially if it is not palpable. Objective. To determine the epidemiological, clinical and ultrasound characteristics and surgical management of thyroid nodules in patients of the Head, Neck and Maxillofacial Surgery Service of the Instituto Nacional de Salud del Niño (INSN) - Breña, 2000-2020. Methods. Observational, descriptive and retrospective study. The Bethesda classification was used for cytological studies. Qualitative variables were expressed in absolute and relative frequencies; and quantitative variables in measures of central tendency and dispersion. Results. There were 66 cases, the average age was 10.94 ± 0.88 years, 48.5% between 11 and 15 years, 78.8% were female, 89.4% had no family history of cancer. The asymptomatic form predominated (74.2%). Ultrasonographically, 43.9% of nodules were located in the right lobe, single presentation (68.2%) and smaller than two centimeters (43.9%). 82.6% of the 23 Bethesda II cases were hemithyroidectomized and 86.4% of 22 Bethesda VI cases were thyroidectomized. Conclusions. In pediatric patients, thyroid nodules occurred mostly in women aged 11 to 15 years, with no family history of cancer, asymptomatic, located in the right lobe, single and less than two centimeters. Grade II of the Bethesda classification was the most frequent. Hemithyroidectomy was the most used surgical intervention

Molecular analyses identifies new domains and structural differences among Streptococcus pneumoniae immune evasion proteins PspC and Hic.

The PspC and Hic proteins of Streptococcus pneumoniae are some of the most variable microbial immune evasion proteins identified to date. Due to structural similarities and conserved binding profiles, it was assumed for a long time that these pneumococcal surface proteins represent a protein family comprised of eleven subgroups. Recently, however, the evaluation of more proteins revealed a greater diversity of individual proteins. In contrast to previous assumptions a pattern evaluation of six PspC and five Hic variants, each representing one of the previously defined subgroups, revealed distinct structural and likely functionally regions of the proteins, and identified nine new domains and new domain alternates. Several domains are unique to PspC and Hic variants, while other domains are also present in other virulence factors encoded by pneumococci and other bacterial pathogens. This knowledge improved pattern evaluation at the level of full-length proteins, allowed a sequence comparison at the domain level and identified domains with a modular composition. This novel strategy increased understanding of individual proteins variability and modular domain composition, enabled a structural and functional characterization at the domain level and furthermore revealed substantial structural differences between PspC and Hic proteins. Given the exceptional genomic diversity of the multifunctional PspC and Hic proteins a detailed structural and functional evaluation need to be performed at the strain level. Such knowledge will also be useful for molecular strain typing and characterizing PspC and Hic proteins from new clinical S. pneumoniae strains.

Physical activity intention and attendance behaviour in Finnish youth with cerebral palsy - results from a physical activity intervention: an application of the theory of planned behaviour.

Physical activity is associated with better health in individuals with cerebral palsy (CP). Numerous physical activity interventions have been designed to promote physical activity among youth with CP. No previous studies have explored the factors contributing to the intention to participate and predicting attendance behaviour for these interventions. Using theory of planned behaviour (TPB), this study explored the prediction of physical activity intention and attendance behaviour in a physical activity intervention aiming to promote physical activity in a sample of young individuals with CP. Males with CP aged 9-21 years were asked to complete measures of attitude, subjective norms, perceived behavioural control and, intentions towards a physical activity intervention. Participants had no cognitive impairments to understand and follow instructions, were categorised into Gross Motor Function Classification System I-III, did not receive any specific lower limbs' medical treatment, or did not participate in a strength training program for lower limbs within 6 months before the study. Subjective norms were found to be the only significant predictor of intention, accounting for 83% of variance in intention. Intention and perceived behaviour control were found to be a nonsignificant predictor of attendance behaviour in youth with CP. The results show that TPB is a relevant tool in the prediction of intention towards a physical activity intervention in Finnish youth with CP.

Permeable reactive surface-biobarriers. Testing and evaluation of an ecotechnology for the removal of agrotoxic compounds carried by agricultural runoffs.

PURPOSE: The objective of the work is to determine the best operating conditions for variants of an ecological engineering tool (permeable reactive surface biobarrier -PRSB-) potentially useful for the protection of water resources, preventing the arrival of sediments and pesticides transported by runoffs and tile drainage from agricultural lands, to water bodies. METHODS: Four PRB-prototypes were constructed as fixed-bed horizontal channels packed with a porous material supporting an enriched microbial biofilm. Their dynamic and stoichiometric performance was evaluated in the presence or absence of granular activated carbon, with limiting or sufficient O2 supply. The removal of the pesticides and their leading catabolic derivatives were determined by HPLC. The most abundant cultivable microorganisms were isolated and identified by the sequencing of 16sDNA amplicons. RESULTS: The pollutant removal efficiencies obtained in the aerobic biobarriers or microaerophilia were similar. In addition, slight differences were observed in the presence of GAC as an adsorbent, meaning that the most economical and straightforward type of biobarrier was adequate to remove the pollutants studied. In addition, among the most abundant microorganisms isolated in the microbial biofilms colonizing the aerobic biobarriers, the microalgae Micractinium sp. showed the capacity to accumulate the insecticides permethrin and cypermethrin. CONCLUSIONS: The main observed role of Micractinium sp. in the aerobic barriers was the bioaccumulation of pyrethroids, meaning that biosorption is also a valuable removal mechanism operating in the aerobic PRBs. In this aspect, they behave analogously to subsurface constructed wetlands but, instead of superficial plant life, aerobic PRSBs host microalgae.

Utilidad de la espirometría en los pacientes hospitalizados por la enfermedad pulmonar obstructiva crónica (EPOC) exacerbada / Usefulness of spirometry in hospitalized patients with Chronic Obstructive Pulmonary Disease (COPD) exacerbated

RESUMEN La espirometría es la prueba estándar para confirmar el diagnóstico de la enfermedad pulmonar obstructiva crónica (EPOC). En los servicios de hospitalización es frecuente encontrar pacientes que ingresan con sospecha de esta condición durante una exacerbación. Con base en las guías actuales, existe controversia sobre si se puede o no realizar la espirometría en este escenario. Objetivo: revisar la utilidad diagnóstica de la espirometría en pacientes hospitalizados con sospecha de EPOC exacerbada. Metodología: se realizó una revisión no sistemática hasta noviembre del 2019. Las bases de datos consultadas fueron PubMed, Cochrane, EMBASE, LILACS, Google Scholar. Se incluyeron estudios prospectivos en pacientes mayores de 18 años, en idioma inglés y español. Se encontraron 573 artículos y se seleccionaron 20 artículos de relevancia para la discusión. Conclusiones: en los pacientes hospitalizados por sospecha de EPOC exacerbada, la espirometría es una herramienta útil para confirmar el diagnóstico. El momento para realizarla es previo al egreso, una vez el paciente está estable. Anotamos que el VEF1 no sería útil para establecer la gravedad.

SUMMARY Spirometry is the standard test to confirm the diagnosis of Chronic Obstructive Pulmonary Disease (COPD). In hospitalization services it is common to find patients admitted with suspicion of this condition during an exacerbation. Based on current guidelines, there is controversy about use or not spirometry in this setting. Objective: To review the diagnostic utility of spirometry in hospitalized patients with suspected exacerbated COPD. Development: A nonsystematic review was carried out until November 2019. The databases consulted were PubMed, Cochrane, EMBASE, LILACS, Google Scholar. Prospective studies were included, in patients over 18 years of age, in English and Spanish. Were found 573 articles and 20 relevant articles were selected for discussion. Conclusions: In hospitalized patients with suspected exacerbated COPD, spirometry is a useful tool to confirm the diagnosis. The time to perform it would be prior to discharge, once the patient is stable. Of note, FEV1 would not be useful for establishing severity.

Costos de los egresos inoportunos de origen no médico en una clínica de alto nivel de complejidad en Colombia / The costs of nonclinical delayed discharge in a tertiary care clinic in Colombia

Resumen Objetivo: describir los pacientes con egreso inoportuno de origen no médico, sus características sociodemográficas, clínicas y estimar los costos derivados, durante el 01 de junio y el 31 de agosto de 2017, en una institución de salud de alto nivel de complejidad. Material y métodos: estudio observacional, retrospectivo. La definición operativa de egreso inoportuno correspondía a pacientes que desde el punto de vista clínico podían ser dados de alta, pero continuaban ocupando una cama por problemas no médicos. Los datos fueron tomados de las historias clínicas y para el análisis de costo, se tuvieron en cuenta el total de los costos generados por la estancia inapropiada, incluyendo el costo día de hospitalización, insumos, medicamentos y evaluaciones médicas. La perspectiva del costo fue desde el pagador. Resultados: de los 3273 egresos durante el periodo analizado, se presentaron 79 casos de egresos inoportunos (2.4%). La edad media de los pacientes fue de 49 años. El 62% eran del régimen subsidiado; 60.7% eran procedentes de otros departamentos o municipios lejanos. El total de días hospitalización perdidos por esta causa fue de 547, con una media de 6.6 días por paciente. Los servicios más afectados fueron medicina interna y cuidados paliativos. Las principales causas fueron: necesidad de tiquetes para retornar al lugar de procedencia, alta temprana por medicamentos o insumos para el egreso y necesidad de oxígeno. El costo total que se generó por egreso inoportuno fue US$83 935, con un valor promedio de US$1062.47 por paciente. Conclusión: el egreso inoportuno es un problema evidente en la atención. Las principales causas que encontramos fueron problemas sociales y traslados regionales. Además genera un gasto adicional importante de recursos materiales y económicos tanto para las instituciones como para el sistema.(Acta Med Colomb 2020; 45. DOI:https://doi.org/10.36104/amc.2020.1255).

Abstract Objective: to describe the sociodemographic and clinical characteristics of patients with nonclinical delayed discharge and estimate the resulting costs in a tertiary healthcare facility from June 1 to August 31, 2017. Materials and methods: a retrospective observational study. The working definition of delayed discharge was patients who, from a clinical perspective, could be discharged but who continued to occupy a bed due to nonclinical problems. The data were taken from the medical charts, and the total costs incurred by the inappropriate length of stay were used for the cost analysis, including the cost per inpatient day, supplies, medications and medical assessments, from the payer's cost perspective. Results: of the 3,273 discharges during the study period, there were 79 cases of delayed discharge (2.4%). The mean age of the patients was 49 years. Sixty-two percent had subsidized medical insurance; 60.7% came from other departments or distant towns. The total number of inpatient days lost due to this cause was 547, with a me of 6.6 days per patient. The departments which were most affected were internal medicine and palliative care. The main causes were: the need for tickets to return to their place of origin, early discharge due to medications or discharge supplies and the need for oxygen. The total cost incurred due to delayed discharge was USD 83,935, with an average of USD 1,062.47 per patient. Conclusion: delayed discharge is an evident healthcare problem. The main causes found in this study were social and regional transport issues. Delayed discharge also causes a significant additional expense in material and financial resources for both the institutions and the system.(Acta Med Colomb 2020; 45. DOI:https://doi.org/10.36104/amc.2020.1255).

Enolase From Aspergillus fumigatus Is a Moonlighting Protein That Binds the Human Plasma Complement Proteins Factor H, FHL-1, C4BP, and Plasminogen.

The opportunistic fungal pathogen Aspergillus fumigatus can cause severe infections, particularly in immunocompromised individuals. Upon infection, A. fumigatus faces the powerful and directly acting immune defense of the human host. The mechanisms on how A. fumigatus evades innate immune attack and complement are still poorly understood. Here, we identify A. fumigatus enolase, AfEno1, which was also characterized as fungal allergen, as a surface ligand for human plasma complement regulators. AfEno1 binds factor H, factor-H-like protein 1 (FHL-1), C4b binding protein (C4BP), and plasminogen. Factor H attaches to AfEno1 via two regions, via short conserved repeats (SCRs) 6-7 and 19-20, and FHL-1 contacts AfEno1 via SCRs 6-7. Both regulators when bound to AfEno1 retain cofactor activity and assist in C3b inactivation. Similarly, the classical pathway regulator C4BP binds to AfEno1 and bound to AfEno1; C4BP assists in C4b inactivation. Plasminogen which binds to AfEno1 via lysine residues is accessible for the tissue-type plasminogen activator (tPA), and active plasmin cleaves the chromogenic substrate S2251, degrades fibrinogen, and inactivates C3 and C3b. Plasmin attached to swollen A. fumigatus conidia damages human A549 lung epithelial cells, reduces the cellular metabolic activity, and induces cell retraction, which results in exposure of the extracellular matrix. Thus, A. fumigatus AfEno1 is a moonlighting protein and virulence factor which recruits several human regulators. The attached human regulators allow the fungal pathogen to control complement at the level of C3 and to damage endothelial cell layers and tissue components.

Identification of coenzyme-binding proteins with machine learning algorithms.

The coenzyme-binding proteins play a vital role in the cellular metabolism processes, such as fatty acid biosynthesis, enzyme and gene regulation, lipid synthesis, particular vesicular traffic, and ß-oxidation donation of acyl-CoA esters. Based on the theory of Star Graph Topological Indices (SGTIs) of protein primary sequences, we proposed a method to develop a first classification model for predicting protein with coenzyme-binding properties. To simulate the properties of coenzyme-binding proteins, we created a dataset containing 2897 proteins, among 456 proteins functioned as coenzyme-binding activity. The SGTIs of peptide sequence were calculated with Sequence to Star Network (S2SNet) application. We used the SGTIs as inputs to several classification techniques with a machine learning software - Weka. A Random Forest classifier based on 3 features of the embedded and non-embedded graphs was identified as the best predictive model for coenzyme-binding proteins. This model developed was with the true positive (TP) rate of 91.7%, false positive (FP) rate of 7.6%, and Area Under the Receiver Operating Characteristic Curve (AUROC) of 0.971. The prediction of new coenzyme-binding activity proteins using this model could be useful for further drug development or enzyme metabolism researches.

Culture-attenuated pathogenic Leptospira lose the ability to survive to complement-mediated-killing due to lower expression of factor H binding proteins.

Several pathogens including Gram-negative bacteria hijack complement regulators to escape host's innate response. Pathogenic Leptospira species bind Factor H, C4b binding protein and vitronectin from the complement system. We evaluated the ability of low passage (LP) and culture-attenuated (CA) pathogenic strains of Leptospira, to bind Factor H. We used LOCaS46 (Leptospira interrogans sv Canicola), LOVe30 (L. interrogans sv Icterohaemorrhagiae) and MOCA45 (L. santarosai sv Tarassovi), and ten high passage strains of Leptospira [used in the microscopic agglutination test (MAT)]. Afterwards, we assessed their survival in normal human serum (NHS). Interestingly, the ability in binding Factor H was higher for LOCaS46 and LOVe30 LP strains, than for the respective CA strains suggesting that the ability of evading the alternative complement pathway is lost after culture attenuation. Accordingly, the level of mRNA expression of the Factor H binding proteins, LigA, LigB and Lsa23 was higher in these LP strains than in the corresponding CA strains. Unexpectedly, no difference in Factor H binding and surviving was observed between LP and CA MOCA45 strains. The high passage MAT-reference strains showed variation in Factor H binding ability, but, in most cases, the ability for capturing Factor H by Leptospira strains correlated with their survival in NHS.

Genome-Wide Patterns of Sequence Divergence of Protein-Coding Genes Between Drosophila buzzatii and D. mojavensis.

Evolutionary rates for protein-coding genes are determined not only by natural selection but also by multiple genomic factors including mutation rates, recombination, gene expression levels, and chromosomal location. To investigate the joint effects of different genomic determinants on protein evolution, we compared the coding sequences of 9017 single-copy orthologs between 2 cactophilic species from the Drosophila subgenus, Drosophila mojavensis and D. buzzatii, whose genomes have been previously sequenced. We assessed the impact of 7 genomic determinants, that is, chromosome type, recombination, chromosomal inversions, expression breadth, expression level, gene length, and the number of exons, on divergence rates of protein-coding genes to understand patterns of evolutionary variation. Integrative analysis of these factors revealed that 1) X-linked and autosomal genes evolve at significantly different rates in agreement with the faster-X hypothesis, 2) genes located on the dot chromosome and pericentromeric regions have higher divergence rates, 3) genes located at chromosomes with more fixed inversions have higher pairwise divergence than those located at nearly collinear chromosomes, and 4) gene expression patterns can be considered the strongest determinant of protein evolution. In addition, the number of exons and protein length had a significant effect on pairwise divergence at synonymous sites. All in all, our results show the relative importance of each genomic factor on the rates of protein evolution and functional constraint in these 2 cactophilic Drosophila species.

Computational Sequence Analysis of Inversion Breakpoint Regions in the Cactophilic Drosophila mojavensis Lineage.

We investigated rates of chromosomal evolution in Drosophila mojavensis using whole-genome sequence information from D. mojavensis, Drosophila buzzatii, and Drosophila virilis. Drosophila mojavensis is a cactophilic species of the repleta group living under extreme ecological conditions in the deserts of the Southwestern United States and Northwestern México. The genome of D. buzzatii, another member of the repleta group, was recently sequenced and the largest scaffolds anchored to all chromosomes using diverse procedures. Chromosome organization between D. mojavensis and D. buzzatii was compared using MUMmer and GRIMM software. Our results corroborate previous cytological analyses that indicated chromosome 2 differed between these 2 species by 10 inversions, chromosomes X and 5 differed by one inversion each, and chromosome 4 was homosequential. In contrast, we found that chromosome 3 differed by 5 inversions instead of the expected 2 that were previously inferred by cytological analyses. Thirteen of these inversions occurred in the D. mojavensis lineage: 12 are fixed and one of them is a polymorphic inversion previously described in populations from Sonora and Baja California, México. We previously investigated the breakpoints of chromosome 2 inversions fixed in D. mojavensis. Here we characterized the breakpoint regions of the 5 inversions found in chromosome 3 in order to infer the molecular mechanism that generated each inversion and its putative functional consequences. Overall, our results reveal a number of gene alterations at the inversion breakpoints with putative adaptive consequences that point to natural selection as the cause for fast chromosomal evolution in D. mojavensis.

Culture-attenuated pathogenic Leptospira lose the ability to survive to complement-mediated-killing due to lower expression of factor H binding proteins

Several pathogens including Gram-negative bacteria hijack complement regulators to escape host's innate response. Pathogenic Leptospira species bind Factor H, C4b binding protein and vitronectin from the complement system. We evaluated the ability of low passage (LP) and culture-attenuated (CA) pathogenic strains of Leptospira, to bind Factor H. We used LOCaS46 (Leptospira interrogans sv Canicola), LOVe30 (L. interrogans sv Icterohaemorrhagiae) and MOCA45 (L. santarosai sv Tarassovi), and ten high passage strains of Leptospira [used in the microscopic agglutination test (MAT)]. Afterwards, we assessed their survival in normal human serum (NHS). Interestingly, the ability in binding Factor H was higher for LOCaS46 and LOVe30 LP strains, than for the respective CA strains suggesting that the ability of evading the alternative complement pathway is lost after culture attenuation. Accordingly, the level of mRNA expression of the Factor H binding proteins, LigA, LigB and Lsa23 was higher in these LP strains than in the corresponding CA strains. Unexpectedly, no difference in Factor H binding and surviving was observed between LP and CA MOCA45 strains. The high passage MAT-reference strains showed variation in Factor H binding ability, but, in most cases, the ability for capturing Factor H by Leptospira strains correlated with their survival in NHS.

Culture-attenuated pathogenic Leptospira lose the ability to survive to complement-mediated-killing due to lower expression of factor H binding proteins

Several pathogens including Gram-negative bacteria hijack complement regulators to escape host's innate response. Pathogenic Leptospira species bind Factor H, C4b binding protein and vitronectin from the complement system. We evaluated the ability of low passage (LP) and culture-attenuated (CA) pathogenic strains of Leptospira, to bind Factor H. We used LOCaS46 (Leptospira interrogans sv Canicola), LOVe30 (L. interrogans sv Icterohaemorrhagiae) and MOCA45 (L. santarosai sv Tarassovi), and ten high passage strains of Leptospira [used in the microscopic agglutination test (MAT)]. Afterwards, we assessed their survival in normal human serum (NHS). Interestingly, the ability in binding Factor H was higher for LOCaS46 and LOVe30 LP strains, than for the respective CA strains suggesting that the ability of evading the alternative complement pathway is lost after culture attenuation. Accordingly, the level of mRNA expression of the Factor H binding proteins, LigA, LigB and Lsa23 was higher in these LP strains than in the corresponding CA strains. Unexpectedly, no difference in Factor H binding and surviving was observed between LP and CA MOCA45 strains. The high passage MAT-reference strains showed variation in Factor H binding ability, but, in most cases, the ability for capturing Factor H by Leptospira strains correlated with their survival in NHS.

Concurrent Duplication of Drosophila Cid and Cenp-C Genes Resulted in Accelerated Evolution and Male Germline-Biased Expression of the New Copies.

Despite their essential role in the process of chromosome segregation in eukaryotes, kinetochore proteins are highly diverse across species, being lost, duplicated, created, or diversified during evolution. Based on comparative genomics, the duplication of the inner kinetochore proteins CenH3 and Cenp-C, which are interdependent in their roles of establishing centromere identity and function, can be said to be rare in animals. Surprisingly, the Drosophila CenH3 homolog Cid underwent four independent duplication events during evolution. Particularly interesting are the highly diverged Cid1 and Cid5 paralogs of the Drosophila subgenus, which are probably present in over one thousand species. Given that CenH3 and Cenp-C likely co-evolve as a functional unit, we investigated the molecular evolution of Cenp-C in species of Drosophila. We report yet another Cid duplication (leading to Cid6) within the Drosophila subgenus and show that not only Cid, but also Cenp-C is duplicated in the entire subgenus. The Cenp-C paralogs, which we named Cenp-C1 and Cenp-C2, are highly divergent. Both Cenp-C1 and Cenp-C2 retain key motifs involved in centromere localization and function, while some functional motifs are conserved in an alternate manner between the paralogs. Interestingly, both Cid5 and Cenp-C2 are male germline-biased and evolved adaptively. However, it is currently unclear if the paralogs subfunctionalized or if the new copies acquired a new function. Our findings point towards a specific inner kinetochore composition in a specific context (i.e., spermatogenesis), which could prove valuable for the understanding of how the extensive kinetochore diversity is related to essential cellular functions.