Improving the production of the denatured recombinant N-terminal domain of rhoptry-associated protein 2 from a Plasmodium falciparum target in the pathology of anemia in falciparum malaria

Rhoptry-associated protein 2 (RAP2) is known to be discharged from rhoptry onto the membrane surface of infected and uninfected erythrocytes (UEs) ex vivo and in vitro and this information provides new insights into the understanding of the pathology of severe anemia in falciparum malaria. In this study, a hexahistidine-tagged recombinant protein corresponding to residues 5-190 of the N-terminal of Plasmodium falciparum RAP2 (rN-RAP2) was produced using a new method of solubilization and purification. Expression was induced with D-lactose, a less expensive alternative inducer to the more common isopropyl-²-D-thio-galactopyranosidase. The recombinant protein was purified using two types of commercially-available affinity columns, iminodiacetic and nitrilotriacetic. rN-RAP2 had immunogenic potential, since it induced high titers of anti-RAP2 antibodies in mice. These antibodies recognized full-length RAP2 prepared from Triton X-100 extracts from two strains of P. falciparum. In fact, the antibody recognized a 29-kDa product of RAP2 cleavage as well as 82 and 70-kDa products of RAP1 cleavage. These results indicate that the two antigens share sequence epitopes. Our expressed protein fragment was shown to contain a functional epitope that is also present in rhoptry-derived ring surface protein 2 which attaches to the surface of both infected and UEs and erythroid precursor cells in the bone marrow of malaria patients. Serum from malaria patients who developed anemia during infection recognized rN-RAP2, suggesting that this protein fragment may be important for epidemiological studies investigating whether immune responses to RAP2 exacerbate hemolysis in falciparum malaria patients.CNP

Imaging Characterization of the Lower Lumbar Spine and Pelvis for Surgical Planning

Funding Information: The present publication was funded by Fundação Ciência e Tecnologia, IP national support through CHRC (UIDP/04923/2020). The remaining authors have no conflicts to report.Objective: To determine and compare pelvic and lumbosacral reference parameters with computed tomography in patients with low back pain (LBP) and a control group of asymptomatic patients to provide quantification data and morphological correlations for L5S1 transforaminal endoscopic approach (L5S1TEA). Methods: We prospectively evaluated 100 patients with LBP and a control group of 100 individuals, with spinopelvic computed tomography. We measured lumbopelvic and L5S1 transforaminal approach parameters: maximum approach angle (maxAA) and minimum approach angle (minAA) and skin incision (maxSI and minSI), iliac crest (IC) projection at intersection point (ICPi), distance between the projected intersection of maxAA with the ilium (ICi) and the posterior limit of the IC (ΔICi-ICpost), and distance between ICi and spinous process (ΔICi-SP). Results: Females and ICPi were increased in the LBP group: maxAA: 48.38° ± 5.09°; minAA:32.5° ± 3.90°; maxSI: 11.39 ± 1.86 cm; and minSI: 8.30 ± 1.48 cm. Ilium intersection was increased in males; IC projection at the highest point (ICPh) was higher than ICPi; maxAA intersected the ilium in 28% and minAA in 1.5% of cases; ICi was positively correlated with facet angle, ICPh, and ICPi and negatively with ΔICi-SP. Conclusions: Our results set preliminary reference values for L5S1TEA surgical planning. Besides higher ICPi, there were no differences between groups in measured parameters. Traditional IC height (ICPh) does not correspond to the point of intersection of the approach and is significantly higher than ICPi. ICi correlated to higher facet angle values, ICPh and ICPi grades, and lower ΔICi-SP. Potential conflict with the ilium is increased in the male population. IC is not impeditive of L5S1TEA in most cases.proofepub_ahead_of_prin

Comparison of Statistical and Machine Learning Models on Road Traffic Accident Severity Classification

Portugal has the sixth highest road fatality rate among European Union members. This is a problem of different dimensions with serious consequences in people’s lives. This study analyses daily data from police and government authorities on road traffic accidents that occurred between 2016 and 2019 in a district of Portugal. This paper looks for the determinants that contribute to the existence of victims in road traffic accidents, as well as the determinants for fatalities and/or serious injuries in accidents with victims. We use logistic regression models, and the results are compared to the machine-learning model results. For the severity model, where the response variable indicates whether only property damage or casualties resulted in the traffic accident, we used a large sample with a small imbalance. For the serious injuries model, where the response variable indicates whether or not there were victims with serious injuries and/or fatalities in the traffic accident with victims, we used a small sample with very imbalanced data. Empirical analysis supports the conclusion that, with a small sample of imbalanced data, machine-learning models generally do not perform better than statistical models; however, they perform similarly when the sample is large and has a small imbalance

Rotavirus and adenovirus in Rondônia

The present work consists of analysis of the Work Routine Management development and implementation, in a process (register) of Operational Units in an Agro-industrial Cooperative, in order to use management tools that facilitate the identification of possible problems that hamper the results and their respective causes, and the execution of routine activities. The analysis consists of process mapping, creation and review of process indicators, as well as the definition of goals for monitoring and activating Routine Management. It also includes the survey of opportunities for improvement and the definition of an action plan in order to optimize the process for those involved, making it possible to act by monitoring the results obtained, tracing future actions and not only for correcting mistakes occurred during work routines

Acute Exposure to Two Biocides Causes Morphological and Molecular Changes in the Gill Ciliary Epithelium of the Invasive Golden Mussel Limnoperna fortunei (Dunker, 1857)

Limnoperna fortunei, the golden mussel, is a bivalve mollusk considered an invader in South America. This species is responsible for ecological and economic damages due to its voluminous fouling capability. Chemical biocides such as MXD-100™ and sodium dichloroisocyanurate (NaDCC) are often used to control L. fortunei infestations in hydraulic systems. Thus, we proposed to investigate the effects of different periods (24, 48 and 72 h) of exposure to MXD-100™ (0.56 mg L−1) and NaDCC (1.5 mg L−1) on the gills of L. fortunei through morphological and molecular analyses. NaDCC promoted progressive morphological changes during the analyzed periods and only an upregulation of SOD and HSP70 expression during the first 24 h of exposure. MXD-100™ led to severe morphological changes from the first period of exposure, in addition to an upregulation of SOD, CAT, HSP70 and CYP expression during the first 24 h. In contrast, MXD-100™ led to a downregulation of CAT transcription between 24 and 48 h. In static conditions, NaDCC causes lethal damage after 72 h of exposure, and that exposure needs to be continuous to achieve the control of the species. Meanwhile, the MXD-100™ treatment presented several effects during the first 24 h, showing acute toxicity in a shorter period of time

Genetic Diversity of MSP1 Block 2 of Plasmodium vivax Isolates from Manaus (Central Brazilian Amazon)

The diversity of MSP1 in both Plasmodium falciparum and P. vivax is presumed be associated to parasite immune evasion. In this study, we assessed genetic diversity of the most variable domain of vaccine candidate N-terminal PvMSP1 (Block 2) in field isolates of Manaus. Forty-seven blood samples the polymorphism of PvMSP1 Block 2 generates four fragment sizes. In twentyeight of them, sequencing indicated seven haplotypes of PvMSP1 Block 2 circulating among field isolates. Evidence of striking exchanges was observed with two stretches flanking the repeat region and two predicted recombination sites were described. Single nucleotide polymorphisms determined with concurrent infections per patient indicated that nonsynonymous substitutions occurred preferentially in the repeat-rich regions which also were predicted as B-cell epitopes. The comprehensive understanding of the genetic diversity of the promising Block 2 associated with clinical immunity and a reduced risk of infection by Plasmodium vivax would be important for the rationale of malaria vaccine designs

Root morphology of Arabica coffee cultivars subjected to different spatial arrangements

O objetivo deste trabalho foi avaliar as respostas morfológicas do sistema radicular de quatro cultivares de café arábica submetidas a diferentes arranjos espaciais. Os espaçamentos adotados foram os de 0,40, 0,50, 0,60, 0,70 e 0,80 m entre plantas na linha, tendo-se mantido fixo 3,8 m na entrelinha. Aos 27, 35 e 39 meses após o transplantio, foram coletadas amostras de solo+raízes em três posições em relação aos caules das plantas e três profundidades. As raízes foram lavadas, coloridas, digitalizadas e processadas com o programa Safira. A cultivar Tupi RN IAC 1669‑13 apresentou sistema radicular com características morfológicas relacionadas à absorção de nutrientes e de água superiores às das cultivares Catuaí Vermelho IAC 144, Catuaí Amarelo IAC 62 e Catuaí Amarelo IAC 32, para todos os arranjos espaciais testados. A redução no espaçamento aumenta a abundância de raízes por volume de solo, sem alterar a qualidade morfológica do sistema radicular, nem aprofundá‑lo. Os sistemas radiculares das cultivares avaliadas são mais abundantes e apresentam superfície e comprimento específico maiores entre plantas adjacentes, seguidas das posições afastadas a 0,25 e 0,50 m do caule. A proporção de raízes na camada de 0,1 m de profundidade é maior que a na de 0,4 m, para todos os espaçamentos. A variação no espaçamento entre plantas na linha altera a morfologia do sistema radicular do café arábica.The objective of this work was to evaluate the morphological responses of the root system of four Arabica coffee cultivars subjected to different spatial arrangements. The adopted plant spacings were 0.40, 0.50, 0.60, 0.70, and 0.80 m between plants within rows, maintaining 3.8 m constant between rows. At 27, 35, and 39 months after transplanting, samples of soil+roots were collected at three surface positions in relation to plant stems and at three depths. Roots were washed, colored, digitalized, and processed with the Safira software. The Tupi RN IAC 1669‑13 cultivar showed a root system with superior morphological traits related to the absorption of nutrients and water, in comparison to those of the Catuaí Vermelho IAC 144, Catuaí Amarelo IAC 62, and Catuaí Amarelo IAC 32 cultivars, for all tested spatial arrangements. The reduction in spacing increases the amount of total root per soil volume unit, without affecting the morphological quality of the root system, nor deepening it. The root systems of the evaluated cultivars are more abundant and show higher surface and specific length between adjacent plants, followed by the 0.25 and 0.50 m positions away from the coffee stem. The proportion of roots at the 0.1 m layer is higher than at the 0.4 m layer, for all plant spacings. Changes in spacings between plants within lines affect the morphology of Arabica coffee root system

Multidrug resistant Pseudomonas aeruginosa survey in a stream receiving effluents from ineffective wastewater hospital plants

Background: Multi-drug resistant forms of Pseudomonas aeruginosa (MDRPA) are a major source of nosocomial infections and when discharged into streams and rivers from hospital wastewater treatment plants (HWWTP) they are known to be able to persist for extended periods. In the city of Manaus (Western Brazilian Amazon), the effluent of three HWWTPs feed into the urban Mindu stream which crosses the city from its rainforest source before draining into the Rio Negro. The stream is routinely used by Manaus residents for bathing and cleaning (of clothes as well as domestic utensils) and, during periods of flooding, can contaminate wells used for drinking water. Results: 16S rRNA metagenomic sequence analysis of 293 cloned PCR fragments, detected an abundance of Pseudomonas aeruginosa (P. aeruginosa) at the stream's Rio Negro drainage site, but failed to detect it at the stream's source. An array of antimicrobial resistance profiles and resistance to all 14 tested antimicrobials was detected among P. aeruginosa cultures prepared from wastewater samples taken from water entering and being discharged from a Manaus HWWTP. Just one P. aeruginosa antimicrobial resistance profile, however, was detected from cultures made from Mindu stream isolates. Comparisons made between P. aeruginosa isolates' genomic DNA restriction enzyme digest fingerprints, failed to determine if any of the P. aeruginosa found in the Mindu stream were of HWWTP origin, but suggested that Mindu stream P. aeruginosa are from diverse origins. Culturing experiments also showed that P. aeruginosa biofilm formation and the extent of biofilm formation produced were both significantly higher in multi drug resistant forms of P. aeruginosa. Conclusions: Our results show that a diverse range of MDRPA are being discharged in an urban stream from a HWWTP in Manaus and that P. aeruginosa strains with ampicillin and amikacin can persist well within it. © 2016 The Author(s)