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Tetracyclines are antibiotics considered emerging pollutants and currently, wastewater treatment plants are not able to remove them efficiently. Laccases are promising enzymes for bioremediation because they can oxidize a wide variety of substrates. The aim of this study was to evaluate the Botrytis aclada laccase for the oxidation of chlortetracycline and its isomers in the absence of a mediator molecule, at a pH range between 3.0 to 7.0, and to characterize the transformation products by LC–MS. Chlortetracycline and three isomers were detected in both, controls and reaction mixtures at 0 h and in controls after 48 h of incubation but in different proportions depending on pH. An additional isomer was also detected, but only in the presence of BaLac. Based on the transformation products identified in the enzymatic reactions and information from literature, we assembled a network of transformation pathways starting from chlortetracycline and its isomers. The spectrometric analysis of the products indicated the probable occurrence of oxygen insertion, dehydrogenation, demethylation and deamination reactions. Four new products were identified, and we also described a novel transformation product without the chloro group. We observed that increasing pH led to higher diversity of main products. This is the first study using the laccase from fungi Botrytis aclada to oxidate chlortetracycline and its isomers and it can be considered as an ecological alternative to be used in bioremediation processes such as wastewater. [ABSTRACT FROM AUTHOR]

The transit service in a city should be the most efficient, least polluting, most accessible, and sustainable means of transportation for its citizens. However, serious shortcomings have been detected, mainly in medium-sized cities in developing countries. These shortcomings are related to a lack of user information, insecurity, low service availability, and repeated stops in inappropriate and/or unauthorized places. Some of these shortcomings contribute to high accident rates and traffic congestion. The development of tools to improve the characteristics and conditions of transit service in cities has become an imperative need to improve the quality of life of citizens and city sustainability. Transit service tracking is relevant in aspects such as online location information to travelers and control by transport companies for compliance with speed limits, schedules, routes, and stops. This research proposes a transit vehicle tracking system based on the Internet of Vehicles (IoV) in Vehicle-to-Roadside (V2R) classification. The proposed system is ideal for the use of electric vehicles due to the low power consumption of the tracking device. This system uses Intelligent Transportation Systems (ITS) tracking service architecture, Long Range (LoRa) communication technology, and its LoRa Wide Area Network (LoRaWAN) protocol. Additionally, the system offers real-time location prediction in the absence of position data. The IoV tracking device integrates a GPS-LoRa module card with an Inertial Measurement Unit (IMU). A location prediction algorithm was implemented to train and store a prediction model with previously collected data from tracking devices. To evaluate the developed model, a case study in the city of Popayán (Colombia) was implemented, using three routes for testing. The results of the system implementation were satisfactory, obtaining an average coverage of 60.4% of the routes in the final field tests through LoRa communication. For the remaining 39.6% of the routes, location data prediction was used, with an average accuracy of 177 m with respect to the real location. Considering the obtained results, a tracking system such as the one proposed in this article can be used in the transit systems of medium-sized cities in developing countries to improve service quality and fleet control. [ABSTRACT FROM AUTHOR]

In Colombia, the first case of COVID-19 was confirmed on 6 March 2020. On 13 March 2023, Colombia registered 6,360,780 confirmed positive cases of COVID-19, representing 12.18% of the total population. The National Administrative Department of Statistics (DANE) in Colombia published in 2020 a COVID-19 vulnerability index, which estimates the vulnerability (per city block) of being infected with COVID-19. Unfortunately, DANE did not consider multiple factors that could increase the risk of COVID-19 (in addition to demographic and health), such as environmental and mobility data (found in the related literature). The proposed multidimensional index considers variables of different types (unemployment rate, gross domestic product, citizens' mobility, vaccination data, and climatological and spatial information) in which the incidence of COVID-19 is calculated and compared with the incidence of the COVID-19 vulnerability index provided by DANE. The collection, data preparation, modeling, and evaluation phases of the Cross-Industry Standard Process for Data Mining methodology (CRISP-DM) were considered for constructing the index. The multidimensional index was evaluated using multiple machine learning models to calculate the incidence of COVID-19 cases in the main cities of Colombia. The results showed that the best-performing model to predict the incidence of COVID-19 in Colombia is the Extra Trees Regressor algorithm, obtaining an R-squared of 0.829. This work is the first step toward a multidimensional analysis of COVID-19 risk factors, which has the potential to support decision making in public health programs. The results are also relevant for calculating vulnerability indexes for other viral diseases, such as dengue. [ABSTRACT FROM AUTHOR]

Traffic accidents are one of the most serious problems worldwide, being one of the leading causes of death and economic loss in the world. Low- and middle-income countries, mainly their medium-sized cities, are among the most affected by this problem. 93% of traffic accidents occur in low and middle-income countries, even though these countries have approximately 60% of the world's vehicles. This occurs mainly because in these types of countries, especially in medium-sized cities (target context), there are no ideal conditions for driving, such as adequate road infrastructure, good condition of vehicles, and rigorous safety policies. Advanced data analysis techniques including machine learning (ML) have increasingly been used to solve this problem. Naturalistic driving (ND) can be applied as a data collection method that provides information on traffic accidents. ND commonly uses a vehicle's kinematic data to detect high-risk driving behaviors that could cause an accident. The objectives of this document are to present a review of different alternatives that help in data collection and creation of intelligent solutions related to detection of possible traffic accidents, principally using ND; and to propose an intelligent collision risk detection system (ICRDS) for identification of areas with a high probability of TA in the target context. Through the review, it was possible to analyze and evaluate the devices, variables and algorithms that help characterize a risk event in driving, considering the target context. The development of a prototype of an ICRDS for a medium-sized city in a developing country is considered viable, considering the identified components, with the aim of identifying risk events in driving, and areas of high probability of accidents in the city. [ABSTRACT FROM AUTHOR]

Introduction: COVID-19 has affected people in several countries around the world. They experience respiratory symptoms that can be mild, moderate, or severe. Several reviews that characterize the risk factors of COVID-19 have been performed, but most address only risk factors associated with medical conditions, ignoring environmental and sociodemographic-socioeconomic factors. Objective: This study aims at characterizing different risk factors in the published literature that influence contagion by COVID-19. Methods: The review consists of three stages, including a systematic mapping with studies found in the Scopus database, an analysis of results, and finally the identification of relevant COVID-19 risk factors. Results: A map of studies id provided considering two main groups: the type of research and context. Most studies consider risk factors associated with medical conditions, while research on other factors is scarce. Conclusions: Medical conditions such as diabetes, obesity, cardiovascular disease, hypertension, and factors such as age and sex, appear to be the ones that increase the risk of contracting COVID-19. Further research is needed on environmental, sociodemographic, and socioeconomic risk f [ABSTRACT FROM AUTHOR]

Computerized techniques for image analysis are critical for progress in cell biology. The complexity of the data in current methods eliminates the need for manual image analysis and usually requires the application of multiple algorithms sequentially to the images. Our aim was to develop a software for immunohistochemical analysis of brain dopaminergic neurons combining several computational approaches to automatically analyze and quantify their number in the substantia nigra after a neurotoxic injury. For this purpose, we used a Parkinson's disease animal model to test our application. The dopaminergic neurotoxin, 6-hydroxydopamine, was administered in adult male rats to damage dopaminergic neurons in substantia nigra and to induce hemiparkinsonism. The lesion was corroborated by behavioral evaluation in response to apomorphine and amphetamine. The animals were euthanized and their brains processed for tyrosine hydroxylase immunohistochemistry for dopamine neuron identification. Neurons positive for tyrosine hydroxylase were evaluated in substantia nigra by light microscopy. The images were used to show quantification applicability. To test our software counting accuracy and validity, automatic dopamine neuron number was correlated with the data obtained by three independent observers. Several parameters were used to depict neuronal function in dataset images from control and lesioned brains. In conclusion, we could perform an automated quantification of dopaminergic neurons and corroborate the validity and accuracy of a freely available software. [ABSTRACT FROM AUTHOR]

Epilepsy diagnosis is a medical care process that requires considerable transformation, mainly in developed countries, to provide efficient and effective care services taking into consideration the low number of available neurologists, especially in rural areas. EEG remains the most common test used to diagnose epilepsy. In recent years, there has been an increase in deep learning techniques to analyze electroencephalograms (EEG) to detect epileptiform events. These types of techniques support the epilepsy diagnostic processes performed by neurologists. There have been several approaches such as biomedical signal processing, analysis of characteristics extracted from the signals, and image analysis to detect epileptiform events. Most of the works reported in the literature, which use images, transformed the signals into a two-dimensional space interpreted as an image. However, only a few of them use the raw EEG image. This paper presents a computational model for detecting epileptiform events from raw EEG images, using convolutional neural networks and a transfer learning approach. To perform this work, 100 pediatric EEGs were collected, noting six characteristics of epileptiform events in each exam: spikes, poly-spikes, spike-and-wave, sharp waves, periodic, and a combination of them. Then, pre-trained convolutional neural networks were used, which, through transfer learning techniques, were retrained to classify possible events. The model's performance was evaluated in terms of precision, accuracy, and Mathews' correlation coefficient. The model offered a performance above 95% accuracy for binary classification and above 87% for multi-class classification. These results demonstrated that identifying epileptiform events from raw EEG images combined with deep learning techniques such as transfer learning is feasible. Significance: The proposed method for the evaluation of EEG tests, as a support tool for the diagnosis of epilepsy, can help to reduce the time of reading EEGs, which is very important, especially in developing countries with a limitation of a specialist in neurology. [ABSTRACT FROM AUTHOR]

Several approaches from different perspectives have been used to solve problems with traffic accidents (TA), which mainly affect low- and middle-income countries. Conditions of certain cities, regarding road infrastructure, enforcement of traffic safety regulations, and motor vehicle numbers, influence the increase in TAs. Therefore, medium-sized cities in developing countries (context of interest), which commonly have worrying conditions, are a relevant scenario. One of the approaches to reduce TAs has been the use of data analysis through Machine Learning (ML); however, these techniques require a large amount of data, and medium-sized cities commonly do not have enough. Techniques such as Naturalistic Driving (ND) can be applied as a data collection method. This work proposes an intelligent collision risk detection system (ICDRS) using ND and ML to improve sustainability and safety of transportation in medium-sized cities. The ICRDS design considered the limitations of the context of interest and uses two data collection devices in the vehicle. The ICRDS validation included the design and execution of tests using ND. This validation identified if the collected data in a certain time interval contained high-risk collision events (sudden acceleration, sudden braking, aggressive left or right turn, aggressive left or right lane change). The system implementation results were satisfactory. The developed ML algorithm obtained an average value 0.98 in all the metrics. Two data sets of driving on routes were collected. In addition, the performed tests were able to identify city areas with high accident rates. [ABSTRACT FROM AUTHOR]

The coupling of PHB generation with NADH reoxidation is required to generate PHB as a fermentation product. A fundamental trait to accomplish this feature is to express a functional NADH-preferring acetoacetyl-CoA reductase, engaged in PHB accumulation. One way to obtain such a reductase is by engineering the cofactor preference of the acetoacetyl-CoA reductase encoded by the phaB1 gene from Cupriavidus necator (AARCn1). Aiming to have a deeper understanding of the structural determinants of the cofactor preference in AARCn1, and to obtain an NADH-preferring acetoacetyl-CoA reductase derived from this protein, some engineered enzymes were expressed, purified and kinetically characterized, together with the parental AARCn1. One of these engineered enzymes, Chimera 5, experimentally showed a selectivity ratio ((kcat/KM)NADH/(kcat/KM)NADPH) ≈ 18, which is 160 times higher than the selectivity ratio experimentally observed in the parental AARCn1. A thermodynamic-kinetic approach was employed to estimate the cofactor preference and flux capacity of Chimera 5 under physiological conditions. According to this approach, Chimera 5 could prefer NADH over NADPH between 25 and 150 times. Being a derivative of AARCn1, Chimera 5 should be readily functional in Escherichia coli and C. necator. Moreover, with the expected expression level, its activity should be enough to sustain PHB accumulation fluxes similar to the fluxes previously observed in these biotechnologically relevant cell factories. [ABSTRACT FROM AUTHOR]

Background: Availability and opportunity of epilepsy diagnostic services is a significant challenge, especially in developing countries with a low number of neurologists. The most commonly used test to diagnose epilepsy is electroencephalogram (EEG). A typical EEG recording lasts for 20 to 30 minutes; however, a specialist requires much more time to read it. Furthermore, no evidence was found in the literature on open-source systems for the cost-effective management of patient information using electronic health records (EHR) that adequately integrate EEG analysis for automatic identification of abnormal signals. Objective: To develop an integrated open-source EHR system for the management of the patients' personal, clinical, and EEG data, and for automatic identification of abnormal EEG signals. Methods: The core of the system is an EHR and telehealth service based on the OpenMRS platform. On top of that, we developed an intelligent component to automatically detect abnormal segments of EEG tests using machine learning algorithms, as well as a service to annotate and visualize abnormal segments in EEG signals. Finally, we evaluated the intelligent component and the integrated system using precision, recall, and accuracy metrics. Results: The system allowed to manage patients' information properly, store and manage the EEG tests recorded with a medical EEG device, and to detect abnormal segments of signals with a precision of 85.10%, a recall of 97.16%, and an accuracy of 99.92%. Conclusion: Digital health is a multidisciplinary field of research in which artificial intelligence is playing a significant role in boosting traditional health services. Notably, the developed system could significantly reduce the time a neurologist spends in the reading of an EEG for the diagnosis of epilepsy, saving approximately 65– 75% of the time consumed. It can be used in a telehealth environment. In this way, the availability and provision of diagnostic services for epilepsy management could be improved, especially in developing countries where the number of neurologists is low. [ABSTRACT FROM AUTHOR]

The present study aimed to determine whether an i.c.v. administration of allopregnanolone (ALLO) rapidly modifies the hypothalamic and ovarian 3β‐hydroxysteroid dehydrogenase (3β‐HSD) enzymatic activity and gene expression in in vivo and ex vivo systems in pro‐oestrus (PE) and dioestrus I (DI) rats. Animals were injected with vehicle, ALLO, bicuculline or bicuculline plus ALLO and were then killed. In the in vivo experiment, the hypothalamus, ovaries and serum were extracted and analysed. In the ex vivo experiment, the superior mesenteric ganglion ‐ ovarian nerve plexus ‐ ovary system was extracted and incubated during 120 minutes at 37 ºC. The serum and ovarian compartment fluids were used to determine progesterone by radioimmunoanalysis. In the in vivo experiments, ALLO caused a decrease in hypothalamic and ovarian 3β‐HSD enzymatic activity during PE. During DI, ALLO increased hypothalamic and ovarian 3β‐HSD activity and gene expression. The ovarian 3β‐HSD activity increased in both stages in the ex vivo system; gene expression increased only during DI. ALLO induced an increase in serum progesterone only in D1 and in the ovarian incubation liquids in both stages. All findings were reversed by an injection of bicuculline before ALLO. Ovarian steroidogenic changes could be attributed to signals coming from ganglion neurones, which are affected by the acute central neurosteroid stimulation. The i.c.v. administration of ALLO via the GABAergic system altered 3β‐HSD activity and gene expression, modulating the neuroendocrine axis. The present study reveals the action that ALLO exerts on the GABAA receptor in both the central and peripheral nervous system and its relationship with hormonal variations. ALLO is involved in the "fine tuning" of neurosecretory functions as a potent modulator of reproductive processes in female rats. [ABSTRACT FROM AUTHOR]

Systems trying to solve mobility issues in cities, such as high levels of accidents and traffic congestion, have been developed worldwide. Intelligent Transportation Systems (ITS) services focused on urban public transport are an option contributing to solve such issues. A few intermediate cities in the Latin American context have developed some of these ITS services, which are mainly based on a tracking system for public transport vehicles. Such tracking systems have great limitations in terms of coverage, availability, and operational cost. In addition, they are commonly isolated mobility solutions, which cannot be easily integrated with other mobility services in the city because they are not based on an ITS architecture. In order to improve public transportation systems in intermediate cities, we proposed the development of an IoT-based public vehicle tracking system, using LoRa (Long Range) and ITS services. In this research, we developed the proposed system as a proof of concept. We designed and executed some experiments, in order to adjust parameters of the LoRa technology and to test its operation. This article presents the methods we followed for developing the proof-of-concept model, a description of the experiments, and their results. The results lead to conclude that the LoRa technology and an IoT-based system are adequate for implementation of a mobility service such as the one we propose, once important technical restrictions related mainly to Line of Sight (LoS) are considered. Key aspects for implementation were also identified for deploying the service (as a prototype) in the city of Popayán. [ABSTRACT FROM AUTHOR]

Inorganic polyphosphate (polyP) and its metabolic enzymes are important in several cellular processes related with virulence and antibiotic susceptibility. Accordingly, bacterial polyP synthesis has been proposed as a good target for designing novel antivirulence molecules as alternative to conventional antibiotics. In most pathogenic bacteria, polyphosphate kinase 1 (PPK1), in charge of polyP synthesis from ATP, is widely conserved. Current colorimetric and radioactive polyP synthesis enzymatic assays are not suitable for high-throughput screening of PPK1 inhibitors. Given the ability of polyP to modify the excitation-emission spectra of DAPI (4ʹ-6-diamidino-2-phenylindole), a fluorescence assay was previously developed by using a purified recombinant PPK1 enzyme from Escherichia coli. In this work we have developed a suitable methodology for high-throughput measurement of E. coli PPK1 activity. This platform can be used for the screening putative antimicrobial molecules for related enteropathogenic bacteria. [ABSTRACT FROM AUTHOR]

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It has been proposed that NADP+-specificity of isocitrate dehydrogenase (ICDH) evolved as an adaptation of microorganisms to grow on acetate as the sole source of carbon and energy. In Escherichia coli, changing the cofactor specificity of ICDH from NADP+ to NAD+ (cofactor swapping) decreases the growth rate on acetate. However, the metabolic basis of this phenotype has not been analyzed. In this work, we used constraint-based modeling to investigate the effect of the cofactor swapping of ICDH in terms of energy production, response of alternative sources of NADPH, and partitioning of fluxes between ICDH and isocitrate lyase (ICL) -a crucial bifurcation when the bacterium grows on acetate-. We generated E. coli strains expressing NAD+-specific ICDH instead of the native enzyme, and bearing the deletion of the NADPH-producing transhydrogenase PntAB. We measured their growth rate and acetate uptake rate, modeled the distribution of metabolic fluxes by Flux Balance Analysis (FBA), and quantified the specific activities of NADPH-producing dehydrogenases in central pathways. The cofactor swapping of ICDH led to one-third decrease in biomass yield, irrespective of the presence of PntAB. According to our simulations, the diminution in growth rate observed upon cofactor swapping could be explained by one-half decrease in the total production of NADPH and a lower availability of carbon for biosynthesis because of a change in the partition at the isocitrate bifurcation. Together with an increased total ATP production, this scenario resulted in a 10-fold increment in the flux of ATP not used for growing purposes. PntAB was identified as the primary NADPH balancing response, with the dehydrogenases of the oxidative branch of the Pentose Phosphate Pathway and the malic enzyme playing a role in its absence. We propose that in the context of E. coli growing on acetate, the NADP+-specificity of ICDH is a trait that impacts not only NADPH production, but also the efficient allocation of carbon and energy. [ABSTRACT FROM AUTHOR]

Abstract: Background: Despite suffering high ultraviolet radiation levels, few data on malignant melanoma (MM) in Macaronesia are available. Methods: Observational study of cutaneous MM cases diagnosed during a period of 12 years at a tertiary hospital in Canary Islands. Results: A total of 532 patients (female/male = 1.4) with an average age of 56 years were included; 5% developed more than one MM, and 7% reported family history of MM. Phototype II (43%), dark eyes (41%), and dark hair (41%) predominated. There was a lower frequency of light‐colored hair and eyes in those born in the Canary Islands. The most frequent locations of MM were on the back for men (37%) and on the lower extremities for women (35%). Among the infiltrating tumors (83%), the (median) thickness was 1.07 mm (women, 0.90 mm; men, 1.21 mm). Anatomopathological ulceration (AU) and a mitotic rate ≥1 mitosis/mm2 (HMR) were recorded in 27% of patients. Patients with regional disease constituted 12% of the population. The most common stage was IA (34%). Melanoma‐specific survival (MSSV) decreased significantly with thickness, presence of AU, HMR, and sentinel lymph node disease. These four variables were independent prognostic factors. The five‐year MSSV varied between 100% (stage IA) and 39% (stage IIIC). Conclusions: The characteristics of the patients were similar to those published in datasets from continental Europe, although the pigmentary features were darker in those originating from Macaronesia. The prognostic parameters described in the 7th edition of the American Joint Committee on Cancer (AJCC) independently predict MSSV in our patients. [ABSTRACT FROM AUTHOR]

Glucose 6-Phosphate Dehydrogenases (G6PDHs) from different sources show varying specificities towards NAD+ and NADP+ as cofactors. However, it is not known to what extent structural determinants of cofactor preference are conserved in the G6PDH family. In this work, molecular simulations, kinetic characterization of site-directed mutants and phylogenetic analyses were used to study the structural basis for the strong preference towards NADP+ shown by the G6PDH from Escherichia coli. Molecular Dynamics trajectories of homology models showed a highly favorable binding energy for residues K18 and R50 when interacting with the 2'-phosphate of NADP+, but the same residues formed no observable interactions in the case of NAD+. Alanine mutants of both residues were kinetically characterized and analyzed with respect to the binding energy of the transition state, according to the kcat/KM value determined for each cofactor. Whereas both residues contribute to the binding energy of NADP+, only R50 makes a contribution (about -1 kcal/mol) to NAD+ binding. In the absence of both positive charges the enzyme was unable to discriminate NADP+ from NAD+. Although kinetic data is sparse, the observed distribution of cofactor preferences within the phylogenetic tree is sufficient to rule out the possibility that the known NADP+-specific G6PDHs form a monophyletic group. While the β1-α1 loop shows no strict conservation of K18, (rather, S and T seem to be more frequent), in the case of the β2-α2 loop, different degrees of conservation are observed for R50. Noteworthy is the fact that a K18T mutant is indistinguishable from K18A in terms of cofactor preference. We conclude that the structural determinants for the strict discrimination against NAD+ in the case of the NADP+-specific enzymes have evolved independently through different means during the evolution of the G6PDH family. We further suggest that other regions in the cofactor binding pocket, besides the β1-α1 and β2-α2 loops, play a role in determining cofactor preference. [ABSTRACT FROM AUTHOR]

There is a growing amount of evidence for a neuroprotective role of progesterone and its neuroactive metabolite, allopregnanolone, in animal models of neurodegenerative diseases. By using a model of hemiparkinsonism in male rats, injection of the neurotoxic 6-OHDA in left striatum, we studied progesterone’s effects on rotational behavior induced by amphetamine or apomorphine. Also, in order to find potential explanatory mechanisms, we studied expression and activity of nigrostriatal 3α-hydroxysteroid oxidoreductase, the enzyme that catalyzes progesterone to its active metabolite allopregnanolone. Coherently, we tested allopregnanolone for a possible neuromodulatory effect on rotational behavior. Also, since allopregnanolone is known as a GABAA modulator, we finally examined the action of GABAA antagonist bicuculline. We found that progesterone, in addition to an apparent neuroprotective effect, also increased ipsilateral expression and activity of 3α-hydroxysteroid oxidoreductase. It was interesting to note that ipsilateral administration of allopregnanolone reversed a clear sign of motor neurodegeneration, that is, contralateral rotational behavior. A possible GABAA involvement modulated by allopregnanolone was shown by the blocking effect of bicuculline. Our results suggest that early administration of progesterone possibly activates genomic mechanisms that promote neuroprotection subchronically. This, in turn, could be partially mediated by fast, nongenomic, actions of allopregnanolone acting as an acute modulator of GABAergic transmission. [ABSTRACT FROM AUTHOR]

Ethanol consumption during pregnancy may induce profound changes in fetal CNS development. We postulate that some of the effects of ethanol on striatal glutamatergic transmission and neurotrophin expression could be modulated by allopregnanolone, a neurosteroid modulator of GABAA receptor activity. We describe the acute pharmacological effect of allopregnanolone (65 μg/kg, s.c.) administered to juvenile male rats (day 21 of age) on the corticostriatal glutamatergic pathway, in both control and prenatally ethanol-exposed rats (two ip injections of 2.9 g/kg in 24% v/v saline solution on gestational day 8). Prenatal ethanol administration decreased the K+-induced release of glutamate regarding the control group. Interestingly, this effect was reverted by allopregnanolone. Regarding BDNF, allopregnanolone decreases the content of this neurotrophic factor in the striatum of control groups. However, both ethanol alone and ethanol plus allopregnanolone treated animals did not show any change regarding control values. We suggest that prenatal ethanol exposure may produce an alteration of GABAA receptors which blocks the GABA agonist-like effect of allopregnanolone on rapid glutamate release, thus disturbing normal neural transmission. Furthermore, the reciprocal interactions found between GABAergic neurosteroids and BDNF could underlie mechanisms operating during the neuronal plasticity of fetal development. [ABSTRACT FROM AUTHOR]

Polyols are enzymatically-produced plant compounds which can act as compatible solutes during periods of abiotic stress. Nicotinamide adenine dinucleotide+-dependent SORBITOL DEHYDROGENASE (SDH, E. C. 1.1.1.14) from Arabidopsis thaliana L. sorbitol dehydrogenase (AtSDH) is capable of oxidizing several polyols including sorbitol, ribitol, and xylitol. In the present study, enzymatic assays using recombinant AtSDH demonstrated a higher specificity constant for xylitol compared to sorbitol and ribitol, all of which are C2 (S) and C4 (R) polyols. Enzyme activitywas reduced by preincubation with ethylenediaminetetraacetic acid, indicating a requirement for zinc ions. In humans, it has been proposed that sorbitol becomes part of a pentahedric coordination sphere of the catalytic zinc during the reaction mechanism. In order to determine the validity of this pentahedric coordination model in a plant SDH, homology modeling, and Molecular Dynamics simulations of AtSDH ternary complexes with the three polyols were performed using crystal structures of human and Bemisia argentifolii (Genn.) (Hemiptera: Aleyrodidae) SDHs as scaffolds. The results indicate that the differences in interaction with structural water molecules correlate very well with the observed enzymatic parameters, validate the proposed pentahedric coordination of the catalytic zinc ion in a plant SDH, and provide an explanation for why AtSDH shows a preference for polyols with a chirality of C2 (S) and C4 (R). [ABSTRACT FROM AUTHOR]

In the last ten years, approximately, urban transit systems of Latin American capital cities have evolved significantly. Colombia, specifically, has concentrated this development in its capital cities, consolidated through digital transformation programs in the transportation sector. However, the same phenomenon does not occur in medium-sized cities for different reasons that are important to analyze. This paper presents an exploratory qualitative study involving eight medium-sized cities in the implementation phase of their strategic urban transit systems. Three main aspects that drive this study were identified: technologies and their cost, functional requirements to implement information technology services in transit systems, and economy and administration associated with this type of implementation. Based on this, a semi-structured interview data collection instrument was designed, with the participation of 15 officials distributed in the eight target cities, and one expert from an intelligent transportation system in a capital city. With the information collected, an exploratory analysis was made contrasting the responses given by each interviewee. The most relevant results show that the interviewees prioritize technologies based on open standards to provide information to users; that the northern medium-sized cities of the country do not have strategies that regularize and motivate the use of public transportation; instead, the southern medium-sized cities of the country consider the use of transportation to be necessary. Finally, it was concluded that the information technology services to be included in the provision of transit services should promote these cities' cultural and economic growth. [ABSTRACT FROM AUTHOR]

Objectives: Progesterone has been reported to have a neuroprotective role in depression-like rats in a hemiparkinsonian model of the disease. In this work, we investigate if this hormone affects the three principal neurochemicals striatal systems (dopaminergic, glutamatergic, and GABAergic) that are involved in the physiopathology of the disease in a hemiparkinsonim male rat model at 8 weeks post-chemical injury. Methods: For this purpose, we design three experimental groups: (1) sham group; (2) hemiparkinsonian group; and (3) hemiparkinsonian group subcutaneously injected with progesterone at 7 days post- chemical injury. Animals were tested in an automated rotational device at 8 weeks post-chemical injury. After behavioral test, K+-evoked [³H]-dopamine, [³H]-glutamate, and [³H]-gamma aminobutyric acid release from striatum slices were analyzed by superfusion experiments. Results: The hemiparkinsonian group showed distinctive alterations that are produced by neurodegenera- tion of left nigrostriatal dopaminergic pathway by 6-hydroxydopamine hydrobromide (6-OHDA). On the other hand, the administration of progesterone 7 days after the injection of the neurotoxin was able to (1) improve the K+-evoked [³H]-dopamine release from the damaged striata (left); (2) avoid significant increase in the K+-evoked [³H]-glutamate release from the left striata; and (3) progesterone does not modify the K+-evoked [³H]-gamma aminobutyric acid release from the left striata. Discussion: These results suggest that progesterone does have neuroprotective and neuromodulatory effects on striatal neurotransmission systems in the hemiparkinsonian male rats. The possible mechanisms would involve genomic and non-genomic actions of this neuroactive steroid which would modulate the activity of dopaminergic, glutamatergic, and GABAergic pathways. [ABSTRACT FROM AUTHOR]

Oestradiol (E2) is a key hormone in the regulation of reproductive processes. The aims of this work were a) to examine the distributions of oestrogen receptor α (ERα) and ERb in the neurons of the superior mesenteric ganglion (SMG) in the oestrus stage by immunohistochemistry, b) to demonstrate whether E2 in the SMG modifies progesterone (P4), androstenedione (A2) and nitrite release in the ovarian compartment on oestrus day and c) to demonstrate whether E2 in the ganglion modifies the activity and gene expression in the ovary of the steroidogenic enzymes 3β-hydroxysteroid dehydrogenase (3β-HSD) and 20α-hydroxysteroid dehydrogenase (20α-HSD). The ex vivo SMG-ovarian nervous plexus-ovary system was used. E2, tamoxifen (Txf) and E2 plus Txf were added in the ganglion to measure ovarian P4 release, while E2 alone was added to measure ovarian A2 and nitrites release. Immunohistochemistry revealed cytoplasmic ERα immunoreactivity only in the neural somas in the SMG. E2 increased ovarian P4 and A2 release at 15, 30 and 60 min but decreased nitrites. The activity and gene expression of 3β-HSD increased, while the activity and gene expression of 20α-HSD did not show changes with respect to the control. Txf in the ganglion diminished P4 release only at 60 min. E2 plus Txf in the ganglion reverted the effect of E2 alone and the inhibitory effect of Txf. The results of this study demonstrate that ERα activation in the SMG has an impact on ovarian steroidogenesis in rats, thus providing evidence for the critical role of periphERαl system neurons in the control of ovarian functions under normal and pathological conditions. [ABSTRACT FROM AUTHOR]

LHRH release from hypothalamus is influenced by the neurotransmitter glutamate that acts, among others, on NMDA receptors present in LHRH neurons. On the other hand, the neurosteroid allopregnanolone can modulate the activity of specific neurotransmitter receptors and affect neurotransmitter release. We examined the role of allopregnanolone on in vitro LHRH and glutamate release from mediobasal hypothalamus and anterior preoptic area of ovariectomized rats with estrogen and progesterone replacement. Moreover, we evaluated whether the neurosteroid might act through modulation of NMDA receptors. Allopregnanolone induced an increase in LHRH release. This effect was reversed when the NMDA receptors were blocked by the NMDA antagonist 2-amino-7-phosphonoheptanoic acid (AP-7) indicating that this neurosteroid would interact with NMDA receptors. Moreover allopregnanolone induced an augment in K evoked [H]-glutamate release from mediobasal hypothalamus-anterior preoptic area explants and this effect was also reversed when NMDA receptors were blocked with AP-7. These results suggest an important physiologic function of allopregnanolone on the regulation of neuroendocrine function in female adult rats. Not only appears to be involved in enhancing LHRH release through modulation of NMDA receptors but also in the release of glutamate which is critical in the control of LHRH release. [ABSTRACT FROM AUTHOR]

It has been shown that a single exposure to amphetamine is sufficient to induce long-term behavioral, neurochemical, and neuroendocrine sensitization in rats. Dopaminergic neurotransmission in the nucleus accumbens and the caudateputamen plays a critical role in the addictive properties of drugs of abuse. Angiotensin (Ang) II receptors are found on the soma and terminals of mesolimbic dopaminergic neurons and it has been shown that Ang II acting through its AT1 receptors facilitates dopamine release. The hypothesis was tested that Ang II AT1 receptors are involved in the neuroadaptative changes induced by a single exposure to amphetamine and that such changes are related to the development of behavioral and neurochemical sensitization. For this purpose, the study examined the expression of amphetamine-enhanced (0.5 mg kg-1 i.p.) locomotor activity in animals pretreated with candesartan, an AT1 blocker, (3 mg kg-1 p.o 3 5 days), 3 weeks after an amphetamine injection (5 mg kg-1 i.p.). Dopaminergic hyperreactivity was tested by measuring the 3H-DA release in vitro from caudate-putamen and nucleus accumbens slices, induced by K+ stimulus. It was confirmed the behavioral sensitization in the two-injection protocoland candesartan pretreatment attenuate this response. It was also found that AT1 blockade pretreatment did not affect the locomotor response to dopamine agonists. In respect to the neurochemical sensitization tested using ex vivo 3H-DA release experiments it was found that AT1 receptor pretreatment blunted the enhanced response induced by K+ stimulus. The results support the idea that the development of neuroadaptive changes induced by amphetamine involves brain AT1 Ang II receptor activation. [ABSTRACT FROM AUTHOR]

The present study evaluated the reproductive compatibility of the crosses between adult ticks of the following three geographically different populations of Amblyomma cajennense: State of São Paulo (SP), southeastern Brazil; State of Rondônia (RO), northern Brazil; and Colombia (CO). In addition, crosses between A. cajennense ticks from Argentina (AR) and SP ticks were also performed. The Argentinean population (AR) was compatible with SP because their crosses resulted in high % egg hatching (mean values ranging from 71.5 to 93.5%), similarly to all homologous (intrapopulational) crosses. In contrast, the tick populations SP, RO, and CO were shown to be incompatible with each other, since their heterologous (interpopulational) crosses always resulted in very low % egg hatching (range: 0-5%). The F larval offspring derived from some of these females that yielded 5% egg hatching were reared until the F adult stage. In all cases, only adult females molted from engorged nymphs. These F females were likely to be a product of thelytokous parthenogenesis of the SP, RO, and CO females that were used in the heterologous crosses. Reproductive incompatibility is not expected to occur between different populations of a single species. Thus, our results suggest that the taxon A. cajennense might be represented by a complex of different species, whereas SP and AR ticks might represent a single species. Further populational genetic studies, coupled with extensive morphological analyses, are needed to clarify and determine a possible complex of valid species that might have been classified under the taxon A. cajennense. [ABSTRACT FROM AUTHOR]

Copyright of Ginecología y Obstetricia de México is the property of Federacion Mexicana de Ginecologia y Obstetricia and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Ginecología y Obstetricia de México is the property of Federacion Mexicana de Ginecologia y Obstetricia and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

In previous studies we have found that blockade of NMDA ( N-Methyl- d-Aspartic-Acid)-type glutamatergic receptor with intracerebroventricular (ICV) selective drugs induces an inhibition of lordosis in ovariectomized (OVX) estrogen primed rats receiving progesterone or luteinizing hormone releasing hormone (LHRH). By the opposite way, stimulation with NMDA in OVX estrogen primed rats induced a significant increase of lordosis. In the present study the action of an α1-noradrenergic antagonist, HEAT (BE 2254/2-beta-4-Hydroxyphenyl-Ethyl-Aminomethyl-1-Tetralone), and Metoprolol, a β-noradrenergic antagonist, were studied injecting them ICV previously to NMDA administration in treated OVX estrogen primed rats. In experiment 1, the enhancing effect on lordosis induced by NMDA at high dose (1 μg) was abolished by HEAT administration ( P < 0.001 for 3 and 6 μg), and the LH plasma levels were decreased only with the higher dose ( P < 0.05), suggesting that behavioral effects are quite more sensitive to the α-blockade than hormonal effects. In experiment 2, enhancing effects on lordosis behavior were not observed with neither the NMDA at low dose (0.5 μg) nor the metoprolol alone (5.71 μg), but a synergism was observed when both were simultaneously administered ( P < 0.001). The LH plasma levels were increased by Metoprolol alone ( P < 0.05), and powered by the combination with NMDA at low dose ( P < 0.01 vs. SAL and NMDA alone); no differences were observed with Metoprolol. LH increase was observed with Metoprolol even without behavioural modifications. These findings strongly suggest that facilitatory and inhibitory effects of NMDA in this model are mediated by α- and β-adrenergic transmission in both, behavioral and hormonal effects. [ABSTRACT FROM AUTHOR]

Thyroid transcription factor 1 (TTF1) [also known as Nkx2.1 (related to the NK-2 class of homeobox genes) and T/ebp (thyroid-specific enhancer-binding protein)], a homeodomain gene required for basal forebrain morphogenesis, remains expressed in the hypothalamus after birth, suggesting a role in neuroendocrine function. Here, we show an involvement of TTF1 in the control of mammalian puberty and adult reproductive function. Gene expression profiling of the nonhuman primate hypothalamus revealed that TTF1 expression increases at puberty. Mice in which the Ttf1 gene was ablated from differentiated neurons grew normally and had normal basal ganglia/hypothalamic morphology but exhibited delayed puberty, reduced reproductive capacity, and a short reproductive span. These defects were associated with reduced hypothalamic expression of genes required for sexual development and deregulation of a gene involved in restraining puberty. No extrapyramidal impairments associated with basal ganglia dysfunction were apparent. Thus, although TTF1 appears to fulfill only a morphogenic function in the ventral telencephalon, once this function is satisfied in the hypothalamus, TTF1 remains active as part of the transcriptional machinery controlling female sexual development. [ABSTRACT FROM AUTHOR]

Abstract: In a previous work, chemical modification of Cys-238 of Escherichia coli Pfk-2 raised concerns on the importance of the dimeric state of Pfk-2 for enzyme activity, whereas modification of Cys-295 impaired the enzymatic activity and the MgATP-induced tetramerization of the enzyme. The results presented here demonstrate that the dimeric state of Pfk-2 is critical for the stability and the activity of the enzyme. The replacement of Cys-238 by either Ala or Phe shows no effect on the kinetic parameters, allosteric inhibition, dimer stability and oligomeric structure of Pfk-2. However, the mutation of Cys-295 by either Ala or Phe provokes a decrease in the k cat value and an increment in the K m values for both substrates. We suggest that the Cys-295 residue participates in intersubunit interactions in the tetramer since the Cys-295-Phe mutant exhibits higher tetramer stability, which in turn results in an increase in the fructose-6-P concentration required for the reversal of the MgATP inhibition relative to the wild type enzyme. [Copyright &y& Elsevier]

1. The neurosteroids are compounds derived from steroid hormones and synthesized in the nervous system. They can modulate different neurotransmitter pathways. In previous work we demonstrated that progesterone modulates dopamine release induced by the glutamatergic agonist N-methyl-D-aspartic acid (NMDA). 2. The aim of this work was to evaluate a possible modulatory role of the progesterone metabolite allopregnanolone on NMDA-evoked [3H]dopamine release from corpus striatum slices obtained from cycling and ovariectomized female rats. 3. We used a dynamic superfusion method to evaluate the release of [3H]dopamine. Allopregnanolone at 50–600 nM was added to the superfusion buffer (Krebs–Ringer–bicarbonate–glucose, pH 7.4, with constant O2/CO2 gassing). The results are expressed as a percentage over basal [3H]dopamine loaded by the tissue. 4. Allopregnanolone (50 and 100 nM) increased the NMDA-evoked[3H]dopamine release from estrus rats. The remaining doses did not show significant changes in the pattern of release. This effect was not observed in diestrus rats. The ovariectomy abolished the facilitatory effect of allopregnanolone on NMDA-evoked 2 [3H]dopamine release. 5. Subcutaneous administration of exogenous estrogen (25 mg/rat) and progesterone (1 mg/rat) restored the facilitatory effect on dopaminergic input. 6. These results suggest that allopregnanolone is a neurosteroid able to modulate dopamine release in an ovarian-hormone-fluctuation-dependent manner and provide further support for a role of allopregnanolone as a modulator of glutamatergic–dopaminergic interaction in the corpus striatum. [ABSTRACT FROM AUTHOR]

The effect of intracerebroventricular (icv) injection of allopregnanolone (5α-pregnan-3α-ol-20-one) on the dopaminergic and reproductive function in ovariectomized rats primed with estrogen and progesterone was investigated. Thirty minutes after icv allopregnanolone injection, the sexual receptivity, luteinizing hormone (LH) release, dopamine content, and release in the medial basal hypothalamus (MBH) and preoptic area (POA) were determined. After allopregnanolone injection, LH serum levels were reduced ( p<0.001) and lordosis behavior was inhibited ( p<0.005). Intracerebroventricular injection of bicuculline (a γ-aminobutyric acid [GABA] antagonist) alone was ineffective. The injection of allopregnanolone plus bicuculline blocked the effects of allopregnanolone on sexual receptivity and on LH serum levels. At the same time, endogenous dopamine concentration in both the MBH and POA was augmented ( p<0.005 and p<0.006, respectively) and the turnover rate decreased in both structures. Moreover, in vitro H-dopamine release from MBH and POA was lower in rats injected with allopregnanolone in comparison with vehicle-treated rats. These results suggest that allopregnanolone influences the dopaminergic mechanisms in female rats, which may, in turn, be responsible for the reduced reproductive activity. Allopreganolone may exert its effects on sexual behavior through GABA receptor modulation and a decrease in dopaminergic activity in the MBH and POA. These actions could explain the inhibition of LH release. [ABSTRACT FROM AUTHOR]

Traffic congestion has several causes, including insufficient road capacity, unrestricted demand and improper scheduling of traffic signal phases. A great variety of efforts have been made to properly program such phases. Some of them are based on traditional transportation assumptions, and others are adaptive, allowing the system to learn the control law (signal program) from data obtained from different sources. Reinforcement Learning (RL) is a technique commonly used in previous research. However, properly determining the states and the reward is key to obtain good results and to have a real chance to implement it. This paper proposes and implements a traffic signal control system (TSCS), detailing its development stages: (a) Intelligent Transportation System (ITS) architecture design for the TSCS; (b) design and development of a system prototype, including an RL algorithm to minimize the vehicle queue at intersections, and detection and calculation of such queues by adapting a computer vision algorithm; and (c) design and development of system tests to validate operation of the algorithms and the system prototype. Results include the development of the tests for each module (vehicle queue measurement and RL algorithm) and real-time integration tests. Finally, the article presents a system simulation in the context of a medium-sized city in a developing country, showing that the proposed system allowed reduction of vehicle queues by 29%, of waiting time by 50%, and of lost time by 50%, when compared to fixed phase times in traffic signals. [ABSTRACT FROM AUTHOR]

The effects of spontaneous circling motor activity on the in vitro phosphorylation of the protein kinase C substrate GAP-43/B-50 was studied on striatal membranes of developing rats (30 days of age). At this time of postnatal development, permanent plastic changes in cholinergic and dopaminergic systems are produced by physiological motor activity. Exercised animals showed a significant reduction of 31% in the level of GAP-43/B-50 endogenous phosphorylation in the contralateral striatum respect to the ipsilateral side (P < 0.01), while control animals did not show asymmetric differences. Compared to controls, the contralateral striatum of exercised animals showed a 33% reduction in the incorporation of 32P-phosphate into GAP-43/B-50 30 minutes post-exercise (P < 0.01). This change in GAP-43/B-50 phosphorylation was correlated with the running speed developed by the animals (r:0.8986, P = 0.015). GAP-43/B-50 immunoblots revealed no changes in the amount of this protein in any group. Moreover, a significant variation of 25% (P < 0.05) in the PKC activity was seen between both exercised striata. Interhemispheric differences were not found in control animals. We conclude that endogenous phosphorylation of this protein is also altered by motor activity in the same period that permanent changes in striatal neuroreceptors are triggered after motor training. [ABSTRACT FROM AUTHOR]

Long-range (LoRa) technology is a low power wide area network (LPWAN) technology that is currently being used for development of Internet of things (IoT)-based solutions. Transit transport, mainly in medium-sized cities where transit vehicles do not have exclusive lanes, is a service that can be improved with a tracking service using technology such as LoRa. Although some proposals exist, there is not enough experimental information to validate the LoRa technology as adequate. This article: (a) evaluates the operation of LoRa technology in a transit vehicle tracking service in a medium-sized city, based on an Intelligent Transportation Systems architecture and IoT; and (b) investigates optimal LoRa technology configuration parameters for the service. Experiments were performed in a semi-controlled environment using LoRa devices and a gateway, by measuring the received packets and the receive signal strength indicator (RSSI) and modifying: (a) distance; (b) number of devices; and (c) the main LoRa transmission parameters. Obtained results show the ideal values of parameters vary considerably with distance and number of devices used. There were very few settings of the experiments in which the RSSI and packet levels were adequate while distance and number of devices were both changed. [ABSTRACT FROM AUTHOR]

In medium-sized cities in developing countries, transit services without dedicated lanes have issues related to route compliance, schedules, speed control, and safety. An efficient way for dealing with this issue is the use of Information and Communication Technologies (ICT), to implement a Fleet Management and Control Systems (FMCS). Such implementation can be performed using Intelligent Transportation Systems (ITSs), which allow integration of services and adequate standardization. This article features: (a) a literature review, related to FMCS based on ITS and enabling technologies, (b) design of the ITS architecture of an FMCS, and (c) some advances in the development of the proposed FMCS in a Colombian city (Popayán). The results of the literature review allowed identifying the most important requirements of FMCS in order to design the ITS architecture and build a prototype featuring the suggested technologies. Finally, some experiments were performed to evaluate the operation of the developed prototype. The results showed evidence of adequate operation in sending and receiving messages from and to four prototypes developed for the vehicles, also complying with the established requirements of location, tracking, exchanged data, and security. This allows continuing the development of the proposed FMCS, with some adjustments. [ABSTRACT FROM AUTHOR]

The continuous search for novel enzyme backbones and the engineering of already well studied enzymes for biotechnological applications has become an increasing challenge, especially by the increasing potential diversity space provided by directed enzyme evolution approaches and the demands of experimental data generated by rational design of enzymes. In this work, we propose a semi-rational mutational strategy focused on introducing diversity in structurally variable regions in enzymes. The identified sequences are subjected to a progressive deletion of two amino acids and the joining residues are subjected to saturation mutagenesis using NNK degenerate codons. This strategy offers a novel library diversity approach while simultaneously decreasing enzyme size in the variable regions. In this way, we intend to identify and reduce variable regions found in enzymes, probably resulting from neutral drift evolution, and simultaneously studying the functional effect of said regions. This strategy was applied to Bacillus. subtilis lipase A (BSLA), by selecting and deleting six variable enzyme regions (named regions 1 to 6) by the deletion of two amino acids and additionally randomizing the joining amino acid residues. After screening, no active variants were found in libraries 1% and 4%, 15% active variants were found in libraries 2% and 3%, and 25% for libraries 5 and 6 (n = 3000 per library, activity detected using tributyrin agar plates). Active variants were assessed for activity in microtiter plate assay (pNP-butyrate), thermal stability, substrate preference (pNP-butyrate, -palmitate), and compared to wildtype BSLA. From these analyses, variant P5F3 (F41L-ΔW42-ΔD43-K44P), from library 3 was identified, showing increased activity towards longer chain p-nitrophenyl fatty acid esters, when compared to BSLA. This study allowed to propose the targeted region 3 (positions 40–46) as a potential modulator for substrate specificity (fatty acid chain length) in BSLA, which can be further studied to increase its substrate spectrum and selectivity. Additionally, this variant showed a decreased thermal resistance but interestingly, higher isopropanol and Triton X-100 resistance. This deletion-randomization strategy could help to expand and explore sequence diversity, even in already well studied and characterized enzyme backbones such as BSLA. In addition, this strategy can contribute to investigate and identify important non-conserved regions in classic and novel enzymes, as well as generating novel biocatalysts with increased performance in specific processes, such as enzyme immobilization. [ABSTRACT FROM AUTHOR]

Escherichia coli contains two phosphofructokinases, Pfk-1 and Pfk-2, which belong to unrelated protein families. In addition to catalytic function, the enzymes have converged in showing substrate inhibition by the nucleotide MgATP. However, although both Pfk-1 and Pfk-2 have been extensively characterized biochemically, only the structure of the former has been solved by X-ray diffraction. In order to fully understand how the same function has evolved on different structural folds, Pfk-2 has been crystallized by the hanging-drop vapour-diffusion method using PEG 6000 as precipitant. Single crystals were grown in the presence of MgATP and diffracted to 1.98 Å. The crystals belong to the orthorhombic system, space group P2221, with unit-cell parameters a = 42.8, b = 86.8, c = 171.3 Å. The calculated Matthews coefficient of 2.45 Å3 Da−1 indicates the presence of two monomers in the asymmetric unit, corresponding to a solvent content of 49%. Structure determination is ongoing. [ABSTRACT FROM AUTHOR]