Antibody-Dependent Respiratory Burst against Plasmodium falciparum Merozoites in Individuals Living in an Area with Declining Malaria Transmission.

Malaria transmission intensity affects the development of naturally acquired immunity to malaria. An absolute correlate measure of protection against malaria is lacking. However, antibody-mediated functions against Plasmodium falciparum correlate with protection against malaria. In children, antibody-mediated functions against P. falciparum decline with reduced exposure. It is unclear whether adults maintain antibody-mediated functions as malaria transmission declines. This study assessed antibody-dependent respiratory burst (ADRB) in individuals from an area with declining malaria transmission. In an age-matched analysis, we compare ADRB activity during high versus low malaria transmission periods. Age significantly predicted higher ADRB activity in the high (p < 0.001) and low (p < 0.001) malaria transmission periods. ADRB activity was higher during the high compared to the low malaria transmission period in older children and adults. Only older adults during the high malaria transmission period had their median ADRB activity above the ADRB cut-off. Ongoing P. falciparum infection influenced ADRB activity during the low (p = 0.01) but not the high (p = 0.29) malaria transmission period. These findings propose that naturally acquired immunity to P. falciparum is affected in children and adults as malaria transmission declines, implying that vaccines will be necessary to induce and maintain protection against malaria.

The role of serum thymidine kinase 1 activity in neoadjuvant-treated HER2-positive breast cancer: biomarker analysis from the Swedish phase II randomized PREDIX HER2 trial.

BACKGROUND: Thymidine kinase 1 (TK1) plays a pivotal role in DNA synthesis and cellular proliferation. TK1 has been studied as a prognostic marker and as an early indicator of treatment response in human epidermal growth factor 2 (HER2)-negative early and metastatic breast cancer (BC). However, the prognostic and predictive value of serial TK1 activity in HER2-positive BC remains unknown. METHODS: In the PREDIX HER2 trial, 197 HER2-positive BC patients were randomized to neoadjuvant trastuzumab, pertuzumab, and docetaxel (DPH) or trastuzumab emtansine (T-DM1), followed by surgery and adjuvant epirubicin and cyclophosphamide. Serum samples were prospectively collected from all participants at multiple timepoints: at baseline, after cycle 1, 2, 4, and 6, at end of adjuvant therapy, annually for a total period of 5 years and/or at the time of recurrence. The associations of sTK1 activity with baseline characteristics, pathologic complete response (pCR), event-free survival (EFS), and disease-free survival (DFS) were evaluated. RESULTS: No association was detected between baseline sTK1 levels and all the baseline clinicopathologic characteristics. An increase of TK1 activity from baseline to cycle 2 was seen in all cases. sTK1 level at baseline, after 2 and 4 cycles was not associated with pCR status. After a median follow-up of 58 months, 23 patients had EFS events. There was no significant effect between baseline or cycle 2 sTK1 activity and time to event. A non-significant trend was noted among patents with residual disease (non-pCR) and high sTK1 activity at the end of treatment visit, indicating a potentially worse long-term prognosis. CONCLUSION: sTK1 activity increased following neoadjuvant therapy for HER2-positive BC but was not associated with patient outcomes or treatment benefit. However, the post-surgery prognostic value in patients that have not attained pCR warrants further investigation. TRIAL REGISTRATION: ClinicalTrials.gov, NCT02568839. Registered on 6 October 2015.

Association between empathy, anxiety and psychological well-being in Mexican medical students. Transversal study.

BACKGROUND: The expression of empathy improves the quality of health care, which is why its instruction is suggested during the years of academic training of medical students. However, expressing empathy can also cause anxiety and impair psychological well-being. OBJECTIVE: To identify profiles that describe the relationship between empathy, anxiety and psychological well-being in Mexican medical students. METHOD: Cross-sectional study in Mexican medical students completed the Multidimensional Empathy Scale, the Beck Anxiety Inventory, and the Psychological Well-Being Scale for Adults. RESULTS: Women presented greater empathy and anxiety and lower psychological well-being (p < 0.01). Anxiety in students of the last year of training was lower compared to students of the first and second year (p < 0.01). Four profiles were detected: 1) men, minimal anxiety, low empathy; 2) first-year students, moderate to severe anxiety, low psychological well-being, high empathy; 3) last year of training, mild anxiety, adequate psychological well-being, medium empathy; 4) women, second and third year of training, very high empathy. CONCLUSIONS: Medical students in their first year of training constitute a population at risk. Attention strategies are necessary for this student group.

ANTECEDENTES: La expresión de empatía mejora la calidad de la atención a la salud, y por ello se sugiere su instrucción durante los años de formación académica de estudiantes de medicina. No obstante, la expresión de empatía también puede ocasionar ansiedad y perjudicar el bienestar psicológico. OBJETIVO: Identificar perfiles que describan la relación entre empatía, ansiedad y bienestar psicológico en estudiantes mexicanos de medicina. MÉTODO: Estudio transversal en estudiantes mexicanos de medicina que completaron la Escala Multidimensional de Empatía, el Inventario de Ansiedad de Beck y la Escala de Bienestar Psicológico para Adultos. RESULTADOS: Las mujeres presentaron mayor empatía y ansiedad, y menor bienestar psicológico (p < 0.01). La ansiedad en estudiantes del último año de formación fue menor que en los estudiantes de primer y segundo año (p < 0.01). Se detectaron cuatro perfiles: 1) hombres, ansiedad mínima, empatía baja; 2) estudiantes de primer año de formación, ansiedad moderada a grave, bienestar psicológico bajo, empatía alta; 3) último año de formación, ansiedad leve, bienestar psicológico adecuado, empatía media; 4) mujeres, segundo y tercer años de formación, muy alta empatía. CONCLUSIONES: Los estudiantes de medicina del primer año de formación constituyen una población de riesgo. Son necesarias estrategias de atención para este grupo estudiantil.

Insight in terms of Behavior in the Clinical Context: Transfer and Rule-governed Behavior / El insight desde el análisis de la conducta: la transferencia y la conducta gobernada por reglas

A redefinition of insight-type events is presented, an initial attempt to view insight in terms of behavior analysis in the clinical context and relevant variables are suggested for their study. This definition assumes the insight as a novel behavior that involves the search for solutions to psychological problems. Solving a problem is discovering an effective behavior pattern, based on the creation of verbal rules that help the client to reach the solution. In this problem-solving process, a new behavior emerges, explained through the solution of a transfer task. The theoretical proposal exposed allows a better understanding of this phenomenon, overcoming the problems of mentalist conceptions about the term of insight, and contributes to a better understanding of some relevant elements of change process. (AU)

Se presenta una redefinición del evento tipo insight, un intento inicial por explicar el insight en términos del análisis de la conducta en el contexto clínico, así como la propuesta de variables relevantes para su estudio. Esta definición asume el insight como una conducta novedosa que involucra la búsqueda de soluciones a problemas de carácter psicológico. Resolver un problema es descubrir un patrón de conducta efectivo, a partir de la creación de reglas verbales que ayudan al cliente a llegar a la solución. En este proceso de resolución de problemas, emerge una conducta novedosa explicada a través de la solución de una tarea de transferencia. La propuesta teórica expuesta permite una mejor comprensión del fenómeno, superando los problemas de concepciones mentalistas y contribuye a un mejor entendimiento de elementos relevantes del proceso de cambio. (AU)

Therapeutic Approaches for ADHD by Developmental Stage and Clinical Presentation.

Attention Deficit Hyperactivity Disorder is a neurodevelopmental disorder with three presentations: inattentive, hyperactive/impulsive and combined. These may represent an independent disease entity. Therefore, the therapeutic approach must be focused on their neurobiological, psychological and social characteristics. To date, there is no comprehensive analysis of the efficacy of different treatments for each presentation of ADHD and each stage of development. This is as narrative overview of scientific papers that summarize the most recent findings and identify the most effective pharmacological and psychosocial treatments by ADHD presentation and age range. Evidence suggests that methylphenidate is the safest and most effective drug for the clinical management of children, adolescents and adults. Atomoxetine is effective in preschoolers and maintains similar efficacy to methylphenidate in adults, whereas guanfacine has proven to be an effective monotherapy for adults and is a worthy adjuvant for the management of cognitive symptoms. The psychosocial treatments with the best results in preschoolers are behavioral interventions that include training of primary caregivers. In adolescents, the combination of cognitive and cognitive-behavioral therapies has shown the best results, whereas cognitive-behavioral interventions are the most effective in adults. Pharmacological and psychosocial treatments must be adjusted to the ADHD presentation and its neurocognitive characteristics through the patient's development.

A robust Bayesian bias-adjusted random effects model for consideration of uncertainty about bias terms in evidence synthesis.

Meta-analysis is a statistical method used in evidence synthesis for combining, analyzing and summarizing studies that have the same target endpoint and aims to derive a pooled quantitative estimate using fixed and random effects models or network models. Differences among included studies depend on variations in target populations (ie, heterogeneity) and variations in study quality due to study design and execution (ie, bias). The risk of bias is usually assessed qualitatively using critical appraisal, and quantitative bias analysis can be used to evaluate the influence of bias on the quantity of interest. We propose a way to consider ignorance or ambiguity in how to quantify bias terms in a bias analysis by characterizing bias with imprecision (as bounds on probability) and use robust Bayesian analysis to estimate the overall effect. Robust Bayesian analysis is here seen as Bayesian updating performed over a set of coherent probability distributions, where the set emerges from a set of bias terms. We show how the set of bias terms can be specified based on judgments on the relative magnitude of biases (ie, low, unclear, and high risk of bias) in one or several domains of the Cochrane's risk of bias table. For illustration, we apply a robust Bayesian bias-adjusted random effects model to an already published meta-analysis on the effect of Rituximab for rheumatoid arthritis from the Cochrane Database of Systematic Reviews.

A suggestion for the quantification of precise and bounded probability to quantify epistemic uncertainty in scientific assessments.

An honest communication of uncertainty about quantities of interest enhances transparency in scientific assessments. To support this communication, risk assessors should choose appropriate ways to evaluate and characterize epistemic uncertainty. A full treatment of uncertainty requires methods that distinguish aleatory from epistemic uncertainty. Quantitative expressions for epistemic uncertainty are advantageous in scientific assessments because they are nonambiguous and enable individual uncertainties to be characterized and combined in a systematic way. Since 2019, the European Food Safety Authority (EFSA) recommends assessors to express epistemic uncertainty in conclusions of scientific assessments quantitatively by subjective probability. A subjective probability can be used to represent an expert judgment, which may or may not be updated using Bayes's rule to integrate evidence available for the assessment and could be either precise or approximate. Approximate (or bounded) probabilities may be enough for decision making and allow experts to reach agreement on certainty when they struggle to specify precise subjective probabilities. The difference between the lower and upper bound on a subjective probability can also be used to reflect someone's strength of knowledge. In this article, we demonstrate how to quantify uncertainty by bounded probability, and explicitly distinguish between epistemic and aleatory uncertainty, by means of robust Bayesian analysis, including standard Bayesian analysis through precise probability as a special case. For illustration, the two analyses are applied to an intake assessment.

Biopsychological correlates of repetitive and restricted behaviors in autism spectrum disorders.

BACKGROUND: Autism Spectrum Disorder (ASD) is considered a neurodevelopmental condition that is characterized by alterations in social interaction and communication, as well as patterns of restrictive and repetitive behaviors (RRBs). RRBs are defined as broad behaviors that comprise stereotypies, insistence on sameness, and attachment to objects or routines. RRBs can be divided into lower-level behaviors (motor, sensory, and object-manipulation behaviors) and higher-level behaviors (restrictive interests, insistence on sameness, and repetitive language). According to the DSM-5, the grade of severity in ASD partially depends on the frequency of RRBs and their consequences for disrupting the life of patients, affecting their adaptive skills, and increasing the need for parental support. METHODS: We conducted a systematic review to examine the biopsychological correlates of the symptomatic domains of RRBs according to the type of RRBs (lower- or higher-level). We searched for articles from the National Library of Medicine (PubMed) using the terms: autism spectrum disorders, ASD, and autism-related to executive functions, inhibitory control, inflexibility, cognitive flexibility, hyper or hypo connectivity, and behavioral approaches. For describing the pathophysiological mechanism of ASD, we also included animal models and followed PRISMA guidelines. RESULTS: One hundred and thirty-one articles were analyzed to explain the etiology, continuance, and clinical evolution of these behaviors observed in ASD patients throughout life. CONCLUSIONS: Biopsychological correlates involved in the origin of RRBs include alterations in a) neurotransmission system, b) brain volume, c) inadequate levels of growth factors, d) hypo- or hyper-neural connectivity, e) impairments in behavioral inhibition, cognitive flexibility, and monitoring and f) non-stimulating environments. Understanding these lower- and higher-level of RRBs can help professionals to improve or design novel therapeutic strategies.

Rule Emission: A Possible Variable for Improved Therapeutic Practice.

It has been suggested that achieving greater effectiveness in psychotherapeutic treatment requires analyzing what therapists actually do and say, how they do this and when it is done. Based on this approach, in this study we focused on the rules emitted by therapists, since providing rules is thought to be of fundamental importance in promoting effective and efficient clinical change. Specifically, we sought to determine whether the experience level of therapists and the brevity of therapy would be related to patterns of therapist rule emission as categorized by the Category System of Rules emitted by the Therapist (SISC-RULES-T) (Vargas-de la Cruz & Pardo-Cebrián, 2014). Greater therapist experience and shorter therapy duration were found to be reliably predictive of more rule emissions across most rule categories (Z values between: Z = -3.68 and Z = -2.05; p values: p < .05 and p < .001). These variables were also predictive of more emissions of rules that specified all three operant contingency elements (situation, behavior, and consequence) rather than fewer elements (Z = -2.59, p < .05; Z = -2.26, p < .05). In the expert therapists and therapist with shorter cases, there was a nonsignificant tendency for the emission of general and conceptual rules to increase over sessions whereas emissions of concrete and particular rules tended to decrease; the explicitness of the three contingency elements also tended to decrease as treatment progressed. These findings may help to identify verbal characteristics of therapists that could lead to improved therapeutic practice.

Una Extensión del Concepto de Regla y su Aplicación a la Terapia Psicológica / An extension of the concept of rule and its application to therapy

Proporcionar y seguir reglas se ha considerado un factor importante en la eficacia de los tratamientos psicológicos; sin embargo, hace falta una clarificación conceptual de lo que se entiende por emitir o seguir una regla y su aplicación en el ámbito clínico. Partiendo de una revisión sobre el concepto original de conducta gobernada por reglas, nuestro propósito fue extender la definición de esta conducta orientada al proceso terapéutico. Esta definición permite su identificación y estudio momento a momento a lo largo del proceso. La extensión de esta definición puede ayudarnos a identificar el papel funcional de las reglas en el cambio terapéutico y las variables involucradas que contribuyen al entendimiento de fenómenos clínicos como la generalización. Los hallazgos que se desprendan del estudio de las reglas desde esta perspectiva podrían contribuir a aumentar la información sobre el proceso de cambio terapéutico y la mejora del ejercicio clínico

Providing and following rules has been considered an important factor in the effectiveness of psychological treatments; however, a conceptual clarification is needed of what is meant by issuing or following a rule and its application in the clinical setting. Based on a review of the original concept of rule-governed behavior, our purpose was to extend the definition of this behavior oriented to the clinical process. This definition allows its identification and study moment by moment throughout the therapeutic process. The extension of this definition can help us to identify the al role of rules in clinical change and the variables involved that contribute to the understanding of clinical phenomena such as generalization. The findings that emerge from the study of rules from this perspective could increase the information on the therapeutic change process and, therefore, contribute to the improvement of clinical practice

The Streptococcus agalactiae Stringent Response Enhances Virulence and Persistence in Human Blood.

Streptococcus agalactiae (group B Streptococcus [GBS]) causes serious infections in neonates. We previously reported a transposon sequencing (Tn-seq) system for performing genomewide assessment of gene fitness in GBS. In order to identify molecular mechanisms required for GBS to transition from a mucosal commensal lifestyle to bloodstream invasion, we performed Tn-seq on GBS strain A909 with human whole blood. Our analysis identified 16 genes conditionally essential for GBS survival in blood, of which 75% were members of the capsular polysaccharide (cps) operon. Among the non-cps genes identified as conditionally essential was relA, which encodes an enzyme whose activity is central to the bacterial stringent response-a conserved adaptation to environmental stress. We used blood coincubation studies of targeted knockout strains to confirm the expected growth defects of GBS deficient in capsule or stringent response activation. Unexpectedly, we found that the relA knockout strains demonstrated decreased expression of ß-hemolysin/cytolysin, an important cytotoxin implicated in facilitating GBS invasion. Furthermore, chemical activation of the stringent response with serine hydroxamate increased ß-hemolysin/cytolysin expression. To establish a mechanism by which the stringent response leads to increased cytotoxicity, we performed transcriptome sequencing (RNA-seq) on two GBS strains grown under stringent response or control conditions. This revealed a conserved decrease in the expression of genes in the arginine deiminase pathway during stringent response activation. Through coincubation with supplemental arginine and the arginine antagonist canavanine, we show that arginine availability is a determinant of GBS cytotoxicity and that the pathway between stringent response activation and increased virulence is arginine dependent.

Beneficial effects of oral administration of C-Phycocyanin and Phycocyanobilin in rodent models of experimental autoimmune encephalomyelitis.

The only three oral treatments currently available for multiple sclerosis (MS) target the relapsing forms of the disease and concerns regarding efficacy, safety and tolerability limit their use. Identifying novel oral disease-modifying therapies for MS, targeting both its inflammatory and neurodegenerative components is still a major goal. AIM: The scope of this study was to provide evidence that the oral administration of C-Phycocyanin (C-PC), the main biliprotein of the Spirulina platensis cyanobacteria and its tetrapyrrolic prosthetic group, Phycocyanobilin (PCB), exert ameliorating actions on rodent models of experimental autoimmune encephalomyelitis (EAE). MAIN METHODS: EAE was induced in Lewis rats using the spinal cord encephalitogen from Sprague Dawley rats and in C57BL6 mice with MOG35-55 peptide. Clinical signs, motor function, oxidative stress markers, cytokine levels by ELISA and transmission electron microscopy analysis were assessed. KEY FINDINGS: Either prophylactic or early therapeutic administration of C-PC to Lewis rats with EAE, significantly improved clinical signs and restored the motor function of the animals. Furthermore, C-PC positively modulated oxidative stress markers measured in brain homogenate and serum and protected the integrity of cerebral myelin sheaths as shown by transmission electron microscopy analysis. In C57BL/6 mice with EAE, PCB orally improved clinical status of the animals and reduced the expression levels of brain IL-6 and IFN-γ proinflammatory cytokines. SIGNIFICANCE: These results, for the first time, support the fact that both C-PC and PCB administered orally could potentially improve neuroinflammation, protect from demyelination and axonal loss, which may be translated into an improved quality of life for MS patients.

Rule emission: a possible variable for improved therapeutic practice

It has been suggested that achieving greater effectiveness in psychotherapeutic treatment requires analyzing what therapists actually do and say, how they do this and when it is done. Based on this approach, in this study we focused on the rules emitted by therapists, since providing rules is thought to be of fundamental importance in promoting effective and efficient clinical change. Specifically, we sought to determine whether the experience level of therapists and the brevity of therapy would be related to patterns of therapist rule emission as categorized by the Category System of Rules emitted by the Therapist (SISC-RULES-T) (Vargas-de la Cruz & Pardo-Cebrián, 2014). Greater therapist experience and shorter therapy duration were found to be reliably predictive of more rule emissions across most rule categories (Z values between: Z = -3.68 and Z = -2.05; p values: p < .05 and p < .001). These variables were also predictive of more emissions of rules that specified all three operant contingency elements (situation, behavior, and consequence) rather than fewer elements (Z = -2.59, p < .05; Z = -2.26, p < .05). In the expert therapists and therapist with shorter cases, there was a nonsignificant tendency for the emission of general and conceptual rules to increase over sessions whereas emissions of concrete and particular rules tended to decrease; the explicitness of the three contingency elements also tended to decrease as treatment progressed. These findings may help to identify verbal characteristics of therapists that could lead to improved therapeutic practice

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Aligner optimization increases accuracy and decreases compute times in multi-species sequence data.

As sequencing technologies have evolved, the tools to analyze these sequences have made similar advances. However, for multi-species samples, we observed important and adverse differences in alignment specificity and computation time for bwa- mem (Burrows-Wheeler aligner-maximum exact matches) relative to bwa-aln. Therefore, we sought to optimize bwa-mem for alignment of data from multi-species samples in order to reduce alignment time and increase the specificity of alignments. In the multi-species cases examined, there was one majority member (i.e. Plasmodium falciparum or Brugia malayi) and one minority member (i.e. human or the Wolbachia endosymbiont wBm) of the sequence data. Increasing bwa-mem seed length from the default value reduced the number of read pairs from the majority sequence member that incorrectly aligned to the reference genome of the minority sequence member. Combining both source genomes into a single reference genome increased the specificity of mapping, while also reducing the central processing unit (CPU) time. In Plasmodium, at a seed length of 18 nt, 24.1 % of reads mapped to the human genome using 1.7±0.1 CPU hours, while 83.6 % of reads mapped to the Plasmodium genome using 0.2±0.0 CPU hours (total: 107.7 % reads mapping; in 1.9±0.1 CPU hours). In contrast, 97.1 % of the reads mapped to a combined Plasmodium-human reference in only 0.7±0.0 CPU hours. Overall, the results suggest that combining all references into a single reference database and using a 23 nt seed length reduces the computational time, while maximizing specificity. Similar results were found for simulated sequence reads from a mock metagenomic data set. We found similar improvements to computation time in a publicly available human-only data set.

Feasibility of screening and prevalence of prenatal depression in an obstetric setting in Spain.

OBJECTIVES: Prenatal depression is a significant public health problem and one of the main risk factors for postpartum depression. Limited research in perinatal depression has been conducted in Spain. The objectives of this study was to: (1) examine the feasibility of integrating a screening program for prenatal depression in an obstetrics setting in a large urban hospital in Spain; and (2) provide an initial estimate of the prevalence rate of the severity of depressive symptoms during pregnancy. STUDY DESIGN: Screening for prenatal depression was conducted during the first trimester in an obstetrics setting in an urban hospital in Madrid, Spain 2014-2015. The Patient Health Questionnaire (PHQ-9) was used as the main screener. RESULTS: Of the 730 women asked to complete the screening protocol, 445 (60.9%) completed the PHQ-9 during the first trimester. Those who did not complete the screening were more likely to be immigrants and did not read Spanish. The prevalence of moderate to high severity of prenatal depressive symptoms prenatal depressive is 14.8% (PHQ-9≥10). CONCLUSION: It is possible to integrate screening for prenatal depression in an urban obstetrics setting, but there are significant structural barriers to implementation. The prevalence of significant prenatal depressive symptoms is similar to previous research in Spain and adds to the limited research in this area.

Las verbalizaciones tipo regla del terapeuta a lo largo del proceso clínico / The Therapist's rule-like Verbalizations throughout the Clinical Process

RESUMEN Se ha reconocido la importancia de las reglas del cliente para explicar un problema psicológico y para lograr superarlo. Sin embargo, no se ha incidido en el papel que podrían tener las reglas del terapeuta en el proceso clínico. Como un primer paso para poder dar una respuesta al respecto, en este estudio realizamos un análisis morfológico de las verbalizaciones en forma de regla emitidas por el terapeuta a lo largo del proceso clínico con el fin de identificar un patrón de emisión de este tipo de verbalizaciones que cambie de acuerdo con los distintos momentos de la terapia. Se analizaron las verbalizaciones en forma de regla emitidas por 9 terapeutas a lo largo de 19 casos clínicos (92 sesiones). Las verbalizaciones se codificaron de acuerdo con un Sistema de Categorización de las Reglas del Terapeuta (SISC-REGLAS-T). Los resultados mostraron diferencias entre el tipo de verbalizaciones en forma de regla y el objetivo que el terapeuta persigue alcanzar en los distintos momentos de la terapia. Estos resultados pueden ayudar a comprender cómo el terapeuta utiliza las verbalizaciones en forma de reglas, lo que contribuye al esclarecimiento del papel de las reglas que emite el terapeuta para conseguir el cambio clínico.

ABSTRACT It has been pointed out the importance of the rules to explain and to treat psychological problems. However it hasn't emphasize the role that the therapist's rules could have in clinical process. As a first step to give an answer, we realize a morphological analysis of verbalization- like-rule emitted by the therapist throughout the clinical process with the aim to identify a patron of emission of this kind of verbalization that changes according to different moments of therapy. This study analyzed the verbalization-like-rules emitted by 9 therapists over 19 clinical cases (92 sessions). The verbalizations were coded according to a system of categorization of the Rules Therapist (SISC-RULES-T). The results showed differences between the type of verbalizations-like-rules and target the therapist aims to achieve at different times of therapy. These results may help to understand how the therapist uses this kind of verbalizations, helping to clarify the functional role of rules issued by the therapist for clinical change.

Efficient Enrichment of Bacterial mRNA from Host-Bacteria Total RNA Samples.

Despite numerous advances in genomics and bioinformatics, technological hurdles remain to examine host-microbe transcriptomics. Sometimes the transcriptome of either or both can be ascertained merely by generating more sequencing reads. However, many cases exist where bacterial mRNA needs to be enriched further to enable cost-effective sequencing of the pathogen or endosymbiont. While a suitable method is commercially available for mammalian samples of this type, development of such methods has languished for invertebrate samples. Furthermore, a common method across multiple taxa would facilitate comparisons between bacteria in invertebrate vectors and their vertebrate hosts. Here, a method is described to concurrently remove polyadenylated transcripts, prokaryotic rRNA, and eukaryotic rRNA, including those with low amounts of starting material (e.g. 100 ng). In a Wolbachia-Drosophila system, this bacterial mRNA enrichment yielded a 3-fold increase in Wolbachia mRNA abundance and a concomitant 3.3-fold increase in the percentage of transcripts detected. More specifically, 70% of the genome could be recovered by transcriptome sequencing compared to 21% in the total RNA. Sequencing of similar bacterial mRNA-enriched samples generated from Ehrlichia-infected canine cells covers 93% of the Ehrlichia genome, suggesting ubiquitous transcription across the entire Ehrlichia chaffeensis genome. This technique can potentially be used to enrich bacterial mRNA in many studies of host-microbe interactions.

The essential genome of Streptococcus agalactiae.

BACKGROUND: Next-generation sequencing of transposon-genome junctions from a saturated bacterial mutant library (Tn-seq) is a powerful tool that permits genome-wide determination of the contribution of genes to fitness of the organism under a wide range of experimental conditions. We report development, testing, and results from a Tn-seq system for use in Streptococcus agalactiae (group B Streptococcus; GBS), an important cause of neonatal sepsis. METHODS: Our method uses a Himar1 mini-transposon that inserts at genomic TA dinucleotide sites, delivered to GBS on a temperature-sensitive plasmid that is subsequently cured from the bacterial population. In order to establish the GBS essential genome, we performed Tn-seq on DNA collected from three independent mutant libraries-with at least 135,000 mutants per library-at serial 24 h time points after outgrowth in rich media. RESULTS: After statistical analysis of transposon insertion density and distribution, we identified 13.5 % of genes as essential and 1.2 % as critical, with high levels of reproducibility. Essential and critical genes are enriched for fundamental cellular housekeeping functions, such as acyl-tRNA biosynthesis, nucleotide metabolism, and glycolysis. We further validated our system by comparing fitness assignments of homologous genes in GBS and a close bacterial relative, Streptococcus pyogenes, which demonstrated 93 % concordance. Finally, we used our fitness assignments to identify signal transduction pathway components predicted to be essential or critical in GBS. CONCLUSIONS: We believe that our baseline fitness assignments will be a valuable tool for GBS researchers and that our system has the potential to reveal key pathogenesis gene networks and potential therapeutic/preventative targets.

Draft Genome Sequence of "Candidatus Phytoplasma oryzae" Strain Mbita1, the Causative Agent of Napier Grass Stunt Disease in Kenya.

Phytoplasmas are bacterial plant pathogens with devastating impact on agricultural production worldwide. In eastern Africa, Napier grass stunt disease causes serious economic losses in the smallholder dairy industry. This draft genome sequence of " ITALIC! CandidatusPhytoplasma oryzae" strain Mbita1 provides insight into its genomic organization and the molecular basis of pathogenicity.

Comparative Neuroregenerative Effects of C-Phycocyanin and IFN-Beta in a Model of Multiple Sclerosis in Mice.

Multiple Sclerosis (MS) therapies approved so far are unable to effectively reverse the chronic phase of the disease or improve the remyelination process. Here our aim is to evaluate the effects of C-Phycocyanin (C-Pc), a biliprotein from Spirulina platensis with anti-oxidant, anti-inflammatory and cytoprotective properties, in a chronic model of experimental autoimmune encephalomyelitis (EAE) in mice. C-Pc (2, 4 or 8 mg/kg i.p.) or IFN-beta (2000 IU, s.c.) was administered daily once a day or every other day, respectively, starting at disease onset, which differ among EAE mice between 11 and 15 days postinduction. Histological and immunohistochemistry (anti-Mac-3, anti-CD3 and anti-APP) assessments were performed in spinal cord in the postinduction time. Global gene expression in the brain was analyzed with the Illumina Mouse WG-6_V2 BeadChip microarray and the expression of particular genes, assessed by qPCR using the Fast SYBR Green RT-PCR Master Mix. Oxidative stress parameters (malondialdehyde, peroxidation potential, CAT/SOD ratio and GSH) were determined spectrophoto-metrically. Results showed that C-Pc ameliorates the clinical deterioration of animals, an effect that expresses the reduction of the inflammatory infiltrates invading the spinal cord tissue, the axonal preservation and the down-regulation of IL-17 expression in brain tissue and serum. C-Pc and IFN-beta improved the redox status in mice subjected to EAE, while microarray analysis showed that both treatments shared a common subset of differentially expressed genes, although they also differentially modulated another subset of genes. Specifically, C-Pc mainly modulated the expression of genes related to remyelination, gliogenesis and axon-glia processes. Taken together, our results indicate that C-Pc has significant therapeutic effects against EAE, mediated by the dynamic regulation of multiple biological processes.