Expression and purification of active shikimate dehydrogenase from Plasmodium falciparum.

Plasmodium falciparum is known to cause severe malaria, current treatment consists in artemisinin-based combination therapy, but resistance can lead to treatment failure. Knowledge concerning P. falciparum essential proteins can be used for searching new antimalarials, among these a potential candidate is shikimate dehydrogenase (SDH), an enzyme part of the shikimate pathway which is responsible for producing endogenous aromatic amino acids. SDH from P. falciparum (PfSDH) is unexplored by the scientific community, therefore, this study aims to establish the first protocol for active PfSDH expression. Putative PfSDH nucleotide sequence was used to construct an optimized expression vector pET28a+PfSDH inserted in E. coli BL21(DE3). As a result, optimal expression conditions were acquired by varying IPTG and temperature through time. Western Blot analysis was applied to verify appropriate PfSDH expression, solubilization and purification started with lysis followed by two-steps IMAC purification. Enzyme activity was measured spectrophotometrically by NADPH oxidation, optimal PfSDH expression occur at 0.1 mM IPTG for 48 hours growing at 37 °C and shaking at 200 rpm, recombinant PfSDH obtained after purification was soluble, pure and its physiological catalysis was confirmed. Thus, this study describes the first protocol for heterologous expression of PfSDH in soluble and active form.

Complications associated with combined direct and indirect bypass in Moyamoya Disease: A meta-analysis.

Bypass revascularization helps prevent complications in Moyamoya Disease (MMD). To systematically review complications associated with combined direct and indirect (CB) bypass in MMD and analyze differences between the adult and pediatric populations. A systematic literature review was conducted per PRISMA guidelines. PUBMED, Cochrane Library, Web of Science, and CINAHL, were queried from January 1980 to March 2022. Complications were defined as any event in the immediate post-surgical period of a minimum 3 months follow-up. Exclusion criteria included lack of surgical complication reports, non-English articles, and CB unspecified or reported separately. 18 final studies were included of 1580 procured. 1151 patients (per study range = 10-150, mean = 63.9) were analyzed. 9 (50.0%) studies included pediatric patients. There were 32 total hemorrhagic, 74 total ischemic and 16 total seizure complications, resulting in a rate of 0.04 (95% CI 0.03, 0.06), 0.7 (95% CI 0.04, 0.10) and 0.03 (95% CI 0.02, 0.05), respectively. The rate of hemorrhagic complications in the pediatric showed no significant difference from the adult subgroup (0.03 (95% CI 0.01-0.08) vs. 0.06 (95% CI 0.04-0.10, p = 0.19), such as the rate of ischemic complications (0.12 (95% CI 0.07-0.23) vs. 0.09 (95% CI 0.05-0.14, p = 0.40). Ischemia is the most common complication in CB for MMD. Pediatric patients had similar hemorrhagic and ischemic complication rates compared to adults.

Maternal exposure to a high-magnitude earthquake during pregnancy influences pre-reading skills in early childhood.

Exposure to an adverse prenatal environment can influence fetal development and result in long-lasting changes in the offspring. However, the association between maternal exposure to stressful events during pregnancy and the achievement of pre-reading skills in the offspring is unknown. Here we examined the association between prenatal exposure to the Chilean high-magnitude earthquake that occurred on February 27th, 2010 and the development of early reading precursors skills (listening comprehension, print knowledge, alphabet knowledge, vocabulary, and phonological awareness) in children at kindergarten age. This multilevel retrospective cohort study including 3280 children, of whom 2415 were unexposed and 865 were prenatally exposed to the earthquake shows substantial evidence that maternal exposure to an unambiguously stressful event resulted in impaired pre-reading skills and that a higher detrimental effect was observed in those children who had been exposed to the earthquake during the first trimester of gestation. In addition, females were more significantly affected by the exposure to the earthquake than their male peers in alphabet knowledge; contrarily, males were more affected than females in print knowledge skills. These findings suggest that early intervention programs for pregnant women and/or children exposed to prenatal stress may be effective strategies to overcome impaired pre-reading skills in children.

Whole Genome Sequencing Identifies CRISPLD2 as a Lung Function Gene in Children With Asthma.

BACKGROUND: Asthma is a common respiratory disorder with a highly heterogeneous nature that remains poorly understood. The objective was to use whole genome sequencing (WGS) data to identify regions of common genetic variation contributing to lung function in individuals with a diagnosis of asthma. METHODS: WGS data were generated for 1,053 individuals from trios and extended pedigrees participating in the family-based Genetic Epidemiology of Asthma in Costa Rica study. Asthma affection status was defined through a physician's diagnosis of asthma, and most participants with asthma also had airway hyperresponsiveness (AHR) to methacholine. Family-based association tests for single variants were performed to assess the associations with lung function phenotypes. RESULTS: A genome-wide significant association was identified between baseline FEV1/FVC ratio and a single-nucleotide polymorphism in the top hit cysteine-rich secretory protein LCCL domain-containing 2 (CRISPLD2) (rs12051168; P = 3.6 × 10-8 in the unadjusted model) that retained suggestive significance in the covariate-adjusted model (P = 5.6 × 10-6). Rs12051168 was also nominally associated with other related phenotypes: baseline FEV1 (P = 3.3 × 10-3), postbronchodilator (PB) FEV1 (7.3 × 10-3), and PB FEV1/FVC ratio (P = 2.7 × 10-3). The identified baseline FEV1/FVC ratio and rs12051168 association was meta-analyzed and replicated in three independent cohorts in which most participants with asthma also had confirmed AHR (combined weighted z-score P = .015) but not in cohorts without information about AHR. CONCLUSIONS: These findings suggest that using specific asthma characteristics, such as AHR, can help identify more genetically homogeneous asthma subgroups with genotype-phenotype associations that may not be observed in all children with asthma. CRISPLD2 also may be important for baseline lung function in individuals with asthma who also may have AHR.

Chronic Fluoxetine Treatment Induces Maturation-Compatible Changes in the Dendritic Arbor and in Synaptic Responses in the Auditory Cortex.

Fluoxetine is a selective serotonin reuptake inhibitor (SSRI) used to treat mood and anxiety disorders. Chronic treatment with this antidepressant drug is thought to favor functional recovery by promoting structural and molecular changes in several forebrain areas. At the synaptic level, chronic fluoxetine induces an increased size and density of dendritic spines and an increased ratio of GluN2A over GluN2B N-methyl-D-aspartate (NMDA) receptor subunits. The "maturation"-promoting molecular changes observed after chronic fluoxetine should also induce structural remodeling of the neuronal dendritic arbor and changes in the synaptic responses. We treated adult rats with fluoxetine (0.7 mg/kg i.p. for 28 days) and performed a morphometric analysis using Golgi stain in limbic and nonlimbic cortical areas. Then, we focused especially on the auditory cortex, where we evaluated the dendritic morphology of pyramidal neurons using a 3-dimensional reconstruction of neurons expressing mRFP after in utero electroporation. With both methodologies, a shortening and decreased complexity of the dendritic arbors was observed, which is compatible with an increased GluN2A over GluN2B ratio. Recordings of extracellular excitatory postsynaptic potentials in the auditory cortex revealed an increased synaptic response after fluoxetine and were consistent with an enrichment of GluN2A-containing NMDA receptors. Our results confirm that fluoxetine favors maturation and refinement of extensive cortical networks, including the auditory cortex. The fluoxetine-induced receptor switch may decrease GluN2B-dependent toxicity and thus could be applied in the future to treat neurodegenerative brain disorders characterized by glutamate toxicity and/or by an aberrant network connectivity.

Bedtime routines for young children: a dose-dependent association with sleep outcomes.

BACKGROUND: Establishment of a consistent bedtime routine (the activities that occur right before lights out) is often recommended as part of healthy sleep habits. However, no studies have investigated the dose-dependent association of a bedtime routine with sleep outcomes, especially in young children for whom they are particularly recommended. Thus, the aim of this study was to examine the associations of a consistent bedtime routine with sleep outcomes in young children (ages 0 through 5 y) in a large global sample and assess whether there is a dose-dependent relationship between the frequency of a bedtime routine both concurrently and retrospectively with sleep outcomes. PARTICIPANTS: Mothers of 10,085 children (Australia-New Zealand, Canada, China, Hong Kong, India, Japan, Korea, Malaysia, Philippines, Singapore, Thailand, United Kingdom, United States) completed the Brief Infant/Child Sleep Questionnaire. RESULTS: A consistent bedtime routine was associated with better sleep outcomes, including earlier bedtimes, shorter sleep onset latency, reduced night wakings, and increased sleep duration. Decreased parent-perceived sleep problems and daytime behavior problems were also related to institution of a regular bedtime routine. Furthermore, there was a dose-dependent relationship, with better outcomes associated with increased "doses" of having a bedtime routine, both currently and retrospectively, and was found within both predominantly Asian and predominantly Caucasian cultural regions. CONCLUSIONS: These results indicate that having a regular nightly bedtime routine is associated with improved sleep in young children, and suggests that the more consistently a bedtime routine is instituted and the younger started the better.

The relative importance of immigrant generation for Mexican Americans' alcohol and tobacco use from adolescence to early adulthood.

The rapidly growing Hispanic population in the US may be at-risk for greater substance use according to immigrant generation status. This study utilized latent growth curve analyses to determine whether trajectories of alcohol and cigarette use vary according to immigrant status in a Mexican-heritage sample (n = 1,274) from the National Longitudinal Study of Adolescent Health. Changes in alcohol and cigarette use were estimated over a 13-year span from adolescence (mean age = 15.9) to early adulthood. Alcohol use did not vary significantly by immigrant status. In contrast, third generation youth experienced greater increases in cigarette use over time compared to their second generation peers. Second generation youth also experienced greater acceleration in cigarette use compared to first generation youth. Immigrants who arrived in the US more recently experienced slower acceleration in cigarette use compared to those who have been in the US for longer periods of time.

Ethnic Identity and Substance Use Among Mexican-Heritage Preadolescents: Moderator Effects of Gender and Time in the United States.

This study examined interactive relationships among ethnic identity, gender, time in the US, and changes in substance use outcomes among a school-based sample of 1,731 Mexican-heritage preadolescents (ages 9-13). Residual change multilevel models adjusting for school clustering and using multiply imputed data assessed changes from beginning to end of fifth grade in use of alcohol, cigarettes, marijuana and inhalants, and four substance use antecedents. Effects of ethnic identity were conditional on time in the US, and in opposite directions by gender. Among males living longer in the US, stronger ethnic identity predicted desirable changes in all but one outcome (substance offers). Among females living longer in the US, stronger ethnic identity predicted undesirable changes in alcohol use, pro-drug norms, and peer substance use. Interpretations focus on differential exposure to substance use opportunities and the erosion of traditional gender role socialization among Mexican-heritage youth having lived longer in the US.

The importance of family factors to protect against substance use related problems among Mexican heritage and White youth.

INTRODUCTION: This study examined the ability of family cohesion, parental control, and parent-child attachment to prevent adolescents with a history of drug or alcohol use from experiencing subsequent problems related to their use. METHODS: Data came from Wave I and Wave II of the National Longitudinal Study of Adolescent Health and included Mexican heritage and White adolescents who reported alcohol use (n = 4894, 25% prevalence) or any other drug use (n = 2875, 14% prevalence) in their lifetime. RESULTS: Logistic regression results indicate greater parent-child attachment predicted lower risk of experiencing drug use problems (OR = 0.87, 95% CI = 0.77-0.98) while stronger family cohesion predicted lower odds of experiencing drug- (OR = 0.82, 95% CI = 0.70-0.97) or alcohol-related (OR = 0.74, 95% CI = 0.65-0.84) problems. Parental control was also negatively associated with odds of problems related to drug use (OR = 0.93, 95% CI = 0.86-0.99) or alcohol use (OR = 0.94, 95% CI = 0.90-0.99). Results also indicated family cohesion was the only protective factor for Mexican heritage youth while family cohesion and parent-child attachment were protective among White youth. Parental control protected White female adolescents from drug use problems more than males. Mexican heritage male adolescents experienced more protection from drug problems compared to females. CONCLUSION: Findings highlight the need for prevention interventions to emphasize parent-child attachment for White youth and family cohesion for both Mexican-heritage and White youth to decrease adolescent substance users' drug- and alcohol-related problems.

Nocturnal enuresis in children: prevalence, correlates, and relationship with obstructive sleep apnea.

OBJECTIVES: To examine the prevalence and correlates of nocturnal enuresis (NE) in primary school children, and to compare the prevalence of NE in children with and those without obstructive sleep apnea (OSA). STUDY DESIGN: Parents of children aged 6-11 years completed a questionnaire eliciting information on sleep-related symptoms, demography, and family and past medical history. Children screened due to high risk for OSA, along with a randomly chosen low-risk group, underwent overnight polysomnography (PSG). RESULTS: A total of 6147 children (3032 girls) were studied. The overall prevalence of NE (≥1 wet night/month) was 4.6% (6.7% of boys and 2.5% of girls). Boys had a significantly greater prevalence across all age groups. In 597 children (215 girls) who underwent PSG, the prevalence of NE was not greater in children with OSA, but was increased with increasing severity of OSA in girls only. Boys with NE had longer deep sleep duration. Sex and sleep-related symptoms were associated with NE. CONCLUSIONS: This community-based study demonstrated a sex-associated prevalence of NE in relation to increasing OSA severity.

Blood pressure is elevated in children with primary snoring.

OBJECTIVES: To compare ambulatory blood pressure (ABP) in nonoverweight, prepubertal children with and without primary snoring (PS), and to investigate whether PS is a part of the dose-response relationship between sleep-disordered breathing (SDB) and BP in children. STUDY DESIGN: This was a cross-sectional community-based study involving 190 children age 6 to 13 years. Each participant underwent an overnight sleep study and ABP monitoring after completing a validated sleep symptoms questionnaire. Individual systolic blood pressure (SBP), diastolic blood pressure (DBP), and mean arterial BP were calculated for wake and sleep periods. Subjects were hypertensive if mean SBP or DBP was > 95th percentile (relative to sex and height) of reference. RESULTS: A total of 56 nonsnoring controls, 46 children with PS, 62 children with an apnea-hypopnea index (AHI) of 1 to 3, and 26 children with an AHI > 3 were identified. The daytime and nighttime BP increased across the severity spectrum of SDB. The dose-response trends for the proportion of subjects with nighttime systolic and diastolic hypertension also were significant. Nighttime DBP was significantly higher in the children with PS compared with controls after adjusting for age, sex, and body mass index. CONCLUSIONS: PS was demonstrated to be an aspect of the dose-response relationship between SDB and BP in children and should not be considered completely benign.

Suicidal behavior in four ethnic groups in the Hague, 2002-2004.

BACKGROUND: Suicidal behavior is a severe public health problem. AIMS: To determine the rates of attempted and completed suicide among ethnic groups in The Hague, The Netherlands (2002-2004). METHODS: By analyzing data on attempted and completed suicide (from the psychiatric department of general medical hospitals; the psychiatric emergency service and the municipal coroners). RESULTS: Turkish and Surinamese females aged 15-24 years were at highest risk for attempted suicide (age-specific rate 545 / 100,000 and 421 / 100,000 person-years, respectively). Both rates were significantly higher than in the same age group of Dutch females (246 / 100,000 person-years). Turkish (2%) and Surinamese (7%) had lower repeat suicide-attempt rates than did Dutch (16%) females aged 15-24. Significantly lower suicide-attempt rates were found for Surinamese than for Dutch females aged 35-54 years. Differences were not explained by socioeconomic living conditions. The ratio fatal/nonfatal events was 4.5 times higher in males than in females and varied across age, gender, and ethnicity strata. Completed suicide was rare among migrant females. No completed suicides were observed in the Turkish and Surinamese females aged 15-24 years. CONCLUSIONS: The study demonstrates a high risk of attempted suicide and a low risk of completed suicide among young Turkish and Surinamese females.

Electrocardiógrafos digitales en función de la telemedicina

En este trabajo se exponen diferentes soluciones que han hecho posible llevar a cabo una primera experiencia de Telemedicina, utilizando electrocardiógrafos digitales desarrollados en el Instituto Central de Investigaciones Digitales. Con estas soluciones se brinda la posibilidad de que estudios realizados en cualquiera de estos electrocardiógrafos, puedan ser transmitidos hacia estaciones remotas donde especialistas de más nivel los revisan y emiten su opinión diagnóstica. Las dos formas de establecer la comunicación son punto a punto a través de módem o vía correo electrónico. Se expone además la forma en que estos sistemas se están llevando a la práctica en algunos policlínicos y hospitales cubanos(AU)

Lack of polymorphism in a Vibrio cholerae O139-specific DNA region encoding the somatic antigen in strains isolated during 1993-1998.

Vibrio cholerae O139 Bengal emerged as a second aetiologic agent of cholera in South Asia in late 1992. This new serogroup arose from a Vibrio cholerae O1 strain by deletion of the chromosomal region encoding O1 specificity and acquisition of a novel 35-kb region encoding the O139 specificity. Previous studies indicated significant phenotypic and genotypic changes in O139 isolates over the years since its first appearance. This prompted us to study possible polymorphism in the 35-kb novel region encoding the O139 specificity. A total of 17 V. cholerae O139 isolates originating from different countries and years in South Asia and China, and a single unrelated V. cholerae O139 isolate from Argentina were studied. The 35-kb chromosomal region was amplified as two fragments of 12 and 23 kb in an extended PCR from all isolates. These amplicons were then treated separately with seven different restriction enzymes and separated by agarose gel electrophoresis. The South Asian and Chinese isolates gave identical patterns for the same enzymes, but different patterns for different enzymes, thus exhibiting no polymorphism in the 35-kb region. However, the Argentine isolate gave distinct patterns for most of the enzymes confirming its different origin. This data indicated that the portion of the chromosome encoding the O139 antigen specificity is highly conserved. As found in previous studies, the early O139 isolates were resistant to trimethoprim-sulfamethoxazole (TMP-SMX) and vibriostatic compound, O/129, and CAMP- haemolysin positive. The isolates of later years diverged exhibiting different patterns by pulsed-field gel electrophoresis (PFGE), and becoming susceptible to TMP-SMX and O/129, and CAMP-haemolysin negative.