Step-by-step optimization of a heterologous pathway for de novo naringenin production in Escherichia coli.

Naringenin is a plant polyphenol, widely explored due to its interesting biological activities, namely anticancer, antioxidant, and anti-inflammatory. Due to its potential applications and attempt to overcome the industrial demand, there has been an increased interest in its heterologous production. The microbial biosynthetic pathway to produce naringenin is composed of tyrosine ammonia-lyase (TAL), 4-coumarate-CoA ligase (4CL), chalcone synthase (CHS), and chalcone isomerase (CHI). Herein, we targeted the efficient de novo production of naringenin in Escherichia coli by performing a step-by-step validation and optimization of the pathway. For that purpose, we first started by expressing two TAL genes from different sources in three different E. coli strains. The highest p-coumaric acid production (2.54 g/L) was obtained in the tyrosine-overproducing M-PAR-121 strain carrying TAL from Flavobacterium johnsoniae (FjTAL). Afterwards, this platform strain was used to express different combinations of 4CL and CHS genes from different sources. The highest naringenin chalcone production (560.2 mg/L) was achieved by expressing FjTAL combined with 4CL from Arabidopsis thaliana (At4CL) and CHS from Cucurbita maxima (CmCHS). Finally, different CHIs were tested and validated, and 765.9 mg/L of naringenin was produced by expressing CHI from Medicago sativa (MsCHI) combined with the other previously chosen genes. To our knowledge, this titer corresponds to the highest de novo production of naringenin reported so far in E. coli. KEY POINTS: • Best enzyme and strain combination were selected for de novo naringenin production. • After genetic and operational optimizations, 765.9 mg/L of naringenin was produced. • This de novo production is the highest reported so far in E. coli.

Tailoring fructooligosaccharides composition with engineered Zymomonas mobilis ZM4.

Zymomonas mobilis ZM4 is an attractive host for the development of microbial cell factories to synthesize high-value compounds, including prebiotics. In this study, a straightforward process to produce fructooligosaccharides (FOS) from sucrose was established. To control the relative FOS composition, recombinant Z. mobilis strains secreting a native levansucrase (encoded by sacB) or a mutated ß-fructofuranosidase (Ffase-Leu196) from Schwanniomyces occidentalis were constructed. Both strains were able to produce a FOS mixture with high concentration of 6-kestose. The best results were obtained with Z. mobilis ZM4 pB1-sacB that was able to produce 73.4 ± 1.6 g L-1 of FOS, with a productivity of 1.53 ± 0.03 g L-1 h-1 and a yield of 0.31 ± 0.03 gFOS gsucrose-1. This is the first report on the FOS production using a mutant Z. mobilis ZM4 strain in a one-step process. KEY POINTS: • Zymomonas mobilis was engineered to produce FOS in a one-step fermentation process. • Mutant strains produced FOS mixtures with high concentration of 6-kestose. • A new route to produce tailor-made FOS mixtures was presented.

Cross-cultural adaptation and validation of the Brazilian Portuguese version of the Female Sexual Distress Scale-Revised questionnaire for women with vaginal laxity.

INTRODUCTION AND HYPOTHESIS: Vaginal laxity (VL) can impair women´s quality of life and there are not many tools aimed at quantitatively addressing this complaint. Sexual distress can be present within this group of patients. The aim of our study is to carry out the cross-cultural adaptation/translation and validation of the Female Sexual Distress Scale-Revised (FSDS-R) for Brazilian Portuguese women with VL. METHODS: Women age ≥ 18 years, with VL (n=82), and without VL (n=53) were included. Continuous variables were described in the form of mean/standard deviation or median/range, and Student's t test was used. The Chi-squared test was used for dichotomous variables. Cronbach's alpha coefficient was used for internal consistency and Spearman's correlation was used to assess construct validity (FSDS-R, Female Sexual Function Index [FSFI], and Incontinence Questionnaire Vaginal Symptoms [ICIQ-VS]). A significance level of 5% was established using a two-tailed test. RESULTS: Women with VL presented more anal/vaginal sexual intercourse than women without VL (p=0.030). All three instruments (FSDS-R, FSFI, and ICIQ-VS) presented discriminant validity between women with and without VL (p<0.001). A high internal consistency (Cronbach´s alpha =0.887) was found in women with VL and without VL (0.917). Regarding construct validity (n=82), there was a strong positive correlation between FSDS-R score and ICIQ-VS scales, except for a weaker correlation between the ICIQ-VS vaginal symptoms subscale (r: +0.2788; p=0.013). A moderate negative correlation was found between FSDS-R and all FSFI domains (p<0.001), except for pain (p<0.062). CONCLUSIONS: The Brazilian version of the FSDS-R showed adequate internal consistency and discriminant validity, and a correlation was found with other instruments such as FSFI and ICIQ-VS.

Higher expression of proline dehydrogenase altered mitochondrial function and increased Trypanosoma cruzi differentiation in vitro and in the insect vector.

The pathogenic protist Trypanosoma cruzi uses kissing bugs as invertebrate hosts that vectorize the infection among mammals. This parasite oxidizes proline to glutamate through two enzymatic steps and one nonenzymatic step. In insect vectors, T. cruzi differentiates from a noninfective replicating form to nonproliferative infective forms. Proline sustains this differentiation, but to date, a link between proline metabolism and differentiation has not been established. In T. cruzi, the enzymatic steps of the proline-glutamate oxidation pathway are catalyzed exclusively by the mitochondrial enzymes proline dehydrogenase [TcPRODH, EC: 1.5.5.2] and Δ1-pyrroline-5-carboxylate dehydrogenase [TcP5CDH, EC: 1.2.1.88]. Both enzymatic steps produce reducing equivalents that are able to directly feed the mitochondrial electron transport chain (ETC) and thus produce ATP. In this study, we demonstrate the contribution of each enzyme of the proline-glutamate pathway to ATP production. In addition, we show that parasites overexpressing these enzymes produce increased levels of H2O2, but only those overexpressing TcP5CDH produce increased levels of superoxide anion. We show that parasites overexpressing TcPRODH, but not parasites overexpressing TcP5CDH, exhibit a higher rate of differentiation into metacyclic trypomastigotes in vitro. Finally, insect hosts infected with parasites overexpressing TcPRODH showed a diminished parasitic load but a higher percent of metacyclic trypomastigotes, when compared with controls. Our data show that parasites overexpressing both, PRODH and P5CDH had increased mitochondrial functions that orchestrated different oxygen signaling, resulting in different outcomes in relation to the efficiency of parasitic differentiation in the invertebrate host.

Avascular necrosis of the femoral head in patients with cerebral palsy after hip surgery-incidence and impact on quality of life.

PURPOSE: This study investigated the incidence of AVN in patients with cerebral palsy who underwent a hip reconstruction surgery (soft tissue release, femur and pelvic osteotomies) and its impact on quality of life. METHODS: Retrospective study, with clinical and radiographic analysis of 104 patients (128 hips) GMFCS IV and V with a minimum two year follow-up. Reimers migration percentage, the amount of abduction, acetabular index, and the neck-shaft angle were collected before and after surgery. Modified Tönnis classification was used to analyze the hips before surgery, and the Bucholz and Ogden classification was used to identify hips with AVN. Function and quality of life were investigated with the CPCHILD questionnaire after surgery. RESULTS: The mean age of participants at surgery was 120.1 months (72-184), and it was not related with AVN (p = 0.946). AVN signs were observed in 62 hips (48.5%). The mean pre-op Reimers value was 68.7% (16-100) in normal hips and 83.1% (0-100) in hips with AVN (p = 0.003). All hips considered as Tönnis IV before surgery developed AVN (p = 0.006). The amount of abduction did not differ between hips that developed AVN and those that did not (p = 0.313). Patients who developed AVN had lower scores of quality of life (p = 0.023) and comfort (p = 0.025) according to the CPCHILD questionnaire. CONCLUSION: We observed a relationship between the greater pre-operative severity according to the Reimers index and the modified Tonnis classification and the development of AVN.

Challenges in the Heterologous Production of Furanocoumarins in Escherichia coli.

Coumarins and furanocoumarins are plant secondary metabolites with known biological activities. As they are present in low amounts in plants, their heterologous production emerged as a more sustainable and efficient approach to plant extraction. Although coumarins biosynthesis has been positively established, furanocoumarin biosynthesis has been far more challenging. This study aims to evaluate if Escherichia coli could be a suitable host for furanocoumarin biosynthesis. The biosynthetic pathway for coumarins biosynthesis in E. coli was effectively constructed, leading to the production of umbelliferone, esculetin and scopoletin (128.7, 17.6, and 15.7 µM, respectively, from tyrosine). However, it was not possible to complete the pathway with the enzymes that ultimately lead to furanocoumarins production. Prenyltransferase, psoralen synthase, and marmesin synthase did not show any activity when expressed in E. coli. Several strategies were tested to improve the enzymes solubility and activity with no success, including removing potential N-terminal transit peptides and expression of cytochrome P450 reductases, chaperones and/or enzymes to increase dimethylallylpyrophosphate availability. Considering the results herein obtained, E. coli does not seem to be an appropriate host to express these enzymes. However, new alternative microbial enzymes may be a suitable option for reconstituting the furanocoumarins pathway in E. coli. Nevertheless, until further microbial enzymes are identified, Saccharomyces cerevisiae may be considered a preferred host as it has already been proven to successfully express some of these plant enzymes.

3-Bromopyruvate: A new strategy for inhibition of glycolytic enzymes in Leishmania amazonensis.

The compound 3-bromopyruvate (3-BrPA) is well-known and studies from several researchers have demonstrated its involvement in tumorigenesis. It is an analogue of pyruvic acid that inhibits ATP synthesis by inhibiting enzymes from the glycolytic pathway and oxidative phosphorylation. In this work, we investigated the effect of 3-BrPA on energy metabolism of L. amazonensis. In order to verify the effect of 3-BrPA on L. amazonensis glycolysis, we measured the activity level of three glycolytic enzymes located at different points of the pathway: (i) glucose kinases, step 1, (ii) glyceraldehyde 3-phosphate dehydrogenase (GAPDH), step 6, and (iii) enolase, step 9. 3-BrPA, in a dose-dependent manner, significantly reduced the activity levels of all the enzymes. In addition, 3-BrPA treatment led to a reduction in the levels of phosphofruto-1-kinase (PFK) protein, suggesting that the mode of action of 3-BrPA involves the downregulation of some glycolytic enzymes. Measurement of ATP levels in promastigotes of L. amazonensis showed a significant reduction in ATP generation. The O2 consumption was also significantly inhibited in promastigotes, confirming the energy depletion effect of 3-BrPA. When 3-BrPA was added to the cells at the beginning of growth cycle, it significantly inhibited L. amazonensis proliferation in a dose-dependent manner. Furthermore, the ability to infect macrophages was reduced by approximately 50% when promastigotes were treated with 3-BrPA. Taken together, these studies corroborate with previous reports which suggest 3-BrPA as a potential drug against pathogenic microorganisms that are reliant on glucose catabolism for ATP supply.

Hydrogen Peroxide Generation as an Underlying Response to High Extracellular Inorganic Phosphate (Pi) in Breast Cancer Cells.

According to the growth rate hypothesis (GRH), tumour cells have high inorganic phosphate (Pi) demands due to accelerated proliferation. Compared to healthy individuals, cancer patients present with a nearly 2.5-fold higher Pi serum concentration. In this work, we show that an increasing concentration of Pi had the opposite effect on Pi-transporters only in MDA-MB-231 when compared to other breast cell lines: MCF-7 or MCF10-A (non-tumoural breast cell line). Here, we show for the first time that high extracellular Pi concentration mediates ROS production in TNBC (MDA-MB-231). After a short-time exposure (1 h), Pi hyperpolarizes the mitochondrial membrane, increases mitochondrial ROS generation, impairs oxygen (O2) consumption and increases PKC activity. However, after 24 h Pi-exposure, the source of H2O2 seems to shift from mitochondria to an NADPH oxidase enzyme (NOX), through activation of PKC by H2O2. Exogenous-added H2O2 modulated Pi-transporters the same way as extracellular high Pi, which could be reversed by the addition of the antioxidant N-acetylcysteine (NAC). NAC was also able to abolish Pi-induced Epithelial-mesenchymal transition (EMT), migration and adhesion of MDA-MB-231. We believe that Pi transporters support part of the energy required for the metastatic processes stimulated by Pi and trigger Pi-induced H2O2 production as a signalling response to promote cell migration and adhesion.

Weight regain and eating behavior in physically active and inactive women after 24 months of bariatric surgery.

PURPOSE: This research has tested the association between anthropometric profile and eating behavior according to the practice of physical activity in women with more than 24 months after bariatric surgery. METHODS: It is a transversal study accomplished with 44 adult women, in which sociodemographic and anthropometric data of eating behavior as well as physical activity practice have been collected. RESULTS: The average of overweight loss was satisfactory (> 50%), the average of weight regain was of 16.4 ± 11.2%, being the weight regain (kg) statistically lower in the physically active group (p = 0.049). There had been predominance of emotional eating in both groups (active p = 0.025; inactive p = 0.040); significant inverse correlation (ß = - 0.286; IC - 0.317; - 0,005; p = 0.044) between food restriction behavior and weight regain (%) depending on postoperative period; and the time of physical activity practice (min) has presented inverse correlation, statistically significant (ß = - 0.311; IC - 0.048; - 0,001; p = 0.039) on the weight regain (%) independent of postoperative period. CONCLUSIONS: The physically active participants have presented the lowest weight regain and predominance of emotional eating domain, evidencing the need to model eating behavior and to encourage the practice of physical activity in these patients. LEVEL III: Case-control analytic study.

Polymyxin B Heteroresistance and Adaptive Resistance in Multidrug- and Extremely Drug-Resistant Acinetobacter baumannii.

Acinetobacter baumannii is an emerging pathogen associated with nosocomial infections and multidrug resistance. Polymyxin B has been used to treat infections caused by multidrug-resistant (MDR) A. baumannii but an increase in polymyxin B resistance has been observed. We aimed to determine the diversity, antimicrobial susceptibility, presence of polymyxin B heteroresistance and adaptive resistance in 72 A. baumannii clinical isolates from two public hospitals in Rio de Janeiro. The isolates were identified by sequencing of rpoB gene. Determination of the genetic diversity of isolates was performed by pulsed-field gel electrophoresis and oxacillinases genes were detected by polymerase chain reaction. The polymyxin B heteroresistance was analyzed by population analysis profile and adaptive resistance was evaluated after serial daily passages of isolates in broth containing increasing polymyxin B concentrations. The results showed that 49% of the isolates were collected from respiratory system and 62% were MDR, while 35% were extensively drug resistant. Additionally, all the isolates carried blaOXA-23-like, blaOXA-51-like genes and ISAba1, while 1% had blaOXA-24-like gene. The association of ISAba1-blaOXA-23 was found in 96% of the isolates. Polymyxin B heteroresistance was found in 36% of the isolates and polymyxin B adaptive resistance was not found in the isolates. Our study demonstrated the high resistance to antimicrobials used in clinical practice and the spread of oxacillinases genes and insertion sequence (IS). We also reported the presence of heteroresistance to polymyxin B used as a last-resort therapy for MDR A. baumannii.

Removal of a mixture of pharmaceuticals sulfamethoxazole and diclofenac from water streams by a polyamide nanofiltration membrane.

Wastewater treatment plants are not specially designed to remove pharmaceutically active compounds (PhACs), since these substances are toxic and bio-refractory. This paper aims to investigate and optimize the performance of the Trisep TS80 nanofiltration (NF) membrane for the removal of a mixture of two of the most detected PhACs in municipal wastewaters worldwide, sulfamethoxazole and diclofenac. Several NF tests were carried out to study the rejections of these contaminants both spiked in demineralized water, filtrated water taken from Mondego River and secondary effluent coming from a municipal wastewater treatment plant. Among the several studied operating variables, pH was the one that most affected the contaminant rejection and membrane permeability. In the case of synthetic effluent, an applied pressure of 10 bar and pH 7 were determined as the best operating conditions, which allowed almost total chemical oxygen demand retention and a global contaminant rejection of 96.3% to be achieved. The application of different water matrices (river water and secondary municipal effluent) had no relevant impact on process efficiency. Vibrio fischeri luminescence inhibition tests revealed that treatment by nanofiltration reduced acute toxicity of all studied effluents.

Latin American Consensus Statement for the Use of Contrast-Enhanced Transcranial Ultrasound as a Diagnostic Test for Detection of Right-to-Left Shunt.

BACKGROUND: The role of patent foramen ovale is a field of debate and current publications have increasing controversies about the patients' management in young undetermined stroke. Work up with echocardiography and transcranial Doppler (TCD) can aid the decision with better anatomical and functional characterization of right-to-left shunt (RLS). Medical and interventional strategy may benefit from this information. SUMMARY: a group of experts from the Latin American participants of the Neurosonology Research Group (NSRG) of World Federation of Neurology created a task force to review literature and describe the better methodology of contrast TCD (c-TCD). All signatories of the present consensus statement have published at least one study on TCD as an author or co-author in an indexed journal. Two meetings were held while the consensus statement was being drafted, during which controversial issues were discussed and voted on by the statement signatories. The statement paper was reviewed and approved by the Executive Committee of the NSRG of the World Federation of Neurology. The main objective of this consensus statement is to establish a standardization of the c-TCD technique and its interpretation, in order to improve the informative quality of the method, resulting in expanding the application of TCD in the clinical setting. These recommendations optimize the comparison of different diagnostic methods and encourage the use of c-TCD for RLS screening and complementary diagnosis in multicenter studies.

The impact generated by publicly and charity-funded research in the United Kingdom: a systematic literature review.

OBJECTIVE: To identify, synthesise and critically assess the empirical evidence of the impact generated by publicly and charity-funded health research in the United Kingdom. METHODS: We conducted a systematic literature review of the empirical evidence published in English in peer-reviewed journals between 2006 and 2017. Studies meeting the inclusion criteria were selected and their findings were analysed using the Payback Framework and categorised into five main dimensions, namely knowledge, benefits to future research and research use, benefits from informing policy and product development, health and health sector benefits, and broader economic benefits. We assessed the studies for risk of selection, reporting and funding bias. RESULTS: Thirteen studies met the inclusion criteria. The majority of the studies (10 out of 13) assessed impact at multiple domains including the main five key themes of the Payback Framework. All of them showed a positive impact of funded research on outcomes. Of those studies, one (8%), six (46%) and six (46%) presented a low, moderate and high risk of bias, respectively. CONCLUSIONS: Empirical evidence on the impact of publicly and charity-funded research is still limited and subject to funding and selection bias. More work is needed to establish the causal effects of funded research on academic outcomes, policy, practice and the broader economy.

From lip to lab: salty tasting skin is the main clue that raises clinical suspicion of cystic fibrosis in young infants.

AIM: This study developed a clinical decision rule (CDR) to rule in or rule out cystic fibrosis (CF) in the first weeks of life. METHODS: We combined a two-step nonconcurrent birth cohort and a cross-sectional controlled study in which observers and mothers were blinded to confirmatory sweat test results. Neonates from uncomplicated pregnancies and deliveries, with two subsequent tests for immunoreactive trypsinogen (IRT) higher than 70 ng/mL, were eligible to take part. RESULTS: We included 49 CF-affected and 177 CF-unaffected infants with an average age of 34 days. CF-affected infants demonstrated statistically significant differences in weight gain, stool frequency and salty tasting skin from birth. Multivariate analysis showed that the main predictors of CF were salty tasting skin (odds ratio 17.2) and weight gain of <10.5 g per day (odds ratio 4.6). Depending on the CDR score, we obtained the following results: sensitivity (42.9-61.2%), specificity (89.6-96.5%), positive (62.5-77.8%) and negative (85.6-89.1%) predictive value and positive (5.8-12.3) and negative (0.4-0.6) likelihood ratio. CONCLUSION: The CDR could provide a reliable index of clinical suspicion and timely referral for sweat testing in settings without newborn screening programmes and may also be applied to false-negative individuals where such programmes already exist.

Symptomatic Cerebral Vasospasm and Delayed Cerebral Ischemia Following Transsphenoidal Resection of a Craniopharyngioma.

BACKGROUND: Vasospasm has been rarely described as a complication associated with craniopharyngioma surgery. Herein we describe a patient who developed symptomatic vasospasm and delayed cerebral ischemia after transsphenoidal surgery for a craniopharyngioma. CASE REPORT: A 67-year-old woman became drowsy 2 weeks after a transsphenoidal resection of a craniopharyngioma. A head computed tomography (CT) was unremarkable except for postoperative findings. Electroencephalogram and laboratory studies were within the normal limits. A repeated CT scan 48 hours after the initial symptoms showed bilateral infarcts in the territory of the anterior cerebral arteries (ACA). Transcranial Doppler (TCD) showed increased blood flow velocities in both anterior cerebral arteries (169 cm/second in the left ACA and 145 cm/second in the right ACA) and right middle cerebral artery (164 cm/second) compatible with vasospasm. A CT angiography confirmed the findings. She was treated with induced hypertension and her level of consciousness improved. TCD velocities normalized after 2 weeks. CONCLUSION: Cerebral vasospasm should be considered in the differential diagnosis of patients with altered neurologic status in the postoperative period following a craniopharyngioma resection.

Cerebral Vasospasm and Delayed Cerebral Ischemia after Warfarin-Induced Subarachnoid Hemorrhage.

BACKGROUND: Subarachnoid hemorrhage (SAH) associated with anticoagulation is a rare event. About 30% of in-hospital patients with aneurysmal SAH develop delayed cerebral ischemia (DCI); however, the occurrence of vasospasm and DCI in patients with nonaneurysmal SAH is still controversial. METHODS: This study is a case report of a patient experiencing an anticoagulation-induced SAH complicated by vasospasm and DCI. RESULTS: A 38-year-old woman presented to our hospital with a sudden onset of severe headache. Head computed tomography (CT) showed bleeding in the posterior fossa subarachnoid space. There was no aneurysm in the CT angiography. The International Normalized Ratio at hospital admission was 9. Anticoagulation was held, and she was treated with fresh frozen plasma (15 mg/kg) and parenteral vitamin K. The patient remained in the intensive care unit and had daily transcranial Doppler (TCD) monitorization. TCD examination detected increased blood flow velocities in the basilar and vertebral arteries, meeting criteria for definitive vasospasm. CT angiography confirmed the presence of posterior circulation vasospasm. Magnetic resonance (MR) imaging 2 weeks after the bleeding showed a small area of restricted diffusion in the left superior cerebellar artery territory. MR angiography showed resolution of the vasospasm at this time point and TCD velocities normalized. CONCLUSIONS: In conclusion, this case report suggests that vasospasm and consequent DCI is a possible mechanism of secondary lesion after anticoagulation-induced SAH. To our knowledge, this is the first report of vasospasm and DCI due to warfarin-associated SAH.

Nox4: a hydrogen peroxide-generating oxygen sensor.

Nox4 is an oddity among members of the Nox family of NADPH oxidases [seven isoenzymes that generate reactive oxygen species (ROS) from molecular oxygen] in that it is constitutively active. All other Nox enzymes except for Nox4 require upstream activators, either calcium or organizer/activator subunits (p47(phox), NOXO1/p67(phox), and NOXA1). Nox4 may also be unusual as it reportedly releases hydrogen peroxide (H2O2) in contrast to Nox1-Nox3 and Nox5, which release superoxide, although this result is controversial in part because of possible membrane compartmentalization of superoxide, which may prevent detection. Our studies were undertaken (1) to identify the Nox4 ROS product using a membrane-free, partially purified preparation of Nox4 and (2) to test the hypothesis that Nox4 activity is acutely regulated not by activator proteins or calcium, but by cellular pO2, allowing it to function as an O2 sensor, the output of which is signaling H2O2. We find that approximately 90% of the electron flux through isolated Nox4 produces H2O2 and 10% forms superoxide. The kinetic mechanism of H2O2 formation is consistent with a mechanism involving binding of one oxygen molecule, which is then sequentially reduced by the heme in two one-electron reduction steps first to form a bound superoxide intermediate and then H2O2; kinetics are not consistent with a previously proposed internal superoxide dismutation mechanism involving two oxygen binding/reduction steps for each H2O2 formed. Critically, Nox4 has an unusually high Km for oxygen (∼18%), similar to the values of known oxygen-sensing enzymes, compared with a Km of 2-3% for Nox2, the phagocyte NADPH oxidase. This allows Nox4 to generate H2O2 as a function of oxygen concentration throughout a physiological range of pO2 values and to respond rapidly to changes in pO2.

Rhodnius prolixus: modulation of antioxidant defenses by Trypanosoma rangeli.

Trypanosoma rangeli is a protozoan parasite of insects and mammals that is challenged by the constant action of reactive oxygen species, generated either by its own metabolism or through the host immune response. The aim of this work was to investigate whether T. rangeli is able to modify the redox state of its insect vector, Rhodnius prolixus, through the modulation of such antioxidant enzymes as superoxide dismutase (SOD), catalase, and GPx present in the midgut of the insect. We verified that in R. prolixus fed with blood infected with T. rangeli there is an increase in SOD activity in the anterior and posterior midguts. However, the activities of enzymes related to hydrogen peroxide and hydroperoxides metabolism, such as catalase and GPx, were decreased in relation to the insect control group, which was only fed blood. These changes in the redox state of the vector led to an increase in lipid peroxidation and thiol oxidation levels in the anterior and posterior midgut tissues. We also verified that the addition of 1 mM GSH in the blood meal of the infected insects increased the proliferation of these parasites by 50%. These results suggest that there is an increase in oxidative stress in the insect gut during T. rangeli infection, and this condition could contribute to the control of the proliferation of these parasites.

Eating Behavior and Nutritional Profile of Children with Autism Spectrum Disorder in a Reference Center in the Amazon.

There is no single pattern for the evolution of the nutritional status of children with autism spectrum disorder (ASD). Previous studies have found a tendency towards food selectivity with food monotony and difficulties with food texture in children with ASD, but studies in this area, especially in Brazil, are still scarce. The nutritional profile and changes in eating behavior were analyzed in patients with autism spectrum disorder assisted at a reference center in Belém. Eating behavior was assessed using the Labyrinth Scale, nutritional status assessment through weight and height (to calculate body mass index-BMI), and consumption food through the 24 h reminder. A total of 80 children of both sexes participated in the study, the majority of whom were male (80%), 47.5% eutrophic, while for the food consumption of the children evaluated, there was an average energy consumption of 1911 kcal daily, with 57.3%, 15.4%, and 27.3% of carbohydrates, proteins, and lipids, respectively. In relation to eating behavior, the highest averages were demonstrated in the domains of food selectivity, behavioral aspects, and mealtime skills. Masticatory motor scores showed a positive correlation with weight, BMI, and the amount of energy consumed by the child. The gastrointestinal symptoms score showed a negative correlation with the child's age. Regarding mealtime skills, a negative correlation was observed with the proportion of carbohydrates in the diet and a positive correlation with the proportion of lipids consumed in the children's diet. Therefore, knowing the main changes in eating behavior is important to ensure a complete and safe approach for each patient.

The Relationship between Anxiety Symptoms and Perceived Quality of Life among Caregivers of Children with Autism Spectrum Disorder in the Amazon.

The relationship between anxiety symptoms and perceived quality of life among caregivers of children with autism was verified. To assess perceived quality of life, the Short Form Healthy Survey Item was used; perception of anxiety symptoms was assessed using the Beck Anxiety Inventory. Eighty caregivers of children with autism participated, with 68.7% of caregivers being mothers. Of the total number of caregivers, 68.8% had a severe level of anxiety. Regarding perceived quality of life, they demonstrated greater impairment in limitation due to physical aspects, limitation due to emotional aspects, vitality, and pain. Caregivers with severe anxiety levels had a worse quality of life in the domains of pain (p = 0.012), social aspects (p < 0.001), limitation due to emotional aspects (p = 0.001), and mental health (p < 0.001). However, in the functional capacity domain, caregivers with a moderate level of anxiety had a better physical capacity score (p = 0.001). There was a negative correlation between the general anxiety score and the general physical (p = 0.029) and general emotional components of perceived quality of life (p < 0.001). It was found that caregivers of children with ASD have a high level of anxiety, which is a predictor of the perception of a worse quality of life.