Extraction, Purification and Electrical Characterization of Gross Galactomannan and Purified Galactomannan Obtained from Adenanthera pavonina L. Seeds.

Aiming of self-sustainable production, the search for biodegradable and biocompatible materials has brought with it the need to know the physicochemical and dielectric characteristics of polysaccharide-based composite structures, which can be used as important and promising raw materials for biotechnology and electronic industries. Galactomannans are polysaccharides, extracted from seeds and microbiological sources, consisting of mannose and galactose. In this context, this work aimed to extract, purify and characterize by XRD, FTIR and impedance spectroscopy galactomannan obtained from seeds of Adenanthera pavonina L. The purification process was made with ethyl alcohol at concentrations of 70, 80 and 90 %. Polymeric films were prepared by solvent slow evaporation at low temperatures. XRD measurements revealed that Galactomannan from Adenanthera pavonina L., after purification, has a semi-crystalline structure due to the identification of two peaks the first between 5.849° and 6.118° and the second between 20.011° and 20.247°. FTIR spectra showed the functional groups associated with monosaccharides of the galactomannan from Adenanthera pavonina L. seeds, as well as the typical polysaccharide bands and peaks, confirmed by literature data. The impedance results give an increment on the state-of-the-art of this biomaterial by showing the existence of dielectric relaxations, independent of the degree of purification, using the dielectric modulus formalism. The permittivity analysis reveals the presence of water in the structure of the film, whose dipoles contribute to the relatively high value of the dielectric constant. From the results obtained, it can be concluded that purified galactomannan has the potential for possible applications in the electronics industry as a green and eco-friendly dielectric material.

Integrative taxonomy of Urocleidoides spp. (Monogenoidea: Dactylogyridae) parasites of characiform and gymnotiform fishes from the coastal drainages of the Eastern Amazon, Brazil.

Eight species (four new) of Urocleidoides are reported from Characiformes and Gymnotiformes fishes of the coastal drainages of the Eastern Amazon. Urocleidoides vanini n. sp. is characterized by having a male copulatory organ (MCO) with three and a half counterclockwise rings, absence of vaginal sclerite, and a V-shaped ventral bar. Urocleidoides atilaiamarinoi n. sp. has MCO with two and a half counterclockwise rings, dumbbell-shaped accessory piece, similar anchors, open V-shaped ventral bar, and open U-shaped dorsal bar. Urocleidoides macrosoma n. sp. exhibits an elongate and robust body, MCO comprising one counterclockwise ring, similar anchors with wavy point, and dumbbell-shaped ventral and dorsal bars. Urocleidoides nataliapasternakae n. sp. has MCO comprising two and a half counterclockwise rings, vaginal canal convoluted, point of the dorsal anchor with ornamentation as sclerotized shredded filaments, elongate dumbbell-shaped ventral bar, and U-shaped dorsal bar. Urocleidoides naris and Urocleidoides brasiliensis from H. malabaricus (Characiformes) and the incertae sedis species, Urocleidoides gymnotus and Urocleidoides carapus, from Sternopygus macrurus (Gymnotiformes) are reported, and their molecular sequences are presented in this study. Phylogenetic analyses based on molecular data (28S rDNA and COI mtDNA) reveal that species of Urocleidoides lacking vaginal sclerite are closely related to species that possess vaginal sclerite, suggesting that the absence of vaginal sclerite in Urocleidoides may be the result of a secondary loss. The relationships between species of Urocleidoides and other Neotropical dactylogyrids are also addressed.

The erythrocyte alloimmunisation in patients with sickle cell anaemia: a systematic review.

Transfusion therapy is a common practice in the treatment of anaemia and can cause erythrocyte alloimmunisation. To systematise data related to erythrocyte alloimmunisation in patients with sickle cell disease (SCD), a bibliographic search was carried out in September 2017 to search for studies in four electronic databases. (i) Referring to the original work, (ii) being cohort or case-control, (iii) having been developed with individuals with SCD and (iv) having evaluated the erythrocyte alloimmunisation. Two reviewers identified the articles for inclusion in the study, extracted the predetermined data and carried out the evaluation of the methodological quality of the work. 21 studies were selected; the studies included data on 20 636 individuals (children and adults), were mostly published in the last 10 years, were developed in the United States and had high methodological quality. The occurrence of erythrocyte alloimmunisation ranged from 4·4 to 76%, and there was a higher rate of alloimmunisation against antigens of the Rh system. The risk factors for alloimmunisation were age; gender (female); red blood cell (RBC) units received; presence of ≥1 autoantibodies, TNF-α, interleukin (IL1B), human leukocyte antigens (HLA)-DRB1 gene polymorphisms; first blood transfusion (BT) after 5 years of age, transfusion episodic, multiple or during inflammatory events, acute chest syndrome (ACS) and vase-occlusive crisis (VOC); increased percentage of CD41 T memory cells; and positive direct antiglobulin test. Transfusion policies should be developed to protect the patient and his or her health based on the main factors associated with its incidence.

Monogenoidean parasites of Acestrorhynchus falcatus (Characiformes: Acestrorhynchidae) from Pará, Brazil: species of Diaphorocleidus and Rhinoxenoides n. gen. (Monogenoidea: Dactylogyridae).

Two new species of Diaphorocleidus and one new species of Rhinoxenoides n. gen. are described from the gills of Acestrorhynchus falcatus (Bloch) from rivers of north-eastern Pará, Brazil. Diaphorocleidus jaymedeloyolai n. sp. is characterized by a male copulatory organ (MCO) possessing three counterclockwise coils; similar anchors with subtriangular superficial roots; a ventral bar with posteromedial projection; and hooks of pairs 1, 4 and 7 approximately three times longer than hook pair 5. Diaphorocleidus sclerocolpus n. sp. differs from its congeners by a dual-branched accessory piece articulated with the MCO and a sclerotized tubular vagina with a bottle-shaped vestibule. Rhinoxenoides n. gen. is proposed and is characterized by possessing: MCO sclerotized with clockwise coils; an accessory piece articulated to the base of MCO; a sinistroventral vaginal aperture; ventral anchor with conspicuous roots; dorsal anchor with superficial root five times longer than deep root; and absence of dorsal bar. The proposal of Rhinoxenoides n. gen. is also supported by its phylogenetic relationship with Protorhinoxenus prochilodi and species of Rhinoxenus, using 16 morphological characters, which resulted in the following hypothesis of sister-group relationships: Rhinoxenoides n. gen. [Protorhinoxenus (Rhinoxenus curimatae (R. nyttus (R. bulbovaginatus (R. guianensis, R. piranhus, R. euryxenus (R. arietinus, R. anaclaudiae)))))].

Photodynamic effect of zinc porphyrin on the promastigote and amastigote forms of Leishmania braziliensis.

Leishmaniasis is a neglected disease present in more than 88 countries. The currently adopted chemotherapy faces challenges related to side effects and the development of resistance. Photodynamic therapy (PDT) is emerging as a therapeutic modality for cutaneous leishmaniasis. Zn(ii) meso-tetrakis(N-ethylpyridinium-2-yl)porphyrin (ZnTE-2-PyP4+, ZnP) is a cationic, water-soluble, zinc porphyrin-based photosensitizer whose photodynamic effect on Leishmania braziliensis was analyzed by evaluating the number of visibly undamaged and motile cells, cell membrane integrity, mitochondrial membrane potential, and ultrastructural damage. Treatment of parasites with ZnP and light induced damage in up to 90% of L. braziliensis promastigote cells. Propidium iodide labeling suggested the loss of plasma membrane integrity. In samples treated with ZnP and light, a hyperpolarization of the mitochondrial membrane potential was also observed. Ultrastructural evaluation of promastigotes after photodynamic treatment indicated a loss of cytoplasmic material and the presence of vacuoles. Scanning electron microscopy showed wrinkling of the plasma membrane and a reduced cell volume. Additionally, the number of amastigotes per macrophage was reduced by about 40% after photodynamic application. The treatment showed no considerable toxicity against mammalian cells. Therefore, the results indicated that PDT associated with ZnTE-2-PyP4+ represents a promising alternative to cutaneous leishmaniasis treatment.

Low-dose estradiol and endothelial and inflammatory biomarkers in menopausal overweight/obese women.

OBJECTIVE: We aimed to investigate the effects of low-dose transdermal estrogen on endothelial and inflammatory biomarkers in menopausal overweight/obese women. METHODS: We recruited 44 menopausal women (47-55 years; body mass index 27.5-34.9 kg/m(2)) and divided them into estradiol (1 mg/day; n = 22) or placebo groups (n = 22). They were double-blinded, followed and treated for 3 months. At baseline and post-intervention, inflammatory biomarkers (hs-CRP, IL-1ß, IL-6, MCP-1 and TNF-α) and of vascular injury (activated circulating endothelial cells, CEC-a) and repair (endothelial progenitor cells, EPC) were quantified. Resting CECs (CEC-r) were also assessed. Microvascular reactivity and vasomotion were analyzed by laser-Doppler flowmetry. RESULTS: Volunteers (51.8 ± 2.3 years; mean body mass index 31.5 ± 2.5 kg/m(2)) had been menopausal for 3 (range 2-5) years. After treatment, no changes were observed in the placebo group, while levels of CEC-r and EPC increased in the estradiol group. In this group, no changes in inflammatory biomarkers were observed but it required a lower cumulative dose of acetylcholine to achieve peak velocity during endothelial-dependent vasodilatation and there was increased endothelial-independent vasodilatation. CONCLUSIONS: The short-term use of low-dose transdermal estradiol therapy in overweight/obese menopausal women increased markers of vascular repair and improved microvascular reactivity without changing the inflammatory biomarkers. CLINICAL TRIAL REGISTRATION: NCT01295892 at www.clinicaltrials.gov .

Fluoxetine reduces periodontal disease progression in a conditioned fear stress model in rats.

BACKGROUND AND OBJECTIVE: Recent evidence suggests that the use of fluoxetine could reduce periodontal disease severity. However, the effect of fluoxetine on periodontal disease has not been tested in the context of conditioned fear stress (CFS). We hypothesized that inhibition of chronic stress by fluoxetine might decrease the levels of bone loss in periodontal disease. The aim of the present study was to analyze the effect of fluoxetine on bone loss in chronic periodontitis. MATERIAL AND METHODS: Fourteen Wistar rats were submitted to ligature-induced periodontal disease and divided into four groups (A-D). Groups A (n = 3) and B (n = 4) were not stressed, while Groups C (n = 3) and D (n = 4) were submitted to a CFS paradigm for 38 d. Daily fluoxetine (20 mg/kg) was administered to Groups B and D from day 20 to day 39, at which point the rats were submitted to an open field test and killed on day 40. Mandibles were removed for histological and immunohistochemical analyses. RESULTS: Stress was associated with a higher level of bone loss in Group C compared with Group A. Additionally, no differences in bone loss were observed among Groups A, B and D. CONCLUSION: We showed that stress is associated with the progression of bone loss in a CFS model in rats and that fluoxetine treatment reduces the bone loss.

Diazepam reverses the alveolar bone loss and hippocampal interleukin-1beta and interleukin-6 enhanced by conditioned fear stress in ligature-induced periodontal disease in rats.

BACKGROUND AND OBJECTIVE: Stress and anxiety have been associated with chronic periodontitis, but few studies examining the effects of psychotropic drugs on periodontal health have been performed. Therefore, we aimed to investigate the effects of diazepam on the progression of periodontitis in chronically stressed rats. MATERIAL AND METHODS: Fourteen Wistar rats were submitted to ligature-induced periodontal disease and were divided into four groups . Two groups were not stressed, whereas two groups were submitted to a conditioned fear stress paradigm for 38 d. Daily diazepam treatment (2 mg/kg, orally) was administered to one unstressed group and to one group submitted to a conditioned fear stress paradigm from day 2 to the day 39, at which point the rats were submitted to an open field test and were killed on day 40. Brains and mandibles were removed for histological and immunohistochemical analyses. RESULTS: Animals exposed to conditioned fear stress presented an increase in freezing behavior, a decrease in locomotor activity, enhanced alveolar bone loss and higher levels of hippocampal interleukin-1beta (IL-1ß) and interleukin-6 (IL-6), compared with the control group. Diazepam, at the dose used in the current study, had no effect on freezing behavior but reversed the decrease in locomotor activity provoked by stress. Additionally, the treatment reduced the levels of hippocampal IL-1ß and IL-6 and alveolar bone loss in Wistar rats. Neither conditioned fear stress nor diazepam treatment had an effect on periodontal IL-1ß or IL-6 levels in animals. CONCLUSION: Our results suggest that diazepam treatment reduces bone loss in rats submitted to conditioned fear stress. In addition, diazepam treatment led to decreased IL-1ß and IL-6 levels in the hippocampus.

Mutations in a novel cochlear gene cause DFNA9, a human nonsyndromic deafness with vestibular dysfunction.

DFNA9 is an autosomal dominant, nonsyndromic, progressive sensorineural hearing loss with vestibular pathology. Here we report three missense mutations in human COCH (previously described as Coch5b2), a novel cochlear gene, in three unrelated kindreds with DFNA9. All three residues mutated in DFNA9 are conserved in mouse and chicken Coch, and are found in a region containing four conserved cysteines with homology to a domain in factor C, a lipopolysaccharide-binding coagulation factor in Limulus polyphemus. COCH message, found at high levels in human cochlear and vestibular organs, occurs in the chicken inner ear in the regions of the auditory and vestibular nerve fibres, the neural and abneural limbs adjacent to the cochlear sensory epithelium and the stroma of the crista ampullaris of the vestibular labyrinth. These areas correspond to human inner ear structures which show histopathological findings of acidophilic ground substance in DFNA9 patients.

Maternal prolactin inhibition during lactation is associated to renal dysfunction in their adult rat offspring.

The renal function of rats whose mothers had hypoprolactinemia at the end of lactation was evaluated during development. Lactating Wistar rats were treated with bromocriptine (BRO, 1 mg twice a day, s.c.) or saline on days 19, 20, and 21 of lactation, and their male offspring were followed from weaning until 180 days old. 1 rat from each of the 12 litters/group was evaluated at 2 time points (90 and 180 days). Body and kidney weights, sodium, potassium, and creatinine were measured. Values were considered significant when p<0.05. Adult BRO-treated offspring presented higher body weight (+10%), lower relative renal weight at 90 and 180 days (-9.2% and -15.7%, respectively), glomerulosclerosis, and peritubular fibrosis. At 90 and 180 days, creatinine clearance was lower (-32% and -30%, respectively), whereas serum potassium was higher (+19% and +29%, respectively), but there were no changes in serum sodium. At 180 days, higher proteinuria (+36%) and serum creatinine levels (+20%) were detected. Our data suggest that prolactin inhibition during late lactation programs renal function damage in adult offspring that develops gradually, first affecting the creatinine clearance and potassium serum levels with further development of hyperproteinuria and higher serum creatinine, without affecting sodium. Thus, precocious weaning programs some components of the metabolic syndrome, which can be a risk factor for further development of kidney disease.

Association between visceral/subcutaneous adipose tissue ratio and plasma inflammatory markers and score for cardiovascular risk prediction in a Brazilian cohort: Pró-Saúde Study.

Visceral adipose tissue (VAT) is associated with various metabolic disorders, and adipokines, secreted by adipose tissue, are involved in their pathogenesis. This study investigated associations between VAT/subcutaneous adipose tissue (SAT) ratio, inflammatory markers, and cardiovascular (CV) risk-score in adults. Plasma levels of adipokines, plasma lipid profile, blood pressure, and body composition (using dual-emission x-ray absorptiometry) were determined. CV risk-score based on the American College of Cardiology and the American Heart Association (ACC/AHA) score was calculated in a sample of 309 Brazilian civil servants aged <60 years. Participants' VAT/SAT ratio were categorized into quartiles. Among males, plasma leptin (2.8 ng/mL) and C reactive protein (CRP) (0.2 mg/dL) (P<0.05) levels were higher at P75 and P50 than P5, and the highest calculated CV risk-score was observed at P75 (7.1%). Among females, higher plasma adiponectin levels were observed at P25 (54.3 ng/mL) compared with P75 (36 ng/mL) (P<0.05). Higher plasma CRP levels were observed at P75 (0.4 mg/dL) compared with P5 (0.1 mg/dL) (P<0.05). Higher CV risk-score was observed at P75 (2.0%) compared with P5 (0.7%). In both sexes, VAT and VAT/SAT ratio were directly associated with plasma leptin, CRP, and CV risk-score, and inversely associated with adiponectin; SAT was directly associated with plasma leptin and CRP (P<0.01); interleukin (IL)-10 and CRP were directly associated with adiponectin and leptin, respectively (P<0.05). Among men only, IL-10 (inversely) and CRP (directly) were associated with CV risk-score (P=0.02). Our results strengthened the relevance of the VAT/SAT ratio in cardiovascular risk.

Leptin treatment during lactation programs leptin synthesis, intermediate metabolism, and liver microsteatosis in adult rats.

Epidemiological and experimental studies have associated development of metabolic syndrome with stressful events (nutritional, hormonal, or environmental) in early life. This phenomenon is known as programing and changes in adipokines levels in early life, especially leptin, seem to be involved with its development. We have shown that neonatal hyperleptinemia on lactation programs for leptin resistance, hyperthyroidism, and higher corticosterone and catecholamines levels with cardiovascular consequences. In the present study, we evaluated the effect of hyperleptinemia during lactation on the glucose and lipid metabolism and liver morphology of adult rats, which were saline or leptin-treated (8 microg/100 g of body weight) daily, for the first 10 days of life. Leptin group had lower body mass during treatment, but higher body mass and hyperleptinemia at adulthood, without difference in fat mass. We showed that the probable source of hyperleptinemia is the higher leptin content in the subcutaneous adipose tissue. The programed rats showed hyperinsulinemia and hypoadiponectinemia with higher expression of the hypothalamic Suppressor of Cytokine Signaling 3 (SOCS3), suggesting insulin resistance. Besides, they presented higher liver glycogen and hypertriglyceridemia. We also observed liver microsteatosis in the leptin-programed adult rats. Our data show that neonatal hyperleptinemia alters glucose metabolism, which seems to be partially compensated by the hyperinsulinemia. However, changes in the lipid metabolism are not compensated. It is probable that these changes induced by neonatal hyperleptinemia result from a selective tissue specific resistance both to insulin and leptin at adulthood, and the increase of SOCS3 may play an important role in this process.

Viability and cell cycle analysis of equine fibroblasts cultured in vitro.

This experiment aimed to study equine fibroblasts in culture analyzing and the cell cycle and viability of cells pre- and post-freezing. Skin fragments were obtained from 6 horses and cultured in DMEM high glucose + 10% FCS in 5% CO(2) until the beginning of confluence. Two passages were performed before freezing. Cells subjected to serum starvation (0.5% FCS) were analyzed for viability and cell cycle at 24, 48, 72, 96, 120, 144 and 168 h of culture. For the confluent groups, cells were analyzed at the moment they achieved confluence. Cellular viability was assisted with Hoescht 33342 and propidium iodide. The analysis of apoptosis/necrosis and cell cycle was performed using a flow cytometer (FACS Calibur BD((R))) after staining the cells with annexin V and propidium iodide. Both optical microscopy and flow cytometry confirmed that cellular viability was similar for serum starvation and confluent groups (average 84%). Similarly, both methods were efficient to synchronize the cell cycle before freezing. However, after thawing, serum starvation, for more than 24 h, was superior to culture for synchronizing cells in G0/G1 (69% x 90%). The results of this experiment indicate that equine fibroblasts can be efficiently cultured after thawing.

Determinants of successful tracheostomy decannulation in children: a multicentric cohort study.

BACKGROUND: Determining prognostic factors for the probability of tracheostomy decannulation is key to an adequate therapeutic plan. METHODS: A retrospective cohort study of 160 paediatric patients undergoing tracheostomy was conducted. Associations between different parameters and eventual tracheostomy decannulation were assessed. RESULTS: Mean follow-up duration was 27.8 months (interquartile range = 25.5-30.2 months). Median age at tracheostomy was 6.96 months (interquartile range = 3.37-29.42 months), with median tracheostomy maintenance of 14.5 months (interquartile range = 3.7-21.5 months). The overall tracheostomy decannulation rate was 22.5 per cent. Factors associated with a higher probability of tracheostomy decannulation included age at tracheostomy (hazard ratio = 1.11, 95 per cent confidence interval = 1.03-1.18) and post-intubation laryngitis as an indication for tracheostomy (hazard ratio = 2.25, 95 per cent confidence interval = 1.09-4.62). Neurological (hazard ratio = 0.30, 95 per cent confidence interval = 0.12-0.80) and pulmonary (hazard ratio = 0.41, 95 per cent confidence interval = 0.18-0.91) co-morbidities were negatively associated with tracheostomy decannulation. The probability of tracheostomy decannulation decreased significantly with increasing numbers of co-morbidities (p < 0.001). CONCLUSION: Age, post-intubation laryngitis, and number and type of co-morbidities influence tracheostomy decannulation rate in the paediatric population.

Temporal evaluation of body composition, glucose homeostasis and lipid profile of male rats programmed by maternal protein restriction during lactation.

Neonatal protein restriction causes lower body weight and hormonal dysfunctions in 6 months-old rats. In this model, we studied the body composition, glycogen content, serum lipid, serum protein, and hormones related to glucose homeostasis in the offspring during development. At birth, lactating rats were divided into: control dams - fed a normal diet (23% protein) and protein restricted dams - fed a diet with 8% protein. After weaning, pups received normal diet. Offspring were killed at 21, 90, and 180 days-old. Protein restricted offspring showed lower visceral fat (90th day: 14%; 180th day: 19%) and lower total fat (90th day: 16%; 180th day: 14%) that explain their lower body weight. They presented lower glycemia (180th day: 17%), lower insulinemia (21st day: 63%; 180th day: 24%), higher adiponectinemia (21st day: 169%), higher liver glycogen (21st day: 104%), and higher muscle glycogen (180th day: 106%), suggesting a higher insulin sensitivity. The higher serum corticosterone (50%), higher adrenal total catecholamines content (98%) as well as in vitro catecholamine secretion (26%) of adult protein restricted offspring, suggest a programming stimulatory effect upon adrenal gland. They also presented several biochemical changes, such as lower serum total protein, albumin and globulin (21st day: 17, 21, 12%, respectively), higher LDL-c (21st day: 69%), lower triglycerides (21st day: 42%; 90th day: 39%), and lower total cholesterol (180th day: 16%). Thus, maternal protein restriction during lactation induces an energy-protein malnutrition, characterized by an impairment of the pup's protein anabolism and, after weaning, the lower adiposity suggests lower lipogenesis and higher lipolytic activity, probably caused by catecholamine and glucocorticoid action.

A comparison of the effect of alendronate and risedronate on bone mineral density in postmenopausal women with osteoporosis: 24-month results from FACTS-International.

OBJECTIVES: To compare alendronate 70 mg once weekly (OW) with risedronate 35 mg OW with respect to change in bone mineral density (BMD), biochemical markers and upper gastrointestinal (UGI) tolerability over 24 months. METHODS: This was a 12-month extension to the Fosamax Actonel Comparison Trial international study (FACTS). Postmenopausal women with osteoporosis randomly assigned to either alendronate 70 mg OW or risedronate 35 mg OW for the 12-month base study continued taking the same double-blind study medication. Efficacy measurements were BMD at the hip trochanter, lumbar spine, total hip, and femoral neck and levels of four bone turnover markers at 24 months. The primary hypothesis was that alendronate would produce a greater mean per cent increase from baseline in hip trochanter BMD at 24 months. RESULTS: Trochanter BMD increased significantly from baseline to month 24 in both groups, with a significantly larger increase with alendronate: adjusted mean treatment difference of 1.50% (95% confidence interval: 0.74%, 2.26%; p < 0.001). Similar results were seen at all BMD sites. Significant geometric mean per cent decreases (p < 0.001) from baseline were seen for all four bone turnover markers in both groups, with significantly larger decreases (p < 0.001) with alendronate: adjusted mean treatment differences ranged from 8.9% to 25.3%. No significant differences were seen in incidence of UGI or other adverse events. CONCLUSIONS: Alendronate 70 mg OW yielded significantly greater BMD gains and larger decreases in bone turnover marker levels than risedronate 35 mg OW over 24 months, with no difference in UGI tolerability.

Diel variation in fish assemblages in tidal creeks in southern Brazil.

Tidal creeks are strongly influenced by tides and are therefore exposed to large differences in salinity and depth daily. Here we compare fish assemblages in tidal creeks between day and night in two tidal creeks in southern Brazil. Monthly day and night, simultaneous collections were carried out in both creeks using fyke nets. Clupeiformes tended to be caught more during the day. Cathorops spixii, Genidens genidens and Rypticus randalli tended to be caught at night. Sciaenidae also tended to be caught more during the night. In general, pelagic species were diurnal, while deep water species were nocturnal. These trends are probably due to a variety of causes, such as phylogeny, predation and net avoidance.

Revealing the needs of children with tracheostomies.

INTRODUCTION: Small children with tracheostomy are at potential risk and have very specific needs. International literature describes the need for tracheostomy in 0.5% to 2% of children following intubation. Reports of children submitted to tracheostomy, their characteristics and needs are limited in developing countries and therefore there is a lack of health programs and government investment directed to medical and non-medical care of these patients. The aim of this study was to describe the characteristics of these children and identify problems related to or caused by the tracheostomy. METHODS: A retrospective cohort study was performed based on a common database applied in four high complexity healthcare facilities to children submitted to tracheostomy from January 2013 to December 2015. Data concerning children's demographics, indication for tracheostomy, early and late complications related to tracheostomy, airway diagnosis, comorbidities and decannulation rates are reported. Patients who did not present a complete database or had a follow-up of less than six months were excluded. RESULTS: A total of 160 children submitted to tracheostomy during the three-year period met the criteria and were enrolled in this study. Median age at tracheostomy was 6.9 months (ranging from 1 month to 16 years, interquartile range of 26 months). Post-intubation laryngitis was the most frequent indication (48.8%). Comorbidities were frequent: neurologic disorders were reported in 40%, pulmonary pathologies in 26.9% and 20% were premature infants. Syndromic children were 23.1% and the most frequent was Down's syndrome. The most common early complication was infection that occurred in 8.1%. Stomal granulomas were the most frequent late complication and occurred in 16.9%. Airway anomalies were frequently diagnosed in follow-up endoscopic evaluations. Subglottic stenosis was the most frequent airway diagnosis and occurred in 29.4% of the cases followed by laryngomalacia, suprastomal collapse and vocal cord paralysis. Decannulation was achieved in 22.5% of the cases in the three-year period. The main cause for persistent tracheostomy was the need for further treatment of airway pathology. Mortality rate was 18.1% during this period but only 1.3% were directly related to the tracheostomy, the other deaths were a consequence of other comorbidities. CONCLUSION: Tracheostomies were performed mostly in very small children and comorbidities were very common. Once a tracheostomy was performed in a child in most cases it was not removed before a year. The most common early complication was stoma infection followed by accidental decannulation. The most frequent late complication was granuloma and suprastomal collapse. Airway abnormalities were very frequent in this population and therefore need to be assessed before attempting decannulation.

Functional and evolutionary characterization of Ohr proteins in eukaryotes reveals many active homologs among pathogenic fungi.

Ohr and OsmC proteins comprise two subfamilies within a large group of proteins that display Cys-based, thiol dependent peroxidase activity. These proteins were previously thought to be restricted to prokaryotes, but we show here, using iterated sequence searches, that Ohr/OsmC homologs are also present in 217 species of eukaryotes with a massive presence in Fungi (186 species). Many of these eukaryotic Ohr proteins possess an N-terminal extension that is predicted to target them to mitochondria. We obtained recombinant proteins for four eukaryotic members of the Ohr/OsmC family and three of them displayed lipoyl peroxidase activity. Further functional and biochemical characterization of the Ohr homologs from the ascomycete fungus Mycosphaerella fijiensis Mf_1 (MfOhr), the causative agent of Black Sigatoka disease in banana plants, was pursued. Similarly to what has been observed for the bacterial proteins, we found that: (i) the peroxidase activity of MfOhr was supported by DTT or dihydrolipoamide (dithiols), but not by ß-mercaptoethanol or GSH (monothiols), even in large excess; (ii) MfOhr displayed preference for organic hydroperoxides (CuOOH and tBOOH) over hydrogen peroxide; (iii) MfOhr presented extraordinary reactivity towards linoleic acid hydroperoxides (k=3.18 (±2.13)×108M-1s-1). Both Cys87 and Cys154 were essential to the peroxidase activity, since single mutants for each Cys residue presented no activity and no formation of intramolecular disulfide bond upon treatment with hydroperoxides. The pKa value of the Cysp residue was determined as 5.7±0.1 by a monobromobimane alkylation method. Therefore, eukaryotic Ohr peroxidases share several biochemical features with prokaryotic orthologues and are preferentially located in mitochondria.

Cyclophosphamide versus methylprednisolone for treating neuropsychiatric involvement in systemic lupus erythematosus.

BACKGROUND: Neuropsychiatric involvement in systemic lupus erythematosus is complex and several clinical presentations are related to this disease such as: convulsions, chronic headache, transverse myelitis, vascular brain disease, psychosis and neural cognitive dysfunction. This systematic review is an update of a review performed in 2000. OBJECTIVES: To assess the efficacy and safety of cyclophosphamide and methylprednisolone in the treatment of neuropsychiatric manifestations of systemic lupus erythematosus. SEARCH STRATEGY: We searched EMBASE, LILACS, Cochrane Central Register of Controlled Trials (CENTRAL) and MEDLINE up to and including May 2005. Additional articles were sought through handsearching in relevant journals. There were no language restrictions. SELECTION CRITERIA: All randomised controlled trials that compared cyclophosphamide to methylprednisolone were included. Patients of any age and gender were included as long as they fulfilled the criterion of the American College of Rheumatology for the diagnosis of systemic lupus erythematosus and presented with any one of the following neuropsychiatric events: convulsions, organic brain syndrome and cranial neuropathy. Outcome measures included the following: a) overall mortality (primary event); b) motor and psychiatric deficit (primary event); c) clinical improvement (secondary event). DATA COLLECTION AND ANALYSIS: Data was independently extracted by two reviewers and cross-checked. The methodological quality of each trial was assessed by the same two reviewers. Details of the randomisation (generation and concealment), blinding, and the number of patients lost to follow-up were recorded. Dichotomous data was presented as relative risks with corresponding 95% confidence intervals and a clinical relevance table was produced. MAIN RESULTS: We found one randomised controlled trial of 32 patients comparing cyclophosphamide versus methylprednisolone for the treatment of neuropsychiatric involvement in the systemic lupus erythematosus. A significantly greater number of people responded to treatment in the cyclophosphamide group. Treatment response was found in 94.7% (18/19) of patients using cyclophosphamide compared with 46.2% (6/13) in the methylprednisolone group at 24 months (RR 2.05, 95% CI 1.13, 3.73) The NNT for response to treatment is 2. Cyclophosphamide use was associated with a reduction in prednisone requirements. A significant decrease in the number seizures per month was observed in the cyclophosphamide group. All the patients in the cyclophosphamide group had electroencephalographic improvement. No significant differences in adverse effects between the groups were found. It was not possible to extract more data from the study because there was a small number of patients in the others clinical subgroups of neurological manifestations and the authors did not provide sufficient information for data extraction. AUTHORS' CONCLUSIONS: This systematic review found one randomised controlled trial with a small number of patients in the different clinical subgroups of neurological manifestation. It seems that cyclophosphamide is more effective in the treatment of neuropsychiatric involvement in systemic erythematosus lupus compared with methylprednisolone. However, properly designed randomised controlled trials that involve large, representative numbers of individuals, with explicit clinical and laboratory diagnosis criteria, sufficient duration of follow-up and description of all relevant outcome measures are necessary to guide practice.