In silico analysis and molecular dynamics simulation of human superoxide dismutase 3 (SOD3) genetic variants.

Oxidative stress is a major factor in aging processes. Superoxide dismutase 3 (SOD3) plays a key role in the protection of extracellular oxidative stress. Missense mutations in SOD3 have been described to be associated with the occurrence of pulmonary, cardiovascular, and neoplastic diseases. This study aims to analyze the effects of missense mutations on the SOD3 structure and function by modeling a complete SOD3 structure as well as analyzing the differences between the wild-types and mutants using computational simulations. Here, ten algorithms were used to predict the structural and functional effects of missense mutations. A complete model of SOD3 protein was made by ab initio and comparative modeling using the Rosetta algorithm and validated by PROCHECK, Verify 3D, QMEAN, and ProSa. Molecular dynamics (MD) simulations were performed and analyzed using the GROMACS package. The deleterious potential of the A58T and R231G mutants was not predicted by the majority of the used algorithms. The analyzed mutations were predicted as destabilizing by at least one algorithm. The MD analyses indicated that protein flexibility may be increased by all of the analyzed mutations, while the protein-ligand stability may be decreased. They also suggested that the variants A91T and R231G increase the overall dimensions of SOD3 and decrease its accessible surface area. Our findings, therefore, indicated that the analyzed mutations could affect the protein structure and its ability to interact with other molecules, which may be related to the functional impairment of SOD3 upon A58T and R231G mutations, as well as their involvement in pathologies.

Effects of season on boar semen parameters and antioxidant enzymes in the south subtropical region in Brazil.

Although boar semen productivity is affected by seasonality, its effects are not equal among different regions which raise concerns regarding the profitability of boar stud farms. Therefore, the goals of this study were (i) to evaluate the seasonal effect on semen production in a commercial boar stud farm located in a subtropical climate region and (ii) to verify whether the activities of superoxide dismutase and glutathione peroxidase in spermatozoa and seminal plasma were associated with seminal traits of fresh and cooled semen. Nine boars were collected twice per season, and routine seminal parameter analyses were performed together with superoxide dismutase and glutathione peroxidase activities in seminal plasma and spermatozoa. Despite a reduction in sperm concentration in spring and summer, most seminal parameters were constant year-round. Temperature-humidity index was higher in the summer compared to spring, autumn and winter (p < .05). Superoxide dismutase activity in spermatozoa was increased in summer compared to autumn and winter (p < .05). The activities of both enzymes in seminal plasma and spermatozoa glutathione peroxidase remained unaltered throughout the seasons. In conclusion, seasonality showed little influence in overall boar seminal parameters despite microclimatic differences among seasons, and spermatozoa collected during summer increased superoxide dismutase activity.

Infrared thermography as a noninvasive method to assess scrotal insulation on sperm production in beef bulls.

This study evaluated the thermoregulation and spermatogenic changes by scrotal temperature gradient using infrared thermography in testicular compromised bulls. Bulls were insulated (n = 6) for 72 hr and control animals (n = 3) remained without insulation during all the experimental period. Seminal evaluation was performed prior, at insult removal and once per week for 13 consecutive weeks. Mean temperature gradient in insulated animals was lower at the time of insulation removal compared to the week prior and after the insult (p < .05). Two weeks after insult, sperm motility was lower in insulated compared to control animals (p < .01) and spermatozoa total defects were higher in insulated compared to control animals (p < .05). Two and seven weeks after insult, the major defects were higher in insulated compared to control animals (p < .05). Scrotal temperature gradient showed a positive correlation with sperm mass motion (p < .01) and a negative correlation with ocular globe temperature (p < .01) in insulated animals. The infrared thermography can be used to evaluate ocular globe temperature in bulls; however, it is only effective to detect changes in scrotal temperature gradient at the insult removal.

Molecular dynamics and protein frustration analysis of human fused in Sarcoma protein variants in Amyotrophic Lateral Sclerosis type 6: An In Silico approach.

Amyotrophic lateral sclerosis (ALS) is the most frequent adult-onset motor neuron disorder. The disease is characterized by degeneration of upper and lower motor neurons, leading to death usually within five years after the onset of symptoms. While most cases are sporadic, 5%-10% of cases can be associated with familial inheritance, including ALS type 6, which is associated with mutations in the Fused in Sarcoma (FUS) gene. This work aimed to evaluate how the most frequent ALS-related mutations in FUS, R521C, R521H, and P525L affect the protein structure and function. We used prediction algorithms to analyze the effects of the non-synonymous single nucleotide polymorphisms and performed evolutionary conservation analysis, protein frustration analysis, and molecular dynamics simulations. Most of the prediction algorithms classified the three mutations as deleterious. All three mutations were predicted to reduce protein stability, especially the mutation R521C, which was also predicted to increase chaperone binding tendency. The protein frustration analysis showed an increase in frustration in the interactions involving the mutated residue 521C. Evolutionary conservation analysis showed that residues 521 and 525 of human FUS are highly conserved sites. The molecular dynamics results indicate that protein stability could be compromised in all three mutations. They also affected the exposed surface area and protein compactness. The analyzed mutations also displayed high flexibility in most residues in all variants, most notably in the interaction site with the nuclear import protein of FUS.

Development and characterization of nanostructured lipid carrier-based gels for the transdermal delivery of donepezil.

Donepezil is one of the main compounds used in the therapy of Alzheimer's disease. Oral administration of this drug presents many drawbacks, resulting in treatment non-adherence among patients. Thus, the development of transdermal formulations for donepezil delivery is important. The aim of this study was to prepare and to evaluate nanostructured lipid carrier-based gels (NLC gel) able to improve the skin delivery of donepezil free base (DPB). The components of nanostructured lipid carriers (NLCs) were selected after evaluating their enhancing effects using in vitro DPB skin delivery assays. DPB-loaded NLC were prepared by a microemulsion technique, by employing stearic acid as a solid lipid, oleic acid as a liquid lipid, lecithin as a surfactant, and sodium taurodeoxycholate as a co-surfactant. The DPB-NLC dispersions were characterized morphologically using atomic force microscopy and physicochemically using dynamic light scattering and surface charge measurements. These data along, with the encapsulation studies, indicated that uniformly nano-sized particles with high drug encapsulation were fabricated. In vitro skin permeation assays were performed, and the results indicated that drug skin permeation from DPB-NLC gel was increased, not only by the enhancing effect of their components, but the lipid nanocarriers also presented an additional enhancing effect to increase drug flux across the skin. Therefore, DPB-NLC gel is an interesting formulation for the enhanced treatment of Alzheimer's disease.

Kinetics of acid-base parameters in conventional hemodialysis.

Details about the acid-base changes in hemodialysis are scarce in the literature but are potentially relevant to adequate management of patients. We addressed the acid-base kinetics during hemodialysis and throughout the interdialytic period in a cross-sectional study of adults undergoing conventional hemodialysis. Samples for blood gas analysis were obtained from the arterial limb of the arteriovenous fistula before the first session of the week (HD1), immediately at the end of HD1, and on sequential collections at 15, 30, 45, 60, and 120 min post-HD1. Additional blood samples were collected after ∼20 h following the end of the first dialysis and immediately prior to the initiation of the second dialysis of the week. Thirty adult patients were analyzed (55±15 years, 50% men, 23% diabetic; dialysis vintage 69±53 months). Mean serum bicarbonate levels increased at the end of HD1 (22.3±2.7 mEq/L vs 17.5±2.3 mEq/L, P<0.001) and remained stable until 20 h after the end of the session. The mean values of pCO2 before HD1 were below reference and at 60 and 120 min post-HD1 were significantly lower than at the start (31.3±2.7 mmHg and 30.9±3.7 mmHg vs 34.3±4.1 mmHg, P=0.041 and P=0.010, respectively). The only point of collection in which mean values of pCO2 were above 35 mmHg was 20 h post-dialysis. Serum bicarbonate levels remained stable for at least 20 h after the dialysis sessions, a finding that may have therapeutic implications. During dialysis, the respiratory response for correction of metabolic acidosis (i.e., pCO2 elevation) was impaired.

Diversity of Anopheles spp. (Diptera: Culicidae) in an Amazonian Urban Area.

The genus Anopheles encompasses several species considered as vectors of human infecting Plasmodium. Environmental changes are responsible for behavior changes in these vectors and therefore the pattern of malaria transmission. To better understand the dynamics of malaria transmission, this study aimed at identify the species of adult anophelines found in a malaria endemic urban area of the Amazon region, Mâncio Lima, located in the Acre State Brazil. Using Shannon-type light traps installed at 11 collection points near fish ponds, a total of 116 anophelines were collected belonging to nine species. Anopheles darlingi Root 1926 and An. albitarsis s.l. Lynch-Arribalzaga 1878 were the most abundant and predominant species. Despite the low number of captured adult anophelines, the occurrence of An. darlingi throughout all urban area and the presence of secondary vectors reinforce the need of a permanent and continuous entomological surveillance.

Neospora caninum DNA distribution in tissues of gerbils as experimental models of chronic neosporosis / [Distribuição do DNA de Neospora caninum em tecidos de gerbilos como modelos experimentais de neosporose crônica]

Neospora caninum is the main etiologic agent of neosporosis in domestic animals and its pathogenesis comprises two characteristic phases: acute and chronic. Rodents are used as experimental models to mimic acute and chronic bovine neosporosis. In this study, we inoculated a total of 27 female gerbils, with different doses of N. caninum tachyzoites aiming to induce chronic disease. DNA was extracted from different organs of each animal after spontaneous death or euthanasia. Encephalic tissues were submitted to a highly sensitive real time PCR aiming to detect chronically infected animals. All the other samples were submitted to standard PCR. A total of 11 gerbils died due to acute neosporosis, as confirmed by N. caninum DNA detection in organs. 5x103 tachyzoites/mL of N. caninum was the dosage of antigen that can induce chronic infection in gerbils. In the encephalon sections of some animals that showed clinical signs of persistent infection, we found 70% positive for the anterior encephalon section, suggesting this area as preferential for cyst formation. Therefore, we determined the doses of tachyzoites that cause acute or chronic infection and detection of positive tissues, preferably, systemic organs during acute and encephalon in chronic phases.(AU)

Neospora caninum é o principal agente etiológico da neosporose em animais domésticos, e sua patogênese compreende duas fases características: aguda e crônica. Roedores são usados como modelos experimentais para simular neosporose bovina aguda e crônica. Neste estudo, foi inoculado um total de 27 gerbilos, fêmeas, com diferentes doses de taquizoítos de N. caninum, visando induzir doença crônica. O DNA foi extraído de diferentes órgãos de cada animal após a morte espontânea ou a eutanásia. Os tecidos encefálicos foram submetidos à PCR em tempo real de alta sensibilidade para detecção de animais com infecção crônica. Todas as outras amostras foram submetidas à PCR padrão. Um total de 11 gerbilos morreu devido à neosporose aguda, como confirmado pela detecção de DNA de N. caninum nos órgãos. A dosagem de antígeno que pode induzir infecção crônica foi de 5x103 taquizoítos/mL de N. caninum. Em seções do encéfalo de alguns animais, que apresentaram sinais clínicos de infecção persistente, encontraram-se 70% de positividade para a seção do encéfalo anterior, sugerindo essa área como preferencial para a formação de cisto. Assim, foram determinadas,, em gerbilos, as dosagens de taquizoítos capazes de induzir infecção crônica ou aguda, bem como foram detectados tecidos positivos, preferencialmente, em órgãos sistêmicos, na fase aguda, e no encéfalo, na crônica.(AU)

Serum myoinositol concentrations in premature infants fed human milk, formula for infants, and parenteral nutrition.

Myoinositol concentration was studied in serum of 65 neonates and their mothers at the time of birth, in samples of various types of feedings for infants, and in serial serum samples of 15 premature infants receiving human milk, formulas for infants, or parenteral nutrition over a 3-wk period. At birth the serum concentration of myoinositol was greater in neonates than in their mothers (108 +/- 10 vs 52 +/- 6 mumol/L, respectively, means +/- SEM, p less than 0.01). In feedings for infants, the concentrations of myoinositol were significantly greater in human milk than in formulas or parenteral nutrition solutions (1840 +/- 451 vs 420 +/- 110 vs 100 +/- 8 mumol/L, respectively, p less than 0.001). Over a 3-wk period the serum concentration of myoinositol increased in infants receiving human milk but not in those receiving formulas or parenteral nutrition. Serum concentrations of myoinositol in neonates are greater than in adults and are directly influenced by myoinositol intake.

Controlled study of transpyloric and intermittent gavage feeding in the small preterm infant.

There were 53 appropriate for gestational age infants with a birth weight of less than 1,700 gm admitted to the Children's Hospital in Denver who were randomly assigned to two groups for gavage or transpyloric (nasojejunal) feedings. The two groups were not significantly different in terms of gestational age, birth weight, and morbidity. No significant differences were observed for caloric intake after 4 days of age, growth parameters (weight, length, and head circumference), serum total protein levels, feeding-related complications, duration of intravenous fluid supplementation, and length of hospitalization. The results indicate that intermittent gastric gavage feedings are as efficacious as transpyloric feedings in supplying nutrition to the low birth-weight infant. Because of inherently greater risks, costs, radiation, and requisite personnel expertise, transpyloric feedings cannot be recommended as a routine method of feeding for the low birth-weight infant.

Nonnutritive sucking during gavage feeding enhances growth and maturation in premature infants.

The effects of nonnutritive sucking (NNS) by means of a pacifier during gavage feeding were studied in 30 premature infants whose birth weight was less than 1,500 g. The addition of NNS accelerated the maturation of the sucking reflex, facilitating a more rapid transition from gavage to oral feedings. Additionally, NNS decreased intestinal transit time and caused a more rapid weight gain despite comparable caloric intake resulting in a shortened hospital stay. Although the physiologic mechanisms resulting from this form of oral stimulation remain to be investigated, our data suggest that NNS may be an important factor to consider in the feeding of premature infants.

Effect of fat infusions on platelet concentration in premature infants.

Critically ill premature infants often require lipid emulsions with parenteral nutrition until enteral feedings can be safely initiated. Because thrombocytopenia has been listed as a potential side effect of fat emulsions, we examined the effect of varying doses of intravenous fat infusions on platelet concentrations in premature infants. An initial validation study demonstrated no artifactual effect of fat infusions on the electronic cell counter method of platelet enumeration. We observed no adverse effect of fat emulsions administered at doses of 0.8 to 3.3 g/kg/day on platelet concentration either during short-term (48 hr) or long-term study periods (4 weeks).

Sustained flexibility in infant feeding tubes containing nonmigrating plasticizer.

Due to medical complications related to stiffness developed in polyvinyl chloride (PVC) feeding tubes presently manufactured with di-octylphthalate (DOP), we have assessed the flexibility of PVC tubes manufactured with a nonmigrating plasticizer, ti-octyltrimellitate (TOTM) (National Catheter Co., Argyle, NY). Two sizes of DOP and TOTM PVC feedings tubes were evaluated, #5 French and #8 French. Number 5 French DOP tubes became significantly stiffer as early as 1 to 2 days after intubation and continued to stiffen up to 8 days of use. Among the #5 French DOP tubes, those placed in the jejunum were found to become significantly stiffer than those placed in the stomach. No correlations were found between the development of stiffness and gestational age, postnatal age, sex, birthweight, or kinds of feedings. In contrast, #5 French TOTM and #8 French tubes (DOP and TOTM) did not develop any significant stiffness up to 5 to 8 days of use. This study demonstrates the sustained flexibility of TOTM tubes, and important factor to increase the safety of enteral feedings in newborn infants.

The effect of 15-hour fat infusions of varying dosage on bilirubin binding to albumin.

Intravenous fat emulsions (1, 2, and 3 g/kg) were administered over 15 hr to 20 appropriate for gestational age premature infants with physiologic hyperbilirubinemia to determine the effect of fat infusions on the serum free fatty acid:albumin molar ratio (F/A) and on unbound bilirubin. Significant increases (p less than 0.05) in F/A occurred with each increase in lipid dose in infants less than 30 wk gestation, but not in infants greater than or equal to 30 wk gestation. There was a direct linear correlation (r = 0.65, p less than 0.001) between F/A ratio and unbound bilirubin (estimated fluorometrically by the ratio of albumin-bound bilirubin/reserve bilirubin binding capacity, B/R). The largest increases in unbound bilirubin (albumin-bound bilirubin/reserve bilirubin binding capacity) were seen in infants with F/A greater than 4.0. The gestational age of infants with F/A greater than 4.0 was significantly less (p less than 0.01) than infants with F/A less than 4.0 (28.7 +/- 0.47 vs. 31.1 +/- 0.40 wk, mean +/- SEM). In 10/58 infusions there was a fall in unbound bilirubin, unrelated to birthweight, gestational age, postnatal age, however, during these infusions the end-infusion F/A was greater than or equal to 3.0. We conclude that 1 g/kg of lipid emulsion infused over a 15-hr period has minimal risk of decreasing bilirubin binding in premature infants less than 30 wk gestation. As doses of 2 or 3 g/kg are used, these infants may be at risk of decreased bilirubin binding, due to elevations in the F/A ratio. Monitoring of the F/A ratio may identify infants at risk for decreased bilirubin binding during lipid infusion and provide guidelines for determining the appropriate lipid dose.

Controversies in neonatal nutrition.

The assessment of growth parameters remains one of the most practical and valuable tools to estimate nutritional status in neonates. Growth assessment in full-term infants is performed by using charts developed by the National Center for Health and Statistics. The assessment of post-natal growth in premature infants is controversial and can be performed by using either intrauterine or extrauterine standards. The selection of appropriate growth charts should be based on clinical, demographic, ethnic, and socioeconomic similarities of the population used for reference. Daily energy intakes ranging from 100 to 120 kcal/kg/day have been recommended for full-term infants, while higher intakes ranging from 114 to 181 kcal/kg/day have been recommended for premature neonates. Full-term infants should be nursed or nipple fed on demand; however, premature infants should ideally be tube fed by intermittent gastric feeding (gavage). Continuous gastric and transpyloric feedings are indicated in selected infants. Human milk is a preferred food for full-term infants during the first six months of life; however, this precept does not suggest that all infants who are exclusively breast-fed will grow adequately. Preterm human milk is also a preferred food for the low birthweight infant, provided nutritional supplements are used. It is unclear whether the supplementation of vitamin D, iron, and fluoride in full-term breast-fed infants should be started at birth, at the time of initiation of solid foods, or at the age of six months. The routine supplementation of multivitamins, folic acid, and vitamin E to all low birthweight infants is controversial. Most investigators suggest vitamin supplementation be given until the intake of formula or breast milk is sufficient to meet daily requirements. Vitamin E appears to exert a protective effect in premature infants against the development of severe retinopathy. The supplementation of vitamin E should be dependent upon the serum vitamin E concentration. It is controversial whether iron supplementation for premature infants should be initiated soon after birth or at two months of age, or whether higher doses of iron should be given to very low birthweight infants. If iron supplementation is started at birth, vitamin E status should be closely monitored. Although the optimal intakes of calcium and phosphorus in infant feedings have not been firmly established, the levels of calcium and phosphorus in human milk appear to be inadequate for the growing low birthweight infant.(ABSTRACT TRUNCATED AT 400 WORDS)

Association of maternal lithium exposure and premature delivery.

Lithium is widely used and the treatment of choice for patients with manic-depressive illness. For pregnant patients with manic-depressive illness, however, the use of lithium during the first trimester of pregnancy may present an increased risk for fetal maldevelopment. We have recently cared for several large-for-gestational-age, prematurely born infants whose mothers were treated with lithium throughout pregnancy. To determine whether maternal lithium use during pregnancy may predispose to the onset of premature labor and fetal macrosomia, we reviewed records from the International Register of Lithium Babies and from a cohort of manic-depressive pregnant women. More than one third (36%) of infants reported to the International Register were born prematurely, and 37% of the premature infants were large for gestational age; 15% of the term infants were born large for gestational age. In the cohort group, manic-depressive mothers who received lithium during pregnancy had a 2.5-fold higher incidence of premature births than manic-depressive pregnant patients who did not receive lithium treatment. The incidence of large-for-gestational-age births in lithium-treated women in the cohort was not different from that of the general population or from manic-depressive women not treated with lithium. In summary, an association between maternal lithium therapy and premature delivery is reported. We recommend that women receiving lithium therapy during pregnancy be closely monitored for the onset of premature labor.

Nutritional care of the extremely premature infant.

In summary, the nutritional care of extremely premature babies includes special attention to glucose homeostasis and fluid and electrolyte balance in the immediate postnatal period. Parenteral nutrition should be started routinely on the 2nd or 3rd day of life, if the infant is metabolically stable, and continued until the infant is receiving sufficient enteral feedings to promote growth. If not clinically contraindicated, minimal enteral feedings should be started before the end of the first week of life. The decision to either advance or maintain minimal enteral feedings at a constant level should take into account the clinical status of the infant. Fortified preterm human milk is the preferred feeding and premature infant formulas, the best substitute. Multivitamin supplements may be necessary, depending on formula intake. Iron supplements can be initiated as early as 2 weeks of age and high levels of intake appear to be necessary during erythropoietin therapy.

Nutritional care of the surgical neonate.

The surgical neonate is at great risk of developing nutritional deficiencies. However, recent advances in nutritional support, including the development of special elemental diets, infant formulas, nutritional supplements and more suitable additives for parenteral nutrition, have allowed health professionals effectively to prevent the occurrence of malnutrition in sick neonates. Strict monitoring of dietary intake, growth velocity, and biochemical parameters are essential while providing nutritional support. The complication of parenteral nutrition can be reduced by following standard recommendations for preparing solutions, caring for the central and the peripheral catheters, and biochemically monitoring the infant. The organization of a hospital nutrition support service composed of physicians, nurses, nutritionists, and pharmacists is essential in establishing guidelines for the safe delivery of nutritional therapy and in educating hospital personnel and parents on how to best meet the nutritional needs of the surgical neonate.

Nutritional deficiencies in the neonate.

This article reviews common nutritional deficiencies that have been identified in newborn infants, including deficiencies of protein; vitamins A, D, K, C, and E; calcium and phosphorus; folate; zinc; and copper. The etiology, clinical presentation, diagnosis, and treatment of these nutritional deficiencies are discussed.