Iron Concentrations in Breast Milk and Selected Maternal Factors of Human Milk Bank Donors

The aim of this study was to evaluate the relationship between iron concentration in mature breast milk and characteristics of 136 donors of a Brazilian milk bank. Iron, vitamin A, zinc, and copper concentrations were assessed in human milk and maternal blood. Data were collected on maternal anthropometrics, obstetric, socioeconomic, demographic, and lifestyle factors. Iron, zinc, and copper in milk and zinc and copper in blood were detected by spectrophotometry. Vitamin A in milk and blood was determined by high-performance liquid chromatography. Hemoglobin was measured by electronic counting and serum iron and ferritin by colorimetry and chemoluminescence, respectively. Transferrin and ceruloplasmin were determined by nephelometry. According to multivariate linear regression analysis, iron in milk was positively associated with vitamin A in milk and with smoking but negatively associated with timing of breast milk donation (P < .001). These results indicate that iron concentration in milk of Brazilian donors may be influenced by nutritional factors and smoking. J Hum Lact. 26(2):175-179

1,4-Diamino-2-butanone, a wide-spectrum microbicide, yields reactive species by metal-catalyzed oxidation

The alpha-aminoketone 1,4-diamino-2-butanone (DAB), a putrescine analogue, is highly toxic to various microorganisms, including Trypanosoma cruzi. However, little is known about the molecular mechanisms underlying DAB`s cytotoxic properties. We report here that DAB (pK(a) 7.5 and 9.5) undergoes aerobic oxidation in phosphate buffer, pH 7.4, at 37 degrees C, catalyzed by Fe(II) and Cu(II) ions yielding NH(4)(+) ion, H(2)O(2), and 4-amino-2-oxobutanal (oxoDAB). OxoDAB, like methylglyoxal and other alpha-oxoaldehydes, is expected to cause protein aggregation and nucleobase lesions. Propagation of DAB oxidation by superoxide radical was confirmed by the inhibitory effect of added SOD (50 U ml(-1)) and stimulatory effect of xanthine/xanthine oxidase, a source of superoxide radical. EPR spin trapping studies with 5,5-dimethyl-1-pyrroline-1-oxide (DMPO) revealed an adduct attributable to DMPO-HO(center dot), and those with alpha-(4-pyridyl-1-oxide)-N-tert-butylnitrone or 3,5-dibromo-4-nitrosobenzenesulfonic acid, a six-line adduct assignable to a DAB(center dot) resonant enoyl radical adduct. Added horse spleen ferritin (HoSF) and bovine apo-transferrin underwent oxidative changes in tryptophan residues in the presence of 1.0-10 mM DAB. Iron release from HoSF was observed as well. Assays performed with fluorescein-encapsulated liposomes of cardiolipin and phosphatidylcholine (20:80) incubated with DAB resulted in extensive lipid peroxidation and consequent vesicle permeabilization. DAB (0-10 mM) administration to cultured LLC-MK2 epithelial cells caused a decline in cell viability, which was inhibited by preaddition of either catalase (4.5 mu M) or aminoguanidine (25 mM). Our findings support the hypothesis that DAB toxicity to several pathogenic microorganisms previously described may involve not only reported inhibition of polyamine metabolism but also DAB pro-oxidant activity. (C) 2011 Elsevier Inc. All rights reserved.

Labile Plasma Iron Generation After Intravenous Iron is Time-dependent and Transitory in Patients Undergoing Chronic Hemodialysis

Iron supplementation in hemodialysis patients is fundamental to erythropoiesis, but may cause harmful effects. We measured oxidative stress using labile plasma iron (LPI) after parenteral iron replacement in chronic hemodialysis patients. Intravenous iron saccharate (100 mg) was administered in patients undergoing chronic hemodialysis (N = 20). LPI was measured by an oxidant-sensitive fluorescent probe at the beginning of dialysis session (T0), at 10 min (T1), 20 min (T2), and 30 min (T3) after the infusion of iron and at the subsequent session; P < 0.05 was significant. The LPI values were significantly raised according to the time of administration and were transitory: -0.02 +/- 0.20 mu mol/L at the beginning of the first session, 0.01 +/- 0.26 mu mol/L at T0, 0.03 +/- 0.23 mu mol/L at T1, 0.09 +/- 0.28 mmol/L at T2, 0.18 +/- 0.52 mmol/L at T3, and -0.02 +/- 0.16 mmol/L (P = 0.001 to 0.041) at the beginning of the second session. The LPI level in patients without iron supplementation was -0.06 +/- 0.16 mmol/L. Correlations of LPI according to time were T1, T2, and T3 vs. serum iron (P = 0.01, P = 0.007, and P = 0.0025, respectively), and T2 and T3 vs. transferrin saturation (P = 0.001 and P = 0.0003, respectively). LPI generation after intravenous saccharate administration is time-dependent and transitorily detected during hemodialysis. The LPI increment had a positive correlation to iron and transferrin saturation.

Proteomic analysis of dorsolateral prefrontal cortex indicates the involvement of cytoskeleton, oligodendrocyte, energy metabolism and new potential markers in schizophrenia

Schizophrenia is likely to be a consequence of serial alterations in a number of genes that, together with environmental factors, will lead to the establishment of the illness. The dorsolateral prefrontal cortex (Brodmann`s Area 46) is implicated in schizophrenia and executes high functions such as working memory, differentiation of conflicting thoughts, determination of right and wrong concepts, correct social behavior and personality expression. We performed a comparative proteome analysis using two-dimensional gel electrophoresis of pools from 9 schizophrenia and 7 healthy control patients` dorsolateral prefrontal cortex aiming to identify, by mass spectrometry, alterations in protein expression that could be related to the disease. In schizophrenia-derived samples, our analysis revealed 10 downregulated and 14 upregulated proteins. These included alterations previously implicated in schizophrenia, such as oligodendrocyte-related proteins (myelin basic protein and transferrin), as well as malate dehydrogenase, aconitase, ATP synthase subunits and cytoskeleton-related proteins. Also, six new putative disease markers were identified, including energy metabolism, cytoskeleton and cell signaling proteins. Our data not only reinforces the involvement of proteins previously implicated in schizophrenia, but also suggests new markers, providing further information to foster the comprehension of this important disease. (C) 2008 Elsevier Ltd. All rights reserved.

Total gastrectomy with substitution of stomach by jejunal pouch with and without duodenal passage. Study in rats

A comparison was done between the F. Paulino jejunal pouch (FP) and a jejunal pouch (JP) as esophagusduodenum interpositional graft, for replacing the stomach after total gastrectomy. It was investigated the effect of the two procedures on esophagus histology, nutritional state and serum gastrin in rats. Methods: Male Wistar rats weighing 282±17g were randomly submitted to sham operation (S), FP and JP after total gastrectomy. After eight weeks the rats were killed with overdose of anesthetic and tissue was taken from the distal esophagus for histology. Serum levels of total proteins, albumin, iron, transferring, folate, cobalamine, calcium, as well as serum gastrin were determined. Survival was considered. Results: Fourty six rats were operated and thirty survived for eight weeks. Five (33.3%) died after FP and 11 (52.3%) after JP (p<0.05). Postoperative esophagitis occurred in 6 JP rats. At 8th week, no difference was observed on body weight when compared FP and JP rats (p>0.05). The JP rats had a significant decrease in serum albumin, glucose, transferrin, iron, folate and calcium, compared to sham (p<0.05). Serum gastrin, iron and calcium were significantly higher in JP rats than in FP rats (p<0.05). In FP rats, transferrin and cobalamine showed significant decrease comparing the preoperative with 8th week levels (p<0.05). Conclusion: F. Paulino pouch in rats had lower mortality than JP, and esophagitis was not detected in it. JP rats had serum gastrin, iron and calcium unaffected, possibly because of preservation of duodenal passage

Tratamento de fístulas enterocutâneas através de sistema a vácuo com alta pressão e dieta oral normal

Enterocutaneous fistulas are associated with prolonged hospital stay, high morbidity/mortality, and increase in hospital costs. This study aims to describe the use of a vacuum system and normal oral diet in dealing with this problem. Methods: Seventy-four consecutive patients with recent and defined external postoperative fistulas were analyzed. Abdominal imaging was used to exclude abscess and distal obstruction. The fistula tract was sealed with Foley catheter, connected to a negative pressure flask, changed daily for 5, 10 or 15 days, as necessary. Normal oral diet was permitted. Results: No patient died. Serum albumin and transferrin showed significantly higher levels at the end of treatment than at the beginning. The moderate and low-output fistulas had the best results (97% closed). Forty-eight (65%) fistulas closed after five days, 16(22%) after 10 days and 4(5%) after 15 days. Treatment failed in 6(8%) patients, who subsequently underwent surgery. Only one patient with low-output did not close her fistula. The cost of the treatment was US$ 41.75/day and it was considered cost effective. Conclusions: The vacuum system demonstrated good results in the treatment of fistulas. It included simplicity, low cost, short hospital stay, absence of skin breakdown, normal eating, good nutrition and activity patterns

Acute phase proteins: a potential approach for diagnosing chronic infection by Trypanosoma vivax

O objetivo do presente estudo foi verificar possíveis alterações nas proteínas de fase aguda em ovinos infectados experimentalmente com Trypanosoma vivax. Para tanto, foram utilizados oito ovinos machos, sendo quatro usados como controle e quatro infectados com 10(5) tripomastigotas de T. vivax. Colheram-se amostras de sangue em dois tempos antes da infecção e, posteriormente, aos 5, 7, 9, 11, 13, 15, 20, 30, 45, 60, 75, 90, 105 e 120 dias após a infecção (dpi); após centrifugação e aliquotização das amostras. As proteínas de fase aguda foram separadas por eletroforese em gel de acrilamida, contendo dodecil sulfato de sódio, e suas concentrações foram determinadas através de densitometria computadorizada. A dosagem de proteína total foi realizada pelo método colorimétrico do biureto. A contagem dos tripanossomas foi realizada diariamente, utilizando-se uma alíquota de 5 µL de sangue disperso em lâmina de microscopia, sob lamínula de 22 × 22 mm, contando-se os parasitos em 100 campos microscópicos, com objetiva de 40×, multiplicados pelo fator de correção do microscópio, e o resultado expresso em parasitos por mL de sangue. Para a análise estatística, empregou-se o teste de Wilcoxon a 5% de probabilidade. Foi observada a diminuição de diversas proteínas de fase aguda e aumento de antitripsina e transferrina que podem ser utilizadas para auxiliar no diagnóstico da infecção por T. vivax, principalmente na fase crônica da infecção.

Proteinogramas séricos de ratos Wistar experimentalmente infectados com Trypanosoma evansi

Estabeleceu-se o perfil eletroforético de proteínas séricas de ratos Wistar experimentalmente infectados com Tripanosoma evansi, utilizando-se 40 ratos, distribuídos em oito grupos de cinco animais cada. Um grupo foi mantido como testemunho (G1), e os demais (G2 a G8) foram inoculados, via intraperitoneal, com cerca de 10³tripomastigota de T. evansi. Amostras de sangue para obtenção de soro foram coletadas no quinto (G2), 10º (G3), 15º (G4), 30º (G5), 45º (G6), 60º (G7) e 75º (G1 e G8) dia após as inoculações. O fracionamento das proteínas foi realizado pela técnica SDS-PAGE. Foram identificadas 31 proteínas, sendo sete de fase aguda: ceruloplasmina (101KD), hemopexina (83KD), transferrina (75KD), albumina (66KD), antitripsina (60KD), haptoglobina (44KD) e glicoproteína ácida (38KD). As proteínas com pesos moleculares 12KD; 22KD; 25KD; 28KD; 32,5KD; 35KD; 53,5KD; 63KD e 72KD apareceram apenas nos ratos inoculados com T. evansi.

Perfil eletroforético do proteinograma sérico de eqüinos com obstrução experimental do cólon menor

Avaliaram-se as alterações do proteinograma sérico de eqüinos submetidos à isquemia e reperfusão do cólon menor por distensão intraluminal. Foram utilizados 10 animais submetidos à laparotomia pelo flanco, em posição quadrupedal, para a indução de obstrução no cólon menor durante um período de quatro horas. Cinco animais foram instrumentados, mas sem distensão (grupo controle - G1). em cinco outros animais, foi realizada isquemia mural por distensão do cólon menor via manguito inflado com 40mmHg (grupo distendido - G2). Foram colhidas amostras de sangue antes da intervenção cirúrgica (M1), com 4 horas da colocação do manguito (M2) e com 3 horas (M3) e 12 horas (M4) de reperfusão. Após centrifugação e fracionamento das amostras, as proteínas de fase aguda foram separadas por eletroforese em gel de poliacrilamida contendo SDS-PAGE, e suas concentrações determinadas por densitometria computadorizada. Foram encontradas 19 proteínas no fracionamento eletroforético, com peso molecular variando de 185.000 a 14.000 Daltons (Da). Os pesos moleculares encontrados, correspondentes às proteínas mais conhecidas, foram ceruloplasmina, 130.000 Da; proteína C-reativa, 122.000 Da; transferrina, 85.000 Da; α1-antitripsina, 61.000 Da; haptoglobina, 47.000 Da; e glicoproteína ácida, 40.000 Da. Os resultados mostram que proteínas de fase aguda se alteram após o trauma cirúrgico.

Proteinograma sérico de bezerros recém-nascidos da raça Holandesa obtido por eletroforese em gel de poliacrilamida

The serum protein concentration of newborn Holstein calves determined by means of sodium dodecyl sulphate-polyacrylamide (SDS-PAGE) was studied. Blood samples from 40 healthy newborn calves were obtained 48 hours after birth. Calves had been given 3 liters of colostrum within 2 hours after birth, following by dose corresponding by 15% of animal weight each 24 hours. The results showed three different proteinograms: 19 calves had 14 proteins with molecular weights (MW) ranging from 28,000 D to 170,000D (proteinogram 1); 11 calves had 14 proteins with MW ranging from 18,000 to 170,000 D (proteinogram 1); and 10 calves had 12 proteins with MW ranging from 28,000 D to 170,000 D (proteinogram 3). The three groups presented similar IgG levels. The highest serum concentration of ceruloplasmin were verified in proteinogram 3, which had the lowest serum level of protein with MW 58,000D. It was verified a1-antitrypsin only in proteinogram 2, which had no proteins with MW of 42,000 D and 37,000D. The highest serum concentrations of IgA and protein with MW 58,000 D, and the lowest serum levels of transferrin, haptoglobin, and acid glycoprotein were verified in proteinogram 3. Measurement of serum protein concentrations by SDS-PAGE may be useful in monitoring the occurrence of hypogammaglobulinemia and the neonatal disease in calves.

Systemic inflammatory response indicators in rabbits (Oryctolagus cuniculus) experimentally infected with sporulated oocysts of Eimeria stiedai (Apicomplexa: Eimeriidae)

Hemograms and acute-phase proteins in adult male New Zealand White rabbits that had been experimentally infected orally with sporulated oocysts of Eimeria stiedai were evaluated over a 28-day period. Fifty animals were used, divided into two groups: group A infected with 1 x 10(4) sporulated oocysts of E. stiedai and group B inoculated with distilled water. on the seventh day after infection, the infected animals presented anemia and leukocytosis with neutrophilia and monocytosis. Protein fractionation by means of electrophoresis identified 19 acute-phase proteins with molecular weights ranging from 24 to 238 kD. Ceruloplasmin, transferrin and haptoglobin showed high levels on the seventh day after infection, with gradual increases in their concentrations until the end of the experimental period. Thus, from the data of the present study, E. stiedai is considered to be a pyogenic etiological agent for which the infection level can be monitored through the leukocyte count and serum concentrations of ceruloplasmin, transferrin and haptoglobin, and these can be recommended as complementary tests.

Influência do puerpério sobre o proteinograma sérico de caprinos da raça Saanen obtido por eletroforese em gel de poliacrilamida

Aiming to evaluate the puerperal influence on the proteinogram of Saanen goats, 108 samples of blood serum from 12 goats were collected, and the results were presented at nine times: just after parturition, 1, 3, 5, 7, 10, 15, 21 and 30 days after parturition. Total amount of serum proteins were determined by the biuret technique, and the sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) was used to the protein fractionation. In this last method, 17 protein bands were observed, from which molecular weights varied between 25 KDa and 275 KDa. In addition, it was possible to identify the following protein fractions: immunoglobulin A (180 KDa), ceruloplasmin (115 KDa), transferrin (79 KDa), albumin (65 KDa), heavy-chain immunoglobulin G (58 KDa), haptoglobin (45 KDa), acid glycoprotein (37 KDa) and light-chain immunoglobulin G (28 KDa). Another 9 nonidentified protein fractions presented, each molecular weights equal to 275 KDa, 140 KDa, 125 KDa, 103 KDa, 95 KDa, 41 KDa, 35 KDa, 30 Kda and 25 KDa. The results allow us to conclude that by the first week of puerperium, an improvement of acid glycoprotein occurs, whereas those others protein fractions do not suffer any puerperal influence.

Polimorfismo da transferrina e albumina e as associações com precocidade sexual em bovinos da raça Nelore doadores de sêmen

O presente estudo utilizou 16 animais Bos taurus indicus da raça Nelore doadores de sêmen. Estes animais foram divididos em grupos de acordo com a idade em que o sêmen congelou pela primeira vez. O grupo I, considerado precoce, apresentou animais com sêmen passível de congelação com idade inferior a 20 meses. O grupo II, composto por animais que tiveram o sêmen congelado com idade entre 21 e 26 meses. E o grupo III, tido como tardio, composto por animais com sêmen congelável com idade superior a 27 meses. Para análise dos padrões eletroforéticos da transferrina e albumina, amostras de sangue foram colhidas em tubos heparinizados e submetidos a centrifugação de 2.500 G por 15 minutos para separação do plasma sangüíneo. As amostras de plasma sangüíneo foram processadas para que a corrida eletroforética em gel de poliacrilamida pudesse ser realizada. Para a coloração do gel, usou-se Coomasie Brilliant Blue. Após análise dos padrões eletroforéticos da transferrina e albumina, observou-se que não houve relação detectável entre os fenótipos da albumina e precocidade sexual de touros doadores. Entretanto, em relação à transferrina, foi possível sugerir uma associação entre o alelo TfD com touros portadores de sêmen congelável precocemente ou medianamente em termos de idade à congelação.

Rhipicephalus (Boophilus) microplus: Distinct acute phase proteins vary during infestations according to the genetic composition of the bovine hosts, Bos taurus and Bos indicus

Tick bites may trigger acute phase responses. Positive and negative acute phase proteins were measured in infested cattle genetically resistant and susceptible to ticks. During heavier infestations levels of haptoglobin increased significantly in susceptible bovines; levels of serum amyloid A increased in resistant bovines; levels of alpha-l-acid glycoprotein decreased significantly in resistant bovines; levels of transferrin decreased significantly in susceptible bovines.In conclusion, tick infestations trigger acute phase responses and enhancement of specific acute phase proteins differs according to the genetic composition of hosts. Acute phase proteins may constitute useful biological signatures for monitoring the stress induced by tick infestations. (c) 2007 Elsevier B.V. All rights reserved.

Influência do tratamento periodontal sobre os níveis da proteína c-reativa ultrassensível em pacientes com insuficiência renal crônica em fase de prétransplante

The present study evaluated the influence of non-surgical periodontal treatment on the levels of C- reactive protein (hsCRP) in patients with chronic renal failure (CRF) in pretransplant. We conducted a controlled and randomized trial to evaluate the periodontal condition and plasma concentrations of hsCRP, albumin and transferrin in 56 dialysis patients divided into two groups: experimental and control. The study was conducted at the dental clinic of Family and Community Health s Unit (USFC), located in Onofre Lopes University Hospital (HUOL), Federal University of Rio Grande do Norte (UFRN), from December 2010 to November 2011. Severe periodontitis was the type of periodontal disease more common, affecting 78.6% of patients. Periodontal conditions, evaluated through the means of probing depth, clinical attachment level, bleeding index and plaque index, proved to be uniform for both groups at the initial examination. There were no differences in levels of inflammatory markers between the two groups. The analysis of the concentrations of hsCRP allowed classifying study participants as at high risk of developing cardiovascular disease. After completion of periodontal treatment in the experimental group, there was a statistically significant reduction of the mean of all periodontal parameters assessed; however this improvement of periodontal health was not accompanied by changes in the levels of hsCRP, albumin and transferrin in the evaluation time. Given this, the periodontal treatment did not promote the reduction of systemic inflammatory burden and risk of cardiovascular complications in patients with CRF