Mineral Disorders in Adult Inpatients Receiving Parenteral Nutrition. Is Older Age a Contributory Factor?

BACKGROUND: Parenteral nutrition (PN)-dependent adults and elderly individuals who are admitted to hospital treatment are potentially susceptible to mineral disorder complications due to depleted physiological reserves, loss of lean body mass, and increased fat mass, thus worsening inflammation. AIM: The purpose of this study is to evaluate the prevalence of hypophosphatemia, hypokalemia, and hypomagnesaemia prior and within the first 7 days of PN infusion. Furthermore, whether malnutrition and old age are associated with these disorders was also investigated. METHODS: This study included a historical cohort of adult patients, and 1,040 patients whose information was prospectively entered in the database were evaluated. RESULTS: Of the 781 patients, 27.3% were ≥65 years, 80.9% had undergone surgical treatment, 74.3% were in the intensive care unit, and 17.9% died during the hospitalization period. About 17.1% patients were malnourished. Protein energy malnutrition (PEM) was observed in 31.9% of the elderly patients and 27.1% of adults in general. Hypophosphatemia, hypokalemia, and hypomagnesemia were more prevalent before the start of PN infusion (D0: 214 [18.4%]), and new events were more common during the first 2 days of PN infusion (D1: 283 [23.1%]; D2: 243 [20.1%]. Elderly patients were more susceptible to developing hypophosphatemia (odds ratio [OR]: 1.69; 95% confidence interval [CI]: 1.29-2.19; p<0.001). Patients with PEM were also more susceptible to hypophosphatemia (OR: 3.75; 95% CI: 1.13-12.47; p=0.036). CONCLUSION: Hypophosphatemia, hypokalemia, and hypomagnesemia were frequently observed in hospitalized adults and elderly patients before and particularly during the first 2 days of PN infusion. Elderly patients and patients with PEM are more susceptible to developing hypophosphatemia.

Open heart surgery for management of right auricular thrombus related to central venous catheterization. / Cirugía a corazón abierto para el tratamiento de trombo auricular derecho relacionado con cateterización venosa central.

Central venous catheters are widely used in critically ill patients; however, they are also associated with increased morbidity and mortality. The literature may underestimate the incidence of catheter-inducible right atrial thrombi that are asymptomatic but potentially life threatening. The recognized risk factors for its development include infections related to the catheter, endothelial injury secondary to mechanical and chemical damage induced by certain medications and infused fluids. The characteristics of the patient and the catheter, such as size, material, type, location and ease of insertion, as well as the duration of placement play an additional role. We report the case of a 38-year-old man, who developed an asymptomatic catheter-inducible right atrial thrombi requiring open heart surgery, after taking a central venous catheter for thirty-five days. The present case highlights existing limitations in making a correct and fast diagnosis, which should be anticipated in patients with multiple risk factors for thrombosis. Given the limited recommendations available, we consider that the most appropriate strategy should be individualized.

Physical performance, quality of life and sexual satisfaction evaluation in adults with cystic fibrosis: An unexplored correlation.

OBJECTIVE: Quality of life (QOL), sexual satisfaction (SS) and physical performance have been assessed in the management of numerous chronic diseases. METHODS: In this study, the following tests and surveys were applied: (i) QOL questionnaire [Cystic Fibrosis Questionnaire (CFQ)]; (ii) SS questionnaire (SSQ) [female sexual quotient (FSQ) and male sexual quotient (MSQ)]; (iii) 6-minute walk test (6MWT). Spearman's correlation was used for comparison between the data; the Mann-Whitney test was applied to analyze the difference between genders. A total of 52 adult patients with CF were included in this study. RESULTS: There was a positive correlation between CFQ domains and SSQ questions. The CFQ showed a positive correlation with peripheral oxygen saturation of hemoglobin (SpO2) and the distance walked in the 6MWT, and a negative correlation with the Borg scale. The SSQ showed positive correlation with the distance walked and a negative correlation with the Borg scale. For some markers evaluated in the 6MWT, there was sometimes association with the evaluated domains and questions. Male patients showed better scores in the emotional CFQ domain, better performance in SSQ and physical performance. CONCLUSIONS: There was a correlation between CFQ, SSQ and 6MWT in CF. Finally; we believe that QOL surveys should assess the domain "sexuality" as well as physical performance tests.

Gelsolin role in microapocrine secretion.

A role of gelsolin in opening the way along the microvilli for secretory vesicles during microapocrine secretion is proposed here. Data obtained with different techniques showed that many digestive enzymes are released by microapocrine secretion in insects. Proteins that might be involved in the machinery of microapocrine secretion were selected from our transcriptomes and literature searches. The proteins were annexin, Complex actin-related proteins 2 and 3 (ARP 2/3) cofilin, fimbrin, gelsolin 1, gelsolin 2, moesin, myosin 1, myosin 6, protein disulphide isomerase 1 (PDI 1), PDI 2 and profilin. The cDNAs coding for annexin, fimbrin, gelsolin 1, myosin 1, PDI 1 and PDI 2 were cloned and their sequences deposited in GenBank. Only gelsolin 1 and myosin 1 are expressed exclusively in the midgut (semiquantitative reverse transcriptase PCR). As myosin 1 may have a structural role in microvilli, gelsolin 1 is the best guess to be involved in the secretory machinery. A truncated recombinant gelsolin 1 was used to generate antibodies with which it was shown labelling inside and around midgut cell microvilli shown in an electron microscope, reinforcing a microvillar role for gelsolin 1. Suppression of gelsolin 1 synthesis by RNA interference prevents secretory vesicles from advancing inside the microvilli, in agreement with its putative role in severing the actin filaments to free the way for the vesicles.

Effect of starch-based supplementation level combined with oil on intake, performance, and methane emissions of growing Nellore bulls on pasture.

Intake of tropical grass forages alone is generally insufficient to avoid nutrition imbalances and reduced animal performance; therefore, supplementation is often recommended. The hypothesis of the present study is that when combined with fat, soybean hulls (SH) could replace corn as a source of energy, reducing methane production without affecting animal performance. This study evaluated the effects of starch-based supplementation level combined with oil on intake, digestibility, performance, and methane emissions of growing Nellore bulls (P = 44; initial BW = 250.69 ± 27 kg) fed cv. Xaraés during the rainy season. There were no interactions between starch level and oil supplementation with regard to intake of DM (P = 0.67), forage DM (P = 0.55), supplement DM (P = 0.14), OM (P = 0.66), CP (P = 0.74), NDF (P = 0.50), ether extract (EE; = 0.47), and GE ( P= 0.68). The intake of EE was greater for animals supplemented with oil than those fed supplements without oil (P < 0.01). There were no interactions between starch level and oil supplementation on digestibility of DM (P= 0.18), OM (P = 0.11), NDF (P= 0.42), and EE (P = 0.14). Moreover, there was interaction between starch and oil supplementation on GE (P < 0.01). Independent of starch level used, the addition of oil decreased the digestibility of OM (P = 0.04) and NDF (P = 0.03). There were no main effects of starch level, oil, or interaction between starch and oil for initial BW (P = 0.10), final BW (P = 0.94), ADG (P = 0.40), feed efficiency (P= 0.37), and carcass gain (P = 0.38). There was no interaction between starch-based supplementation level and oil on methane emissions when expressed in grams per day (P = 0.77), kilograms per year (P = 0.77), grams per kilogram DMI (P = 0.53), and grams per kilogram carcass gain (P= 0.31). There was, however, an interaction (P = 0.04) between starch level and oil on methane emissions when corrected for NDF intake. Additionally, oil decreased enteric methane emission for intake of GE (P = 0.04) and EE (P < 0.01) of animals fed with starch level. Soybean hulls have an estimated feeding value similar to that of corn. The use of oil supplementation may be effective to reduce enteric methane emission of Nellore bulls raised on pasture.

Spatial cognitive deficits in an animal model of Wernicke-Korsakoff syndrome are related to changes in thalamic VDAC protein concentrations.

Proteomic profiles of the thalamus and the correlation between the rats' performance on a spatial learning task and differential protein expression were assessed in the thiamine deficiency (TD) rat model of Wernicke-Korsakoff syndrome. Two-dimensional gel-electrophoresis detected 320 spots and a significant increase or decrease in seven proteins. Four proteins were correlated to rat behavioral performance in the Morris Water Maze. One of the four proteins was identified by mass spectrometry as Voltage-Dependent Anion Channels (VDACs). The association of VDAC is evident in trials in which the rats' performance was worst, in which the VDAC protein was reduced, as confirmed by Western blot. No difference was observed on the mRNA of Vdac genes, indicating that the decreased VDAC expression may be related to a post-transcriptional process. The results show that TD neurodegeneration involves changes in thalamic proteins and suggest that VDAC protein activity might play an important role in an initial stage of the spatial learning process.

Fatty acid profile, carcass and quality traits of meat from Nellore young bulls on pasture supplemented with crude glycerin.

The aim of this study was to evaluate the carcass traits and meat quality of Nellore bulls (279.5±16.3 initial body weight) raised on pasture supplemented with crude glycerin at 0%, 7%, 14%, 21% or 28% (DM basis). The diets were similar in energy and protein levels, and the glycerin replaced corn in the supplement. After slaughter, the carcass characteristics were measured, and the longissimus muscle was collected to determine the meat quality. The inclusion of crude glycerin in the supplement did not change (P>0.05) any of the carcass characteristics and meat quality assessed; however, the pH decreased linearly (P=0.03), and margaric acid (17:0) increased (P=0.02) in the longissimus muscle with the addition of glycerin in the diet. Our conclusion is that the inclusion of crude glycerin up to the level of 28% of dry matter in the supplement does not alter the carcass characteristics or the meat quality from animals raised on pasture.

Insect midgut carboxypeptidases with emphasis on S10 hemipteran and M14 lepidopteran carboxypeptidases.

We compared the whole complement of midgut carboxypeptidases from 10 insects pertaining to five orders based on transcriptomes obtained by deep sequencing and biochemical data. Most of the carboxypeptidases were metallocarboxypeptidases from family M14, with carboxypeptidase A (CPA) predominating over carboxypeptidase B (CPB). They were found in all of the insects studied except for the hemipterans and a bruchid beetle. M14 carboxypeptidases were expressed only in the midgut of Spodoptera frugiperda (Lepidoptera). The most expressed CPA from this insect (SfCPA) was cloned, sequenced and expressed as a recombinant enzyme. This enzyme was used to generate antibodies used to demonstrate that SfCPA is secreted by an exocytic route. Serine carboxypeptidases from family S10 were found in all of the insects studied here. In S. frugiperda, they are expressed in all tissues besides the midgut, in accordance with their presumed lysosomal role. In the hemipteran Dysdercus peruvianus, S10 carboxypeptidases are expressed only in midgut, suggesting that they are digestive enzymes. This was confirmed by enzyme assays of midgut contents. Furthermore, the substrate specificity of D. peruvianus S10 carboxypeptidases are predicted to be one CPC (preferring hydrophobic residues) and one CPD (preferring basic residues), thus able to hydrolyse the peptides formed by their digestive cathepsin D and cathepsin L, respectively. The role of S10 carboxypeptidases in bruchid beetles are suggested to be the same as in hemipterans.

Fatty acid profile, carcass and meat quality traits of young Nellore bulls fed crude glycerin replacing energy sources in the concentrate.

Carcass and meat quality traits of 60 Nellore young bulls fed diets without crude glycerin (CG); with CG replacing corn (CGc; 10% of dry matter - DM) in the concentrate; and with CG replacing soybean hull (CGsh; 10% of DM) in the concentrate were evaluated. Diets were evaluated at two concentrate levels (CLs). The CL did not affect cold carcass weight (CCW; P=0.6074), cold carcass dressing (CCD; P=0.9636), rib fat thickness (RFT; P=0.8696) and longissimus muscle area (LMA; P=0.7524). Animals fed diets with CGc or CGsh showed meat with greater deposition of monounsaturated fatty acid (MUFA; P=0.0022) and CLA (18:2 cis-9, trans-11) contents (P=0.0001) than animals fed diets without CG. The inclusion of 10% of CG in diets CGc or CGsh does not affect the carcass and meat quality traits; however, it increases the MUFA and CLA contents in beef, although these changes are very small in nutritional terms.

Use of inflammatory molecules to predict the occurrence of fever in onco-hematological patients with neutropenia.

Febrile neutropenia remains a frequent complication in onco-hematological patients, and changes in the circulating level of inflammatory molecules (IM) may precede the occurrence of fever. The present observational prospective study was carried out to evaluate the behavior of plasma tumor necrosis factor alpha (TNF-α), soluble TNF-α I and II receptors (sTNFRI and sTNFRII), monocyte chemoattractant protein-1 [MCP-1 or chemokine (c-c motif) ligand 2 (CCL2)], macrophage inflammatory protein-1α (MIP-1α or CCL3), eotaxin (CCL11), interleukin-8 (IL-8 or CXCL8), and interferon-inducible protein-10 (IP-10 or CXCL10) in 32 episodes of neutropenia in 26 onco-hematological patients. IM were tested on enrollment and 24-48 h before the onset of fever and within 24 h of the first occurrence of fever. Eight of 32 episodes of neutropenia did not present fever (control group) and the patients underwent IM tests on three different occasions. sTNFRI levels, measured a median of 11 h (1-15) before the onset of fever, were significantly higher in patients presenting fever during follow-up compared to controls (P = 0.02). Similar results were observed for sTNFRI and CCL2 levels (P = 0.04 for both) in non-transplanted patients. A cut-off of 1514 pg/mL for sTNFRI was able to discriminate between neutropenic patients with or without fever during follow-up, with 65% sensitivity, 87% specificity, and 93% positive predictive value. Measurement of the levels of plasma sTNFRI can be used to predict the occurrence of fever in neutropenic patients.

Use of inflammatory molecules to predict the occurrence of fever in onco-hematological patients with neutropenia

Febrile neutropenia remains a frequent complication in onco-hematological patients, and changes in the circulating level of inflammatory molecules (IM) may precede the occurrence of fever. The present observational prospective study was carried out to evaluate the behavior of plasma tumor necrosis factor alpha (TNF-α), soluble TNF-α I and II receptors (sTNFRI and sTNFRII), monocyte chemoattractant protein-1 [MCP-1 or chemokine (c-c motif) ligand 2 (CCL2)], macrophage inflammatory protein-1α (MIP-1α or CCL3), eotaxin (CCL11), interleukin-8 (IL-8 or CXCL8), and interferon-inducible protein-10 (IP-10 or CXCL10) in 32 episodes of neutropenia in 26 onco-hematological patients. IM were tested on enrollment and 24-48 h before the onset of fever and within 24 h of the first occurrence of fever. Eight of 32 episodes of neutropenia did not present fever (control group) and the patients underwent IM tests on three different occasions. sTNFRI levels, measured a median of 11 h (1-15) before the onset of fever, were significantly higher in patients presenting fever during follow-up compared to controls (P = 0.02). Similar results were observed for sTNFRI and CCL2 levels (P = 0.04 for both) in non-transplanted patients. A cut-off of 1514 pg/mL for sTNFRI was able to discriminate between neutropenic patients with or without fever during follow-up, with 65% sensitivity, 87% specificity, and 93% positive predictive value. Measurement of the levels of plasma sTNFRI can be used to predict the occurrence of fever in neutropenic patients.

A possible role of a cerebral energy gene in alcoholism.

We examined a possible relationship between genes responsible for energy metabolism of the brain and addictive behavior in an animal model. We used non-inbred, Swiss mice exposed to a three-bottle free-choice model [water, 5% (v/v) ethanol, and 10% (v/v) ethanol] over a 16-week period, consisting of four phases: acquisition, withdrawal, reexposure, and quinine-adulteration. The mice were then behaviorally classified into three groups: loss-of-control-drinker (preference for ethanol and high levels of consumption during all phases, N = 6), heavy-drinker (preference for ethanol and high levels of consumption during acquisition and reduction during quinine-adulteration, N = 7), and light-drinker (preference for water during all phases, N = 10). Another group only received tap water (ethanol-naive control mice, N = 9). Further analysis using quantitative real-time PCR showed that in mice behaviorally classified as loss-of-control-drinkers, there was a significant inverse correlation between transcript levels of the Hadh gene and those of other energy metabolism genes in the nucleus of the amygdala, suggesting that this pathway may contribute to ethanol consumption in these mice. We conclude that cerebral energy metabolism is involved with ethanol addiction, meriting further study.

Perfil bioquímico sanguíneo na intoxicação experimental com extrato de Mascagnia rigida (A. Juss.) Griseb. (Malpighiaceae) em coelhos / Biochemistry profile after experimental intoxication by extract of Mascagnia rigida (A. Juss.) Griseb. (Malpighiaceae) in rabbits

Avaliou-se o perfil bioquímico sanguíneo na intoxicação por Mascagnia rigida, uma planta tóxica que gera problema econômico para a pecuária, por causar morte súbita. Nove coelhos Nova Zelândia, machos, com massa corporal média de 3,54kg, foram distribuídos em três grupos (G) (n=3). Os animais receberam, durante oito dias consecutivos, o equivalente a 30g/kg de matéria seca da planta em dois tipos de extratos: solúvel em água (GS) e insolúvel em água (GI), e formou-se também o grupo-controle (GC). Os exames bioquímicos foram realizados previamente ao início do experimento até o nono dia. A administração dos extratos da Mascagnia rigida causou alterações eletrolíticas que podem justificar alguns sinais clínicos observados e atuar de forma significativa na causa mortis.

A study was carried out to evaluate the biochemical profile in Mascagnia rigida poisoning, a toxic plant that generates a significant economic problem to livestock, causing "sudden death". Nine New Zealand rabbits, male, 3,54kg mean body weight were divided into three groups (G) (n = 3). The animals received the equivalent of 30g/kg of dry matter in two types of extracts: water-soluble (GS) and insoluble in water (GI), and the control group (CG) (ultra-pure water) for eight consecutive days. Biochemical exams were done prior to the beginning of the experiment until the ninth day. It was concluded that the administration of extracts of Mascagnia rigida cause electrolyte imbalances that may justify some clinical signs and act significantly in the cause of death.

Considerações sobre a determinação quantitativa de alendronato de sódio: titulometria x cromatografia / Observations on the quantitative determination of alendronate sodium: titrimetry vs. chromatography

O alendronato de sódio é um composto aminodifosfonado capaz de se fixar à matriz óssea e inibir a reabsorção mediada por osteoclastos. A escassez de metodologias oficiais para a determinação quantitativa deste fármaco levou ao desenvolvimento de diversos métodos, os quais empregam, em sua maioria, a cromatografia líquida de alta eficiência (CLAE), com a derivatização do fármaco para poder empregar detectores de ultravioleta. Também há relatos sobre metodologias mais simples para a análise do alendronato, utilizando titulometria ou análise espectrofotométrica. Neste trabalho foi avaliado o emprego da titulometria de neutralização na determinação quantitativa do alendronato de sódio em três lotes de matéria-prima, utilizando NaOH 0,1 M como titulante. Os resultados obtidos na titulometria foram comparados aos encontrados em método cromatográfico de referência (CLAE com derivatização por 9-fluorenilmetilcloroformato ou FMOC), descrito na Farmacopéia Americana (United States Pharmacopeia), os quais apresentaram valores estatisticamente diferentes. Ensaios para a caracterização das amostras também foram realizados e foi observado comportamento distinto das 3 matérias-primas em relação à substância de referência (padrão secundário). O método titulométrico apresentou adequada precisão, mas não mostrou especificidade para a determinação das matérias-primas, embora possa ser validado para determinação do fármaco em produto acabado.

Alendronate sodium is an aminobisphosphonate compound that can bind to the bone matrix and inhibit its osteoclast-mediated resorption. The lack of official monographs on the quantitative analysis of this drug has led to the proposal of a number of different methods for its determination, most of which employ high performance liquid chromatography (HPLC), performed after derivatization of the drug to enable ultraviolet detection. Simpler methodologies based on titrimetry and spectrophotometry have also been described. In this paper, the results obtained by acid-base titration of three batches of bulk alendronate, with 0.1 M sodium hydroxide as titrant, were compared with those achieved by a chromatographic reference method (HPLC of an FMOC derivative of the drug) published in the United States Pharmacopeia. The two methods gave statistically different results for the analysis of the three samples. Also, physical and chemical characterization of these samples showed differences between them and the reference substance (brand-name drug). The acid-base titrimetry was precise but not specific for determination of the bulk drug. However, it may be employed in the quality control of alendronate sodium dosage forms, since the excipients do not interfere with the analysis.

Sequencing of Spodoptera frugiperda midgut trehalases and demonstration of secretion of soluble trehalase by midgut columnar cells.

Both soluble (SfTre1) and membrane-bound (SfTre2) trehalases occur along the midgut of Spodoptera frugiperda larvae. Released SfTre2 was purified as a 67 kDa protein. Its K(m) (1.6 mM) and thermal stability (half life 10 min at 62 degrees C) are different from the previously isolated soluble trehalase (K(m)= 0.47 mM; 100% stable at 62 degrees C). Two cDNAs coding for S. frugiperda trehalases have been cloned using primers based on consensus sequences of trehalases and having as templates a cDNA library prepared from total polyA-containing RNA extracted from midguts. One cDNA codes for a trehalase that has a predicted transmembrane sequence and was defined as SfTre2. The other, after being cloned and expressed, results in a recombinant trehalase with a K(m) value and thermal stability like those of native soluble trehalase. This enzyme was defined as SfTre1 and, after it was used to generate antibodies, it was immunolocalized at the secretory vesicles and at the glycocalyx of columnar cells. Escherichia coli trehalase 3D structure and sequence alignment with SfTre1 support a proposal regarding the residue modulating the pKa value of the proton donor.

Neonatal screening for cystic fibrosis in São Paulo State, Brazil: a pilot study

Cystic fibrosis is one of the most common autosomal recessive hereditary diseases in the Caucasian population, with an incidence of 1:2000 to 1:3500 liveborns. More than 1000 mutations have been described with the most common being F508del. It has a prevalence of 23-55 percent within the Brazilian population. The lack of population-based studies evaluating the incidence of cystic fibrosis in São Paulo State, Brazil, and an analysis concerning the costs of implantation of a screening program motivated the present study. A total of 60,000 dried blood samples from Guthrie cards obtained from April 2005 to January 2006 for neonatal screening at 4 reference centers in São Paulo State were analyzed. The immunoreactive trypsinogen (IRT)/IRT protocol was used with the cut-off value being 70 ng/mL. A total of 532 children (0.9 percent) showed IRT >70 ng/mL and a 2nd sample was collected from 418 (80.3 percent) of these patients. Four affected children were detected at two centers, corresponding to an incidence of 1:8403. The average age at diagnosis was 69 days, and 3 of the children already showed severe symptoms of the disease. The rate of false-positive results was 95.2 percent and the positive predictive value for the test was 8 percent. The cost of detecting an affected subject was approximately US$8,000.00 when this cystic fibrosis program was added to an existing neonatal screening program. The present study clearly shows the difficulties involved in cystic fibrosis screening using the IRT/IRT protocol, particularly in a population with no long-term tradition of neonatal screening.

Neonatal screening for cystic fibrosis in São Paulo State, Brazil: a pilot study.

Cystic fibrosis is one of the most common autosomal recessive hereditary diseases in the Caucasian population, with an incidence of 1:2000 to 1:3500 liveborns. More than 1000 mutations have been described with the most common being F508del. It has a prevalence of 23-55% within the Brazilian population. The lack of population-based studies evaluating the incidence of cystic fibrosis in São Paulo State, Brazil, and an analysis concerning the costs of implantation of a screening program motivated the present study. A total of 60,000 dried blood samples from Guthrie cards obtained from April 2005 to January 2006 for neonatal screening at 4 reference centers in São Paulo State were analyzed. The immunoreactive trypsinogen (IRT)/IRT protocol was used with the cut-off value being 70 ng/mL. A total of 532 children (0.9%) showed IRT >70 ng/mL and a 2nd sample was collected from 418 (80.3%) of these patients. Four affected children were detected at two centers, corresponding to an incidence of 1:8403. The average age at diagnosis was 69 days, and 3 of the children already showed severe symptoms of the disease. The rate of false-positive results was 95.2% and the positive predictive value for the test was 8%. The cost of detecting an affected subject was approximately US$8,000.00 when this cystic fibrosis program was added to an existing neonatal screening program. The present study clearly shows the difficulties involved in cystic fibrosis screening using the IRT/IRT protocol, particularly in a population with no long-term tradition of neonatal screening.

Structure, processing and midgut secretion of putative peritrophic membrane ancillary protein (PMAP) from Tenebrio molitor larvae.

A cDNA coding for a Tenebrio molitor midgut protein named peritrophic membrane ancillary protein (PMAP) was cloned and sequenced. The complete cDNA codes for a protein of 595 amino acids with six insect-allergen-related-repeats that may be grouped in A (predicted globular)- and B (predicted nonglobular)-types forming an ABABAB structure. The PMAP-cDNA was expressed in Pichia pastoris and the recombinant protein (64kDa) was purified to homogeneity and used to raise antibodies in rabbits. The specific antibody detected PMAP peptides (22kDa) in the anterior and middle midgut tissue, luminal contents, peritrophic membrane and feces. These peptides derive from PMAP, as supported by mass spectrometry, and resemble those formed by the in vitro action of trypsin on recombinant PMAP. Both in vitro and in vivo PMAP processing seem to occur by attack of trypsin to susceptible bonds in the coils predicted to link AB pairs, thus releasing the putative functional AB structures. The AB-domain structure of PMAP is found in homologous proteins from several insect orders, except lepidopterans that have the apparently derived protein known as nitrile-specifier protein. Immunocytolocalization shows that PMAP is secreted by exocytosis and becomes entrapped in the glycocalyx, before being released into midgut contents. Circumstantial evidence suggests that PMAP-like proteins have a role in peritrophic membrane type 2 formation.

Assessment of antimutagenic and genotoxic potential of senna (Cassia angustifolia Vahl.) aqueous extract using in vitro assays.

Senna (Cassia angustifolia Vahl.) is widely used as a laxative, although potential side effects, such as toxicity and genotoxicity, have been reported. This study evaluated genotoxic and mutagenic effects of senna aqueous extract (SAE) by means of four experimental assays: inactivation of Escherichia coli cultures; bacterial growth inhibition; reverse mutation test (Mutoxitest) and DNA strand break analysis in plasmid DNA. Our results demonstrated that SAE produces single and double strand breaks in plasmid DNA in a cell free system. On the other hand, SAE was not cytotoxic or mutagenic to Escherichia coli strains tested. In effect, SAE was able to avoid H(2)O(2)-induced mutagenesis and toxicity in Escherichia coli IC203 (uvrA oxyR) and IC205 (uvrA mutM) strains, pointing to a new antioxidant/antimutagenic action of SAE.