Spatio-temporal variability in the Cladocera assemblage of a subtropical hypersaline lagoon / Variabilidade espaço-temporal da assembleia de Cladocera de uma lagoa hipersalina subtropical

Cladocera represent an important zooplankton group because of their seasonal prominence in terms of abundance and their contribution in controlling primary production (phytoplankton). On a global scale, there are few studies on Cladocera in hypersaline environments. The present work aims to evaluate the spatio-temporal variation of the Cladocera assemblage across a salinity gradient in the habitats of the Araruama Lagoon. Samples were collected in random months over a period of four years at 12 fixed stations in the Araruama Lagoon using a WP2 plankton net equipped with a flow meter. Our results do not reveal significant influence of the tide and seasonal variation as factors affecting the Cladocera assemblage. Five Cladocera species were found in the Araruama Lagoon, only in stations 11 and 12 where they reached an average of 1,799 ± 3,103 ind. m-³. The mean of the Shannon Diversity Index was 0.45 ± 0.2. The species that stood out in terms of frequency and abundance were: Penilia avirostris (frequency of occurrence: 71%), followed by Pseudevadne tergestina (41%). The same species also stood out in terms of relative abundance, Penilia avirostris (87%) and Pseudevadne tergestina (11%). The absence of Cladocera in the innermost parts of the lagoon suggests that their entrance to these locations is possibly inhibited by the salinity and temperature gradient of the lagoon, being the main factors influencing the dynamics of the Cladocera assemblages.

Os cladóceros representam um importante grupo de zooplâncton, devido ao seu destaque sazonal em termos de abundância e à sua contribuição no controlo da produção primária (fitoplâncton). Em escala global, há poucos estudos com Cladocera em ambientes hipersalinos. O presente trabalho tem como objetivo avaliar a variação espaço-temporal da assembleia de Cladocera em um gradiente de salinidades da Lagoa de Araruama. As amostras foram coletadas em meses aleatórios, durante um período de quatro anos, em 12 estações fixas na Lagoa de Araruama, utilizando uma rede de plâncton WP2 equipada com um fluxômetro. Os nossos resultados não revelam uma influência significativa da maré e da variação sazonal como fatores que afetam a assembleia de Cladocera. Cinco espécies de Cladocera foram encontradas na Lagoa de Araruama. A assembleia apresentou maior densidade absoluta estações 12 e 11, atingindo uma a média de 1.799 ± 3.103 inds-³. A média do Índice de Diversidade de Shannon foi de 0,45 ± 0,2. As espécies que se destacaram em termos de frequência e abundância foram: Penilia avirostris (frequência de ocorrência: 71%), seguida de Pseudevadne tergestina (41%). As mesmas espécies também se destacaram em termos de abundância relativa Penilia avirostris (87%) e Pseudevadne tergestina (11%). A ausência de Cladocera nas partes mais interiores da lagoa sugere que a sua entrada nestes locais é possivelmente inibida pela salinidade e gradiente de temperatura da lagoa, sendo os principais fatores que influenciam a dinâmica da assembleia de Cladocera.

Spatio-temporal variability in the Cladocera assemblage of a subtropical hypersaline lagoon

Abstract Cladocera represent an important zooplankton group because of their seasonal prominence in terms of abundance and their contribution in controlling primary production (phytoplankton). On a global scale, there are few studies on Cladocera in hypersaline environments. The present work aims to evaluate the spatio-temporal variation of the Cladocera assemblage across a salinity gradient in the habitats of the Araruama Lagoon. Samples were collected in random months over a period of four years at 12 fixed stations in the Araruama Lagoon using a WP2 plankton net equipped with a flow meter. Our results do not reveal significant influence of the tide and seasonal variation as factors affecting the Cladocera assemblage. Five Cladocera species were found in the Araruama Lagoon, only in stations 11 and 12 where they reached an average of 1,799 ± 3,103 ind. m-3. The mean of the Shannon Diversity Index was 0.45 ± 0.2. The species that stood out in terms of frequency and abundance were: Penilia avirostris (frequency of occurrence: 71%), followed by Pseudevadne tergestina (41%). The same species also stood out in terms of relative abundance, Penilia avirostris (87%) and Pseudevadne tergestina (11%). The absence of Cladocera in the innermost parts of the lagoon suggests that their entrance to these locations is possibly inhibited by the salinity and temperature gradient of the lagoon, being the main factors influencing the dynamics of the Cladocera assemblages.

Resumo Os cladóceros representam um importante grupo de zooplâncton, devido ao seu destaque sazonal em termos de abundância e à sua contribuição no controlo da produção primária (fitoplâncton). Em escala global, há poucos estudos com Cladocera em ambientes hipersalinos. O presente trabalho tem como objetivo avaliar a variação espaço-temporal da assembleia de Cladocera em um gradiente de salinidades da Lagoa de Araruama. As amostras foram coletadas em meses aleatórios, durante um período de quatro anos, em 12 estações fixas na Lagoa de Araruama, utilizando uma rede de plâncton WP2 equipada com um fluxômetro. Os nossos resultados não revelam uma influência significativa da maré e da variação sazonal como fatores que afetam a assembleia de Cladocera. Cinco espécies de Cladocera foram encontradas na Lagoa de Araruama. A assembleia apresentou maior densidade absoluta estações 12 e 11, atingindo uma a média de 1.799 ± 3.103 inds-3. A média do Índice de Diversidade de Shannon foi de 0,45 ± 0,2. As espécies que se destacaram em termos de frequência e abundância foram: Penilia avirostris (frequência de ocorrência: 71%), seguida de Pseudevadne tergestina (41%). As mesmas espécies também se destacaram em termos de abundância relativa Penilia avirostris (87%) e Pseudevadne tergestina (11%). A ausência de Cladocera nas partes mais interiores da lagoa sugere que a sua entrada nestes locais é possivelmente inibida pela salinidade e gradiente de temperatura da lagoa, sendo os principais fatores que influenciam a dinâmica da assembleia de Cladocera.

Spatio-temporal variability in the Cladocera assemblage of a subtropical hypersaline lagoon / Variabilidade espaço-temporal da assembleia de Cladocera de uma lagoa hipersalina subtropical

Cladocera represent an important zooplankton group because of their seasonal prominence in terms of abundance and their contribution in controlling primary production (phytoplankton). On a global scale, there are few studies on Cladocera in hypersaline environments. The present work aims to evaluate the spatio-temporal variation of the Cladocera assemblage across a salinity gradient in the habitats of the Araruama Lagoon. Samples were collected in random months over a period of four years at 12 fixed stations in the Araruama Lagoon using a WP2 plankton net equipped with a flow meter. Our results do not reveal significant influence of the tide and seasonal variation as factors affecting the Cladocera assemblage. Five Cladocera species were found in the Araruama Lagoon, only in stations 11 and 12 where they reached an average of 1,799 ± 3,103 ind. m-3. The mean of the Shannon Diversity Index was 0.45 ± 0.2. The species that stood out in terms of frequency and abundance were: Penilia avirostris (frequency of occurrence: 71%), followed by Pseudevadne tergestina (41%). The same species also stood out in terms of relative abundance, Penilia avirostris (87%) and Pseudevadne tergestina (11%). The absence of Cladocera in the innermost parts of the lagoon suggests that their entrance to these locations is possibly inhibited by the salinity and temperature gradient of the lagoon, being the main factors influencing the dynamics of the Cladocera assemblages.

Os cladóceros representam um importante grupo de zooplâncton, devido ao seu destaque sazonal em termos de abundância e à sua contribuição no controlo da produção primária (fitoplâncton). Em escala global, há poucos estudos com Cladocera em ambientes hipersalinos. O presente trabalho tem como objetivo avaliar a variação espaço-temporal da assembleia de Cladocera em um gradiente de salinidades da Lagoa de Araruama. As amostras foram coletadas em meses aleatórios, durante um período de quatro anos, em 12 estações fixas na Lagoa de Araruama, utilizando uma rede de plâncton WP2 equipada com um fluxômetro. Os nossos resultados não revelam uma influência significativa da maré e da variação sazonal como fatores que afetam a assembleia de Cladocera. Cinco espécies de Cladocera foram encontradas na Lagoa de Araruama. A assembleia apresentou maior densidade absoluta estações 12 e 11, atingindo uma a média de 1.799 ± 3.103 inds-3. A média do Índice de Diversidade de Shannon foi de 0,45 ± 0,2. As espécies que se destacaram em termos de frequência e abundância foram: Penilia avirostris (frequência de ocorrência: 71%), seguida de Pseudevadne tergestina (41%). As mesmas espécies também se destacaram em termos de abundância relativa Penilia avirostris (87%) e Pseudevadne tergestina (11%). A ausência de Cladocera nas partes mais interiores da lagoa sugere que a sua entrada nestes locais é possivelmente inibida pela salinidade e gradiente de temperatura da lagoa, sendo os principais fatores que influenciam a dinâmica da assembleia de Cladocera.

Spatio-temporal variability in the Cladocera assemblage of a subtropical hypersaline lagoon.

Cladocera represent an important zooplankton group because of their seasonal prominence in terms of abundance and their contribution in controlling primary production (phytoplankton). On a global scale, there are few studies on Cladocera in hypersaline environments. The present work aims to evaluate the spatio-temporal variation of the Cladocera assemblage across a salinity gradient in the habitats of the Araruama Lagoon. Samples were collected in random months over a period of four years at 12 fixed stations in the Araruama Lagoon using a WP2 plankton net equipped with a flow meter. Our results do not reveal significant influence of the tide and seasonal variation as factors affecting the Cladocera assemblage. Five Cladocera species were found in the Araruama Lagoon, only in stations 11 and 12 where they reached an average of 1,799 ± 3,103 ind. m-3. The mean of the Shannon Diversity Index was 0.45 ± 0.2. The species that stood out in terms of frequency and abundance were: Penilia avirostris (frequency of occurrence: 71%), followed by Pseudevadne tergestina (41%). The same species also stood out in terms of relative abundance, Penilia avirostris (87%) and Pseudevadne tergestina (11%). The absence of Cladocera in the innermost parts of the lagoon suggests that their entrance to these locations is possibly inhibited by the salinity and temperature gradient of the lagoon, being the main factors influencing the dynamics of the Cladocera assemblages.

A novel SDS-stable dimer of a heterogeneous nuclear ribonucleoprotein at presynaptic terminals of squid neurons.

The presence of mRNAs in synaptic terminals and their regulated translation are important factors in neuronal communication and plasticity. Heterogeneous nuclear ribonucleoprotein (hnRNP) complexes are involved in the translocation, stability, and subcellular localization of mRNA and the regulation of its translation. Defects in these processes and mutations in components of the hnRNP complexes have been related to the formation of cytoplasmic inclusion bodies and neurodegenerative diseases. Despite much data on mRNA localization and evidence for protein synthesis, as well as the presence of translation machinery, in axons and presynaptic terminals, the identity of RNA-binding proteins involved in RNA transport and function in presynaptic regions is lacking. We previously characterized a strongly basic RNA-binding protein (p65), member of the hnRNPA/B subfamily, in squid presynaptic terminals. Intriguingly, in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), p65 migrated as a 65-kDa protein, whereas members of the hnRNPA/B family typically have molecular masses ranging from 35 to 42kDa. In this report we present further biochemical and molecular characterization that shows endogenous p65 to be an SDS-stable dimer composed of ∼37-kDa hnRNPA/B-like subunits. We cloned and expressed a recombinant protein corresponding to squid hnRNPA/B-like protein and showed its propensity to aggregate and form SDS-stable dimers in vitro. Our data suggest that this unique hnRNPA/B-like protein co-localizes with synaptic vesicle protein 2 and RNA-binding protein ELAV and thus may serve as a link between local mRNA processing and presynaptic function and regulation.

Exercise training decreases adipose tissue inflammation in cachectic rats.

Bearing in mind that cancer cachexia is associated with chronic systemic inflammation and that endurance training has been adopted as a nonpharmacological anti-inflammatory strategy, we examined the effect of 8 weeks of moderate intensity exercise upon the balance of anti- and pro-inflammatory cytokines in 2 different depots of white adipose tissue in cachectic tumour-bearing (Walker-256 carcinosarcoma) rats. Animals were assigned to a sedentary control (SC), sedentary tumour-bearing (ST), sedentary pair-fed (SPF) or exercise control (EC), exercise tumour-bearing (ET), and exercise pair-fed (EPF) group. Trained rats ran on a treadmill (60% VO(2)max) 60 min/day, 5 days/week, for 8 weeks. The retroperitoneal (RPAT) and mesenteric (MEAT) adipose pads were excised and the mRNA (RT-PCR) and protein (ELISA) expression of IL-1ß, IL-6, TNF-α, and IL-10 were evaluated. The number of infiltrating monocytes in the adipose tissue was increased in cachectic rats. TNF-α mRNA in MEAT was increased in the cachectic animals (p<0.05) in relation to SC. RPAT protein expression of all studied cytokines was increased in cachectic animals in relation to SC and SPF (p<0.05). In this pad, IL-10/TNF-α ratio was reduced in the cachectic animals in comparison with SC (p<0.05) indicating inflammation. Exercise training improved IL-10/TNF-α ratio and induced a reduction of the infiltrating monocytes both in MEAT and RPAT (p<0.05), when compared with ST. We conclude that cachexia is associated with inflammation of white adipose tissue and that exercise training prevents this effect in the MEAT, and partially in RPAT.

Sugar intake is correlated with adiposity and obesity indicators and sedentary lifestyle in Brazilian individuals with morbid obesity.

Obesity is a chronic disease characterized by increased accumulation of body fat. We evaluated the socioeconomic aspects, body composition, risk of metabolic complications associated with obesity, eating habits and lifestyle in both women and men adults and elderly with body mass index (BMI) > 40 kg/m². Among the subjects studied, 79% (n = 32) are female, 5% (n = 2) smokers, 39% (n = 16) use alcohol and only 24% (n = 10) are practitioners of physical exercise. The higher food intake was breads, followed by rice. The daily intake of fruits and vegetables is low. Positive correlation between consumption of sugar and BMI and abdominal circumference (AC) was observed. In summary, was found that morbidly obese patients that looking for nutritional counseling presents increased body fat, poor eating habits and sedentary lifestyle.

Purification and characterization of Ts15, the first member of a new α-KTX subfamily from the venom of the Brazilian scorpion Tityus serrulatus.

Voltage-gated potassium channel toxins (KTxs) are basic short chain peptides comprising 23-43 amino acid residues that can be cross-linked by 3 or 4 disulfide bridges. KTxs are classified into four large families: α-, ß-, γ- and κ-KTx. These peptides display varying selectivity and affinity for K(v) channel subtypes. In this work, a novel toxin from the Tityus serrulatus venom was isolated, characterized and submitted to a wide electrophysiological screening on 5 different subtypes of Na(V) channels (Na(V)1.4; Na(V)1.5; Na(V)1.6; Na(V)1.8 and DmNa(V)1) and 12 different subtypes of K(V) channels (K(V)1.1 - K(V)1.6; K(V)2.1; K(V)3.1; K(V)4.2; K(V)4.3; Shaker IR and ERG). This novel peptide, named Ts15, has 36 amino acids, is cross-linked by 3 disulfide bridges, has a molecular mass of 3956 Da and pI around 9. Electrophysiological experiments using patch clamp and the two-electrode voltage clamp techniques show that Ts15 preferentially blocks K(V)1.2 and K(V)1.3 channels with an IC50 value of 196 ± 25 and 508 ± 67 nM, respectively. No effect on Na(V) channels was observed, at all tested concentrations. Since Ts15 shows low amino acid identity with other known KTxs, it was considered a bona fide novel type of scorpion toxin. Ts15 is the unique member of the new α-Ktx21 subfamily and therefore was classified as α-Ktx21.1.

Exercise training reduces PGE2 levels and induces recovery from steatosis in tumor-bearing rats.

The effects of endurance training on PGE (2) levels and upon the maximal activity of hepatic carnitine palmitoyltransferase (CPT) system were studied in rats bearing the Walker 256 carciosarcoma. Animals were randomly assigned to a sedentary control (SC), sedentary tumor-bearing (ST), exercised control (EC), and as an exercised tumor-bearing (ET) group. Trained rats ran on a treadmill (60% VO (2) max) for 60 min/day, 5 days/week, for 8 weeks. We examined the mRNA expression (RT-PCR) and maximal activity (radioassay) of the carnitine palmitoyltransferase system enzymes (CPT I and CPT II), as well as the gene expression of fatty-acid-binding protein (L-FABP) in the liver. PGE (2) content was measured in the serum, in tumor cells, and in the liver (ELISA). CPT I and CPT II maximal activity were decreased (p<0.01) in ST when compared with SC. In contrast, serum PGE (2) was increased (p<0.05) in cachectic animals as compared with SC. In the liver, PGE (2) content was also increased (p<0.05) when compared with SC. Endurance training restored maximal CPT I and CPT II activity in the tumor-bearing animals (p<0.0001). Exercise training induced PGE (2) levels to return to control values in the liver of tumor-bearing training rats (p<0.05) and decreased the eicosanoid content in the tumor (p<0.01). In conclusion, endurance training was capable of reestablishing liver carnitine palmitoyltransferase (CPT) system activity associated with decreased PGE (2) levels in cachectic tumor-bearing animals, preventing steatosis.

Characterization and mRNA expression analysis of PI31, an endogenous proteasome inhibitor from Schistosoma mansoni.

The proline-rich inhibitor of 31 kDa (PI31) is highly conserved through metazoan evolution, and its activity in the proteasome inhibition is well-established although the precise mechanism of inhibition is unclear. The coding DNA sequence of Schistosoma mansoni PI31 (SmPI31) was cloned, and the recombinant protein was expressed in bacterial system. The correct amino acid sequence was confirmed by mass spectrometry and circular dichroism suggests that SmPI31 contains both α-helix and non-structured regions. Inhibition assays, using the Suc-Leu-Leu-Val-Tyr-4-MCA substrate for proteasome degradation, showed that the S. mansoni proteasome may be regulated by the inhibitory activity of SmPI31. A gene expression assay using qRT-PCR at various stages during the S. mansoni life cycle has shown that SmPI31 transcripts are expressed in all studied stages, suggesting that PI31 plays an important role during the developmental processes of the parasite. In this study first evidence is presented that PI31 has a conserved structure and plays a role as proteasome inhibitor in adult worms and it is expressed through life cycle.

Exercise training changes IL-10/TNF-alpha ratio in the skeletal muscle of post-MI rats.

Heart failure (HF) is associated with changes in the skeletal muscle (SM) which might be a consequence of the unbalanced local expression of pro- (TNF-alpha) and anti- (IL-10) inflammatory cytokines, leading to inflammation-induced myopathy, and SM wasting. This local effect of HF on SM may, on the other hand, contribute to systemic inflammation, as this tissue actively secretes cytokines. Since increasing evidence points out to an anti-inflammatory effect of exercise training, the goal of the present study was to investigate its effect in rats with HF after post-myocardial infarction (MI), with special regard to the expression of TNF-alpha and IL-10 in the soleus and extensor digitorum longus (EDL), muscles with different fiber composition. Wistar rats underwent left thoracotomy with ligation of the left coronary artery, and were randomly assigned to either a sedentary (Sham-operated and MI sedentary) or trained (Sham-operated and MI trained) group. Animals in the trained groups ran on a treadmill (0% grade at 13-20 m/min) for 60 min/day, 5 days/week, for 8-10 weeks. The training protocol was able to reverse the changes induced by MI, decreasing TNF-alpha protein (26%, P<0.05) and mRNA (58%, P<0.05) levels in the soleus, when compared with the sedentary MI group. Training also increased soleus IL-10 expression (2.6-fold, P<0.001) in post-MI HF rats. As a consequence, the IL-10/TNF-alpha ratio was increased. This "anti-inflammatory effect" was more pronounced in the soleus than in the EDL, suggesting a fiber composition dependent response.

A novel 65 kDa RNA-binding protein in squid presynaptic terminals.

A polyclonal antibody (C4), raised against the head domain of chicken myosin Va, reacted strongly towards a 65 kDa polypeptide (p65) on Western blots of extracts from squid optic lobes but did not recognize the heavy chain of squid myosin V. This peptide was not recognized by other myosin Va antibodies, nor by an antibody specific for squid myosin V. In an attempt to identify it, p65 was purified from optic lobes of Loligo plei by cationic exchange and reverse phase chromatography. Several peptide sequences were obtained by mass spectroscopy from p65 cut from sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) gels. BLAST analysis and partial matching with expressed sequence tags (ESTs) from a Loligo pealei data bank indicated that p65 contains consensus signatures for the heterogeneous nuclear ribonucleoprotein (hnRNP) A/B family of RNA-binding proteins. Centrifugation of post mitochondrial extracts from optic lobes on sucrose gradients after treatment with RNase gave biochemical evidence that p65 associates with cytoplasmic RNP complexes in an RNA-dependent manner. Immunohistochemistry and immunofluorescence studies using the C4 antibody showed partial co-labeling with an antibody against squid synaptotagmin in bands within the outer plexiform layer of the optic lobes and at the presynaptic zone of the stellate ganglion. Also, punctate labeling by the C4 antibody was observed within isolated optic lobe synaptosomes. The data indicate that p65 is a novel RNA-binding protein located to the presynaptic terminal within squid neurons and may have a role in synaptic localization of RNA and its translation or processing.

Physico-chemical and antifungal properties of protease inhibitors from Acacia plumosa.

This study was aimed at investigating the purification, biological activity, and some structural properties of three serine protease inhibitors isoforms, denoted ApTIA, ApTIB, and ApTIC from Acacia plumosa Lowe seeds. They were purified from the saline extract of the seeds, using Superdex-75 gel filtration and Mono-S ion exchange chromatography. They were further investigated by mass spectrometry, spectroscopic measurements, surface plasmon resonance, and inhibition assays with proteases and phytopathogenic fungi. The molecular mass of each isoform was estimated at ca. 20 kDa. Each contained two polypeptide chains linked by a disulfide bridge, with different isoelectric points that are acidic in nature. The N-terminal sequences of both chains indicated that they were Kunitz-type inhibitors. Circular dichroism (CD) analyses suggested the predominance of both disordered and beta-strands on ApTI isoforms secondary structure, as expected for beta-II proteins. In addition, it was observed that the proteins were very stable, even at either extreme pH values or at high temperature, with denaturation midpoints close to 75 degrees C. The isoinhibitors could delay, up to 10 times, the blood coagulation time in vitro and inhibited action of trypsin (Ki 1.8 nM), alpha-chymotrypsin (Ki 10.3 nM) and kallikrein (Ki 0.58 microM). The binding of ApTIA, ApTIB, and ApTIC to trypsin and alpha-chymotrypsin, was investigated by surface plasmon resonance (SPR), this giving dissociation constants of 0.39, 0.56 and 0.56 nM with trypsin and 7.5, 6.9 and 3.5 nM with alpha-chymotrypsin, respectively. The growth profiles of Aspergillus niger, Thielaviopsis paradoxa and Colletotrichum sp. P10 were also inhibited by each isoforms. These three potent inhibitors from A. plumosa may therefore be of great interest as specific inhibitors to regulate proteolytic processes.

Endurance training induces depot-specific changes in IL-10/TNF-alpha ratio in rat adipose tissue.

White adipose tissue (WAT) is the source of pro- and anti-inflammatory cytokines and recently, it has been recognized as an important source of interleukin 10 (IL-10). Acute physical exercise is known to induce an anti-inflammatory cytokine profile, however, the effect of chronic physical exercise on the production of IL-10 by WAT has never been examined. We assessed IL-10 and TNF-alpha concentration in WAT of rats engaged in endurance training. Animals were randomly assigned to either a sedentary control group (S, n=7) or an endurance trained group (T, n=8). Trained rats ran on a treadmill 5 days/wk for 8 wk (55-65% VO(2max)). Detection of IL-10 and TNF-alpha protein and mRNA expression, as well as the gene expression of PPAR-gamma, and immunocytochemistry to detect mononuclear phagocytes were carried out. A reduction in absolute retroperitoneal adipose tissue (RPAT) weight in T (44%; p<0.01), when compared with S was observed. IL-10 concentration was increased (1.5-fold, p<0.05), to a higher extent than that of TNF-alpha (66%, p<0.05) in the mesenteric adipose tissue (MEAT) of the trained group, while no change related to training was observed in RPAT. In MEAT, IL-10/TNF-alpha ratio was increased in T, when compared with S (30%; p<0.05). PPAR-gamma gene expression was increased in T (1.1-fold; p<0.01), when compared with S in the same adipose depot. No monocyte infiltration was found. In conclusion, exercise training induced increased IL-10 expression in the mesenteric depot, resulting in a modified IL-10/TNF-alpha ratio. We also conclude that WAT presents a depot-specific response to endurance training regarding the studied aspects.

Acute-phase protein alpha-1-acid glycoprotein mediates neutrophil migration failure in sepsis by a nitric oxide-dependent mechanism.

The reduction of circulating neutrophil migration to infection sites is associated with a poor outcome of severe sepsis. alpha-1-Acid glycoprotein (AGP) was isolated from the sera of severely septic patients by HPLC and acrylamide gel electrophoresis and identified by mass spectrometry. Both the isolated protein and commercial AGP inhibited carrageenin-induced neutrophil migration into the rat peritoneal cavity when administered i.v. at a dose of 4.0 microg per rat (95 pmol per rat). Analysis by intravital microscopy demonstrated that both proteins inhibited the rolling and adhesion of leukocytes in the mesenteric microcirculation. The inhibitory activity was blocked by 50 mg/kg aminoguanidine, s.c., and was not demonstrable in inducible nitric oxide synthase (iNOS) knockout mice. Incubation of AGP with neutrophils from healthy subjects induced the production of NO and inhibited the neutrophil chemotaxis by an iNOS/NO/cyclic guanosine 3,5-monophosphate-dependent pathway. In addition, AGP induced the l-selectin shedding by neutrophils. The administration of AGP to rats with mild cecal ligation puncture sepsis inhibited neutrophil migration and reduced 7-day survival from approximately 80% to 20%. These data demonstrate that AGP, an acute-phase protein, inhibits neutrophil migration by an NO-dependent process and suggest that AGP also participates in human sepsis.

Isolation of bovine immunoglobulins resistant to peptic digestion: new perspectives in the prevention of failure in passive immunization of neonatal calves.

In calves, neonatal mortality and disease susceptibility are greatly influenced by failure in passive immunization, normally provided by colostrum ingestion just after birth. Formulations projected to replace natural colostrum have not been successful, and one of the possible reasons for such failure is that orally administered Ig are probably digested in the gastrointestinal tract, so they are not absorbed as intact functional molecules. With the aim of finding an adequate colostrum substitute, we used columns of immobilized jacalin, a lectin known by its ability to bind O-linked oligosaccharides, to obtain a colostral Ig population putatively protected against enzymatic cleavage by the presence of sugar chains. Immunoglobulin G1 is a major constituent of colostrum Ig bound to jacalin (JB-Ig). This preparation contains 10% of the total colostral Ig and is typically 3 to 6 times more resistant to pepsin digestion than the Ig contained in the fraction that is not bound to jacalin, which presumably does not contain O-glycans. Mass spectrometry analysis demonstrated that the tryptic peptides obtained from JB-Ig and unbound Ig were similar, indicating that their distinct susceptibility to enzyme hydrolysis was associated with differences in their sugar chains. Therefore, the present research suggests that the bovine colostrum JB-Ig has potential application in the immunotherapy of neonatal calves that have not been supplied with colostrum.

Influence of dissolved oxygen and shear conditions on clavulanic acid production by Streptomyces clavuligerus.

Clavulanic acid (CA), a potent beta-lactamase inhibitor, is produced by a filamentous bacterium. Here, the effect of DO and shear, expressed as impeller tip velocity, on CA production was examined. Cultivations were performed in a 4 L fermentor with speeds of 600, 800 and 1,000 rpm and a fixed air flow rate (0.5 vvm). Also, cultivation with automatic control of dissolved oxygen, at 50% air saturation, by varying stirrer speed and using a mixture of air and O(2) (10% v/v) in the inlet gas, and a cultivation with fixed stirrer speed of 800 rpm and air flow rate of 0.5 vvm, enriched with 10% v/v O(2), were performed. Significant variations in CA titer, CA production rate and O(2) uptake-rate were observed. It was also found that the DO level has no remarkable effect on CA production once a critical level is surpassed. The most significant improvement in CA production was related to high stirrer speeds.

Pleomorphic adenoma and carcinoma ex pleomorphic adenoma: immunohistochemical demonstration of the association between tenascin expression and malignancy.

AIMS: To investigate tenascin expression in salivary gland tumours. Tenascin is a matricellular protein that has been studied in several tumour types. Its expression has been correlated with tumour morphogenesis as well as with local invasiveness and tumour metastatic behaviour. METHODS AND RESULTS: The distribution pattern of tenascin in a series of 63 pleomorphic adenomas (PA) and 20 carcinomas ex- pleomorphic adenoma (Ca ex PA) was studied immunohistochemically. Ten normal adult salivary glands were used as controls. Tenascin surrounded the excretory ducts of normal adult salivary gland tissue. It was absent in the basement membrane compartment of both benign and malignant mixed tumours. In the interstitial compartment of the extracellular matrix, the fibro-hyaline type expressed tenascin in a statistically significantly (P < 0.001) lower number of PA cases (25%) in comparison with both malignant and benign areas of Ca ex PA (75% and 90%, respectively). In the Ca ex PA group, a statistically significantly difference (P < 0.001) was found in the frequency of tenascin deposits around aggregates of neoplastic cells between metastasizing (73%) and non-metastasizing neoplasms (0%). CONCLUSIONS: These findings strongly support the hypothesis that tenascin deposition is involved in the mechanisms of malignant transformation of pleomorphic adenomas into carcinomas as well as being associated with clinical disease progression.