Evaluation of Phytase Impact on Protein and Phosphorus Bioaccessibility of Two Lupin Species for Rainbow Trout ().

Legumes are an important source of protein, lipids, and other essential nutrients. As the demand for protein and lipids continues to surge on a global scale, there is a growing interest in incorporating legumes into aquafeeds. This shift is driven not only by the escalating growth of the aquaculture sector in recent years but also by the imperative to diminish the dependency on traditional resources like fishmeal (FM) and fish oil.

Multispecies biofilm removal by XP-endo Finisher and passive ultrasonic irrigation: A scanning electron microscopy study.

The aim was to assess the effect of XP-endo Finisher (XPF) on multispecies biofilm removal, in comparison with passive ultrasonic irrigation (PUI) and conventional syringe irrigation (CSI), by scanning electron microscope (SEM). Fifty mandibular first premolars were instrumented, longitudinally sectioned. The split halves were incubated for 4 days with a broth obtained from three bacteria strains: Enterococcus faecalis, Eikenella corrodens and Streptococcus anginosus.

Effectiveness of different disinfection techniques of the root canal in the elimination of a multi-species biofilm.

Background: The purpose of the study was to evaluate the effectiveness of different root canal disinfection techniques in the elimination of a multi-species biofilm from inside the root canal.

Material And Methods: Fifty mandibular first premolars were used in the present study, standardized to 11mm of root length, and instrumented with a reciprocation system Reciproc, (VDW GmbH, Munich, Germany) to a #50. Longitudinally sectioned halves of the roots were obtained and washed with NaOCl 4%, EDTA 17% and 5% sodium thiosulfate, and sterilized by autoclaving for 15 minutes at 121°C.

Estimation of type-specific anti-streptococcal M-protein antibodies with biotinylated peptides in human sera.

The design of a novel enzyme-linked immunosorbent assay for the estimation of antibodies directed against the N-terminus of the M-protein of Streptococcus pyogenes is described. The ELISA employs biotinylated peptide antigens of the types 1, 4, 12 and 19 immobilized by (strept-)avidin on the surface of polystyrene microtiter wells. In rabbit hyperimmune sera and in human serum samples, antibodies against the corresponding serotype could be detected with high sensitivity and specificity.

The effect of neuropeptides from Limulus on its circadian rhythm in retinal sensitivity.

Retinal responses of the Limulus lateral eyes to light are greater at night than during the day. A circadian clock in the brain of the horseshoe crab controls these rhythmic changes of light sensitivity. The increase in sensitivity (as measured by the amplitude of the electroretinogram) is mediated at least in part by octopamine that is released from efferent axons terminating in the visual cells.

Interaction of guanosine 5'-triphosphate binding protein Gq from Sepia officinalis with illuminated rhodopsin bound to concanavalin A.

The heterotrimeric guanosine 5'-triphosphate (GTP)-binding protein Gq was suggested to couple the light receptor rhodopsin with the effector phospholipase C in visual cells of invertebrates. We indirectly linked Gq from Sepia officinalis to a concanavalin A-sepharose column via rhodopsin. Rhodopsin had been solubilized previously with 10 mM n-dodecyl-beta-maltoside from the purified photosensory membrane under illumination.

Epitope mapping of a C5a neutralizing mAb using a combined approach of phage display, synthetic peptides and site-directed mutagenesis.

Background: The anaphylatoxin C5a is a powerful proinflammatory protein generated on activation of the complement system. Recently, we described an anti-hC5a neoepitope specific mAb, mAb 2925, which was raised against the nonapeptide ISHKDMQLG (C5a-(65-73). This mAb is unique in that it recognizes both hC5a and hC5adesArg, even when it is denatured.

Cardioinhibitory peptides from Limulus polyphemus: modulation of the neurogenic heart.

Four neuropeptides have been isolated and sequenced from acetone extracts of brains of the horseshoe crab Limulus polyphemus. They belong to a newly discovered peptide family in invertebrates. A possible role of the four peptides from Limulus as cardioregulatory neurotransmitters has been tested on the isolated Limulus heart.

Rapid quantification of C3a and C5a using a combination of chromatographic and immunoassay procedures.

Monoclonal antibodies were isolated which reacted specifically with the complement cleavage products C3a, C3adR, C5a, and C5adR but not with the parent molecules C3 or C5. In both cases the mAbs showed a higher affinity towards the desArg forms. These mAbs were used as capture antibodies in immunoassays for C3a/C3adR and C5a/C5adR.

Evaluation of the C-terminal C5a effector site with short synthetic C5a analog peptides.

Biological activities have been determined for a series of 18 peptides based on the C-terminal sequence of human or rat C5a. Lysosomal enzyme release was tested in two cell types, the promyelotic leukemia cell line U937 and polymorphonuclear leukocytes. In addition, an ATP-release assay with guinea pig platelets was performed.

Synthetic peptides as antagonists of the anaphylatoxin C3a.

Peptide compounds resembling the receptor-binding C-terminal domain of the anaphylatoxic peptide C3a were synthesized to examine two kinds of C3a antagonism: (a) specific desensitization of C3a-sensitive cells and (b) competitive binding to the C3a receptor. We used guinea-pig platelets, which express a C3a receptor and specifically release ATP upon stimulation, to evaluate the actions of the C3a analogues. The ATP liberation can be inhibited by pretreatment (i.

Enhanced biopotency of synthetic C3a analogues by membrane binding. A fluorescence anisotropy decay study.

The biological activity of oligopeptide analogues of C3a is markedly increased by N-terminal attachment of a hydrophobic group as, for instance, 9-fluorenylmethoxycarbonyl (Fmoc), either direct or via a flexible 6-aminohexanoyl (Ahx) spacer. This study presents evidence from fluorescence anisotropy decay measurements that the hydrophobic appendix mediates non-specific binding of the synthetic peptide analogues to phospholipid vesicles. According to quantitative considerations no alternative or additional rate-enhancing mechanisms other than surface diffusion are required to account for the gain in biopotency.

Molecular epitope identification by limited proteolysis of an immobilized antigen-antibody complex and mass spectrometric peptide mapping.

Sequences of antigenic determinants were identified by limited proteolysis of peptide antigens bound to an immobilized monoclonal antibody and direct molecular weight determination of the monoclonal antibody-bound peptide fragments by 252Cf plasma desorption mass spectrometry. The epitope peptides to the monoclonal antibody h453 [Burger, R., Zilow, G.

Studies on the total synthesis of an A7,B7-dicarbainsulin. III. Assembly of segments and generation of biological activity.

As a further contribution to the synthesis of an insulin analogue with a stable A7-B7 interchain bond, the synthesis of A(8-21) by solution methods, and of B(9-25) as well as [7-(2,7-diaminosuberic acid)]B(1-8) by solid phase methods is described. In the latter compound, the amino group of the diaminosuberic acid residue was acylated with A(1-6), and the resulting "U-peptide" sequentially elongated with the C-terminal A- and finally B-chain sequences. The conversion of the product into the disulfide moiety gave a mixture which could not be resolved by currently available methods.

Identification of residues in the insulin molecule important for binding to insulin-degrading enzyme.

Insulin-degrading enzyme (IDE) hydrolyzes insulin at a limited number of sites. Although the positions of these cleavages are known, the residues of insulin important in its binding to IDE have not been defined. To this end, we have studied the binding of a variety of insulin analogues to the protease in a solid-phase binding assay using immunoimmobilized IDE.

Reevaluation of the C3a active site using short synthetic C3a analogues.

In 1980 the C-terminal pentapeptide LGLAR (C3a 73-77) was described (Caporale, L. H. et al.

Characterization of C3a receptor-proteins on guinea pig platelets and human polymorphonuclear leukocytes.

The expression of specific membrane receptors for C3a was determined on guinea pig C3a-sensitive (gp R+) platelets and human polymorphonuclear leukocytes (hu PMNL). Binding studies with 125I-labeled C3a from gp or hu sources and Scatchard analysis applied to the binding data revealed the existence of two receptor classes on gp R+ platelets; a high-affinity class with about 200 binding sites/cell and Kd = 1.7 x 10(-9) M, and a relatively low-affinity class with Kd = 10(-8) M and about 500 sites/cell.

Peptide analogues of the anaphylatoxin C3a; syntheses and properties.

The chemical syntheses of C-terminally shortened analogues of C3a, which is the best investigated anaphylatoxin and derives from the third component of complement system, is reported. The peptide assembly was performed with the solid-phase technique using a polyamide support and an orthogonal protection strategy. The base-labile Fmoc group was chosen for N alpha protection in combination with acid-labile side-chain protection.

Design and biological activity of a new generation of synthetic C3a analogues by combination of peptidic and non-peptidic elements.

Based on published X-ray crystallographic data of the anaphylatoxic complement peptide C3a, we have synthesized a series of peptides with appropriate amino acid exchanges and a maximal length of 13 amino acids. N-terminal acylation of these optimized structures with epsilon-aminohexanoic acid and complex aromatic structures like fluorenylmethoxycarbonyl, 2-nitro-4-azidophenyl, fluoresceinyl and rhodaminyl leads to a dramatic increase in biological activity. The culmination of our synthetic efforts is a C3a analogue with 13 amino acid residues and a biological activity six times that of native C3a.

Shortened insulin with enhanced in vitro potency.

After it has been shown that removal of residues B26-B30 leaves insulin with full biological activity, provided the new C-terminus is amidated (Fischer et al. (1985) Biol. Chem.

Immunostimulating hexapeptide from human casein: amino acid sequence, synthesis and biological properties.

A hexapeptide obtained from human casein by enzymatic digestion has been purified, sequenced and synthesized; its structure is: Val-Glu-Pro-Ile-Pro-Tyr. In vitro this hexapeptide stimulates the phagocytosis of opsonized sheep red blood cells by murine peritoneal macrophages. Administered intravenously to adult mice, it enhances the resistance to infection with Klebsiella pneumoniae.

Characterization of two anti-human parathyrin antisera.

Two anti-human parathyrin antisera were raised in sheep. These were characterized by radioimmunoassay using two commercially available bovine parathyrin preparations and one synthetic human parathyrin fragment (sequence 42-55 (42-Tyr)) for radioiodination. In addition, four synthetic human parathyrin fragments (sequences 1-34, 32-43, 434-68, 53-84), one bovine parathyrin peptide (sequence 28-48) and a human parathyrin standard from a tissue culture containing the intact hormone were utilized in a competitive inhibition assay against the two radiolabelled bovine parathyrin preparations.

[Synthesis of two median segments of human parathyrin (author's transl)].

Human parathyrin-(32-43)-dodecapeptide (H-His-Asn-Phe-Val-Ala-Leu-Gly-Ala-Pro-Leu-Ala-Pro-OH) and tyrosyl-[human parathyrin-(43-55)-tridecapeptide] (H-Tyr-Pro-Arg-Asp-Ala-Gly-Ser-Gln-Arg-Pro-Arg-Lys-Lys-Glu-OH) were prepared, using the strategy of segment condensation in combination with acid-labile protecting groups. The peptides will be utilized for radioimmunoassay and receptor-binding studies.

Characterisation of the binding sites of anti-parathyroid hormone antisera using synthetic parathyroid hormone peptides.

Four antisera raised against partly purified PTH preparations all showed a wide range of specificities when reacting with radioiodinated PTH peptides representing several different portions of the intact hormone sequence. In contrast, antisera raised against individual peptides were only able to cross-react with other peptides that contained all or part of their amino acid sequence in common. Cross-reacting peptides were seen to contain one or more amino acid residues having high interspecies variability in common.

Homologous radioimmunoassay for human mid-regional parathyroid hormone.

A radioimmunoassay, selective for the clinically important mid region of human parathyroid hormone (hPTH), is reported. The synthetic 44-68 amino acid sequence (h44-68PTH) was used as both the standard and tracer material. This eliminated many of the undesirable characteristics associated with PTH assays the employ hormone of biological origin.