Yellow field peas were subjected to soaking (SY), germination (SGY), fermentation (SFY), fermentation and pasteurization (SFPY), combined germination and fermentation (SGFY), as well as SGFY upon ...pasteurization (SGFPY). Soaking generally had a negligible impact on the proximate composition and nutritive quality of field peas except that a 23.4% reduction in trypsin inhibitor activity (TIA) was observed. Both germination and fermentation led to a significant reduction in starch content by 11.2–11.7%. Processed peas including SFY, SFPY, SGFY, and SGFPY exhibited higher in vitro protein digestibility (IVPD) values (P < 0.05) of 87.80–89.16%, which was further strengthened by the negative correlation between IVPD and antinutrients including tannin and TIA. The electrophoretic profile of their protein hydrolysates confirmed the IVPD results where the above four treatments resulted in an extensive degradation and breakdown of pea seed's storage proteins with the peptide subunits appeared at molecular weight <16 kDa. SFY, SFPY, SGFY, and SGFPY also showed higher levels of digestible starch (114.1–257.7 g/kg) which agreed with their DSC gelation temperatures and the most distinguishable alternations on their microstructure. The fermentation of germinated peas led to an enhanced nutritional property, the following pasteurization process further changed the starch crystalline organization and protein resembling structure of field peas.
•Germination is less effective than fermentation in affecting pea composition.•Combined germination and fermentation greatly enhanced nutritional profile of peas.•The fermentation of germinated peas upon pasteurization led to highest digestibility.•Variation in protein digestibility was verified by electrophoresis.•Peas with higher digestible starch exhibit distinguishable microscopical changes.
Previous research has demonstrated the effect of enzymatic hydrolysis on the emulsifying properties of soy protein isolate (SPI). The present authors previously reported on the preparation of stable ...emulsions by the addition of oil and vinegar to a commercial SPI hydrolysate. In this paper, emulsions prepared by the direct introduction of fruit juices from several tropical fruits to SPI were examined. The purpose of this research was to elucidate whether the production of stable emulsions such as those made with the proteolytic enzyme papain was possible by this method. Stable emulsions were obtained from papain and pineapple, kiwi, melon, fig, and unripe papaya fruit juices. The appearance of the emulsions, the SDS-PAGE patterns of hydrolyzed SPI, and the pseudo-plastic behavior of the emulsions varied depending on the type of fruit, fruit production area, fruit ripeness, the reactive conditions of the fruit juices, and SPI. A relationship between the decrease in 11S acid subunits and emulsion stability was suggested.
Saskatchewan grown yellow field pea was subjected to different processing conditions including dehulling, micronization, roasting, conventional/microwave cooking, germination, and combined ...germination and conventional cooking/roasting. Their nutritional and antinutritional compositions, functional properties, microstructure, thermal properties, in vitro protein and starch digestibility, and protein composition were studied. Processed field peas including conventional cooked yellow peas (CCYP), microwave cooked yellow peas (MCYP), germinated-conventional cooked yellow peas (GCCYP), and germinated-roasted yellow peas (GRYP) exhibited the significantly higher in vitro protein digestibility (IVPD), which was in accordance with their significantly lower trypsin inhibitor activity and tannin content. The SDS-PAGE and size exclusion HPLC profiles of untreated pea proteins and their hydrolysates also confirmed the IVPD result that these four treatments facilitated the hydrolysis of pea proteins to a greater extent. The CCYP, MCYP, GCCYP, and GRYP also exhibited significantly higher starch digestibility which was supported by their lower onset (To), peak (Tp), and conclusion (Tc) temperatures obtained from DSC thermogram, their lower pasting properties and starch damage results, as well as their distinguished amorphous flakes' configuration observed on the scanning electron microscopic image. LC/ESI-MS/MS analysis following in-gel digests of SDS-PAGE separated proteins allowed detailed compositional characterization of pea proteins. The present study would provide fundamental information to help to better understand the functionality of field peas as ingredients, and particularly in regards to agri-food industry to improve the process efficiency of field peas with enhanced nutritional and techno-functional qualities.
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•Partial removing of antinutrients in pea lead to higher protein/starch digestibility•SDS-PAGE profile of pea proteins is altered due to hydrothermal treatments applied.•Peas with improved starch digestibility after heating exhibit amorphous flakes on SEM.•LC/ESI-MS/MS analysis allows detailed compositional characterization of pea proteins.
Tests for measuring immunoglobulins include quantitative immunoglobulin assays, serum protein fractionation tests, immunoelectrophoresis, and immunofixation electrophoresis. The presence of abnormal ...immunoglobulins is suspected when abnormal bands are detected in electrophoresis or when discrepant data appear between tests. Abnormal immunoglobulins include heavy chain disease proteins and half-molecule immunoglobulins that show molecular defects. Such abnormal proteins can be molecularly qualified by SDS-PAGE or immunoblotting. Bence Jones proteins with microheterogenicity can also be demonstrated as the migration changes by immunofixation electrophoresis of samples before and after enzyme treatment. Furthermore, after agarose membrane electrophoresis, the protein can be spontaneously transferred to a polyvinylidene difluoride (PVDF) membrane for easy immunoblotting. In other words, electrophoresis can be used for both detection and analysis of abnormal immunoglobulins.
Methicillin resistant Staphylococcus aureus (MRSA) is a pathogen to humans causing life-threatening infections. MRSA have the capability to grow resistance to many antibiotics, and phage therapy is ...one treatment option for this infection.
The aim of the present study was to isolate and characterize the lytic bacteriophages specific to MRSA from domestic sewage water at a tertiary care hospital in Egypt.
Thirty MRSA strains were isolated from different clinical samples admitted to the microbiology lab at Theodor Bilharz Research institute (TBRI) hospital, Giza, Egypt. They were confirmed to be MRSA through phenotypic detection and conventional PCR for mecA gene. They were used for the isolation of phages from sewage water of TBRI hospital. Plaque assay was applied to purify and quantify the titer of the isolated phages. The host range of the isolated phages was detected using the spot test assay. The morphology of phages was confirmed using transmission electron microscope (TEM). Digestion of DNA extracted from phages with endonuclease enzymes including EcoRI and SmaI was performed. SDS-PAGE was performed to analyze MRSA specific phage proteins. As a positive control prophages were isolated from a mitomycin C (MitC) treated culture of S. aureus strain ATCC25923. Further characterization using conventional polymerase chain reaction (PCR) was used to select three known Staphylophages by detecting the endolysin gene of phage K, the polymerase gene of phage 44AHJD, and the minor tail gene of phage P68.
Isolated phages in this research displayed a wide host range against MRSA using the spot test, out of thirty tested MRSA isolates 24 were sensitive and got lysed (80%). The titer of the phages was estimated to be 1.04 × 106 pfu/ml using plaque test. Identification of head and tail morphology of the phages was achieved using TEM and they were designated to tailed phages of order Caudovirales, they composed an icosahedral capsid. Prophages were isolated through MitC induction. DNA of phages was digested by endonuclease enzymes. Conventional PCR yielded 341 bp of phage K endolysin gene and phage P68 minor tail protein gene 501 bp. Protein analysis using SDS-PAGE showed 4 proteins of sizes between 42 kDa and 140 kDa.
Phages isolated here are alike to others mentioned in previous studies. The high broad host range of the isolated phages is promising to control MRSA and can be in the future commercially suitable for treatment as lysate preparations. Animal models of phage-bacterial interaction will be our next step that may help in resolving the multidrug resistant crisis of MRSA in Egypt.
Previous research has demonstrated the effect of enzymatic hydrolysis on the emulsifying properties of soy protein isolate (SPI). The present authors previously reported on the preparation of stable ...emulsions by the addition of oil and vinegar to a commercial SPI hydrolysate. In this paper, emulsions prepared by the direct introduction of fruit juices from several tropical fruits to SPI were examined. The purpose of this research was to elucidate whether the production of stable emulsions such as those made with the proteolytic enzyme papain was possible by this method. Stable emulsions were obtained from papain and pineapple, kiwi, melon, fig, and unripe papaya fruit juices. The appearance of the emulsions, the SDS-PAGE patterns of hydrolyzed SPI, and the pseudo-plastic behavior of the emulsions varied depending on the type of fruit, fruit production area, fruit ripeness, the reactive conditions of the fruit juices, and SPI. A relationship between the decrease in 11S acid subunits and emulsion stability was suggested.
Excessive infection and inflammation are the most common complications associated with castration. The objective of this study was to compare the efficacy of flunixin meglumine (FM), meloxicam (MX), ...or firocoxib (FX) for inflammation control after castration in horses using acute-phase proteins (APP) as markers of inflammation. Thirty healthy, unbroken, mixed-breed horses (body weight 358.62±45.57kg and age 4.99±2.63 years) were randomly (n=10 animals/group) allocated to receive one of three different post-castration anti-inflammatory medicines: Group 1 (FM 1.1mg/kg bwt, IV, s.i.d for 5 days); Group 2 (MX 0.6mg/kg bwt, IV, s.i.d for 5 days); and Group 3 (FX 0.1mg/kg bwt, IV, s.i.d for 5 days). All horses were castrated in standing position, using the open technique. Serum and peritoneal APP concentrations were measured by sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis (PAGE) and determined before castration (0), and 3, 5, 24, 48, 72, 120 and 168 hours after castration. The results were submitted to analysis of variance using the SAS statistical program, and means were compared by the Student-Newman-Keuls test (p<0.05). Three animals from the MX group developed hyperthermia (with rectal temperatures of 39.8, 39.3 and 38.9°C on day 4, 5 and 6, respectively) and showed local clinical signs of inflammation (inguinal and excessive scrotal edema) and reluctance to walk, as well as a rigid gait of the hind limbs. The same complications were observed in one FX horse. No complications were observed among the FM animals. The castration resulted in significant changes in serum and peritoneal values of total proteins, ceruloplasmin (Cp), transferrin (Tf), albumin (Alb), haptoglobin (Hp) and α1-acid glycoprotein (Gp) in animals of all experimental groups. However, the animals of the MX and FX groups presented more intense acute phase response compared to the animals of the FM group. Changes in the APP were associated with the surgical trauma of castration, but the differences between groups were associated with the ability of the nonsteroidal anti-inflammatory drug to control the inflammation. In conclusion, and based on the findings of acute phase proteins, flunixin is more efficient to control the magnitude of inflammation following castration as compared to meloxicam and firocoxib.
RESUMO: Infecção e inflamação excessivas são as complicações mais comuns associadas à castração. O objetivo deste estudo foi comparar a eficácia do flunixin meglumine (FM), meloxicam (MX) ou firocoxib (FX) no controle da inflamação após a castração em cavalos usando proteínas da fase aguda (APP) como marcadores de inflamação. Trinta equinos saudáveis (358,62±45,57kg; 4,99±2,63 anos) foram em função dos anti-inflamatórios utilizados após as castrações aleatoriamente (n= 10 animais/grupo) alocados em três diferentes grupos: Grupo 1 (FM 1,1mg/kg de peso, IV, sid por 5 dias); Grupo 2 (MX 0,6mg/kg de peso, IV, s.i.d por 5 dias); e Grupo 3 (FX 0,1mg/kg de peso, IV, s.i.d por 5 dias). Todos os cavalos foram castrados em posição quadrupedal, utilizando a técnica aberta. As concentrações de APP sérica e peritoneal foram separadas por eletroforese em gel de poliacrilamida (PAGE) com dodecil-sulfato de sódio (SDS) e determinadas no momento 0 (antes da castração) e com 3, 5, 24, 48, 72, 120 e 168 horas após a castração. Os resultados foram submetidos à análise de variância pelo programa estatístico SAS e as médias foram comparadas pelo teste de Student-Newman-Keuls (p<0,05). Três animais do grupo MX desenvolveram hipertermia (com temperatura retal de 39,8, 39,3 e 38,9° C nos dias 4, 5 e 6, respectivamente) e mostraram sinais clínicos locais de inflamação (edema inguinal e escrotal excessivo) e relutância em andar, bem como marcha rígida dos membros posteriores. As mesmas complicações foram observadas em um cavalo do FX. Não foram observadas complicações entre os animais do FM. Independente do grupo, a castração resultou em alterações significativas nos valores séricos e peritoneais de proteínas totais, ceruloplasmina (Cp), transferrina (Tf), albumina (Alb), haptoglobina (Hp) e glicoproteína ácida α1 (Gp). No entanto, os animais dos grupos MX e FX apresentaram resposta de fase aguda mais intensa quando comparados aos animais do FM. Alterações na resposta de fase aguda deveram-se ao trauma cirúrgico da castração, mas as diferenças entre os grupos foram associadas à capacidade do anti-inflamatório em controlar a inflamação. Em conclusão, baseado da resposta de fase aguda, o flunixin em comparação com o meloxicam e o firocoxib é mais eficiente no controle da inflamação após a castração em equinos.
The acute-phase response (APR) is a rapid, nonspecific, systemic response occurring secondary to skeletal muscle damage and might be a protective physiological mechanism. The response has a number of ...components including increases in serum concentrations of the acute-phase proteins (APPs), which are primarily produced in the liver. This study was designed to evaluate the influence of marcha gait exercise on serum concentrations of the immunoglobulin A (IgA), immunoglobulin G (IgG), ceruloplasmin, transferrin, albumin (Alb), α1-antitripsin, haptoglobin (Hp), and α1-acid glycoprotein in 35 Mangalarga Marchador horses (18 geldings and 17 mares). The study also aimed to evaluate the differences between genders of the response to marcha gait exercise. Blood samples were assessed before and after official marcha contest. Acute-phase proteins were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The results were submitted to analysis of variance using the SAS statistical program, and means were compared by Student-Newman-Keuls test (P < .05). In both geldings and mares, APP concentrations did not increase in response to exercise. Significant differences of total serum protein concentration, Alb, Hp, IgA, and IgG between geldings and mares were noticed. In conclusion, this study provides evidence that marcha gait exercise performed during official contest is not intense enough to stimulate an APR, demonstrated by no change in serum concentrations of APPs in Mangalarga Marchador horses. The differences between genders in the present study warrant further investigation.
•Physical exercise induces various stress responses and metabolic adaptations.•Exercise elicits a short-lived acute-phase response in horses.•The marcha exercise does not change the levels of acute-phase proteins in Mangalarga Marchador horses.•In athletic horses, gender differences should be taken into consideration.
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