Lower Total Protein Concentrations Research Articles

Comparison of hematologic variables among Warmbloods, Thoroughbreds, and Western stock horse breeds.

Hematology is a diagnostic tool used to evaluate the health status of horses. However, breed differences are often not considered. The objective was to compare complete blood count variables among Warmbloods, Thoroughbreds, and stock horses (SH). Ninety-six healthy horses were grouped by breed (Warmbloods, Thoroughbreds, and SH). Samples were collected through venipuncture for complete blood count analysis. One-way ANOVA with Tukey's tests or Kruskal-Wallis with Dunn's post hoc tests were used to compare hematologic variables among groups. Warmbloods had a significantly lower total white blood cell (WBC) count (6.08 ± 1.11 × 109/L) and lymphocyte count (1.76 ± 0.41 × 109/L) than Thoroughbreds (7.28 ± 1.45; 2.28 ± 5.16 × 109/L, respectively; P < .001) and SH (7.21 ± 1.18 × 109/L, P < .01; 2.10 ± 5.17 × 109/L; P < .05). Warmbloods had a significantly lower red blood cell count (7.7 ± 0.8 × 1012/L) and higher mean corpuscular volume (MCV, 49.4 ± 2.2 fL) than Thoroughbreds (8.42 ± 1.2 × 1012/L, P < .01; 47.3 ± 3.0 fL). Warmbloods had lower MCVs than SH (49.4 ± 2.2 vs 51.2 ± 2.6 fL). The mean cell hemoglobin concentration (MCHC) was higher in Warmbloods (35.0, 33.8-36.2 g/dL) and Thoroughbreds (34.9, 33.4-35.7 g/dL) than in SH breeds (34.0, 33.4-35.4 g/dL; P < .001, both). Total protein concentrations were significantly lower in Thoroughbreds (67, 59-80 g/L) compared with SH (71, 64-83 g/dL) (P < .05). Warmbloods had decreased WBC and lymphocyte counts compared with Thoroughbreds and SH, and Thoroughbreds had increased red blood cell counts. Thoroughbreds had lower total protein concentrations than SH. Clinicians should consider breed differences when interpreting hematologic values.

Efeitos dos fatores maternos e da estação do ano sobre a saúde, o crescimento e os parâmetros reprodutivos de novilhas Holandesas jovens

ABSTRACT This observational study describes the short- and long-term effects of maternal factors in 226 Holstein heifers and their offspring. We obtained maternal data and information from the dairy register. Young heifers were assessed in the window of 60 to 105 days of life to perform the Calf Health Scoring System and performance index, and blood samples to measure inflammatory biomarkers. Prospective data on reproduction were obtained from dairy farm registers. The eutocic weight was higher than the dystocic calving heifers. Plasma total protein and weight were higher in the offspring of multiparous dams than in those of primiparous dams. Calves that were born lighter and in summer had lower concentrations of total protein and performance indices. The punctuation of the fecal score was higher in calves born in summer and cold months for BRD. Offspring from multiparous dams born at term exhibited greater precocity in terms of age at first artificial insemination and first parturition. Finally, seasonality at birth affected pregnancy loss and age at first calving. Heifers born in winter had lower rates of pregnancy loss than those born in summer. In addition, these animals were more precocious than animals born during the cold seasons.

A-111 There is More to Diluting Icterus Interference than Meets the Eye

Abstract Background Total protein is measured in serum and body fluids as part of routine evaluations of general nutritional status and differentiating exudative and transudative effusions. Spectral interference thresholds for bilirubin (e.g., icterus) is set by assay manufacturers to prevent inaccurate results from being reported. The Roche Cobas serum total protein assay package insert states no significant interference from bilirubin up to an icterus (I) index of 20. Body fluids tend to have lower total protein concentrations; therefore, thresholds were derived during prior validation studies and set to I = 10, a threshold that is exceeded routinely. Laboratories can mitigate some interferences by recollecting (e.g., hemolysis) or ultracentrifuging (e.g., lipemia) specimens. Serial dilutions are another option used; however, it is imperative that the laboratory understands the mechanism of interference and can demonstrate that it is mitigated by sample dilution. The aim of this study was to investigate whether bilirubin interference can be mitigated by sample dilution in pleural fluid, peritoneal fluid, and serum specimens. Methods Residual clinically ordered pleural fluid (n = 3), peritoneal fluid (n = 3), and serum (n = 3) samples submitted for protein testing were split into two equal volume aliquots with concentrations ranging 2.7 to 4.3 g/dL in pleural fluids, 3.9 to 5.4 g/dL in peritoneal fluids, and 3.8 to 11.0 g/dL in serum. One aliquot was spiked (&amp;lt;10% by volume) with a bilirubin conjugate solution (∼1000 mg/dL) to surpass the icterus index threshold (spiked). The second aliquot was spiked with an equal volume of 0.9% saline (control). Total protein was measured on the Roche Cobas c701 instrument in both samples to calculate difference (Spiked-Control). The mean(SD) difference was calculated for each sample type. Each spiked sample was serially diluted (2-fold, 4-fold and 8-fold) with 0.9% saline. Total protein was measured for each diluted sample. Mean(SD) difference was calculated (DilutedX-Control), where X = 2-fold, 4-fold, 8-fold. Linear regression analysis was performed by plotting the measured (x-axis) vs expected (y-axis) total protein concentration. Slope, y-intercept, and R2 were determined from serially diluted samples where the expected concentrations were derived using the spiked concentration to replicate the practice of diluting an icteric sample. Results Bilirubin spiking (average icterus index = 33.5) into pleural fluid, peritoneal fluid, and serum, respectively demonstrated mean(SD) bias (g/dL) = −1.0(0.2), −0.9(0.0), and −0.7(0.0). Subsequent 2-fold dilution demonstrated mean(SD) bias (g/dL) = −1.0(0.2), −1.0(0.0), and −0.8(0.1), 4-fold dilution with mean(SD) bias (g/dL) = −1.1(0.2), −1.0(0.0), and -1.0(0.1), and 8-fold dilution with mean(SD) bias (g/dL) = −1.3(0.2), −1.0(0.0), and −1.4(0.1). Diluting pleural, peritoneal, and serum samples, respectively spiked with bilirubin resulted in slope = 0.99, 1.00, and 0.99; y-intercept = 0.04, 0.01, and 0.07; R2 = 0.99, 0.99, 0.99. Conclusion The dilutions exhibited a linear dilution response, however the dilution results never returned to baseline (non-spiked) concentration. Laboratories should exercise caution when conducting serial dilutions to diminish bilirubin interference in samples when measuring total protein as the mechanism of interference does not appear to be reversible by dilution. This behavior was demonstrated in both body fluids and serum.

Physicochemical properties and antioxidant potential of protein isolate from camellia cake (Camellia oleifera Abel.): Effect of different processing techniques on industrial scale

This study aimed to assess the effects of industrial-scale oil-pressing techniques (hydraulic pressing (HP), twin-screw pressing (TP)), and organic solvent extraction (OE) on the physicochemical properties and antioxidant activities of protein isolates derived from Camellia (Camellia oleifera Abel.) seed cakes. The predominant protein fractions in camellia seed cake were albumin (32–37 mg/g) and glutelin (19–27 mg/g), and the twin-screw pressed cake had a lower total protein content than the others. Spectral analysis showed that the α-helix content increased at the expense of random coil content in proteins from HP and TP cakes, indicating that extrusion altered the structure of the protein. The camellia seed protein particle size from HP and TP cakes was significantly reduced compared to that from the OE cake. The proteins obtained by HP and TP had stronger antioxidant activities than those obtained by OE. This research presents the potential for further applications of camellia seed cake protein to be utilized as antioxidants and emulsifiers in the food industries.

Temporal variability in foliar protein content and trypsin inhibitory levels in two host trees of tropical Tasar silkworm Antheraea mylitta, Drury

Aim: Variation in total soluble proteins and protease inhibitor levels were investigated to determine nutritional and plant defense status in different leaf types of Terminalia arjuna (Roxb.) Wight and Arn., and Schleichera oleosa (Lou.) Oken, two host tree species of the economically important, non-mulberry Tasar silkworm, A. mylitta. Methodology: Quantitative spectrophotometric assays based on diagnostic amidolytic substrates were conducted to assess serine protease inhibitor activities in young, semi-mature and mature leaf types. A microplate quantification assay for total protein estimations was used with leaf types sampled over a year. Efficacy of total trypsin inhibitor and chymotrypsin inhibitor activities detected in T. arjuna (a primary host plant) was further evaluated on gut extracts of fourth instar A. mylitta and Pieris brassicae L. (a Pierid pest of crucifers) to assess the physiological adaptation of larvae to dietary antifeedants. Molecular provenances with rbcL genes were obtained that are available as NCBI accessions #MN460810 for T. arjuna and #MT010554 for S. oleosa. Results: Intra-specific variations were evident in leaf phenology of two tree species. Generally, young leaf type of both tree species had high levels of total protein and trypsin inhibitory activities, while mature leaf type of T. arjuna had low total protein content and trypsin inhibitor levels. Mature leaf type of S. oleosa had low trypsin inhibitor levels during the months of July and August. Midgut proteases of A. mylitta and P. brassicae were significantly more susceptible (p≤0.05) to both trypsin and chymotrypsin inhibitors detected in young and semi-mature leaf types of T. arjuna than the mature leaf type. Interpretation: In this study, differential inhibition of digestive proteases in A. mylitta and P. brassicae by protease inhibitors from different leaf types of T. arjuna, suggested adaptation to dietary antifeedants. Such reports on nutritional quality, foliar antifeedants, phenology and host plant utilization are relevant for strategies to domesticate the tropical Tasar silkworm, A. mylitta. Key words: Antheraea mylitta, Protease Inhibitors, Terminalia arjuna, Total protein, Tree phenology

Performance of dairy cows offered high or low digestibility grass silages at a high or low total diet crude protein content

Performance of dairy cows offered high or low digestibility grass silages at a high or low total diet crude protein content

Assessment of M-protein quantification using capillary electrophoresis and immunosubtraction-based integration in clinical samples with low M-protein concentrations

ObjectivesQuantification of monoclonal protein (M-protein) by serum protein electrophoresis (SPE) is indispensable for diagnosing and monitoring monoclonal gammopathies. However, quantification of small and beta migrating M-proteins is challenging because of overlapping non-immunoglobulin and/or polyclonal immunoglobulin protein fractions. We compared a new integration method based on immunosubtraction (IS-CE) using capillary zone electrophoresis (CZE) against the routine method, which includes a combination of perpendicular drop (0.4%), corrected perpendicular drop (1%) and tangent skimming (98.5%). Design & methodsThe proposed method of M-protein quantification involves calculating the difference in area under the curve between the SPE and a class-specific IS-CE trace. We analyzed the difference in estimated M-protein concentrations obtained with the new method and routine integration methods using 913 samples. For IgA M-proteins at < 10 g/L, the estimated M-protein concentrations were compared with the total IgA concentration. ResultsThe median M-protein concentration of 913 consecutive samples was 6.2 g/L (IQR 2.1–14.3 g/L). The median and median % difference between the two integration methods was 0.68 g/L (IQR 0.01–1.55 g/L) and 10.9% (IQR 0.18–38.7%), showing a larger estimated M-protein concentration with the new method. More than 25% difference was observed in 38% of the samples and was associated with lower total protein concentration, lower M-protein concentration, IgA and IgM heavy chain isotypes, and beta- or beta-gamma migration. When 161 samples with IgA M-protein < 10 g/L were compared against total IgA concentration, the median bias of the new method was smaller compared to that of the existing method (-0.95 g/L vs. −1.3 g/L, P < 0.0001). ConclusionsThe use of IS based integration using CZE and IS-CE is promising especially for small and beta migrating M-proteins.

Fecal PCR testing for detection of Clostridium perfringens and Clostridioides difficile toxin genes and other pathogens in foals with diarrhea: 28 cases

Clostridium perfringens and Clostridioides difficile cause significant morbidity and mortality in foals. Antemortem diagnosis of C. perfringens infection has been complicated by a paucity of tests available for toxin detection. Fecal PCR panels have assays for a variety of C. perfringens toxin gene sequences as well as for several other foal gastrointestinal pathogens. We evaluated results of a comprehensive fecal diarrhea PCR panel in 28 foals that had been presented to a referral hospital because of diarrhea. Sixteen (57%) foals were positive for C. perfringens and/or C. difficile toxin gene sequences on fecal PCR, including 3 foals positive for NetF toxin. These foals were younger (p = 0.0029) and had higher hematocrits (p = 0.0087), hemoglobin (p = 0.0067), and red blood cell concentrations (p = 0.028) than foals with diarrhea that tested negative for clostridial toxins. The foals had lower total protein concentrations (p = 0.045) and were more likely to have band neutrophils on a CBC (p = 0.013; OR: 16.2). All 3 foals with NetF toxin gene sequences detected in feces survived to discharge, indicating that diarrhea caused by NetF toxigenic C. perfringens isolates is not uniformly fatal.

Heat sensitive protein-heat stable protein interaction: Synergistic enhancement in the thermal co-aggregation and gelation of lactoferrin and α-lactalbumin

The synergistic enhancement in the thermal co-aggregation and gelation of lactoferrin (LF), a heat sensitive protein, and α-lactalbumin (ALA), a heat stable protein, was investigated at pH 7.0. Heating temperatures (70 °C and 90 °C; 30 min) and ALA concentrations (0–0.5 mM) significantly affected the structural characteristics of the resultant thermal aggregates and gels. The turbidity and size of LF-ALA thermal aggregates were increased with increasing ALA concentration; at a low total protein concentration of 0.57 mM, LF-ALA gels can be formed. The presence of ALA led to some hydrophobic residues originally located in the interior of LF to be exposed further during heating. New intermolecular disulfide bonds, mainly unstable ones, were formed between LF molecules and/or ALA molecules during co-heating. The aggregation of LF and ALA was a gradual denaturation process, accompanied by an increase in β-sheet content and decrease in α-helix content. Random spherical aggregates with large size (1–5 μm) were observed by transmission electron microscopy, clearly confirming the nucleation and growth of LF with ALA. There existed strong rheological synergism between LF and ALA, thereby leading to a large reduction in gelation times (4–11 min) with increasing ALA concentration and heating temperature. Considering these data, LF and ALA played different and indispensable roles in thermal aggregation and gelation: LF engaged readily in thermal aggregation, while ALA mainly assisted the LF thermal aggregation. Three types of mechanisms (co-fusion, nucleation and growth) involved in the aggregation and gelation processes. In all, the data of the current study has enhanced the comprehension of heat sensitive protein-heat stable protein thermal aggregation and gelation, and may help to design LF-based new ingredients for the control of food textures and delivery systems for food and pharmaceutical applications.

Human Placental Hematopoietic Stem Cell Derived Natural Killer Cells (CYNK-001) Mediate Protection Against Influenza A Viral Infection

Background: Influenza A virus (IAV) infections are associated with a high healthcare burden around the world and there is an urgent need to develop more effective therapies. Natural killer (NK) cells provide the first line of innate defense against IAV by killing infected epithelial cells, by producing antiviral cytokines and affecting adaptive immunity. Preclinical studies have demonstrated that NK cells play a pivotal role in reducing IAV-induced pulmonary infection; however, little is known about the therapeutic potential of adoptively transferred NK cells for IAV infections. Celularity Inc. is developing human placental hematopoietic stem cell-derived allogeneic, off-the-shelf NK cell therapy (CYNK-001) for the treatment of viral infections, including coronavirus disease of 2019. Here, we report the evaluation of antiviral activities of CYNK-001 against IAV infection.Methods:In vitro antiviral activities of CYNK-001 were evaluated using human alveolar epithelial cell line A549, infected with IAV strain A/PR/8/34 (H1N1) at variable multiplicity of infection (MOI). The expression of ligands for NK cell receptors was analyzed on infected A549 cells using Fc-coupled recombinant proteins. CYNK-001 was added to A549 cells 16 hours post infection. CYNK-001 degranulation was measured after 4 hours of coculture, and CYNK-001 cytotoxicity against IAV-infected A549 was measured real-time using impedance-based xCELLigence platform. In vivo antiviral and immunomodulatory activities of CYNK-001 were assessed in A/PR/8/34 (H1N1)-induced severe acute lung injury mouse model. Mice were intranasally infected with 2500 PFU IAV. PBS or 1 x 107 CYNK-001 cells were intravenously administered twice at 1 and 3 days post infection (dpi). At 6 dpi, lungs were collected for the evaluation of viral load by qPCR, lung injury and immune cell profiling by histology. Bronchoalveolar lavage fluid (BALF) was collected at 6 dpi for cytokine analysis by multiplex assays, total protein concentration by ELISA and immune cell profiling by flow cytometry.Results:In vitro, IAV infection corresponded with dose-dependent expression of ligands to NK cell-activating receptors, including NKp44, NKp46 and NKG2D. CYNK-001 cells exhibited increased IFNγ, TNFα and GM-CSF production, and elevated level of degranulation upon coculture with IAV-infected A549 cells. Cytokines in culture supernatant and CD107a expression in CYNK-001 cells were upregulated in a virus dose-dependent manner. Consistent with this finding, CYNK-001 cytotoxicity against IAV-infected A549 cells increased from 35% at 0 MOI to 50%, 60% and 75% at 0.001, 0.01 and 0.1 MOI, respectively. These data indicate that CYNK-001 cells recognize virally infected cells, resulting in specific cytotoxic elimination of the source of infection. In vivo, treatment of IAV-infected mice with CYNK-001 reduced weight loss and increased their likelihood of survival. PBS control group developed a severe disease and 37.5% mortality was observed as early as day 4. In the group treated with CYNK-001, disease onset was delayed by 2 days. qPCR analysis of viral RNA showed that CYNK-001-treated mice had lower viral load in the lung than vehicle-treated mice, demonstrating antiviral function of CYNK-001 in vivo. CYNK-001-treated mice had reduced lung injury as assessed by lower total protein concentration in BALF. Moreover, CYNK-001 reduced BALF murine cytokines and chemokines, including IFNγ (p<0.001), IL-6, TNFα, MCP-1 (p<0.05), CXCL2 and CXCL9. Lastly, immunohistochemical analysis of the lung showed that CYNK-001-treated mice had an altered immune response to IAV with higher number of CD68+ macrophages and CD8+ T cells at 6 dpi.Conclusions: Our in vitro and in vivo data show the promising antiviral activities of CYNK-001 against IAV infection. In a severe IAV infection mouse model, CYNK-001 treatment demonstrates lower mortality rate, lower weight loss, lower lung viral load and reduced lung injury along with reduced inflammation. These results support our hypothesis that the adoptive transfer of CYNK-001 could reduce the burden of viral infection through the elimination of infected epithelial cells, coordinate a more effective immune response, and result in a clinical benefit in patients with severe viral infection.DisclosuresHe:Celularity Inc.: Current Employment. Mahlakõiv:Celularity Inc.: Current Employment. Gleason:Celularity Inc.: Current Employment, Current equity holder in private company. Van Der Touw:Celularity Inc.: Current Employment. Kang:Celularity Inc.: Current Employment. Hariri:Celularity Inc.: Current Employment, Current equity holder in private company. Zhang:Celularity Inc.: Current Employment, Current equity holder in private company.

Genotypic and environmental factors influence the proximate composition and quality attributes of sweetpotato (Ipomoea batatas L.)

BackgroundSweetpotato is an important staple food crop worldwide. The genotype mainly influences the nutritional quality of its storage roots, but environmental conditions could produce significant variations in chemical composition and quality. The aim of this study was to characterize sweetpotato diversity of 20 selected genotypes and identify harvest time (90, 120, and 150 days after planting-DAP) and environmental effects on quality attributes estimated by proximate analysis (dry matter, ash, crude fiber, total protein, and total soluble solids). Red (R), green (G), and blue (B) (RGB) analysis was used to characterize the genotypes phenotypically.ResultsThe results of the current study revealed that flesh color was associated with proximate composition. RGB analysis showed that low B pixel values were present in yellow–orange- and purple-fleshed genotypes, which simultaneously exhibited high total protein content (TPC), ash content (AC), and crude fiber (CF), while cream- and white-fleshed genotypes showed high B pixel values and were related to high dry matter content (DMC). In these genotypes, the high DMC was maintained or increased through harvest time, however, a reduction in the proportion of accumulated AC and TPC was observed. On the other hand, in the pigmented genotypes, DMC increased up to 120 DAP, showing major stability in terms of AC and TPC. Regardless of harvest time, more intense rain events 30 days before harvest, affected the storage of DMC in roots negatively, while AC, TPC, and CF were accumulated more efficiently, or their proportion increased. High temperatures increased total soluble solids (TSS) accumulation and reduced AC, TPC, and CF accumulation. The combined analysis, according to the additive main effects and multiplicative interaction (AMMI) model, confirmed these results.ConclusionsThese findings indicated that white-fleshed genotypes contain high DMC, although, with lower TPC, AC, and CF contents compared to yellow–orange- and purple-fleshed genotypes. Although there is an increase in DMC in extended harvests (in orange-fleshed genotypes up to 120 DAP), the presence of pre-harvest rain promotes its translocation and loss. On the contrary, TPC, AC, and CF can be kept stable or even increase, except in environments with high temperatures that induce low accumulation. The current study provides a better understanding of the nutritional response of sweetpotato diversity under several growing conditions, which can be recognized and used integrally to improve food quality.

Effects of Aqueous Leaf Extract of Acalypha wilkesiana on some Serum ‎Biochemical Profiles of Mice in Sub-Acute Toxicity Studies

Acalypha wilkesiana has been used for various medicinal purposes such as in the treatment of hypertension, fungal and bacterial infections. In this study, the effects of prolonged oral administration of the aqueous leaf extract on some biochemical profiles of mice were evaluated. A total of 80 mice were allocated to 4 groups (A, B, C and D) comprising of 20 mice each. Mice in group A (control) were given (1 mL) of distilled water orally while 1 mL of the extract was given to the mice in groups B, C and D daily at the concentrations of 800, 1000 and 1200 mg/kg, respectively. This was continued for three weeks and at the end of each week, 4 mice from each group were humanely sacrificed and sera from blood samples collected were used for determination of activities of biochemical parameters. The results showed that administration of the extract resulted in significantly (p &lt; 0.05) lower concentrations of total protein and cholesterol in the treatment groups at the 3rd week of treatment when compared to control values. Glucose level was significantly (p &gt; 0.05) higher in the extract-treated groups by the 3rd week of treatment, while activities of AST, ALT and ALP were not significantly (p &gt; 0.05) altered by the extract. There were no significant (p &gt; 0.05) differences between the levels of calcium, total and conjugated bilirubin in treatment and control groups, while sodium concentration was significantly (p &lt; 0.05) higher at day 14 after withdrawal of treatment, compared to the control. This study suggests that the extract of Acalypha wilkesiana has a hyperglycaemic effect.

Clinicopathological abnormalities associated with increased animal triage trauma score in cats presenting for vehicular trauma: 75 cases (1998-2009).

To document the clinical and clinicopathological changes in cats presenting with vehicular trauma (VT) and to determine whether the calculated animal trauma triage (ATT) score was associated with any clinicopathological abnormalities. Retrospective descriptive study conducted between 1998 and 2009. University veterinary teaching hospital. Seventy-five client-owned cats that presented for VT to an urban veterinary hospital. None. Pertinent history, physical examination findings, results of biochemical testing, and outcome were extracted from medical records. ATT score was calculated based on physical examination. Patients were classified as having either a low (<5) ATT (n=30) or a high (≥5) ATT (n=45) score. Male cats were overrepresented (60.0%), and a majority of cats had outdoor access (65.3%). Low PCV (P= 0.024), low total plasma protein concentration (P= 0.032), low venous blood pH (P= 0.047), high plasma lactate concentration (P= 0.047), low plasma bicarbonate concentration (P= 0.047), low base excess (P= 0.047), and high plasma glucose concentration (P= 0.047) were associated with higher ATT scores. In addition, low noninvasive blood pressure measurements (P= 0.008) were associated with higher ATT scores. There was a significant association between lower PCV, lower total plasma protein concentration, lower venous blood pH, higher plasma lactate concentration, lower plasma bicarbonate concentration, lower base excess, higher whole blood glucose concentration, and lower noninvasive blood pressure and higher ATT scores at presentation in feline patients suffering from VT. Prospective evaluation of these values may prove useful in furthering understanding of the pathophysiology of trauma in cats.

Nutritional Characteristics of Teak Grasshopper (Valanga nigricornis Burmeister), Cricket (Brachytrupes portentosus L.), and Mealworm (Tenebrio molitor) as Alternative Food Sources in Indonesia

Insect is one of the alternative food sources which can be easily obtained and cultivated at a lower cost. Food sources must also provide daily nutrition; therefore, this study aims to compare the nutrition of edible insects in Indonesia, namely teak grasshopper (Valanga nigricornis Burmeister), cricket (Brachytrupes portentosus L.), and mealworm (Tenebrio molitor), with cork fish (Chana striata) that has been commonly consumed by the Indonesian people. The highest total protein content (Kjeldahl method) was found for fish (82.69%), while the lower ones were found for grasshopper (72.50%), cricket (59.00%) and mealworm (53.08%). This result was also in line with the total protein content found for fish (11984.00 ppm as the highest) and grasshopper (1016.33 ppm as the lowest). SDS-PAGE result also confirmed the highest protein content for fish where it showed the thickest band intensity for fish sample. Although cricket and mealworm were found to have lower total protein content (proximate assay), their fat and calorie levels were higher with calorie from fat values of 235.35 kcal/100 g for cricket and 195.93 kcal/100 g for mealworm. Proteolytic assay was carried out using papain to compare protein digestibility. The absorbance values from the proteolytic assay of cork fish and mealworm were higher than grasshopper and cricket, indicating that proteins of fish and mealworm were easier to hydrolyze. This could be explained so, that adult insects have a chitin structure that limits the hydrolysis process, or that papain might have a specific recognition sites and it fitted better to the fish and mealworm protein. Thus, each insect sample showed its characteristic nutritional profile and therefore, can be used as alternative food ingredient to meet the needs of protein and fat.

Ethylene Induced Flower Opening, Expression of Receptor Genes, and Antioxidant Defensive Responses in Cut Roses

The present research aimed to investigate the effect of ethylene on the physiological and molecular responses of the cut roses, Rosa hybrida L. Two cut rose cultivars (‘Amada +’ and ‘Brillante’) with different ethylene sensitivity were treated by 10 ppm exogenous ethylene for 24 h under identical vase life room conditions at partially open stage. The cultivars were then examined at both partially and fully open stage by gas chromatography, UV/Vis spectroscopy, and qRT-PCR. The cultivar with high sensitivity had lower total protein contents and showed higher ethylene production and antioxidant activity. Either exogenously applied ethylene or endogenously produced one resulted in increased antioxidant activity and decreased total protein contents. This observation is possibly because of enhanced free radical generation and protease activation with ethylene-induced flower opening and senescence. The qRT-PCR analysis showed a significant increase in the expression of ethylene signal transduction genes (RhETR1,3 and RhCTR1,2) of the treated cultivars with high sensitivity at commercial stage. However, there was a significant up-regulation of the RhETR1,3 genes in the cultivars with low sensitivity at fully-open stage. The ethylene induced responses were found highly dependent on the cultivar ability in both ethylene synthesis and perception.

Determination of serum aflatoxin B1-lysine and biochemical parameters in broiler chicks fed an aflatoxin B1-contaminated diet

In the present trial, the levels of serum aflatoxin B1 (AFB1)-lysine and their relationship with biochemical parameters in broiler chicks fed an AFB1-contaminated diet were determined. The experimental design was completely randomized with two treatments (control and 222.17 µg/kg AFB1) and 20 bird per treatment. Feeds were offered to broiler chicks for 14 days, from 28 to 42 days of age. Animals were vaccinated against Newcastle’s and Marek’s diseases on the 14th day of life, and were killed at 42 days of age. Broilers receiving AFB1 did not demonstrate any sign of toxicity. Compared with controls, aspartate aminotransferase and globulin levels were not affected in the AFB1-treated group. However, higher levels of gamma-glutamyl transferase and lower concentrations of total protein and albumin were observed in the group receiving AFB1 on days 35 and 42. AFB1-lysine were detected in the serum of all broilers fed the AFB1-contaminated diet, at mean levels of 56.52–77.83 ng/mg albumin on days 35 and 42 of age, respectively. These values indicated the internal dose of AFB1 in birds, which negatively correlated with total protein, albumin, and globulin levels. Data indicated that AFB1-lysine shows the potential to be a sensitive and specific biomarker for the evaluation of broiler exposure to dietary aflatoxin, as well as for diagnostic purposes. Further studies are necessary to determine physiologically-based toxicokinetics of serum AFB1-lysine in broilers.

Study of the Physicochemical and Biological Properties of a Protein–Peptide Complex Isolated from Cattle Ovarian Tissue

A protein–peptide complex isolated from cattle ovarian tissues was studied. It was shown that it contains polypeptides with molecular weights from 800 to 6000 Da and a protein of the mammalian albumin family with a molecular weight of 66 690 Da containing the 16-member N-terminal amino acid sequence DTHKSEIAHRFKDLGE. The use of a model of the roller organotypic cultivation of rat ovaries demonstrated that the effect of this protein–peptide complex at low total protein concentrations of 10–8–10–15 mg/mL contributes to increases in the viability and maturation of secondary follicles in rat ovarian tissues in vitro.

Development of a cell‐based assay for Tau strain discrimination

Alzheimer's Disease and other related tauopathies are characterized by the accumulation of the microtubule‐associated protein Tau into fibrillary deposits. Interestingly, there is evidence supporting amyloid structures, or strains, formed as a consequence of Tau aggregation are unique to each pathology, suggesting different conformations play an important role in the development of specific tauopathies. The potential critical role of Tau in disease progression, together with the isolation of 19 different Tau strains, led us to work in developing a panel of cell‐based fluorescence resonance energy transfer (FRET) biosensors. Each biosensor carried a different mutation on Tau's aggregation‐prone repeat domain, based on the hypothesis that each mutant would have unique interactions with Tau strains and, thus, alter Tau aggregation kinetics within the cell. We hoped that by studying these differences in FRET signal we would be able to discriminate between distinct amyloid conformations. In order to do this, we first created the biosensors by transducing HEK 293 cells with lentivirus coding Tau's repeat domain with specific mutations, fused to both, cyan florescent protein (CFP) or yellow florescent protein (YFP). These biosensors were later treated with different sources of Tau seeds and FRET flow cytometry was used to describe their seeding activity. Experiments with the 19 strains in our Tau library and the engineered biosensors revealed noticeable differences in percent FRET positivity, such that discrimination was achieved for most of the strains. Interestingly, the highest seeding activity among all the biosensors was obtained from strain 15 (DS15), even at low total protein concentrations. Additionally, biosensors expressing the wild‐type four‐repeat domain of Tau had higher percent FRET positivity only when seeded with strains derived from Corticobasal Degeneration (CBD) patients. Furthermore, treatment of our panel of biosensors with brain homogenates of multiple tauopathy patients correlated with our previous results and revealed more distinct patterns of FRET behavior. Together, these results suggest that measuring proteopathic seeding activity while using our panel of biosensors is a sensitive and reliable tool to discriminate between different amyloid conformations, which could potentially lead the development of improved diagnostic methods for neurodegenerative disorders.Support or Funding InformationThis research project was supported by the NIH‐Maximizing Access to Research Careers (Grant # 5T34GM0072821‐38), the Summer Undergraduate Research Fellowship at UT Southwestern Medical Center and the Center for Alzheimer's and Neurodegenerative Diseases at UT Southwestern Medical Center.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Inflammatory bowel disease (IBD) in horses: a retrospective study exploring the value of different diagnostic approaches

BackgroundDiagnosing IBD in horses is challenging and requires a multimodal approach, since no conclusive diagnostic test is available.The objectives of this study were to provide an overview of population characteristics, results of applied diagnostic tests, treatment modalities and outcome in a large group of horses thought to have IBD and that were presented to four large equine referral hospitals, and to provide an exploratory investigation of possible associations between results of applied diagnostic tests, applied treatment modalities and outcome.A retrospective case series was performed across four large equine referral hospitals. Seventy-eight horses, thought to have IBD were included. Case history, clinical findings, diagnostic test results including oral glucose tolerance test (OGTT) and enteral biopsies (both duodenal and rectal), applied therapy and outcome were studied. A Chi-Square test was used to identify associations between results of diagnostic tests, treatment and outcome. P-values < 0.05 were considered significant.ResultsLethargy, diarrhoea, recurrent colic and weight loss were recorded in respectively 21,8%; 14,1%; 28,2% and 78,2% of cases. Over 70% of horses thought to have IBD had an abnormal OGTT. Only weight loss was significantly associated with aberrant enteral biopsy results, but not with abnormal OGTT results or low blood total protein. There was no association between an aberrant biopsy result and a disturbed OGTT. There was no association between either OGTT results or enteral biopsy results and a low blood total protein content, presence of gastric ulcer disease or an aberrant endoscopic aspect of the duodenal entrance.ConclusionsWeight loss is a highly prevalent symptom in IBD suspected horses. Enteral biopsies may be a useful diagnostic aid in the work-up of horses thought to suffer from IBD, however further research is required to demonstrate their true diagnostic value. Until more standardized scientific research is available, one should be careful with the interpretation of enteral biopsy results There is a need for better standardization of enteral biopsy procedures and the histopathological scoring of biopsies.

Carobs in productive animal nutrition

The carob tree belongs to the nutrient plants and it is known since antiquity as a native plant of Greece. Its scientific name Ceratonia siliqua, originates from the Greek word "keraton" which means "horn", and which indicates the shape of its fruit. The carob tree is an evergreen, long-lived, polygamous, monoecious or dioecious. It is easily cultivated and thrives in all types of soil, except the humid and non-affluent. The wood, the bark and the leaves of carob have different uses. The fruit of the carob tree, the carob, is a lobe and it is 10-30 cm long and 2-3 cm wide with a brown and leathery exocarp and a fleshy and juicy mesocarp, in which 8 to 16 oval, flattened, hard and shiny reddish seeds are enclosed. The collection of the fruits must be finished before the winter rains. The fruits fall from the tree by themselves or by caning. A 6 years old carob tree can produce 2.25 kg carobs, while a 12 years old tree can produce 45 kg or more. The annual Greek carob production for the year 2005 was 14,816 tons, while the average yield was 9.8 kg/tree. The world carob production is about 315,000 tons/year. The carobs have low total fat and crude protein content, but high sugar content. The carobs' characteristic odor is due to their isobutyric acid content (1.3%). Furthermore, they include an unusually large amount of tannins, 16-20% of which are polyphenols and 27-50% of which are bound with cellulose. The tannins are a complex group of secondary metabolites of plants and are separated from other polyphenol compounds due to their ability to create sediment with proteins. The presence of tannin in feed reduces the nutrients' digestibility and has a negative effect on body weight gain. However, tannins can also have positive effects depending on their biological potency, because they have the ability to prevent bacterial, fungal and yeast growth. Carobs and their by-products have a considerable fibre content (average 8%) and therefore they can be included in supplementary ruminant rations up to 20% - 30%. Experiments carried out by adding carobs in broiler feeds resulted in decreased body weight gain and increased feed conversion ratio, probably because of the high cellulose and tannin content of the carob. Nevertheless, if the broiler's ration is enriched with fats or oils and synthetic amino acids, these birds show normal weight gain, but with a high feed conversion ratio. The carobs can be included up to 10% - 20% in the ration of pigs. Also, since the carobs have high sugar content, they can increase the feed consumption of the piglets and they can replace other sugar feeds with high sugar content (dextrose, starch, milk powder) without adversely affecting the feed intake. Greece produces a considerable quantity of carobs every year, therefore a part of this production could be utilized as animal feed