Bioactive Peptides from Algae: Traditional and Novel Generation Strategies,Structure-Function Relationships,and Bioinformatics as Predictive Tools for Bioactivity

Over the last decade, algae have been explored as alternative and sustainable protein sources for a balanced diet and more recently, as a potential source of algal-derived bioactive peptides with potential health benefits. This review will focus on the emerging processes for the generation and isolation of bioactive peptides or cryptides from algae, including: (1) pre-treatments of algae for the extraction of protein by physical and biochemical methods; and (2) methods for the generation of bioactive including enzymatic hydrolysis and other emerging methods. To date, the main biological properties of the peptides identified from algae, including anti-hypertensive, antioxidant and anti-proliferative/cytotoxic effects (for this review, anti-proliferative/cytotoxic will be referred to by the term anti-cancer), assayed in vitro and/or in vivo, will also be summarized emphasizing the structure–function relationship and mechanism of action of these peptides. Moreover, the use of in silico methods, such as quantitative structural activity relationships (QSAR) and molecular docking for the identification of specific peptides of bioactive interest from hydrolysates will be described in detail together with the main challenges and opportunities to exploit algae as a source of bioactive peptides.     

Physiologic assessment of blood glucose homeostasis via combined intravenous glucose and insulin testing in horses

OBJECTIVE: To characterize the physiologic response to i.v. bolus injection of glucose and insulin for development of a combined glucose-insulin test (CGIT) in horses. ANIMALS: 6 healthy mares and 1 mare each with pituitary adenoma and urolithiasis. PROCEDURE: Horses were given a CGIT (glucose, 150 mg/kg; insulin, 0.1 U/kg); results were compared with a singular i.v. glucose tolerance test (GTT; 150 mg/kg) and a singular i.v. insulin sensitivity test (IST; 0.1 U/kg). Healthy horses were also given a CGIT after receiving xylazine and undergoing stress. RESULTS: Physiologically, the CGIT resulted in a 2-phase curve with positive (hyperglycemic) and negative (hypoglycemic) portions; the positive phase came first (250% of baseline at 1 minute). The descending segment declined linearly to baseline by approximately 30 minutes and to a nadir at 58% of baseline by 75 minutes. After a 35-minute valley, a linear ascent to baseline began. Addition of insulin in the CGIT increased glucose utilization by approximately 4.5 times during the positive phase but not during the negative phase. The diseases' effects and experimental inhibition of insulin secretion with xylazine and stress were detectable by use of the 2 phases of the CGIT. Only a single positive phase resulted from the GTT and a single negative phase from the IST CONCLUSIONS AND CLINICAL RELEVANCE: The CGIT resulted in a consistent, well-defined glycemia profile, which can be disrupted experimentally or by a disease process. The CGIT has clinical potential because it provides integrated information and more information than either the singular GTT or IST.     

Lack of Native Vegetation Recovery Following Biological Control of Leafy Spurge

Leafy spurge (Euphorbia esula L.) is an aggressive exotic species that has been successfully suppressed in a variety of situations using classical biological control (flea beetles; Aphthona spp.). This 9-yr study investigated patterns of vegetation responses following significant reductions in leafy spurge cover and density by flea beetles in southeastern Montana. We hypothesized that the vegetation following leafy spurge suppression would be dominated by species and plant functional groups able to persist through heavy infestations. Flea beetles were first released in 1998, and by 2006 leafy spurge foliar cover was reduced 80% to 90% compared to 1998 values on both release and nonrelease plots. Although total cover of the resident vegetation, excluding leafy spurge, increased 72% to 88%, relative cover of the functional groups (native forbs, native sedges, native grasses, and non-native species) was similar among years and between release and nonrelease plots. Mean diversity and mean species richness values did not differ among years or between release and nonrelease plots (P < 0.05), but mean diversity on both release and nonrelease plots was significantly less than noninfested plots, although richness was similar (P < 0.05). Indicator species analysis revealed that non-native Poa spp. replaced leafy spurge as the dominant species on release and nonrelease plots. Conversely, noninfested plots contained a variety of native species with high indicator values. Although total abundance of the resident vegetation in 2006 was significantly greater than 1998, plant species composition and relative cover showed little change for the duration of the study. Failure of the native vegetation to recover to a community that approached nearby noninfested conditions may be attributed to a variety of interacting scenarios, some of which may be ameliorated by treating infestations as soon as possible to avoid long-term residual effects.     

Activation of the ovine hypothalamic-pituitary-adrenal axis and febrile response by interleukin-6: A comparative study with bacterial lipopolysaccharide endotoxin

Sheep were subjected to immune challenge with either recombinant human interleukin-6 (rhIL-6; 2.0 μg/kg; n = 5), Escherichia coli lipopolysaccharide (LPS) endotoxin (400 ng/kg; n = 7), or saline (n = 6) to determine if IL-6 activates the febrile and hypothalamic-pituitary-adrenal axis (HPAA) responses in sheep, and to compare these responses with those associated with endotoxemia. Blood was collected over time to measure plasma adrenocorticotropic hormone (ACTH) and serum cortisol concentrations as indicators of HPAA activity. Unlike LPS, rhIL-6 was not pyrogenic in sheep at this challenge dose. In contrast, rhIL-6 elicited ACTH and cortisol responses that peaked earlier than those induced by LPS. These results suggest that this dose of IL-6, alone, is not sufficient to elicit the febrile response in sheep, however, it is a potent activator of the ovine HPAA response.     

Evaluation of a commercially available hyperimmune plasma product for prevention of naturally acquired pneumonia caused by Rhodococcus equi in foals

OBJECTIVE: To determine efficacy of a commercially available hyperimmune plasma product for prevention of naturally acquired pneumonia caused by Rhodococcus equi in foals. DESIGN: Randomized clinical trial. ANIMALS: 165 foals. PROCEDURE: Foals were randomly assigned to 1 of 2 groups (hyperimmune plasma or nontreated controls). Foals with failure of passive transfer (FPT) of immunity were treated with hyperimmune plasma and evaluated as a third group. Foals that received plasma were given 950 ml between 1 and 10 days of age and between 30 and 50 days of age. A tracheobronchial aspirate was obtained from foals with clinical signs of respiratory tract disease for bacteriologic culture. RESULTS: A significant difference in incidence of pneumonia caused by R equi in foals with adequate passive transfer was not detected between foals that received plasma (19.1%) and nontreated foals (30%). Of 13 foals without FPT that received plasma and developed pneumonia caused by R equi, 12 developed disease prior to administration of the second dose of hyperimmune plasma. Incidence of undifferentiated pneumonia of all causes was not different between groups. CONCLUSION AND CLINICAL RELEVANCE: Intravenous administration of the commercially available hyperimmune plasma was safe, and the product contained high concentrations of anti-R equi antibodies. However, within this limited foal population, the difference in incidence of pneumonia caused by R equi observed between foals that received plasma and control foals was not significant.     

Characterization of the DNA nucleotide sequences in the genome of red sea bream iridoviruses isolated in Korea

The nucleotide sequences of DNA fragments amplified by polymerase chain reaction (PCR) from four different genomic regions of nine red sea bream iridoviruses (RSIVs) isolated from different species of fish, different areas and in different years in Korea were compared with the reported reference sequences. One isolate, RSIV Namhae, showed 100% homology to the reference sequences, while the other eight isolates, which appeared to contain identical nucleotide sequences, showed 96.6–98.9% homology with reference sequences depending upon the target regions of PCR gene amplification. However, differences in nucleotide sequences were not apparent between the RSIVs isolated in different locations, in different years or in different host species. We also cloned and sequenced the 3′ end flanking region (K1) of the DNA polymerase (DPOL) gene using the cassette ligation-mediated PCR method. This sequence was 4436-bp long and possessed two open reading frames (ORF-1 and ORF-2) oriented in opposite directions. The putative proteins encoded by these two ORFs could not be characterized by comparison with the proteins of other species in the data banks. The presence of the ribonucleotide reductase small subunit (RNRS) gene at the 3′ end of the K1 region allowed us to determine that these two genes, RNRS and DPOL, are separated 5508 bp and oriented in the same direction in the genome of RSIV. Moreover, it is of interest that a PstI-restriction fragment, of which the sequence but not the location within the RSIV genome had previously been reported, is located at nucleotide positions from 1096 to 2054, extending from within the ORF-1 region, spanning the intervening sequence between ORF-1 and ORF-2, and extending into the ORF-2 region. Various repeating sequences up to 86 bp were present at the 3′ ends of ORFs, especially within the nucleotide sequences at the 3′ terminus of ORF-2. No similarities were detected when the DNA sequences of the K1 region were compared to the DNA sequences of a repetitive element in the genome of other iridoviruses.     

Use of infrared thermography to detect injections and palmar digital neurectomy in horses

Thermography is a non-contact, non-invasive technique that detects surface heat emitted as infrared radiation. Because skin temperature reflects the status of underlying tissue metabolism and blood circulation, abnormal thermal patterns can signify areas of superficial inflammation. The objective of this study was to determine if thermography could detect the injection of analgesic and neurolytic agents and surgical palmar digital neurectomy. Procedures evaluated include injection of the lumbar region, suspensory ligaments, tibial nerve, palmar digital nerves, and palmar digital neurectomy. Thermographic images were obtained before and after the procedures until a significant difference was no longer detected. Local injection of the lumbar region and the suspensory ligament produced detectable thermal patterns for two days, and tibial nerve infiltration with a neurolytic agent was significant for two days. Analgesia of the palmar nerves was significant for 24h with bupivicaine, compared to five days for ammonium chloride. Palmar digital neurectomy produced more variable thermal patterns. While sensitive enough to detect changes in heat patterns from control regions, thermography is not specific enough to discriminate between procedures and injury inducing an inflammatory response.