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.     

Acute cortisol responses of calves to scoop dehorning using local anaesthesia and/or cautery of the wound

Objective To measure plasma cortisol responses in calves dehorned using a scoop after administration of local anaesthesia and/or cautery of the wounds.
Design A physiological study with controls.
Procedure There were six treatments: control handling with and without local anaesthesia, dehorning, dehorning after local anaesthesia, dehorning followed by wound cautery, and dehorning after local anaesthesia followed by wound cautery. Blood samples were taken before and after dehorning.
Results Dehorning caused an increase in plasma cortisol concentrations, which decreased a little to plateau values and then declined to pretreatment values 3 to 4 h after dehorning. The peak was smaller after local anaesthesia was administered but when its effects wore off, cortisol concentrations increased and thereafter were similar to those in the dehorned animals. The combination of local anaesthesia and cautery resulted in a plasma cortisol response similar to those in control calves with or without local anaesthesia.
Conclusions If plasma cortisol concentrations reflect the distress being experienced by the calves, then local anaesthesia reduces the acute distress for about 3 h after dehorning but not during the subsequent 3 to 4 h. Combining local anaesthetic and cautery prevented the significant increase in plasma cortisol following dehorning and may eliminate the acute distress caused by scoop dehorning.     

One-step RT-PCR for the Detection of Infectious Bursal Disease Virus in Clinical Samples

A single-tube, non-interrupted, one-step RT-PCR has been standardized to amplify the hypervariable region of the VP2 gene sequence of infectious bursal disease virus (IBDV). The technique standardized on purified viral RNA was successfully applied to the detection of the virus directly in clinical samples. The amplified products were confirmed to be IBDV specific by their size in ethidium bromide-stained agarose gel, nested PCR and restriction enzyme digestion. Digestion of the amplicons with StyI restriction enzyme also differentiated classical virus from six very virulent field isolates. The sensitivity of the one-step RT-PCR was found to be 0.2 pg of viral RNA.     

Protection of mice against <Emphasis Type="Italic">Brucella abortus 544</Emphasis> challenge by vaccination with recombinant OMP28 adjuvanted with CpG oligonucleotides

Brucella abortus, a gram negative, facultative intracellular pathogen causes brucellosis in many animal species and humans. Although live, attenuated vaccines are available against this infection, they suffer from certain limitations. Therefore, the development of an effective subunit vaccine against brucellosis is an area of intense research. The outer membrane proteins (OMPs) of Brucella species have been extensively studied for its immunogenicity and protective ability. We have investigated the potential of CpG ODN to enhance the immunogenicity and protective efficacy of recombinant 28 kDa outer membrane protein (rOMP28) of Brucella melitensis. The study demonstrated vigorous immunoglobulin G (IgG) response of OMP28. The administration of rOMP28 with CpG caused increased cell mediated immune response in terms of induced IgG2a, T-cell proliferation and up-regulation of type I cytokine expression. In contrast, the free antigen suppressed the interferon gamma (type I cytokine) production on in-vitro stimulation of spleenocytes. The result indicates the role of OMP28 in the down regulation of IFN-γ production. Moreover, the B. abortus S-19 vaccinated mice showed highest production of IL-4 and IFN-γ. The protective ability of the antigen was evaluated by systemic bacterial clearance after challenging the mouse with B. abortus 544 pathogen. The level of protection was significant in rOMP28+CpG treated mice but was lower than the required level. The results of the present study indicate that rOMP28 could be an immunogen capable of inducing both humoral and cellular immune response. The humoral response was biased towards Th1 type when it was co-administered with CpG.     

Antifungal properties of Ocimum gratissimum essential oil (ethyl cinnamate chemotype).

Largely widespread in tropical countries, Ocimum gratissimum has been claimed for many uses in folk medicine. Recent research on its essential oils showed five chemotypes. An Indian chemotype, with a high level of ethyl cinnamate, presents, in vitro, an interesting spectrum of antifungal properties.     

Regeneration of tree species in the sub-tropical forest of Alaknanda Valley, Garhwal Himalaya, India

We studied the regeneration of tree species in the sub-tropical forest of Alaknanda Valley in Garhwal Himalaya, India. The overall regeneration status was fairly good in the study area. Seedling density ranged between 520 and 1,240 seedlings per ha while the density of saplings varied between 400 and 800 saplings per ha. Out of eight sites studied, five sites, viz., A1 , A2 , B1 , B2and C2contained the highest number of seedlings (280-480 per ha) and saplings (200-440 per ha) for Pinus roxburghii and remaining three sites viz., C1, D1and D2represented the highest number of seedlings (240-400 per ha) and saplings (200-240 per ha) for Anogeissus latifolius. The DBH class distribution of the tree species revealed that the highest number of individuals was concentrated in the lower diameter classes while smallest numbers were found in the higher diameter classes. Species such as Acacia catechu, Anogeissus latifolius, Dalbergia sissoo, Engelhardtia spicata, Lannea coromandelica, Mallotus philippensis and Pinus roxburghii have the largest number of saplings and seedlings in the lower DBH classes, suggesting that they have good regeneration potential. Other species such as Aegle marmelos, Bauhinia variegata, Bombax ceiba, Cassia fistula, Erythrina variegata, Haldinia cordifolia, Mangifera indica, Ougeinia oojeinensis, Phyllanthus emblica, Syzygium cumini, Terminalia alata and Toona hexandra have either no or very small number of saplings in the lower DBH classes, which indicates that the status of these species implies poor regeneration.     

Carbonic anhydrase in Tectona grandis: kinetics, stability, isozyme analysis and relationship with photosynthesis

Carbonic anhydrase (CA, EC: 4.2.1.1) activity in teak (Tectona grandis L.f.) was studied to determine its characteristics, kinetics and isozyme patterns. We also investigated effects of leaf age, plant age and genotype on CA activity and gas exchange parameters. Carbonic anhydrase extracted from leaves in 12 mM veronal buffer, pH 7.8, had a K(m) for CO(2) of 15.20 mM and a V(max) of 35,448 U mg(-1) chlorophyll min(-1), which values declined by 50 and 70%, respectively, after 1 week of storage at 4 degrees C. A 15% native polyacrylamide gel revealed the absence of CA isozymes in teak, with only a single CA band of 45 kD molecular mass observed across 10 segregating half-sib families and groups of trees ranging in age from 10 to 25 years. Activity remained stable during the first month in storage at 0 degrees C, but gradually declined to 25% of the initial value after 1 year in storage. During the period of active growth (February-May), maximal CA activity was observed in fully expanded and illuminated leaves. Significant variation was observed in CA activity across 10 1-year-old half-sib families and 21 5-year-old half-sib families. There was a positive correlation between CA activity and photosynthetic rate in a population of 10-year-old trees (P < 0.005). Positive correlations between CA activity and photosynthetic rate were found in 10 of 21 5-year-old half-sib families (P < 0.005 to P < 0.05), which showed greater diversity in CA activity than in photosynthetic characteristics. Thus, CA may serve as a biochemical marker for photosynthetic capacity in teak genotypes.     

Path-Coefficient Analysis of Leaf-Anatomical Characters Affecting Frost Injury in Potato

Correlation and path-coefficient analysis for leaf-anatomical characters affecting frost injury was carried out in twenty eight varieties and accessions belonging to several wild and cultivated tuber-bearing species of Solanum in order to identify selection criteria in breeding for frost resistance in potato Palisade thickness and palisade proportion showed a significant negative correlation with frost injury Path-coefficient analysis revealed that palisade thickness had the highest direct negative effect on frost injury whereas leaf thickness, not conforming to its behaviour as found in simple correlation analysis, emerged ax the next highest factor to have direct positive effect on frost injury. Direct and indirect effects of these factors on frost injury vis a vis their interdependence were found to be in support ot the ‘palisade thickness method’ for screening frost resistant genotypes in potato breeding programmes.