Spectrum of celiac disease in paediatric population: experience of tertiary care center from Pakistan

Objective: To determine clinical features and relevant laboratory investigations of patient with celiac disease (CD) and comparing classical celiac disease (CCD) with Non-diarrheal celiac disease (NDCD).Methods: This is a five years retrospective study conducted at The Aga Khan University Hospital Karachi, Pakistan from January 2010 to December 2015, enrolling children from one year to 15 years of either gender diagnosed as celiac disease in accordance with revised ESPGHAN criteria. Biopsy samples with grade 2 or more on Modified Marsh Classification were considered as consistent with celiac disease. Celiac patients were categorized into Classical celiac disease (with Chronic Diarrhea) and non-diarrheal celiac disease (Atypical celiac) and their clinical features and relevant laboratory investigations were documented.Results: Total 66 patients were selected with celiac disease according to inclusion criteria, 39 (59.09%) patients were labeled as CCD and 27 (40.91%) patients were labeled as NDCD. Marsh grading 3a and above were more marked in CCD as compared to NDCD. Mean titer for Tissue transglutaminase antibodies (TTG) were higher in CCD group in comparison to NDCD group. In CCD, the most common clinical presentations were abdominal distension whereas in NDCD, the most remarkable features were recurrent abdominal pain (62.9%). Frequency of failure to thrive is significantly high in CCD (82.05%) but patients merely with short stature were more common in NDCD (33.3%). Refractory anemia was present in 66.6% patients in NDCD group and 41.1% patients in CCD group. 74.3% patients in CCD group were vitamin D deficient whereas 85% patient had vitamin D deficiency in NDCD group (p= 0.03). Conclusion: NDCD is not uncommon in our population. Recurrent abdominal pain, failure to thrive or patients only with short stature and refractory anemia are prominent features in NCDC group whereas abdominal distension, failure to thrive and recurrent abdominal pain were noticeable features in CCD. High grade histopathology and raised antibodies titer is hallmark of CCD. Vitamin D deficiency is almost equally present in both groups

Functional properties of Australian blue lupin (Lupinus angustifolius) protein and biological activities of protein hydrolysates

Lupin is an undervalued legume despite its high protein content with known health benefits. In this research, Australian blue lupin protein was isolated and hydrolysed enzymatically to produce bioactive peptides with a view to assess their potential for nutraceutical and therapeutic applications. Pepsin, pancreatin and flavourzyme were used to enzymatically hydrolyse blue lupin protein, and the hydrolysates were subjected to molecular weight cut‐off (MWCO) fractionation. Measurement of biological activities led to the identification of angiotensin converting enzyme (ACE) inhibitory fractions in the molecular weight range of 2–3 and 3–5 kDa. For the most active fractions in this range, the ACE inhibitory activities were very significant with IC50 values from 450 to 600 μg/ml. Blue lupin protein‐derived MWCO fractions were significantly active against Gram‐positive bacteria and only a little inhibition was observed against Gram‐negative bacteria. Pancreatin hydrolysed fractions showed the best antimicrobial activities with several fractions exhibiting ≥85% inhibition against Bacillus cereus and Staphylococcus aureus. These properties reveal the potential of lupin protein hydrolysates for developing antihypertensive and host defence agents. In order to demonstrate the potential of isolated blue lupin protein in food industry, functional properties including water and oil absorption capacity, gelling properties, solubility and emulsifying properties were evaluated and found to be extremely suitable for developing functional foods with enhanced health benefits

Enzymatic hydrolysis of lupin (Lupinus angustifolius) protein : isolation and characterization of bioactive peptides

Blue lupin (L. angustifolius) is one of few sweet lupin species that is most commonly grown in Australia as soil nitrogen fixing crop. Australia is the largest producer and exporter on sweet lupins in the world. Despite the fact that lupin is rich of protein and fibre, and the lupin based foods offer several health benefits, the grain is mostly used for animal consumption. Hence, it is a low priced commodity. It is therefore; very beneficial to demonstrate the possibility of deriving nutraceutical, functional and therapeutic agents from sweet lupin grain/flour that is most abundant in Australia. Research in this direction will benefit Australian farmers, food industry as well as consumers. This project aims to derive bioactive peptides from lupin protein. It is well known that bioactive peptides can be derived from many food proteins (both animal and plant sources); such as milk, eggs, cheese, meat, fish, soy, rice, wheat and legumes. The process of protein hydrolysis to produce bioactive peptides is of several types, namely, (i) gastrointestinal digestion by digestive enzymes (ii) fermentation by microbial enzymes and (iii) in vitro hydrolysis by commercial enzymes. Many proteinases such as alcalase, trypsin, chymotrypsin, pancreatin, pepsin and flavourzyme have been employed in the literature to produce important bioactive peptides from food proteins. This study aims to: • Enzymatically hydrolyse sweet lupin protein using pepsin, pancreatin and flavourzyme (lupin hydrolysates). • Separate the lupin hydrolysates by molecular weight cut-off (MWCO) membrane. • Further purification of MWCO fractions by bioactivity guided purification methods. • Size exclusion chromatography (SEC) is used in this research for bioactivity guided purification. • Several bioactivities were determined at different stages of purification. These include; antioxidant, immunomodulatory, ACE inhibitory and antimicrobial activities. • The thesis also aims to evaluate the MRI contrast potential of bioactive lupin peptides isolated in this research

Bioactive peptides from legumes : functional and nutraceutical potential

Protein from legumes are excellent sources of bioactive peptides. Enzymatically derived bioactive peptides from legume proteins display a spectrum of biological activities ranging from nutraceutical to therapeutic potential. Consequently, these peptides possess excellent health promoting properties and potentially prevent many diseases making them extremely suitable for nutraceutical applications. Literature also revealed that the legume proteins: such as soy, lupin, chickpea, yellow pea, common bean and lentil proteins display direct health promoting properties and/or their consumption leads to the production of bioactive peptides by digestive enzymes. Therefore, the direct consumption of legume protein and the food containing such protein provides functional benefits. Literature reviewed in this article showed significant bioactivities of soy protein hydrolysates including the prevention of liver, lung and colon cancer. Soybean peptides are widely used as functional foods to improve the health benefits of many food products. Lupin, yellow pea and chickpea protein hydrolysates have also displayed potent cholesterol lowering, anti-inflammatory, ACE inhibitory, immunomodulatory and antitumor properties. This review highlights the functional applications of legume protein and the nutraceutical applications of legume protein derived bioactive peptides. In this arena of developing extremely beneficial bioactive peptides from food protein, it is necessary to understand the structure-activity relationship of these important biomolecules. A thorough knowledge of such a relationship will provide cues for the selection of suitable enzymes to produce extremely active peptide structures from the known food protein sequences. This review has therefore devoted significant attention to structure-activity relationship of bioactive peptides. This review also highlights the importance of structure determination tools such as NMR spectroscopy as many of the food derived peptide structures are yet to be determined

Application of NMR spectroscopy for structural characterization of bioactive peptides derived from food protein

Traditionally, food has not only been the source of human nutrition but also an excellent factor for deriving nutraceuticals and pharmaceuticals. The biopolymers such as protein and polysaccharides derived from food sources have special significane as they are important part of human diet. Several important bioactive peptides have been discovered from food protein hydrolysates and fermented products that have shown to possess a spectrum of beneficial pharmacological activities. This chapter deals with food protein derived bioactive peptides and their structural characterization employing NMR spectroscopy. It is well known from the vast literature on bioactive peptides that NMR spectroscopy is an indispensable tool to fully characterize the structures of bioactive peptides in their inherent physiological state. However, limited literature exists on detailed structural characterization of food derived bioactive peptides. It is therefore very important to review the literature on structural aspects of food peptides and provide an appraisal of NMR techniques suitable for comprehensive structural characterization of this class of bioactive peptides. This chapter is aimed to introduce NMR spectroscopy and its applications to a wide range of readers in the field of food chemistry in general and to the food peptide researchers in particular. The main objective is to review the literature on food derived bioactive peptides and highlight the potential of NMR spectroscopy to understand structure-activity relationship of bioactive peptides in order to boost further developments in this important field. With a view to make this chapter readable to beginners in the field, basic principles of NMR spectroscopy will also be introduced. Application of solution NMR spectroscopy to bioactive peptides will then be described. Salient features including the size and structural differences of food bioactive peptides when compared to the other classes of bioactive peptides will then be discussed in order to make an informed assessment of choice of NMR techniques suitable for their structural charaterization

Antioxidant activities of lupin (Lupinus angustifolius) protein hydrolysates and their potential for nutraceutical and functional foods

Bioactive peptides are the compounds used as medicines, nutraceuticals and food preservatives. In this study, lupin protein hydrolysates were produced by enzymatic hydrolysis using pepsin, pancreatin and flavourzyme and antioxidative activities of the hydrolysates were measured. Protein was isoelectrically isolated from the lupin seed flour and enzymatically hydrolysed. The hydrolysates were ultrafiltered using molecular weight cut-off (MWCO) membranes. Fractions with masses of <2 kDa, 2–3 kDa, 3–5 kDa and 5–10 kDa were separated from the hydrolysates obtained at different hydrolysis times and then subjected to further fractionation by Size Exclusion Chromatography. Radical scavenging activities against DPPH·, ABTS·+, OH· and Fe2+ chelating abilities of these peptide fractions were measured. The best iron chelating, OH•, DPPH• and ABTS•+scavenging activities (IC50) were 30 ± 5.3, 40 ± 3.9, 60 ± 3.9 and 90 ± 8.2 µg/mL respectively. Pancreatin and Flavourzyme have produced more fractions with best scavenging activities after 3–4 h of hydrolysis (IC50 values in the range of 40–110 µg/mL). Generally Size Exclusion purified fractions displayed superior activities than the parent fractions. Several lupin peptide fractions showed comparable antioxidant capacities to those derived from soybean protein and displayed superior activities when compared with chickpea derived peptide fractions. Therefore, lupin protein hydrolysates are a potential source for nutraceuticals and functional foods

Genetic Behavior of Tomato (Solanum lycopersicum L.) Germplasm Governing Heavy Metal Tolerance and Yield Traits under Wastewater Irrigation

Wastewater irrigation is a substitute for surface water scarcity, but traces of heavy metals (HMs) result in deleterious implications for soil, crop productivity, and in humans. Crops presenting HMs tolerance in genetic behavior are important for producing tolerant genotypes cultivated under wastewater irrigation. In the first part of this experiment, the results obtained previously are re-assessed in a hydroponic system and similar patterns and concentrations of HMs are found in different tomato organs. Following this trial, the tomato&rsquo;s (Solanum&nbsp;lycopersicum L.) genetic basis of traits conferring HMs tolerance and yield are assessed when irrigated with waste or canal water. The North Carolina Mating II analysis illustrate the amount of gene action, nature, and inheritance pattern. Genetic components depict the involvement of non-additive, additive, and maternal genetic effects in HMs tolerance inheritance and yield. A noticeable increase in cumulative additive variance for the number of flowers (11,907.2) and the number of fruits (10,557.9) is recorded for tomato plants irrigated with wastewater, illustrating additive gene action. However, female and male (MSf/MSm) square ratios also show an association with cytoplasmic inheritance. For HMs tolerance, both additive and dominant variances appeared to be significant; cumulative dominance variance (4.83, 16.1, 4.69, 76.95, and 249.37) is higher compared to additive variance (0.18, 2.36, 0.19, &minus;0.27, and 14.14) for nickel (Ni), chromium (Cr), lead (Pb), manganese (Mn), and zinc (Zn), respectively, indicating dominance gene action. The genotype RIOGRANDI accumulated and translocated fewer HMs to the aerial part of the plant compared to CLN-2418A and PB-017906, thus presenting a tolerant tomato genotype according to the hydroponic experiment. This also exhibited a differential pattern of gene action for HMs tolerance, suggesting that genotypes possess significant differences for HMs tolerance

Nutritional and bioactive characteristics of buckwheat, and its potential for developing gluten-free products: An updated overview

In the present era, food scientists are concerned about exploiting functional crops with nutraceutical properties. Buckwheat is one of the functional pseudocereals with nutraceutical components used in the treatment of health-related diseases, malnutrition, and celiac diseases. As a preferred diet as a gluten-free product for celiac diseases, buckwheat is a good source of nutrients, bioactive components, phytochemicals, and antioxidants. The general characteristics and better nutritional profile of buckwheat than other cereal family crops were highlighted by previous investigations. In buckwheats, bioactive components like peptides, flavonoids, phenolic acids, d-fagomine, fagopyritols, and fagopyrins are posing significant health benefits. This study highlights the current knowledge about buckwheat and its characteristics, nutritional constituents, bioactive components, and their potential for developing gluten-free products to target celiac people (1.4% of the world population) and other health-related diseases