Publications by authors named "Baljit Singh"

293 Publications

Engineering and characterization of human β-defensin-3 and its analogues and microcin J25 peptides against Mannheimia haemolytica and bovine neutrophils.

Vet Res 2021 Jun 10;52(1):83. Epub 2021 Jun 10.

Department of Veterinary Biomedical Science, Western College of Veterinary Medicine, University of Saskatchewan, Saskatoon, SK, S7N 5B4, Canada.

Mannheimia haemolytica-induced bovine respiratory disease causes loss of millions of dollars to Canadian cattle industry. Current antimicrobials are proving to be ineffective and leave residues in meat. Antimicrobial peptides (AMPs) may be effective against M. haemolytica while minimizing the risk of drug residues. Cationic AMPs can kill bacteria through interactions with the anionic bacterial membrane. Human β-Defensin 3 (HBD3) and microcin J25 (MccJ25) are AMPs with potent activity against many Gram-negative bacteria. We tested the microbicidal activity of wild-type HBD3, three HBD3 peptide analogues (28 amino acid, 20AA, and 10AA) derived from the sequence of natural HBD3, and MccJ25 in vitro against M. haemolytica. Three C-terminal analogues of HBD3 with all cysteines replaced with valines were manually synthesized using solid phase peptide synthesis. Since AMPs can act as chemoattractant we tested the chemotactic effect of HBD3, 28AA, 20AA, and 10AA peptides on bovine neutrophils in Boyden chamber. Minimum bactericidal concentration (MBC) assay showed that M. haemolytica was intermediately sensitive to HBD3, 28AA and 20AA analogues with an MBC of 50 µg/mL. The 10AA analogue had MBC 6.3 µg/mL which is likely a result of lower final inoculum size. MccJ25 didn't have significant bactericidal effect below an MBC < 100 µg/mL. Bovine neutrophils showed chemotaxis towards HBD3 and 20AA peptides (P < 0.05) but not towards 28AA analogue. Co-incubation of neutrophils with any of the peptides did not affect their chemotaxis towards N-formyl-L-methionyl-L-leucyl-phenylalanine (fMLP). The data show that these peptides are effective against M. haemolytica and are chemotactic for neutrophils in vitro.
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http://dx.doi.org/10.1186/s13567-021-00956-4DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8194028PMC
June 2021

Localization of Nucleobindin2/Nesfatin-1-Like Immunoreactivity in Human Lungs and Neutrophils.

Ann Anat 2021 May 31:151774. Epub 2021 May 31.

Department of Veterinary Biomedical Sciences, University of Saskatchewan, Saskatoon, Canada. Electronic address:

Nucleobindin2 (NUCB2)/nesfatin-1 expression in human plasma positively correlates with the expression of pro-inflammatory cytokines in patients with chronic obstructive pulmonary disease (COPD), implicating its potential role in neutrophilic lung inflammation. There are no data on the localization of nucleobindin2 (NUCB2)/nesfatin-1 in human lungs and inflammatory cells. We examined the localization of NUCB2/nesfatin-1-immunoreactivity in normal and inflamed human lungs obtained from COPD patients and neutrophils with light and immunoelectron microscopy. Immunohistology showed localization of NUCB2/nesfatin-1-like immunoreactivity in the bronchiolar epithelium, alveolar septa, vascular endothelium and various immune cells of normal and inflamed lungs. Further, NUCB2/nesfatin-1-like immunoreactivity accumulated within 0.5 μm of the plasma membrane in human neutrophils following 90 min. of 1 ng/mL LPS stimulation. NUCB2/nesfatin-1-like immunoreactivity was also found to localize in euchromatic portions of neutrophilic nuclei at five times the mean concentration compared to heterochromatin. Finally, our results indicate that NUCB2/nesfatin-1-like immunoreactivity is predominantly cytoplasmic including that in the Golgi complex and vesicles as it localizes at two times the concentration in neutrophilic cytoplasm compared to nucleus. Our study is the first to detail the localization of NUCB2/nesfatin-1-like immunoreactivity in lungs and neutrophils, and nuclear localization of NUCB2/nesfatin-1 also implicates its potential role in transcriptional regulation.
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http://dx.doi.org/10.1016/j.aanat.2021.151774DOI Listing
May 2021

Correction to: Exposures to 2,4-Dichlorophenoxyacetic acid with or without endotoxin upregulate small cell lung cancer pathway.

J Occup Med Toxicol 2021 May 4;16(1):17. Epub 2021 May 4.

Department of Animal Biotechnology, College of Animal Biotechnology, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana, Punjab, 141004, India.

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http://dx.doi.org/10.1186/s12995-021-00307-1DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8097843PMC
May 2021

Pentraxin 3 expression in lungs and neutrophils of calves.

Vet Immunol Immunopathol 2021 Jun 22;236:110251. Epub 2021 Apr 22.

Department of Veterinary Biomedical Sciences, Western College of Veterinary Medicine, University of Saskatchewan, Saskatoon, S7N 5B4, Canada. Electronic address:

Bacterial lung disease caused by Mannheimia haemolytica inflict significant mortality and morbidity resulting in enormous economic losses to cattle industry. The use of antibiotics is becoming more challenging because of development of anti-microbial resistance. The innate immune system plays a critical role in the initiation of immune response in the lung. Pentraxin 3 (PTX3), a pattern-recognition receptor is produced at sites of inflammation by many cell types, recognizes and binds to many pathogens, activates the complement cascade, and has a role in the clearance of apoptotic and necrotic cells. Because there are very few data on the expression of PTX3 in the lungs, we examined PTX3 expression in lungs of normal and M. haemolytica-infected calves and normal and E. coli lipopolysaccharide-treated cattle neutrophils using light and electron microscopic immunochemistry and Western blots. Immunohistology showed the presence of PTX3 in airway epithelial cells, alveolar septa and macrophages in normal and inflamed lungs of calves and the blots showed a significant increase in the expression of PTX3 in lungs from infected calves. Immuno-gold electron microscopy showed PTX3 in the nuclei, cytoplasm, and vesicular organelles of alveolar macrophages, endothelial cells and pulmonary intravascular macrophages (PIMs). Immunohistochemical staining for PTX3 in peripheral blood neutrophils shows an altered staining pattern in neutrophils stimulated with lipopolysachharide (LPS). However, western blots no significant change in PTX3 amount in LPS-treated neutrophils compared to the controls. These are the first data on the expression of PTX3 in the lungs and the neutrophils of cattle which may add to our understanding of innate immunity in cattle lungs.
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http://dx.doi.org/10.1016/j.vetimm.2021.110251DOI Listing
June 2021

Exposures to 2,4-Dichlorophenoxyacetic acid with or without endotoxin upregulate small cell lung cancer pathway.

J Occup Med Toxicol 2021 Apr 17;16(1):14. Epub 2021 Apr 17.

Department of Animal Biotechnology, College of Animal Biotechnology, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana, Punjab, 141004, India.

Background: Pesticide residues in food and environment along with airborne contaminants such as endotoxins pose health risk. Although herbicide 2,4-Dichlorophenoxyacetic acid (2,4-D) has been associated with increased risk of lung cancers such as small cell lung cancer (SCLC) among agricultural workers, there are no data on the SCLC signaling pathway upon 2,4-D exposure without LPS or in combination with endotoxin.

Methods: We exposed Swiss albino mice (N = 48) orally to high (9.58 mg kg) and low (5.12 mg kg) dosages of 2,4-D dissolved in corn oil for 90 days followed by E. coli lipopolysaccharide (LPS) or normal saline solution (80 μl/animal). Lung samples and broncho-alveolar fluid (BALF) were subjected to Total histological score (THS) and total leucocyte count (TLC) and differential leucocytes count (DLC) analyses, respectively. We used microarray and bioinformatics tools for transcriptomic analyses and differentially expressed genes were analyzed to predict the top canonical pathways followed by validation of selected genes by qRT-PCR and immunohistochemistry.

Results: Total histological score (THS) along with BALF analyses showed lung inflammation following long term dietary exposure to high or low doses of 2,4-D individually or in combination with LPS. Microarray analysis revealed exposure to high dose of 2,4-D without or with LPS upregulated 2178 and 2142 and downregulated 1965 and 1719 genes, respectively (p < 0.05; minimum cut off 1.5 log fold change). The low dose without or with LPS upregulated 2133 and 2054 and downregulated 1838 and 1625 genes, respectively. Bioinformatics analysis showed SCLC as topmost dysregulated pathway along with differential expression of Itgb1, NF-κB1, p53, Cdk6 and Apaf1. Immunohistological and quantitative real time PCR (qRT-PCR) analyses also supported the transcriptomic data.

Conclusions: Taken together, the data show exposures to high and low dose of 2,4-D with/without LPS induced lung inflammation and altered pulmonary transcriptome profile with the involvement of the SCLC pathway. The data from the study provide the insights of the potential damage on lungs caused by 2,4-D and help to better understand the mechanism of this complex relation.
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http://dx.doi.org/10.1186/s12995-021-00304-4DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8052721PMC
April 2021

Pulmonary inflammatory response from co-exposure to LPS and glyphosate.

Environ Toxicol Pharmacol 2021 Mar 31;86:103651. Epub 2021 Mar 31.

University of Saskatchewan, 52 Campus Drive, Saskatoon, Saskatchewan, S7N 5B4, Canada.

Agricultural airborne work exposures are complex in nature and workplace exposures are a risk for respiratory outcomes in workers. Endotoxin and glyphosate are two common agents in agricultural exposures. While endotoxin (lipopolysaccaride, LPS) is a potent inflammatory agent it explains only a portion of the respiratory inflammatory response. The inflammatory potential when LPS is presented with another common agricultural respiratory agent, glyphosate, is not known.

Methods: Mice were assigned to four treatment groups: control, LPS alone, glyphosate alone, glyphosate and LPS combined. Treatments were for 1, 5 or 10 days.

Results: Five days of repeated exposure to the comintation of LPS and glyphosate resulted in higher neutrophil counts, myloperoxidase, TNF-α, IL-6, KC levels, and ICAM-1 and TLR-2 expression compared to the same length of treatment to LPS or glyphosate alone. After 10-days of exposure, inflammatory responses decreased, however leukocyte infiltration persisted along with increases in IL-4.

Conclusions: Glyphosate exposure modified LPS induced lung inflammatory responses and TLR-2 may be important in the modulated inflammatory response.
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http://dx.doi.org/10.1016/j.etap.2021.103651DOI Listing
March 2021

Loss of Nucleobindin-2/Nesfatin-1 increases lipopolysaccharide-induced murine acute lung inflammation.

Cell Tissue Res 2021 Mar 30. Epub 2021 Mar 30.

Department of Veterinary Biomedical Sciences, Western College of Veterinary Medicine, University of Saskatchewan, Saskatoon, Canada.

NUCB2/nesfatin-1 is expressed in variety of tissues. Treatment with nesfatin-1 reduces inflammation in rat models of subarachnoid hemorrhage-induced oxidative brain damage and traumatic brain injury as well as myocardial injury. There is only one study showing anti-inflammatory actions of nesfatin-1 on acute lung inflammation. To more precisely determine the role of NUCB2/nesfatin-1 in acute lung inflammation, we conducted a study using NUCB2/nesfatin-1 knockout (NKO) mice as well as neutrophils isolated from the bone marrows of WT and NKO mice. Our findings suggest that the absence of NUCB2/nesfatin-1 significantly increases the accumulation of adherent neutrophils by approximately 3 times compared with WT within LPS-treated lungs. Integrating this with observations from both BALF and neutrophil cytokine expression, we propose that although neutrophils lacking NUCB2/nesfatin-1 individually secrete less pro-inflammatory cytokines compared with stimulated WT cells, the result of knocking out NUCB2/nesfatin-1 is net pro-inflammatory. No change was found in NUCB2/nesfatin-1 mRNA or protein expression comparing WT LPS and PBS-treated samples. Taken together, our results show that NUCB2/nesfatin-1 is constitutively expressed in mouse lungs and neutrophils and demonstrates anti-inflammatory properties in mouse lungs during acute lung injury, by inhibiting adherent neutrophil accumulation and inflammatory cytokine expression.
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http://dx.doi.org/10.1007/s00441-021-03435-6DOI Listing
March 2021

Enhancement of Digestibility of Nutrients (In vitro), Antioxidant Potential and Functional Attributes of Wheat Flour Through Grain Germination.

Plant Foods Hum Nutr 2021 Mar 26;76(1):118-124. Epub 2021 Feb 26.

Department of Food Science and Technology, Punjab Agricultural University, Ludhiana, Punjab, 141004, India.

Wheat grains were germinated at different time (12, 24, 36, and 48 h) and temperature (25, 30, and 35°C) to enhance the functionality of resultant flour. Results revealed that an increase in germination time and temperature enhanced the in vitro digestibility of starch (10.35-42.30 %) and proteins (6.31-44.02 %) owing to their depolymerization by hydrolytic enzymes. Total phenolic and flavonoid content of wheat during germination at variable conditions were enhanced significantly (p < 0.05) from 3.62 to 5.54 mg GAE/g and 32.06 to 54.33 mg QE/100 g, respectively. Germination at elevated temperature (35°C) for a prolonged time (48 h) increased the DPPH RSA by 58.85 %, reducing power by 80.40 % and metal chelating activity by 112.26 % as a result of the structural breakdown of bound phenolics. Increased activity of hydrolytic enzymes also results in a continuous reduction in the viscosity and lightness values of wheat flour. Tailored germination, therefore, can be offered as a tool to increase the nutrient digestibility and bioactive potential of wheat thus resulting in producing the naturally modified flour with enhanced functionality.
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http://dx.doi.org/10.1007/s11130-021-00881-zDOI Listing
March 2021

Lack of CD34 delays bacterial endotoxin-induced lung inflammation.

Respir Res 2021 Feb 25;22(1):69. Epub 2021 Feb 25.

Veterinary Biomedical Sciences, Western College of Veterinary Medicine, University of Saskatchewan, Saskatoon, SK, Canada.

Background: CD34, a pan-selectin binding protein when glycosylated, has been shown to be involved in leukocyte migration to the site of inflammation. However, only one report is available on the expression and role of CD34 in neutrophil recruitment during acute lung inflammation.

Methods: We proceeded to study the role of CD34 in lung neutrophil migration using mouse model of endotoxin induced acute lung inflammation and studied over multiple time points, in generic CD34 knock-out (KO) strain.

Results: While there was no difference in BAL total or differential leukocyte counts, lung MPO content was lower in LPS exposed KO compared to WT group at 3 h time-point (p = 0.0308). The MPO levels in CD34 KO mice begin to rise at 9 h (p = 0.0021), as opposed to an early 3 h rise in WT mice (p = 0.0001), indicating that KO mice display delays in lung neutrophil recruitment kinetics. KO mice do not loose endotoxin induced lung vascular barrier properties as suggested by lower BAL total protein at 3 h (p = 0.0452) and 24 h (p = 0.0113) time-points. Several pro-inflammatory cytokines and chemokines (TNF-α, IL-1β, KC, MIP-1α, IL-6, IL-10 and IL-12 p70 sub-unit; p < 0.05) had higher levels in WT compared to KO group, at 3 h. Lung immunofluorescence in healthy WT mice reveals CD34 expression in the bronchiolar epithelium, in addition to alveolar septa.

Conclusion: Thus, given CD34's pan-selectin affinity, and expression in the bronchiolar epithelium as well as alveolar septa, our study points towards a role of CD34 in lung neutrophil recruitment but not alveolar migration, cytokine expression and lung inflammation.
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http://dx.doi.org/10.1186/s12931-021-01667-2DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7908703PMC
February 2021

Cumulative 5-year Results of a Randomized Controlled Trial Comparing Biological Mesh with Primary Perineal Wound Closure after Extralevator Abdominoperineal Resection (BIOPEX-study).

Ann Surg 2021 Feb 1. Epub 2021 Feb 1.

Department of Surgery, Cancer Center Amsterdam, Amsterdam UMC, University of Amsterdam, Amsterdam, the Netherlands LEXOR, Center for Experimental and Molecular Medicine, Oncode Institute, Cancer Center Amsterdam, Amsterdam UMC, University of Amsterdam, Amsterdam, the Netherlands Department of Radiology and Nuclear Medicine, Amsterdam UMC, University of Amsterdam, Amsterdam, Amsterdam Gastroenterology Endocrinology Metabolism, the Netherlands Department of Sexology and Psychosomatic Gynecology, Amsterdam UMC, University of Amsterdam, Amsterdam, The Netherlands Department Surgery, Deventer Hospital, Deventer, the Netherlands Department of Surgery, Catharina Hospital, Eindhoven, the Netherlands Department of Surgery, University Hospitals Leicester, Leicester, United Kingdom Department of Surgery, Gelre Hospital, Apeldoorn, the Netherlands Department of Surgery, University Medical Center Groningen, University of Groningen, Groningen, the Netherlands Department of Surgery, Tergooi Hospital, Hilversum, the Netherlands Department of Surgery, IJsselland Hospital, Capelle aan de IJssel, the Netherlands Department of Surgery, Medical Center Leeuwarden, Leeuwarden, the Netherlands Department of Surgery, St. Laurentius Hospital, Roermond, the Netherlands Department of Surgical Oncology, Erasmus MC Cancer Institute, Rotterdam, the Netherlands Department of Surgery, Spaarne Gasthuis, Haarlem, the Netherlands Department of Surgery, Radboud University Medical Center, Nijmegen, the Netherlands Department of Epidemiology and Data Science, Amsterdam UMC, University of Amsterdam, Amsterdam, the Netherlands.

Objective: To determine long-term outcomes of a randomized trial (BIOPEX) comparing biological mesh and primary perineal closure in rectal cancer patients following extralevator abdominoperineal resection (APR) and preoperative radiotherapy, with a primary focus on symptomatic perineal hernia.

Summary Background Data: BIOPEX is the only randomized trial in this field, which was negative on its primary endpoint (30-day wound healing).

Methods: This was a post-hoc secondary analysis of patients randomized in the BIOPEX trial to either biological mesh closure (n = 50; 2 dropouts) or primary perineal closure (n = 54; 1 dropout). Patients were followed for five years. Actuarial 5-year probabilities were determined by the Kaplan-Meier statistic.

Results: Actuarial 5-year symptomatic perineal hernia rates were 7% (95% CI, 0-30) after biological mesh closure versus 30% (95% CI, 10-49) after primary closure (P = 0.006). One patient (2%) in the biomesh group underwent elective perineal hernia repair, compared to seven patients (13%) in the primary closure group (P = 0.062). Reoperations for small bowel obstruction were necessary in 1/48 patients (2%) and 5/53 patients (9%), respectively (P = 0.208). No significant differences were found for chronic perineal wound problems, locoregional recurrence, overall survival, and main domains of quality of life and functional outcome.

Conclusions: Symptomatic perineal hernia rate at five-year follow-up after APR for rectal cancer was significantly lower after biological mesh closure. Biological mesh closure did not improve quality of life or functional outcomes.
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http://dx.doi.org/10.1097/SLA.0000000000004763DOI Listing
February 2021

Is there really a shortage of veterinarians in Canada? If so, what are we going to do?

Authors:
Baljit Singh

Can Vet J 2021 01;62(1):75-76

Faculty of Veterinary Medicine, University of Calgary, Calgary.

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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7739382PMC
January 2021

SARS-CoV2 infectivity is potentially modulated by host redox status.

Comput Struct Biotechnol J 2020 20;18:3705-3711. Epub 2020 Nov 20.

Faculty of Veterinary Medicine, University of Calgary, Calgary, Alberta T2N 1N4, Canada.

The current coronavirus disease (COVID-19) outbreak caused by Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV2) has emerged as a threat to global social and economic systems. Disparity in the infection of SARS-CoV2 among host population and species is an established fact without any clear explanation. To initiate infection, viral S-protein binds to the Angiotensin-Converting Enzyme 2 (ACE2) receptor of the host cell. Our analysis of retrieved amino acid sequences deposited in data bases shows that S-proteins and ACE2 are rich in cysteine (Cys) residues, many of which are conserved in various SARS-related coronaviruses and participate in intra-molecular disulfide bonds. High-resolution protein structures of S-proteins and ACE2 receptors highlighted the probability that two of these disulfide bonds are potentially redox-active, facilitating the primal interaction between the receptor and the spike protein. Presence of redox-active disulfides in the interacting parts of S-protein, ACE2, and a ferredoxin-like fold domain in ACE2, strongly indicate the role of redox in COVID-19 pathogenesis and severity. Resistant animals lack a redox-active disulfide (Cys133-Cys141) in ACE2 sequences, further strengthening the redox hypothesis for infectivity. ACE2 is a known regulator of oxidative stress. Augmentation of cellular oxidation with aging and illness is the most likely explanation of increased vulnerability of the elderly and persons with underlying health conditions to COVID-19.
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http://dx.doi.org/10.1016/j.csbj.2020.11.016DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7678423PMC
November 2020

Reply to comments on prone CPR for COVID-19 patients.

Indian J Anaesth 2020 Sep 1;64(9):828-829. Epub 2020 Sep 1.

Department of Anaesthesiology, Gian Sagar Medical College, Patiala, Punjab, India.

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http://dx.doi.org/10.4103/ija.IJA_981_20DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7641069PMC
September 2020

Where do deans of veterinary medicine in the United States and Canada come from?

Can Vet J 2020 11;61(11):1187-1196

Faculty of Veterinary Medicine, 3280 Hospital Drive, NW, University of Calgary, Calgary, Alberta T2N 4Z6.

Deans use passion, integrative thinking, communication skills, charisma, and other leadership skills to build collaborations to advance academic innovation, promote societal awareness of veterinary medicine, and enhance diversity and inclusion. This study analyzed the educational and experiential backgrounds as well as the ethnicity and gender of veterinary medical college deans in the United States and Canada. Data were collected on the deans using public sources from 1966 when the Association of American Veterinary Medical Colleges was started, until 2018. It was found that the advent of specialty colleges led to more visibility of clinical credentials of deans; about 17% of the deans were pathologists, and few deans came from basic science disciplines. The data show that an overwhelming majority of deans have been Caucasian male and very few were racialized/non-Caucasian academics. There are growing numbers of women veterinarians becoming deans. These data may provide some insights on how to assemble leadership training programs to create a more diverse pool of academic veterinary leaders so that more women and ethnic minorities can aspire to become deans.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7560764PMC
November 2020

Depletion of pulmonary intravascular macrophages rescues inflammation-induced delayed neutrophil apoptosis in horses.

Am J Physiol Lung Cell Mol Physiol 2021 01 4;320(1):L126-L136. Epub 2020 Nov 4.

Department of Veterinary Biomedical Sciences, Western College of Veterinary Medicine, University of Saskatchewan, Saskatoon, Saskatchewan, Canada.

The objective of this study was to determine the effect of pulmonary intravascular macrophage depletion on systemic inflammation and ex vivo neutrophil apoptosis using an experimental model of intestinal ischemia and reperfusion injury in horses. Neutrophils were isolated before and after surgery from horses that were randomized to three treatment groups, namely, sham celiotomy (CEL, = 4), intestinal ischemia and reperfusion (IR, = 6), and intestinal ischemia and reperfusion with gadolinium chloride treatment to deplete pulmonary intravascular macrophages (PIMs, IRGC, = 6). Neutrophil apoptosis was assessed with Annexin V and propidium iodide staining quantified with flow cytometry and caspase-3, caspase-8, and caspase-9 activities in neutrophil lysates. All horses experienced a systemic inflammatory response following surgery. Following surgery, ex vivo neutrophil apoptosis was significantly delayed after 12 or 24 h in culture, except in IRGC horses (12 h: CEL: = 0.03, IR: = 0.05, IRGC: = 0.2; 24 h: CEL: = 0.001, IR: = 0.004, IRGC: = 0.3). Caspase-3, caspase-8, and caspase-9 activities were significantly reduced in neutrophils isolated after surgery and cultured for 12 h in IR horses, but not in IRGC horses (IR caspase-3: = 0.002, IR caspase-8: = 0.002, IR caspase-9: = 0.04). Serum TNF-α concentration was increased in IRGC horses for 6-18 h following jejunal ischemia. Following surgery, ex vivo equine neutrophil apoptosis was delayed via downregulation of caspase activity, which was ameliorated by PIM depletion potentially via upregulation of TNF-α.
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http://dx.doi.org/10.1152/ajplung.00392.2019DOI Listing
January 2021

Integrin alpha-v/beta3 expression in equine lungs and jejunum.

Can J Vet Res 2020 Oct;84(4):245-251

Western College of Veterinary Medicine (Le, Gerdts, Singh) and Vaccine and Infectious Disease Organization - International Vaccine Centre (Gerdts), University of Saskatchewan, Saskatoon, Saskatchewan S7N 5B4; Faculty of Animal Science and Veterinary Medicine, Nong Lam University, Ho Chi Minh City, Vietnam (Le); Faculty of Veterinary Medicine, University of Calgary, Calgary, Alberta T2N 4Z6 (Singh).

Integrin alpha-v/beta3 (αvβ3) recognizes arginine-glycine-aspartic acid (RGD) sequences and has important functions in cell adhesion, signaling, and survival. However, the expression of integrin αvβ3 in the equine lungs and jejunum is not well understood. The objective of this study was to explore the hitherto unknown expression of integrin αvβ3 in the lungs and jejuna of the horse using light and electron immunocytochemistry. Immunohistochemistry showed integrin αvβ3 on the epithelium, the immune cells in Peyer's patches, the smooth muscle, and the endothelium of equine jejuna. In equine lungs, we recognized integrin αvβ3 on the endothelium of blood vessels, the alveolar septa, the bronchial lymph nodes, and the cartilages, although the expression of integrin αvβ3 was weak on the epithelium of bronchioles. In conclusion, these are the first data to show the expression of integrin αvβ3 in equine lungs and jejuna.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7491005PMC
October 2020

Public health interventions slowed but did not halt the spread of COVID-19 in India.

Transbound Emerg Dis 2020 Oct 4. Epub 2020 Oct 4.

Department of Community Health Sciences, Cumming School of Medicine, University of Calgary, Calgary, Canada.

The government of India implemented social distancing interventions to contain the COVID-19 epidemic. However, effects of these interventions on epidemic dynamics are yet to be understood. Rates of laboratory-confirmed COVID-19 infections per day and effective reproduction number (R ) were estimated for 7 periods (Pre-lockdown, Lockdown Phases 1 to 4 and Unlock 1-2) according to nationally implemented interventions with phased relaxation. Adoption of these interventions was estimated using Google mobility data. Estimates at the national level and for 12 Indian states most affected by COVID-19 are presented. Daily case rates ranged from 0.03 to 285.60/10 million people across 7 discrete periods in India. From 18 May to 31 July 2020, the NCT of Delhi had the highest case rate (999/10 million people/day), whereas Madhya Pradesh had the lowest (49/10 million/day). Average R was 1.99 (95% CI 1.93-2.06) and 1.39 (95% CI 1.38-1.40) for the entirety of India during the period from 22 March 2020 to 17 May 2020 and from 18 May 2020 to 31 July 2020, respectively. Median mobility in India decreased in all contact domains during the period from 22 March 2020 to 17 May 2020, with the lowest being 21% in retail/recreation, except home which increased to 129% compared to the 100% baseline value. Median mobility in the 'Grocery and Pharmacy' returned to levels observed before 22 March 2020 in Unlock 1 and 2, and the enhanced mobility in the Pharmacy sector needs to be investigated. The Indian government imposed strict contact mitigation, followed by a phased relaxation, which slowed the spread of COVID-19 epidemic progression in India. The identified daily COVID-19 case rates and R will aid national and state governments in formulating ongoing COVID-19 containment plans. Furthermore, these findings may inform COVID-19 public health policy in developing countries with similar settings to India.
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http://dx.doi.org/10.1111/tbed.13868DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7675717PMC
October 2020

Impact of different processing treatments on techno and biofunctional characteristics of dhaincha ().

Food Sci Technol Int 2021 Apr 20;27(3):251-263. Epub 2020 Aug 20.

Department of Food Science and Technology, Punjab Agricultural University, Ludhiana, India.

Impact of different processing techniques (wet heating, microwave processing, dry heating, soaking, and extrusion) on the techno-biofunctional characteristics of dhaincha () was investigated to ascertain the most effective method for processing dhaincha to improve its potentiality for utilization. All the processing techniques improved the cooking quality of dhaincha seeds. Thermal processing techniques were effective in reducing antinutrients and enhancing the antioxidant activity in comparison to soaking. Microwave and extrusion processing completely inactivated lectin and trypsin inhibitor activity and reduced 13.48% and 23.68% saponins, respectively. Extrusion treatment resulted in the maximum increase in total phenols (21.94 to 28.48 GAE mg/g) and flavonoids (0.94 to 1.41 QE mg/g) with consequent highest increase in ABTS· + RSA (161.21 to 261.27 TEAC μmol/100g), FRAP (34.97 to 39.04 TEAC mol/g), reducing power (12.18 to 17.05 AAE mg/g), and metal chelating activity (2.65 to 3.76 mmol EDTAE/100g). Furthermore, extrusion treatment improved techno-functional characteristics and exhibited improvement in the freeze and refrigeration thaw stability in comparison to other methods of processing. Extrusion processing was the most effective method to process dhaincha for improving its techno-biofunctionality.
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http://dx.doi.org/10.1177/1082013220949812DOI Listing
April 2021

Aerosol containment device for use on suspected COVID-19 patients.

Indian J Anaesth 2020 May 23;64(Suppl 2):S154-S156. Epub 2020 May 23.

Department of Anaesthesiology and Critical Care, Faculty of Medicine and Health Sciences, SGT University, Gurugram, Haryana, India.

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http://dx.doi.org/10.4103/ija.IJA_586_20DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7293369PMC
May 2020

Indian Resuscitation Council (IRC) suggested guidelines for Comprehensive Cardiopulmonary Life Support (CCLS) for suspected or confirmed coronavirus disease (COVID-19) patient.

Indian J Anaesth 2020 May 23;64(Suppl 2):S91-S96. Epub 2020 May 23.

Department of Anaesthesiology, Gian Sagar Medical College, Patiala, Punjab, India.

Management of the recent outbreak of the novel coronavirus disease (COVID-19) caused by the severe acute respiratory syndrome coronavirus 2(SARS-CoV-2) remains challenging. The challenges are not only limited to its preventive strategies, but also extend to curative treatment, and are amplified during the management of critically ill patients with COVID-19. Older persons with comorbidities like diabetes mellitus, cardiac diseases, hepatic impairment, renal disorders and respiratory pathologies or immune impairing conditions are more vulnerable and have a higher mortality from COVID-19. Earlier, the Indian Resuscitation Council (IRC) had proposed the Comprehensive Cardiopulmonary Life Support (CCLS) for management of cardiac arrest victims in the hospital setting. However, in patients with COVID-19, the guidelines need to be modified,due to various concerns like differing etiology of cardiac arrest, virulence of the virus, risk of its transmission to rescuers, and the need to avoid or minimize aerosolization from the patient due to various interventions. There is limited evidence in these patients, as the SARS-CoV-2 is a novel infection and not much literature is available with high-level evidence related to CPR in patients of COVID-19. These suggested guidelines are a continuum of CCLS guidelines by IRC with an emphasis on the various challenges and concerns being faced during the resuscitative management of COVID-19 patients with cardiopulmonary arrest.
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http://dx.doi.org/10.4103/ija.IJA_481_20DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7293361PMC
May 2020

Perineal wound closure using gluteal turnover flap or primary closure after abdominoperineal resection for rectal cancer: study protocol of a randomised controlled multicentre trial (BIOPEX-2 study).

BMC Surg 2020 Jul 23;20(1):164. Epub 2020 Jul 23.

Department of Surgery, OLVG Hospital, Amsterdam, The Netherlands.

Background: Abdominoperineal resection (APR) for rectal cancer is associated with high morbidity of the perineal wound, and controversy exists about the optimal closure technique. Primary perineal wound closure is still the standard of care in the Netherlands. Biological mesh closure did not improve wound healing in our previous randomised controlled trial (BIOPEX-study). It is suggested, based on meta-analysis of cohort studies, that filling of the perineal defect with well-vascularised tissue improves perineal wound healing. A gluteal turnover flap seems to be a promising method for this purpose, and with the advantage of not having a donor site scar. The aim of this study is to investigate whether a gluteal turnover flap improves the uncomplicated perineal wound healing after APR for rectal cancer.

Methods: Patients with primary or recurrent rectal cancer who are planned for APR will be considered eligible in this multicentre randomised controlled trial. Exclusion criteria are total exenteration, sacral resection above S4/S5, intersphincteric APR, biological mesh closure of the pelvic floor, collagen disorders, and severe systemic diseases. A total of 160 patients will be randomised between gluteal turnover flap (experimental arm) and primary closure (control arm). The total follow-up duration is 12 months, and outcome assessors and patients will be blinded for type of perineal wound closure. The primary outcome is the percentage of uncomplicated perineal wound healing on day 30, defined as a Southampton wound score of less than two. Secondary outcomes include time to perineal wound closure, incidence of perineal hernia, the number, duration and nature of the complications, re-interventions, quality of life and urogenital function.

Discussion: The uncomplicated perineal wound healing rate is expected to increase from 65 to 85% by using the gluteal turnover flap. With proven effectiveness, a quick implementation of this relatively simple surgical technique is expected to take place.

Trial Registration: The trial was retrospectively registered at Clinicaltrials.gov NCT04004650 on July 2, 2019.
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http://dx.doi.org/10.1186/s12893-020-00823-7DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7376711PMC
July 2020

Graft copolymerization of polyvinylpyrollidone onto Azadirachta indica gum polysaccharide in the presence of crosslinker to develop hydrogels for drug delivery applications.

Int J Biol Macromol 2020 Sep 15;159:264-275. Epub 2020 May 15.

Department of Chemistry, Himachal Pradesh University, Shimla 171005, India.

In this work, graft-copolymerization of poly vinylpyrollidone onto Azadirachta indica gum polysaccharide in the presence of crosslinker has been carried out to prepare the hydrogel for use in drug delivery. The polymers were characterized by cryo-SEM, AFM, FTIR, and C NMR. The gel strength, cross-link density, mesh size, thrombogenicity, antioxidant and mucoadhesion properties of the gum-PVP hydrogels were determined along with the evaluation of drug release profile of methyl prednisolone, a colonic anti-inflammatory agent, from the drug loaded hydrogels. Cryo SEM images showed the porous crosslinked structure of the polymer network. The drug release from the polymer followed non-Fickian diffusion mechanism. The polymers showed 71.47 ± 4.63% haemo-compatibility and 05.52 ± 0.59 Nmm gel strength. The value of DPPH radical scavenging assay (73.16 ± 04.85%) indicated that the gum-PVP polymers are antioxidant. The results of biocompatibility, antioxidant activity, mucoadhesion and drug release properties of the polymers inferred the use of this drug carrier for colonic drug delivery.
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http://dx.doi.org/10.1016/j.ijbiomac.2020.05.091DOI Listing
September 2020

Animal models to study the role of pulmonary intravascular macrophages in spontaneous and induced acute pancreatitis.

Cell Tissue Res 2020 May 22;380(2):207-222. Epub 2020 Apr 22.

Faculty of Veterinary Medicine, University of Calgary, Calgary, AB, T2N 4Z6, Canada.

Acute necrotizing pancreatitis (ANP) is a common gastrointestinal cause of emergency admissions in dogs and humans and can lead to a systemic inflammatory response syndrome resulting in multiple organ dysfunction syndrome. Among the various complications associated with ANP, acute lung injury (ALI) or acute respiratory distress syndrome (ARDS) is a major contributor leading to high mortality rates associated with severe acute pancreatitis (AP) in human patients. The incidence of ALI/ARDS in ANP dogs is not well-characterized in spontaneous AP and there are no models to study it in rodent models. Most of the data related to AP comes from rodent models of AP, which may not always represent the true mechanisms occurring in the lungs of dogs or humans with ANP. Therefore, this manuscript provides a review of current and potential models to study the role of pulmonary intravascular macrophages (PIMs) in acute pancreatitis. Recently, we characterized lung inflammation in clinical cases of AP in dogs and found significant recruitment of PIMs which have been credited as pro-inflammatory cells in species such as cattle, horse, pigs, and sheep that normally have them. Considering the pro-inflammatory roles of constitutive or induced PIMs, we investigated whether a well-established mouse model of ANP has induced PIMs. We found induced PIMs in L-arginine-induced ANP in mouse and that MCP-1 is important in PIM induction in this model. Taken together, now we summarize information on spontaneous ANP in dog and a mouse model of induced ANP to study mechanisms of lung dysfunction and the role of PIMs during ANP.
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http://dx.doi.org/10.1007/s00441-020-03211-yDOI Listing
May 2020

Graft and crosslinked polymerization of polysaccharide gum to form hydrogel wound dressings for drug delivery applications.

Carbohydr Res 2020 Mar 6;489:107949. Epub 2020 Feb 6.

Department of Chemistry, Himachal Pradesh University, Shimla, 171005, India.

The present article discusses the exploration of the potential of moringa gum (MOG) polysaccharides for developing a hydrogel dressing as a slow drug carrier to the enhanced wound healing. The polymer films were formed by grafting of carbomer onto MOG by radiation induced crosslinking technique. The polymers were characterized by cryo-SEM, AFM, FTIR, C NMR spectroscopy, and swelling studies. Drug delivery and biomedical properties of the dressings were also determined. Polymer dressing absorbed 4.20 ± 0.09 g/g simulated wound fluid. The release of levofloxacin was observed without burst effect and followed non-Fickian diffusion mechanism and best fitted in Higuchi kinetic model. The hydrogel films were permeable to O and HO vapour and impermeable to microbes and showed antioxidant activity.
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http://dx.doi.org/10.1016/j.carres.2020.107949DOI Listing
March 2020

Synthesis and characterization of alginate and sterculia gum based hydrogel for brain drug delivery applications.

Int J Biol Macromol 2020 Apr 16;148:248-257. Epub 2020 Jan 16.

Department of Chemistry, Himachal Pradesh University, Shimla 171005, India.

The present work deals with the design of the alginate, sterculia gum polysaccharide and PVP based hydrogel for brain drug delivery applications. The release dynamics of citicoline drug, a nerve regenerating agent, was evaluated. The polymers were investigated by Cryo-SEMs, AFM, FTIR, XRD, C NMR, and swelling studies. The drug release occurred slowly without burst effect and followed mechanism that was approaching the Fickian diffusion mechanism and first order kinetic model. The polymer matrix showed drug loading 40.0 ± 0.8%, thrombose percentage 68.70 ± 8.95%, hemolytic index value 3.66 ± 1.65%, detachment force from the intestinal mucosa = 0.124 ± 0.04 N, and tensile strength 7.67 ± 0.40 N/mm. These films were found biocompatible, antioxidant and mucoadhesive and could be explored for brain drug delivery.
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http://dx.doi.org/10.1016/j.ijbiomac.2020.01.147DOI Listing
April 2020

Exploration of arabinogalactan of gum polysaccharide potential in hydrogel formation and controlled drug delivery applications.

Int J Biol Macromol 2020 Mar 15;147:482-492. Epub 2020 Jan 15.

Department of Chemistry, Himachal Pradesh University, Shimla 171005, India.

Chronic diarrhea is the most common problem in most of the countries with low socio-economic conditions. Hence, efficient therapeutic formulations are required. The present article explores the potential of the gum polysaccharide that itself has an anti-diarrheal activity, to develop the antibiotic drug 'meropenem' carrier, to improve the pharmacotherapy of the diarrhea. The gum based pure and sterile polymeric drug delivery device was prepared by radiation induced crosslinking method. Polymer matrix was characterized by cryo-SEM, AFM, FTIR, C NMR, swelling and drug release studies, gel strength, along with some biomedical properties. The slow release was found without a burst effect with non-Fickian diffusion mechanism. It has also been found that polymer was having pore size = 21.73 nm and crosslink density = 5.28 × 10mol cm which were synthesized with [HEMA] 4.70 × 10 mol/L and irradiation dose 24.62 kGy during copolymerization reaction. The arabinogalactan crosslinked poly(HEMA) polymers were biocompatible, antioxidant and mucoadhesive.
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http://dx.doi.org/10.1016/j.ijbiomac.2020.01.087DOI Listing
March 2020

Electrochemical enzyme-linked immunosorbent assay (e-ELISA) for parasitic nematode Ostertagia ostertagi (brown stomach worm) infections in dairy cattle.

Analyst 2019 Sep;144(19):5748-5754

MiCRA Biodiagnostics Technology Gateway, Technological University Dublin (TU Dublin), Tallaght, Dublin 24, D24 FKT9, Ireland.

A sensitive electrochemical immunoassay (e-ELISA) has been developed for the detection of the gastrointestinal parasitic nematode Ostertagia ostertagi (brown stomach worm) in infected and control serum samples. An antigen-indirect immunoassay format was employed to detect the presence of O. ostertagi antibodies, coupled with an anti-species monoclonal horseradish peroxidase (HRP) conjugate. ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid)) and TMB (3,3',5,5'-tetramethylbenzidine/hydrogen peroxide) were investigated as both chromogenic visualising reagents for optical ELISA and electroactive substrates for electrochemical ELISA in the HRP catalysed oxidation reaction. Coulometry was applied for the detection of O. ostertagi antibodies (via TMB electrochemistry) and compared with the commercial optical ELISA (ABTS based SVANOVIR® O. ostertagi-Ab ELISA kit). Cost-effective in-house sensors were designed and fabricated using polyester and chemical adhesive materials with the aid of stencil printing and laser machining techniques. The performance of the electrochemical ELISA and sensor was evaluated by investigating redox mediators (ABTS vs. TMB), stop solutions (sodium dodecyl sulfate vs. sulfuric acid) and incubation times (150 min vs. 70 min vs. 25 min). For a total assay incubation time of 70 minutes, the TMB/H2SO4 based e-ELISA was able to differentiate between positive (P) and negative (N) control serum samples, with a P/N70 control ratio 1.6 times higher than that of optical ELISA (TMB/H2SO4 combination) and 2.9 times higher than that of the commercial ELISA kit (ABTS/SDS combination). Furthermore, the e-ELISA approach is quicker and required only 25 min (total incubation time) with even better response (P/N25 = 14.7), which is approximately 4-fold higher than the optical immunoassay (P/N25 = 3.8). The proposed e-ELISA is specific (selective Ab-Ag interactions) and highly sensitive - capable of detecting up to 16-fold dilutions of a positive control serum sample. The electrochemical ELISA approach has the potential for rapid sample screening in a portable, disposable format, contributing to the quest for effective prevention and control of parasitic Ostertagia ostertagi infections in cattle.
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http://dx.doi.org/10.1039/c9an00982eDOI Listing
September 2019

Impact of Feed Moisture on Microstructure, Crystallinity, Pasting, Physico-Functional Properties and In Vitro Digestibility of Twin-Screw Extruded Corn and Potato Starches.

Plant Foods Hum Nutr 2019 Dec;74(4):474-480

Department of Food Science and Technology, Punjab Agricultural University, Ludhiana, 141004, India.

The effect of extrusion feed moisture (FM) on the microstructure, pasting, physico-functional properties and in vitro starch digestibility (IVSD) of corn and potato starches was investigated using scanning electron microscopy (SEM), X-ray diffractometry and a rapid visco-analyser. Starches were extruded at 14, 18 and 22% FM with an extrusion temperature of 100 °C and a screw speed of 100 rpm. Extruded starches showed lower L* (lightness) values and higher a* and b* values than native starches. An increase in FM increased the L* values and decreased the a* and b* values of extruded starches. Extrusion resulted in complete destruction and reduced crystallinity of the starch structure. Extruded starches showed a lower water absorption index (WAI), peak viscosity (PV), final viscosity (FV), breakdown viscosity (BDV) and setback viscosity (SBV) with a higher water solubility index (WSI) and IVSD than native starches. FM showed a negative correlation with the WSI and IVSD and a positive correlation with the WAI, PV, FV, BDV and SBV of extruded starches.
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http://dx.doi.org/10.1007/s11130-019-00762-6DOI Listing
December 2019

Characterization of in vitro antioxidant activity, bioactive components, and nutrient digestibility in pigeon pea (Cajanus cajan) as influenced by germination time and temperature.

J Food Biochem 2019 02 8;43(2):e12706. Epub 2018 Nov 8.

Department of Food Science and Technology, Punjab Agricultural University, Ludhiana, India.

Germination of legumes is potential bioprocessing technique to improve the nutrient digestibility and enhancement of bioactive components. The present investigation studies the effects of different germination conditions on the bioactive components, antioxidant activity, and in vitro nutrient digestibility of pigeon pea. Results obtained indicated that increase in germination time and temperature modifies the bioactive components and nutritional digestibility of the pigeon pea. Studies have shown that increase in germination time from 12 to 48 hr and temperature from 25 to 35°C, results in significant increase in accumulation of total pheolics and flavonoid content as a result of cell wall degrading enzymes. Germination for prolonged time at higher temperature also significantly increases the antioxidant potential and reducing power of the germinated pigeon pea. Increased activity of hydrolytic enzymes alters the structure of starch and proteins and thus enhanced in vitro starch and protein digestibility and also lowers down the hot paste viscosity of germinated pigeon pea. PRACTICAL APPLICATIONS: Currently, consumers are increasingly interested in high-quality natural health foods with high biological value and better nutrient digestibility. Germinated grains legumes become popular among people engrossed in improving and maintaining their health by changing dietary habits. Germinated pigeon pea exhibited better nutrient digestibility with improved texture and flavor, higher antioxidant activity and more bioactive components with lower pasting viscosity due to enzymatic modification of starch and protein in the grain. For this reason, use of germinated pigeon pea flour can provide an excellent example of functional food with high dietary protein value. Germinated pigeon pea flour can be utilized as a functional ingredient in the preparation of novel function foods, and it would intensify metabolism, strengthen immunity, reimburse deficiencies of vitamins and mineral, lower the risk of various diseases and exert health-promoting effects.
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http://dx.doi.org/10.1111/jfbc.12706DOI Listing
February 2019