Complete Digestion Research Articles

157 Starch and Dry Matter Digestibility Values Are Not Associated with Improved Growth and Feed Efficiency in Weanling Pigs Fed a Therapeutic Multi-antimicrobial-supplemented Diet

Abstract The porcine terminal small intestinal starch digestive activity and capacity undergo further developmental changes during the acute and adaptive weaning transition. Objective of this study was to address the hypothesis that weanling porcine growth promotion associated with therapeutic antimicrobials was mediated via improving intestinal starch digestion. A total of 170 crossbred barrows (n = 34 pens; 5 barrows/pen), weaned on d 19–21 with an average initial BW of 6.9±0.1 kg, were randomly assigned to two dried whey, corn and SBM-based diets, i.e., control vs. antimicrobials, for 21 d according to a randomized complete block design. The therapeutic multi-antimicrobial diet was supplemented with aureomycin 220, tiamulin 31.2 and ZnO 2,358 at mg/kg diet. The apparent ileal, cecal and total tract digestibility values of total starch and DM were determined by using the titanium dioxide as the digestibility marker (0.30% in the diets). There were no differences (P > 0.05) in the starch digestibility at the ileal (control, 98.3±0.9% vs. antimicrobials, 98.3±0.5%), cecal (control, 96.3±0.6% vs. antimicrobials, 97.1±0.6%) and total tract (control, 98.5±0.3% vs. antimicrobials, 98.7±0.3%) levels between the two diets. Furthermore, there were no differences (P > 0.05) in the DM digestibility at the ileal (control, 75.7±2.2% vs. antimicrobials, 72.7±2.0%), cecal (control, 80.6±1.2% vs. antimicrobials, 81.3±1.2%) and total tract (control, 85.8±0.8% vs. antimicrobials, 84.7±0.9%) levels between the two diets. The Spearman’s correlation analyses revealed no significant associations (P > 0.05) between the growth performance endpoints of ADG, ADFI and F: G and the digestibility values of total starch and DM at the levels of ileal, cecal and total tract. Our results suggest that starch is near complete digestion at the distal small intestine, thus not limiting growth and feed conversion efficiency of weanling pigs fed corn and SBM-based diets. Hence, therapeutic multi-antimicrobial effects are unlikely channeled through modulating digestive utilization efficiency of starch and DM in weanling pigs.

S2596 Glycogenic Hepatopathy: A Rare Cause of Reversible Severe Hepatitis

Introduction: Glycogenic hepatopathy (GH) is a rare reversible complication of poorly controlled diabetes mellitus. Its presentation can range from mild to severe hepatitis and diagnosis can be challenging. Here we present a case of GH presenting as severe hepatitis. Case Description/Methods: A 51-year-old male with a history of hypertension and poorly uncontrolled T1DM presented with two days of intractable nausea and vomiting. Physical examination revealed cachectic male only oriented to name. Abdominal exam was normal without tenderness to palpation or organomegaly. Laboratory results were significant for bicarbonate < 5.0 mmol/L, glucose 1169 mg/dL, total bilirubin (T bili.) 0.4 mg/dL, alkaline phosphatase (ALKP) 141 U/L, alanine aminotransferase (ALT) 161 U/L, aspartate aminotransferase (AST) 107 U/L, albumin 3.6 g/dL, INR 1.0, hemoglobin A1c 13.7%. Head computed tomography showed no acute intracranial pathology. He was admitted to the intensive care unit, diagnosed with DKA and treated with insulin infusion therapy. On day 4, laboratory results showed T bili. 0.4 mg/dL, ALKP 295 U/L, ALT 1570 U/L, AST 5250 U/L, INR 1.0. An ultrasound showed mild hepatomegaly 14cm, hyperechoic liver parenchyma, no intra or extra hepatic biliary dilation, and patent hepatic and portal veins. Workup of viral hepatitis A, B and C, autoimmune hepatitis, Wilson’s disease, alpha 1 antitrypsin deficiency, and hemochromatosis was negative. An ultrasound guided liver biopsy showed diffuse swollen hepatocytes, minimal focal fibrosis and mild inflammation composed of scattered lymphocytes, plasma cells and few polymorphonuclear leukocytes. Periodic acid-Schiff (PAS) stain with and without diastase highlighted abundant cytoplasmic glycogen. Patient was diagnosed with GH and a rapid downtrend in liver enzymes was noted with DKA treatment. Discussion: GH was first described by Peter Mauriac in 1930 in children with uncontrolled T1DM. Its incidence is unknown as there have been less than 100 cases reported so far in English literature. Pathophysiology of GH remains unclear but rapid fluctuations in glucose and insulin leading to excessive glycogen deposition in hepatocytes has been suggested. Distinguishing GH from NASH, DILI and other chronic liver disease is challenging without a liver biopsy. With proper glycemic control, GH is quickly reversible and has a good prognosis. No long term hepatic complications are known. GH remains underdiagnosed due to a combination of diagnostic challenges and lack of awareness amongst specialists.Figure 1.: Histopathological section of the liver showing: A) Hepatocytes with clear and fine granular cytoplasm under hematoxylin and eosin stain, B) PAS stain showing large amount of dark red-colored glycogen in hepatocytes, C) Complete digestion of glycogen after adding diastase stain and reappearance of clear hepatocytes.

First report of interspecific transmission of sarcoptic mange from Iberian ibex to wild boar

BackgroundSarcoptic mange is a globally distributed parasitic disease caused by the burrowing mite Sarcoptes scabiei. This mite has a certain degree of host specificity, although interspecific transmission can occur among phylogenetically related species or through prey–predator mediated exposure. In 2018, a wild boar (Sus scrofa) with lesions compatible with sarcoptic mange was hunted in Ports de Tortosa i Beseit Natural Park (PTB, north-eastern Spain), where an active epizootic outbreak of sarcoptic mange is affecting Iberian ibexes (Capra pyrenaica) since 2014.MethodsA complete necropsy, skin scrapings and skin digestions with hydroxide potassium were performed to confirm the diagnosis. Routine histopathological analysis, toluidine blue staining and immunohistochemistry were used to characterize the lesions and the inflammatory infiltrate. Finally, 10 specific S. scabiei microsatellites were molecularly genotyped through polymerase chain reactions in mites obtained from the affected wild boar. For phylogenetic comparison, mites obtained from sympatric Iberian ibexes and allopatric wild boars and Iberian ibexes from southern Spain were analysed.ResultsSarcoptes scabiei was visually and molecularly identified in the infested wild boar from PTB, causing skin lesions with dermal inflammatory infiltrate rich in T and B cells, which indicate an adaptive immune response. Three S. scabiei genetic clusters were identified: one included mites from southern Iberian ibexes, another included mites from southern wild boars, and a third one distinctively grouped the wild boar from PTB with the sympatric ibexes.ConclusionsTo the authors’ knowledge, this is the first reported case of sarcoptic mange in wild boar in Spain and the first documented case of S. scabiei cross-transmission from a wild ruminant host to a wild boar. The wild boar presented an ordinary scabies type reaction, which is typical of the self-limiting infestations reported in other cases of interspecific transmission.Graphical abstract

Structural requirements of flavonoids for the selective inhibition of α-amylase versus α-glucosidase

In the present study, 14 structurally unique flavonoids were screened to systematically investigate structural requirements for selectively inhibiting human α-amylase versus α-glucosidase to obtain a slow but complete starch digestion for health benefit. The selective inhibition property of three flavonoids chosen against the two classes of starch digestive enzymes was confirmed through various analytical techniques - in vitro inhibition assay, fluorescence quenching, kinetic study, and molecular modeling. Considering the chemical structure of flavonoids, the double bond between C2 and C3 and OH groups at A5 and B3 are critical for the inhibition of α-amylase allowing flavonoids to lie parallel on the α-amylase catalytic active site, whereas the OH groups at B3 and C3 are important for α-glucosidase inhibition causing B-ring specific entry into the catalytic active site of α-glucosidase. Our findings provide insights into how to apply flavonoids to effectively control digestion rate for improving physiological responses.

Dietary starch is weight reducing when distally digested in the small intestine

Nowadays, carbohydrate-based foods have a negative consumer connotation and low carb diets have become a popular way to lose weight. Here, we show how digestible starch and flavonoids can be used as a dietary approach to manage food intake and weight gain through elevation of glucagon-like peptide-1 (GLP-1) secretion for gut-brain axis communication. This was achieved by extending the digestion of cooked starch to the distal small intestine using luteolin or quercetin as α-amylase-specific inhibitors with competitive inhibition mechanism. In a mouse model, extended and complete digestion produced a signature blunted glycemic profile that induced elevation of GLP-1 and positive regulation of hypothalamic neuropeptides with significantly reduced food intake and weight gain (p < 0.05). These findings represent a shift in paradigm of dietary carbohydrates from weight increasing to reducing, and have implications for industry and public health related to the design of carbohydrate-based foods/ingredients for managing obesity and diabetes.

Gut Microbiota and Their Role in Health and Metabolic Disease of Dairy Cow.

Ruminants are mostly herbivorous animals that employ rumen fermentation for the digestion of feed materials, including dairy cows. Ruminants consume plant fibre as their regular diet, but lack the machinery for their digestion. For this reason, ruminants maintain a symbiotic relation with microorganisms that are capable of producing enzymes to degrade plant polymers. Various species of microflora including bacteria, protozoa, fungi, archaea, and bacteriophages are hosted at distinct concentrations for accomplishing complete digestion. The ingested feed is digested at a defined stratum. The polysaccharic plant fibrils are degraded by cellulolytic bacteria, and the substrate formed is acted upon by other bacteria. This sequential degradative mechanism forms the base of complete digestion as well as harvesting energy from the ingested feed. The composition of microbiota readily gets tuned to the changes in the feed habits of the dairy cow. The overall energy production as well as digestion is decided by the intactness of the resident communal flora. Disturbances in the homogeneity gastrointestinal microflora has severe effects on the digestive system and various other organs. This disharmony in communal relationship also causes various metabolic disorders. The dominance of methanogens sometimes lead to bloating, and high sugar feed culminates in ruminal acidosis. Likewise, disruptive microfloral constitution also ignites reticuloperitonitis, ulcers, diarrhoea, etc. The role of symbiotic microflora in the occurrence and progress of a few important metabolic diseases are discussed in this review. Future studies in multiomics provides platform to determine the physiological and phenotypical upgradation of dairy cow for milk production.

Agile fusion method for the determination of Pu isotopes in diverse sediments

Atmospheric nuclear testing has occurred worldwide resulting in trace amounts of plutonium (Pu) isotopes in soils and sediments across the globe. Soils and sediments can vary greatly in both physical and chemical properties within close geographic regions presenting a major challenge for sample preparation in the analysis of fallout radionuclides. It is vital to have a rapid, agile method for the complete digestion of samples containing refractory Pu particles to address this challenge. A radioanalytical procedure for the analysis of Pu isotopes in soils/sediments has been developed improving sample preparation using fusion. This is done by eliminating the need for pre-dissolution combustion to remove organic material and providing greater application to a diverse range of samples including those of metal-bearing mineralogy. The integration of borate and peroxide flux materials creates a sample dissolution procedure that is greater than the sum of its parts. This method is applicable to the analysis of 239/240Pu in soil/sediment samples with a sample mass of 3 g, ranging from 0.05 to 5000 mBq/g achieving 242Pu tracer recoveries between 50 and 94%, with a trace-level detection limit of 0.04 mBq/g and measurement uncertainties down to 19%; adjustable to measurement priorities with variation in count time, background counts, chemical recovery and total activity of the sample. This sample preparation technique can be utilized with complementary analysis of alpha spectrometry or ICP-MS. Achieving these parameters provides a highly adaptable standard method for 239/240Pu in soil/sediment analysis applicable to samples from radioecological research at trace environmental levels to emergency response scenarios with 48-h turn-around producing highly repeatable, succinct and high-quality analysis.

Efficiency of acid digestion procedures for geochemical analysis of tungsten mining wastes

There is an increasing global demand for tungsten, which is a critical element used in various industries. There are millions of tons of current and legacy mineral processing tungsten tailings worldwide that can potentially contaminate the environment and pose human health risks. These tailings could also become valuable resources if we thoroughly characterize their geochemical composition. In this study, an innovative method was developed to achieve the complete digestion of tungsten tailings. We tested three different digestion methods (hotplate digestion, bomb digestion and ColdBlock TM digestion) and compared the results. Additionally, an alkali fusion for major element analysis was applied and tested. The results showed that alkali fusion is the best method for major element analysis, while bomb digestion is best for tungsten and trace element analysis, although volatile chlorite loss was also observed. The hot plate digestion method was not recommended, owing to poor recovery of trace elements compared to the bomb digestion method. The quicker and safer ColdBlock TM digestion method can be used for bismuth, molybdenum, and several rare-earth element analyses, as indicated by their recovery being close to that from the bomb digestion method.

Development and validation of a fast ultra-high-performance liquid chromatography tandem mass spectrometry method for determining carbonic anhydrase inhibitors and their metabolites in urine and hair.

A new, rapid, sensitive, and comprehensive ultra‐high‐performance liquid chromatography tandem mass spectrometry (UHPLC–MS/MS) method for quantifying diuretics (acetazolamide, brinzolamide, dorzolamide, and their metabolites) in human urine and hair was developed and fully validated. Twenty‐five milligrams of hair were incubated with 500‐μl M3® buffer reagent at 100°C for 1 h for complete digestion. After cooling, 1‐μl supernatant was injected onto chromatography system. Urine samples were simply diluted before injection. The chromatographic run time was short (8 min) through a column with a mobile phase gradient. The method was linear (determination coefficients always higher than 0.99) from limit of quantification (LOQ) to 500 ng/ml in urine and from LOQ to 10 ng/mg in hair. LOQs ranged from 0.07 to 1.16 ng/ml in urine and from 0.02 to 0.15 ng/mg in hair. No significant ion suppression due to matrix effect was observed, and process efficiency was always higher than 80%. Intra‐ and inter‐assay precision was lower than 15%. The suitability of the methods was tested with six urine and hair specimens from patients treated with acetazolamide, dorzolamide, or brinzolamide for ocular diseases or systemic hypertension. Average urine concentrations were 266.32 ng/ml for dorzolamide and 47.61 ng/ml for N‐deethyl‐dorzolamide (n = 3), 109.27 ng/ml for brinzolamide and 1.02 ng/ml for O‐desmethyl‐brinzolamide (n = 2), and finally, 12.63 ng/ml for acetazolamide. Average hair concentrations were 5.94 ng/mg for dorzolamide and 0.048 ng/mg for N‐deethyl‐dorzolamide (n = 3), 3.26 ng/mg for brinzolamide (n = 2), and 2.3 ng/mg for acetazolamide (n = 1). The developed method was simple and fast both in the extraction procedures making it eligible in high‐throughput analysis for clinical forensic and doping purposes.

STUDY OF GIT RELATED KOSHTHANGA W.S.R. TO PAKWASHAYA

Human anatomy is the science where human body structures are designed to enable complete physiological action thus establishing homeostasis of the human body. Acharya Sushruta had a keen observation about the human body reflecting that without the absolute knowledge of Rachana Sharir. Chikitsak cannot be considered an expert. In Ayurveda, the part of G.I.T. is mentioned in Koshthanga by various Acharya. These Koshthanga are Aamashaya, Pakwashaya, Purishdhara, Uttarguda, Adharguda, Kshudrantra etc. which are situated within the koshtha. In modern anatomy, G.I.T. or Alimentary canal includes all the structures between the mouth and anus, forming a continuous passageway that includes the main organ of digestion, namely the stomach small intestine and large intestine, each part of the Gastric intestinal tract is adapted to its specific function. The 'oesophagus' function primarily to conduct food rapidly from the pharynx to the stomach mixing along with the digestive juic- es, carrying out partial digestion and then propelling the food into the duodenum is the function of the stomach small intestine is designed for complete digestion and absorption of nutrients. Absorption of water and electrolyte from the chyme to form solid faeces is the function of the large intestine. Pakwashaya is the main organ related to the site of Vata Dosha, Purishvaha srotas, Purishdhara kala, Koshthanga and Aashaya. Pakwashaya plays an important role in formation of urine and digestion of food. Keywords: Pakwashaya, Purish

Development of a liquid biopsy based purely quantitative digital droplet PCR assay for detection of MLH1 promoter methylation in colorectal cancer patients

BackgroundMutL Homolog 1 (MLH1) promotor methylation is associated with microsatellite instability high colorectal cancer (CRC). The strong correlation between methylation status and cancer development and progression has led to a growing interest in the use of methylation markers in circulating tumor DNA (ctDNA) for early cancer detection and longitudinal monitoring. As cancer-specific DNA methylation changes in body fluids are limited, it is particularly challenging to develop clinically applicable liquid biopsy methodologies with high sensitivity and specificity. The purpose of this study was to develop a fit-for-purpose methylation sensitive restriction enzyme (MSRE) based digital droplet PCR (ddPCR) assay to examine MLH1 promoter methylation in ctDNA in advanced CRC.MethodsPrimers and probes were designed to amplify CpG sites of the MLH1 promoter. Methylated and unmethylated control genomic DNA were sheared to mimic ctDNA and subjected to MSRE HpaII digestion. Plasma samples from 20 healthy donors and 28 CRC patients were analyzed with the optimized MSRE procedure using ddPCR.ResultsUsing methylated and unmethylated controls, we optimized the conditions for HpaII enzyme digestion to ensure complete digestion and avoid false positives. Based on the results from the ddPCR assay using 1 ng circulating cell-free DNA (cfDNA) input from healthy donors or CRC samples, ROC curves were generated with an area under the curve (AUC) value of 0.965 (95% CI: 0.94, 0.99). The statistically optimal assay sensitivity and specificity was achieved when 8 positive droplets were used as acceptance criteria (78% sensitivity and 100% specificity, 95% CI: 0.45, 0.95). A tiered-based cutoff (20, 50, 80% percentile based) was applied to distinguish CRC samples with different methylation level.ConclusionsOur study demonstrated that the liquid biopsy assay for MLH1 promoter methylation detection using purely quantitative ddPCR is a simple and highly sensitive procedure that provides reliable methylation detection in ctDNA. The MSRE ddPCR approach can also be applied to other genes of interest where methylation patterns could reveal clinically relevant information for future clinical biomarker and/or companion diagnostic development.

Multielemental Analysis and In Vitro Evaluation of Free Radical Scavenging Activity of Natural Phytopigments by ICP-OES and HPTLC.

The phytopigments derived from ethnomedicinal plants employed as traditional medicines appear to be the simplest alternative for artificial radical colorants. This can be because of persistent use of synthetic dyes and their harmful impacts linked to human lives as well as to the ecosystem. The literature evidences clearly reveal the complications from growing demands of radical colorants from artificial origin. The planned analysis work hence focuses on screening of the fundamental composition of phytopigments, obtained from plant sources by subtle technique of ICP-OES, with axial plasma combined with nebulizer motor–assisted gas flow approach, utilizing microwave digester for complete digestion of phytopigments, thereby establishing the pigments being safe for consumption. Additionally, the observations from free radical scavenging activity using DPPH by HPTLC concluded that the natural pigments obtained from plant sources are rich in flavonoids with potent antioxidant property. Thus, an effort has been made through the developed ICP-OES methodology, to beat the distinct imprecise practice of food labeling, once natural pigments are utilized in a variety of additives, as food colorants with amounts of components detected as arsenic, lead, and metal, within specified limits of FSSAI, demonstrate and establish safety of natural foodstuff agents, as compared over hazardous synthetic azo dyes.

Polyenzyme preparations in feeding of monogastric animals

In animal feeding enzyme preparations are mainly used, which are not produced in the body of animals, although in chickens in the first weeks of life and piglets after weaning an insufficient amount of their own proteases and amylases is formed, so their inclusion in feed is also useful. By now highly active enzyme preparations with various specific activities have been created and are being produced. In feeds prepared using traditional raw materials up to 30% of nutrients are not available for digestion by enzymes secreted in the gastrointestinal tract. The inclusion of exogenous enzymes in the feed provides additional use of the indigestible fraction. It is impossible to achieve complete digestibility of feed, although in vitro exogenous enzymes digest almost 100% of the target substrates. The problem is due to the fact that natural feed does not contain pure substrates — they are associated with other substances that limit the availability of enzymes to them. The properties of natural substrates, their concentration and availability even in the same raw material are variable, which excludes reliable prediction of the effectiveness of the action of feed enzymes. Partially these difficulties are overcomed by the use of polyenzyme drugs. It is necessary to further study the specificity of native substrates and factors that increase their availability for the action of enzymes.

MiRNA identification by nuclease digestion in ELISA for diagnosis of osteosarcoma.

Osteosarcoma is a bone cancer formed by the cells of the bone. Children, young adults, and teens are highly affected by osteosarcoma. Early identification of osteosarcoma is mandatory to improve the treatment and increase the lifespan of the patients. MicroRNA-195 (miR-195) was shown to be a suitable biomarker for osteosarcoma, and the present study describes a sensitive method of miR-195 identification by nuclease digestion in ELISA to detect and quantify the level of miR-195. S1 nuclease catalyzed endo- and exonucleolytic digestion of single-stranded (ss) RNA and DNA on ELISA polystyrene substrate, which helped to identify duplexed miR-195. This method selectively and specifically identified miR-195 without any biofouling interactions and reached the limit of detection at 10fM within the range from 10fM to 10nM. Due to complete digestion of ssDNA, single- and triple-mismatched sequences failed to increase the ELISA signal, indicating specific miRNA detection. Furthermore, human serum spiked with miR-195 did not interfere with the detection, confirming selective identification. This method identified miR-195 at a lower level and will help to diagnose earlier stages of osteosarcoma.

Occurrence and Distribution of Technology-Critical Elements in Recent Freshwater and Marine Pristine Lake Sediments in Croatia: A Case Study.

The occurrence and vertical distribution of ten technology critical elements (TCEs) (Li, Nb, Sc, Ga, Y, La, Sb, Ge, Te, and W) were studied in sediment cores collected from remote freshwater and marine lakes (Plitvice, Visovac and Mir Lakes) in three protected areas of Croatia. These environmental archives were used to assess natural TCE levels in lake sediments and temporal trends in historical anthropogenic atmospheric deposition. TCE was determined after complete sediment digestion using high-resolution inductively coupled plasma mass spectrometry (HR ICP-MS). The measured TCE concentrations spanned a wide range, which can be attributed to the varying input of terrigenous material into the studied lake systems. All obtained TCE concentrations were close to natural conditions and therefore could be used as a reference for other equivalent sediment systems in the coming years. The evaluation of anthropogenic influence on TCE concentrations showed a slight anthropogenic enrichment with Sb and Te in the upper sediment layers of some lakes (Plitvice and Mir Lakes), indicating a widespread atmospheric deposition, which, however, cannot be related to the recent increase in the use of TCE in modern technology.

From simple and specific zymographic detections to the annotation of a fungus Daldinia caldariorum D263 that encodes a wide range of highly bioactive cellulolytic enzymes

BackgroundLignocellulolytic enzymes are essential for agricultural waste disposal and production of renewable bioenergy. Many commercialized cellulase mixtures have been developed, mostly from saprophytic or endophytic fungal species. The cost of complete cellulose digestion is considerable because a wide range of cellulolytic enzymes is needed. However, most fungi can only produce limited range of highly bioactive cellulolytic enzymes. We aimed to investigate a simple yet specific method for discovering unique enzymes so that fungal species producing a diverse group of cellulolytic enzymes can be identified.ResultsThe culture medium of an endophytic fungus, Daldinia caldariorum D263, contained a complete set of cellulolytic enzymes capable of effectively digesting cellulose residues into glucose. By taking advantage of the unique product inhibition property of β-glucosidases, we have established an improved zymography method that can easily distinguish β-glucosidase and exoglucanase activity. Our zymography method revealed that D263 can secrete a wide range of highly bioactive cellulases. Analyzing the assembled genome of D263, we found over 100 potential genes for cellulolytic enzymes that are distinct from those of the commercially used fungal species Trichoderma reesei and Aspergillus niger. We further identified several of these cellulolytic enzymes by mass spectrometry.ConclusionsThe genome of Daldinia caldariorum D263 has been sequenced and annotated taking advantage of a simple yet specific zymography method followed by mass spectrometry analysis, and it appears to encode and secrete a wide range of bioactive cellulolytic enzymes. The genome and cellulolytic enzyme secretion of this unique endophytic fungus should be of value for identifying active cellulolytic enzymes that can facilitate conversion of agricultural wastes to fermentable sugars for the industrial production of biofuels.

Methacrylate-functionalized proanthocyanidins as novel polymerizable collagen cross-linkers – Part 1: Efficacy in dentin collagen bio-stabilization and cross-linking

ObjectiveThe aim of the study was to investigate the effects of methacrylate-functionalized proanthocyanidins (MAPAs) on dentin collagen’s bio-stabilization against enzymatic degradation and crosslinking capability. MethodsThree MAPAs were synthesized via varying methacrylate (MA) to proanthocyanidins (PA) feeding ratios of 1:2, 1:1, and 2:1 to obtain MAPA-1, MAPA-2, and MAPA-3, respectively. The three MAPAs were structurally characterized by proton nuclear magnetic resonance (1H NMR) and Fourier transform infrared (FTIR) spectroscopic methods. 5-μm-thick dentin films were microtomed from dentin slabs of third molars. Following demineralization, films or slabs were treated with 1% MAPAs or PA in ethanol for 30 s. Collagen bio-stabilization against enzymatic degradation was analyzed by weight loss (WL) and hydroxyproline release (HYP) of films, as well as scanning electron microscopy (SEM) on dentin slabs. Crosslinking capacity and interactions of MAPAs with collagen were investigated by FTIR. Data were analyzed by ANOVA and Tukey’s test (α = 0.05%). ResultsMA:PA feeding ratios affected MAPAs’ chemical structures which in turn led to different collagen stabilization efficacy against degradation and varied collagen crosslinking capabilities. Higher collagen stabilization efficacy was detected using MAPA-1 (WL 10.52%; HYP 13.53 μg/mg) and MAPA-2 (WL 5.99%; HYP 11.02 μg/mg), which was comparable to that using PA (WL 8.79%; HYP 13.17 μg/mg) (p > 0.05), while a lower collagen stability occurred in MAPA-3 (WL 38.48%; HYP 29.49 μg/mg), indicating excessive MA-functionalization would compromise its stabilization efficacy. In comparison, complete digestion was detected for untreated collagen (WL 100%; HYP 102.76 μg/mg). The above results were consistent with collagen crosslinking efficacy of the three MAPAs revealed by SEM and FTIR. SignificanceA new class of novel polymerizable collagen cross-linkers MAPAs was synthesized and shown that, when appropriate MA:PA ratios were applied, the resulting MAPAs could render high collagen stability and the ability to copolymerize with resin monomers, overcoming the drawbacks of PA. These new polymerizable crosslinkers, when included in adhesives, could lead to long-lasting dentin bonding.

20 Effect of Adding Bioactive Peptide in Combination of Pharmaceutical Zinc Oxide or Organic Acids on Growth Performance, Hematology Profile, and Nutrient Digestibility in Nursery Pigs

Abstract To evaluate the effect of bioactive peptide (P) in combination with high level of zinc (HZ) or acidifiers on growth performance, complete blood cell counts (CBC) and nutrient digestibility in nursery pigs, a total of 288 weaned pigs (PIC1050xDNA600) were stratified by initial BW within gender and allotted to 1of 7 treatments. Treatments for phase 1&amp;2 were: 1) nutrient adequate positive control with HZ (PC), 2) nutrient deficient negative control with HZ (NC, -0.13% SID Lysine by reducing fish meal), 3) NC+0.25% peptide (0.25PZ), 4) NC+0.5% peptide (0.5PZ), 5) NC+0.25% peptide with standard zinc (0.25P), 6) NC+0.5% peptide with standard zinc (0.5P), 7) as 5 + 0.1% sodium butyrate and 0.5% benzoic acid (PSB). All pigs were fed a common low Zn diet (197 ppm) during phase 3. The whole blood was obtained from a close-to-average pen-BW pig repeatedly at weaning, and at the end of phase 2 and 3 to determine CBC. Titanium dioxide was used as an indigestible marker to determine nutrient digestibility. Data were analyzed using the Mixed procedures of SAS as a RCBD with treatment as fixed effect, and BW block as random effect. In overall phase 1&amp;2, pigs fed PSB had similar ADG and BW when compared to pigs fed 0.25PZ and both were greater than NC pigs (Table 1). With the same inclusion rate of peptide, pigs fed a high zinc diet had greater BW and ADG than pigs fed a standard zinc diet. PSB pigs had the greatest G:F ratio and nitrogen digestibility among treatments. Increasing peptide in high zinc diets gradually decreased Neutrophil-to-lymphocyte ratio. This study indicates that the improvement in growth performance from pigs fed peptide is pharmaceutical zinc dependent and acidifiers can be an alternative to replace ZnO without affecting growth performance.

Assessment of Bioactive Compounds under Simulated Gastrointestinal Digestion of Bee Pollen and Bee Bread: Bioaccessibility and Antioxidant Activity.

Bee pollen and bee bread have always been regarded as excellent natural resources for application in food and pharmaceutical fields due to their rich nutrient content and diversity of bioactive compounds with health-improving properties. Extensive studies on both bee products as ingredients for a healthy diet were reported, although the data concerning their metabolization on the gastrointestinal tract is quite limited. Here, we report, at each digestive stage, the bioactive profile for both bee products, their bioaccessibility levels and the antioxidant activity evaluation. The findings indicated that the average bioaccessibility level of total phenolic and total flavonoid content for bee pollen was 31% and 25%, respectively, while it was 38% and 35% for bee bread. This was reflected in a decrease of their antioxidant capacity at the end of in vitro gastrointestinal digestion, both in free radicals scavenging capacity and in reducing power. Moreover, within the 35 phytochemicals identified, the most affected by gastrointestinal digestion were phenylamides, with a complete digestibility at the end of the intestinal phase. Overall, our results highlight that bioactive compounds in both raw products do not reflect the real amount absorbed in the intestine, being bee bread more accessible in bioactive content than bee pollen.

Glacial Dust Surpasses Both Volcanic Ash and Desert Dust in Its Iron Fertilization Potential

AbstractThe subarctic Pacific Ocean and Bering Sea comprise the second‐largest high nitrate, low chlorophyll region in the world, where primary production is limited by the availability of iron (Fe). To estimate the potential impact of different terrestrial aerosol Fe sources on marine ecosystems, we performed a suite of laboratory assessments following established protocols, including: (1) leaching with Milli‐Q water, (2) sequential extractions, (3) complete acid digestions, (4) X‐ray diffraction, and (5) grain size analysis. Measurements were performed on 20 fine‐grained (&lt;5 μm) glacier‐derived sediments from Alaska and the Yukon, two fresh, never‐wetted volcanic ashes (Redoubt 2009 and Pavlof 2016), and six weathered ashes (Redoubt and Augustine) which span the past ∼8,700 years. We compared results to published data on Asian desert‐derived sediments, finding that the glacier‐derived sediments have five times higher easily reducible Fe (median 2.3 ± 0.6 wt.%) than desert‐derived samples (0.49 ± 0.1 wt.%) and fourteen times higher easily reducible Fe than fresh ash (0.16 ± 0.1 wt.%). In addition, fractional Fe solubility was higher in glacial sediment (median cumulative 0.31 ± 0.11% FeS) than volcanic ash (0.04 ± 0.02% FeS). Glacial sediments contained higher concentrations of other bioactive metals including Co, Ni, Cu, Zn, Mo, Cd, and Pb. Results suggest that glacier‐derived dust may provide the subarctic Pacific with more bioavailable iron per unit mass than either volcanic ash or Asian desert‐derived dust.