Pepsin Activity Research Articles

Ontogenic Development of Digestive Enzymes in Mealworm Larvae (Tenebrio molitor) and Their Suitable Harvesting Time for Use as Fish Feed

Mealworm larvae (Tenebrio molitor) are edible insects consumed in feed and food. In the current study, the optimal harvesting time of mealworm larvae for use as aquafeed was investigated during the ages of 30–90 days after hatching (DAH). Development of digestive enzymes, proximate composition, and in vitro protein digestibility using digestive enzymes from African catfish (Clarias gariepinus) and Nile tilapia (Oreochromis niloticus), were used as criteria. The specific activities of pepsin and trypsin significantly decreased with age (p < 0.05) from the first harvesting time until 50 and 45 DAH, respectively, while steadiness in these enzyme activities was observed onwards. Chymotrypsin specific activity appeared constant across all harvesting times. The specific activity of amylase significantly decreased in the later stages of development, while cellulase exhibited a different pattern suggesting it has a major role in dietary fiber utilization relative to starch. Regarding proximate compositions of the mealworm larvae, the moisture and ash contents decreased significantly with age, while the protein content exhibited the opposite trend with the highest contents from 60 to 90 DAH. Crude lipid was generally fairly constant, but its lowest value was observed in the earliest stage. In vitro protein digestibility was not significantly different across all harvesting times for both fish species, except for the significantly decreased digestibility value at 65 DAH relative to 30 and 35 DAH for Nile tilapia. However, based on the economic benefits of time for growth increment and proximate chemical composition, approximately 60 DAH is proposed as suitable for harvesting mealworm larvae to be used in fish feed.

Exploring the impact of Crohn's disease on the intragastric environment of fasted adults.

We explored the potential impact of Crohn’s disease on the intragastric environment of fasted adults with a view to potential effects on intragastric performance of orally administered drugs in the fasted state. Data were collected from 15 healthy individuals and 15 patients with Crohn’s disease. All subjects remained fasted for at least 12h prior to gastroscopy. Intragastric resting volume and pH were measured upon aspiration. Osmolality, surface tension, pepsin activity, and content of six bile acids were measured within 4 months upon sample collection. Unlike intragastric volumes, intragastric osmolality was significantly increased by Crohn’s disease. However, mean osmolality value in patients was only slightly higher than in healthy individuals (293 vs. 257 mOsmol/kg, respectively), therefore, unlikely to affect intragastric drug product performance. Primarily due to the high variability of data in healthy individuals, the potential effects on intragastric pH and surface activity could not be evaluated on a statistical basis. However, based on average (mean and median) values, even if they are statistically significant, it seems unlikely to be of clinical significance. Inter-subject variability of pepsin activity, and total bile acids content was high in both the healthy and the patients’ groups. Statistical investigation of the potential impact of Crohn’s disease on these parameters requires prior designation of the minimum differences to be detected; such differences will determine the minimum sample size required of relevant investigations.

Effects of different feeding frequencies on growth, feed utilisation, digestive enzyme activities and plasma biochemistry of gilthead sea bream (Sparus aurata) fed with different fishmeal and fish oil dietary levels

In the context of Mediterranean aquaculture many efforts have been made in terms of reducing marine-derived ingredients in aquafeed formulation. On the other hand, little attention has been paid to the manipulation of feeding frequency at the on-growing phase, where the high costs related to feeding procedures and the optimisation of feed efficiency and fish health are key aspects for the economic and environmental sustainability of the production cycle. The effects of different feeding frequencies (F) (1F) one meal day−1, (2F) two meals day−1, (3F) three meals day−1 on growth, digestive enzyme activity, feed digestibility and plasma biochemistry were studied in gilthead sea bream (Sparus aurata, L. 1758) fed with high (FM30/FO15, 30% fishmeal FM, 15% fish oil, FO) and low (FM10/FO3; 10% FM and 3% FO) FM and FO levels. Isonitrogenous and isolipidic extruded diets were fed to triplicate fish groups (initial weight: 88.3 ± 2.4 g) by a fixed ration over 109 days. No significant effects of feeding frequency on overall performance, feed efficiency and feed digestibility during the on-growing of gilthead sea bream fed high or low fishmeal and fish oil dietary level were observed. Pepsin activity showed an apparent decrease in fish receiving more than one meal a day which was not compensated by an increased production of alkaline proteases (either trypsin or chymotrypsin), particularly in fish fed on low FM. Although there were no effects on growth and feed utilisation at increasing feeding frequency, trypsin decreased significantly with an increasing number of meals only under low FMFO diet. Thus, it seemed that consecutive meals could have amplified the potential trypsin inhibitor effect of the vegetable meal-based diet adopted. Most of the plasma parameters related to nutritional and physiological conditions were not affected by feeding frequency, however an effect on electrolytes (Na+, Cl), cortisol and creatinine was observed. The higher level of plasma creatinine detected in fish fed a single daily meal with high FMFO level seems to be within physiological values in relation to the higher protein efficiency observed with this diet. However, it will require further attention to exclude a possible overload of kidney functionality. According to the results, gilthead sea bream seems able to maximize feed utilisation regardless of the number of meals, and this could be a useful indicator for planning feeding activity at farm level to optimise growth of fish and costs of feeding procedures.

Growth and ontogenetic development of digestive functionality in black Amur bream (Megalobrama terminalis)

The function of digestive physiology during ontogenetic development is essential to ensure high survival and growth rates. In order to evaluate the digestive physiological capacity of the black Amur bream (Megalobrama terminalis), changes of morphology and digestive enzyme activity (trypsin, lipase, amylase, pepsin, leucine aminopeptidase and alkaline phosphatase) in larvae were examined from hatching to 40 days after hatching (DAH). Results indicated that fluctuation patterns differed between the total and specific activities of the digestive enzymes. The total activities of these six enzymes gradually increased throughout the fish growth. The specific activity of trypsin peaked at 5 DAH and then decreased dramatically, while it increased remarkably again from 8 to 10 DAH and remained stable level after 20 DAH. Pepsin activity was first examined in M. terminalis at 15 DAH and gradually elevated towards the end of the experiment. The specific activity of lipase displayed obvious peaks at 5 and 20 DAH. For the amylase, its specific activity reached plateau at 4 DAH, underwent sharp decrease, and remained stable after 20 DAH. In addition, we found that the specific activity of alkaline phosphatase raised significantly from hatching to 5 DAH, and tended to keep slight fluctuation after 15 DAH. From the above, we concluded that the specific activities of digestive enzymes in the larvae varied constantly from 3 to 20 DAH and turned relatively stable after 20 DAH. The present study provides effective information that is useful to improve the seedling cultivation and the technology for healthy breeding.

Drinking water temperature affects cognitive function and progression of Alzheimer's disease in a mouse model.

Lifestyle factors may affect mental health and play a critical role in the development of neurodegenerative diseases including Alzheimer's disease (AD). However, whether the temperatures of daily beverages have any impact on cognitive function and AD development has never been studied. In this study, we investigated the effects of daily drinking water temperatures on cognitive function and AD development and progression in mice and the underlying mechanisms. Cognitive function of mice was assessed using passive avoidance test, open field test, and Morris water maze. Wild-type Kunming mice receiving intragastric water (IW, 10 mL/kg, 2 times/day) at 0 °C for consecutive 15 days displayed significant cognitive defects accompanied by significant decrease in gain of body weight, gastric emptying rate, pepsin activity, and an increase in the energy charge in the cortex when compared with mice receiving the same amount of IW at 25 °C (a temperature mimicking most common drinking habits in human), suggesting the altered neuroenergetics may cause cognitive decline. Similarly, in the transgenic APPwse/PS1De9 familial AD mice and their age- and gender-matched wild-type C57BL/6 mice, receiving IW at 0 °C, but not at 25 °C, for 35 days caused a significant time-dependent decrease in body weight and cognitive function, accompanied by a decreased expression of PI3K, Akt, the glutamate/GABA ratio, as well as neuropathy with significant amyloid lesion in the cortex and hippocampus. All of these changes were significantly aggravated in the APPwse/PS1De9 mice than in the control C57BL/6 mice. These data demonstrate that daily beverage at 0 °C may alter brain insulin-mediated neuroenergetics, glutamate/GABA ratio, cause cognitive decline and neuropathy, and promote AD progression.

Ontogeny of Expression and Activity of Digestive Enzymes and Establishment of gh/igf1 Axis in the Omnivorous Fish Chelon labrosus.

Simple SummaryThick-lipped grey mullet (Chelon labrosus) feeds on the lowest trophic levels during adult stages, for which it is considered a viable candidate for an economically and environmentally sustainable aquaculture. Similar to most of marine fish species, C. labrosus produce a large number of eggs, leading to morphologically and anatomically larvae that are not completely mature and have to pass through substantial differentiation and development in their functional systems to acquire adult features. Therefore, the study of the development of digestive tract and of the growth regulation can provide useful information to adapt the feeding protocols and rearing conditions to the physiological requirements at each stage. This work aimed to evaluate the early ontogeny of key digestive enzymes and somatotropic factors at biochemical and/or transcriptional levels. Our results evidenced that maturation of the digestive system and acquisition of the adult mode of digestion occurs around 60 to 70 days post hatch (dph), when starch or other low-cost carbohydrate-based compounds could be used in formulated diets at increasing levels. Furthermore, our results implied an independent expression of the studied somatotropic genes during the first 40 dph and establishment of a functional growth hormone/insulin-like growth factor 1 axis from 50 dph onward.Thick-lipped grey mullet (Chelon labrosus) is a candidate for sustainable aquaculture due to its omnivorous/detritivorous feeding habit. This work aimed to evaluate its digestive and growth potentials from larval to early juvenile stages. To attain these objectives the activity of key digestive enzymes was measured from three until 90 days post hatch (dph). Expression of genes involved in digestion of proteins (try2, ctr, pga2, and atp4a), carbohydrates (amy2a), and lipids (cel and pla2g1b), together with two somatotropic factors (gh and igf1) were also quantified. No chymotrypsin or pepsin activities were detected. While specific activity of trypsin and lipase were high during the first 30 dph and declined afterward, amylase activity was low until 57 dph and increased significantly beyond that point. Expression of try2, ctr, amy2a, and cel increased continuously along development, and showed a peak at the end of metamorphosis. Expression of pla2g1b, pga2 and atp4a increased until the middle of metamorphosis and decreased afterwars. Most of these trends contrast the usual patterns in carnivorous species and highlight the transition from larvae, with high protein requirements, to post-larvae/juvenile stages, with omnivorous/detritivorous feeding preferences. Somatotropic genes, gh and igf1, showed approximately inverse expression patterns, suggesting the establishment of the Gh/Igf1 axis from 50 dph.

Is gastrointestinal motility related to alkaloids of Charred Semen Arecae?

Ethnopharmacological relevanceSemen Arecae (SA) is one of the most commonly used Traditional Chinese Medicine. Charred Semen Arecae (CSA) is the processed product of SA. Alkaloids are considered as pharmacological mechanisms of SA and CSA on gastrointestinal motility. Recent studies have shown alkaloids decreased quickly after procession. However, the promoting on gastrointestinal motility were not decreased.Is gastrointestinal motility related to alkaloids of CSA? This study explored the effects of SA, CSA, Semen Arecae-Removal (SA-R), and Charred Semen Arecae-Removal (CSA-R) on gastrointestinal motility, Gastric Inhibitory Polypeptide (GIP), Glucagon Like Peptide-1 (GLP-1), gastric juice and bile in rats. Material and methodsRats were randomly divided into six groups, including the Control group, SA group, CSA group, SA-R group, CSA-R group, and Positive drug group (Mosapride). Alkaloids of samples were knocked out by using the “target constituent removal” strategy. Gastric residue and intestinal propulsion rate were evaluated in rats. Serum levels of GIP and GLP-1 were measured by Enzyme-Linked Immunosorbent Assay (ELISA). Gastric juice and bile were examined, respectively. ResultsCSA-R and SA-R have been investigated by the Preparative Thin-layer Chromatography (PTLC) method. Intestinal propulsion and gastric residue assessments confirmed the effectiveness of CSA and CSA-R. CSA-R was higher than SA-R in the GLP-1, pepsin activity, the secretion of bile, Bilirubin (BIL), and Cholesterol (CHO). The statistical comparison demonstrated that there is no difference between the CSA group and CSA-R group. ConclusionsAfter processing, the promoting gastrointestinal motility might be not related to alkaloids. Maillard reaction could be produced to promote the secretion of GLP-1, bile, and CHO for gastrointestinal motility. Our findings provide a pharmacological reference for the clinical application of SA and CSA in the treatment of digestive diseases.

Effects of fishmeal replacement with cottonseed protein concentrate on growth, digestive proteinase, intestinal morphology and microflora in pearl gentian grouper (♀ Epinephelus fuscoguttatus × ♂ Epinephelus lanceolatu )

A 56‐day feeding trial was conducted to study the effects of cottonseed protein concentrate (CPC) as fishmeal (FM) replacement on growth, digestive proteinase, intestinal morphology and microflora in pearl gentian grouper (♀Epinephelus fuscoguttatus × ♂Epinephelus lanceolatu). FM of 0% (FM), 12.00% (R12), 24.00% (R24), 36.00% (R36), 48.00% (R48) and 60.00% (R60) was replaced with CPC, to prepare six diets and feed groupers. Weight gain (WG) and specific growth rate (SGR) were highest in the R24 group, and the lowest feed coefficient rate (FCR) was observed in this group compared with the other groups (p < .05). Pepsin activity of the R12 group and chymotrypsin activity of the R36 group reached significant maximum (p < .05). With increasing CPC replacement, the plica height and width and the lamina propria thickness of intestine decreased significantly (p < .05). 16S rDNA amplicon sequencing suggested dominant intestinal microflora of grouper subjected to different treatments was similar. At the phylum level, the abundance of Proteobacteria, Firmicutes, Verrucomicrobia, Bacteroidetes and Actinobacteria was high, while Photobacterium, Pseudoxanthomonas, Romboutsia, Akkermansia, Vibrio and Bifidobacterium were the dominant taxa at the genus level. The abundance of probiotics (Bifidobacterium) decreased, whereas the abundance of opportunistic pathogen (Proteobacteria, Actinobacteria and Vibrio) increased. These results suggested that CPC replacement affected growth, digestive proteinase activity, intestinal morphology and microflora structure in grouper. Under the conditions of this experiment, the fishmeal replacement with CPC (24% level) yielded the most satisfactory results, providing reference for sustainable aquaculture.

Effects of live microalgae and algae powder on microbial community, survival, metamorphosis and digestive enzyme activity of Penaeus monodon larvae at different growth stages

Most hatcheries use live microalgae and algae powder for artificial rearing; however, the effects of these different diets on the rearing process and bacterial communities is not clear. In order to investigate the effects of live algae and algal powder on the larval culture of Penaeus monodon. Two kinds of live algae (Chaetoceros muelleri, Thalassiosira weissflogii) and Spirulina powder were used to feed larvae in this study, and the effects on the environmental parameters of water, the digestive enzymes activity of larvae and the bacterial community from the zoea stage to the post larval (PL) stage were explored. The results revealed that feeding larvae with C. muelleri significantly improved the enzymatic activity of amylase and the survival rate of the larvae, and significantly reduce the concentrations of NH4-N, NO2-N and the quantity of Vibrio in the water (P < 0.05). Feeding with T. weissflogii significantly improved the enzymatic activity of lipase and pepsin, and the percentage of metamorphosis (P < 0.05). It suggested that adding live microalgae was beneficial in increasing the diversity of the bacterial community in larvae. Cyanobacteria and Patescibacteria were dominant bacteria in the algal solutions of C. muelleri and T. weissflogii. The entire rearing environment was dominated by two phyla: Bacteroidetes and Proteobacteria. Virgulinella fragilis, the bacteria present in the liquid of C. muelleri and T. weissflogii, accounts for a high proportion of bacteria in the rearing water at the zoea stage. The bacterial community in the zoea stage showed significant negative correlations with NH4-N and NO2-N (P < .05), whereas the bacterial community in the PL stage showed significant positive correlations with NH4-N and NO2-N (P < .05). Saprospiraceae and Microbacteriaceae in the water were significantly positively correlated with NH4-N and NO2-N (P < .05). Feeding larvae with C. muelleri and T. weissflogii and Spirulina powder could increased the abundance of Microbacteriaceae, Devosiaceae and Saprospiraceae at the family level, and increased the abundance of Pseudoalteromonas, Roseovarius and Nautella at genus level. In conclusion, feeding larvae with live microalgae could improve the percentage of metamorphosis and survival rates of larvae by improving the external water environment, enhancing digestion, and regulating bacterial community.

A formulated diet improved digestive capacity, immune function and intestinal microbiota structure of juvenile hybrid grouper (Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂) when compared with chilled trash fish

The effects of two diets on the growth, intestinal microbiota, digestive enzymes, immunity-related enzymes and gene expression were assessed in the juvenile hybrid grouper (Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂). Hybrid grouper were fed either a formulated diet (FM group) or chilled trash fish (CTF group). Following a 10-week feeding study, the results showed that the final weight, weight gain rate, specific growth rate and survival rate of fish were not significantly different between the groups. However, bio-economic analysis of the feeds showed that the profit from fish in the FM group was significantly higher than from the CTF group. In addition, digestive enzyme (trypsin and pepsin) activities in fish were significantly higher in the FM group, while lysozyme activity was significantly higher in the CTF group. Levels of complement component 4 (C4) and immunoglobulin M (IgM) in fish serum were significantly lower in FM group fish, while complement component 3 (C3) levels were higher in FM group fish. In the fish intestine, IgM levels were significantly higher in the FM group, while lysozyme activity and C3 and C4 levels were higher in the CTF group. The expression of innate immune genes (MHC II β and TLR22) and inflammatory cytokines (IL-1β, IL-6, IL-10, TNF α1 and IFN γ) were significantly lower in the FM group. The bacterial community of the distal intestine was analyzed by Next-Generation Sequencing. At the phylum level, the relative abundance of Acidobacteria was significantly lower in the FM group, while the abundance of Bacteroidetes was lower in the CTF group. At the family level, the relative abundance of Flavobacterium was significantly lower in the FM group, while the abundance of Bacteroidales_S24-7_group and Lachnospiraceae was lower in the CTF group. The most significantly changed pathways between the two groups were metabolism and genetic information processing. The overall results showed that feed bio-economics, digestive capacity, and immunity of hybrid grouper were superior in the FM group compared to the CTF group.

Reduction of Purification Time of Polyspecific Equine F(ab´)2 Antivenom against Scorpion Envenomation

Background and Aims: In this study we improved the purification of immunoglobulins from equine antiserum raised against the venom of 6 types of scorption species. Caprylic acid (octanoic acid), a fatty acid, was found to have no effect on the activity of the enzymes pepsin, which is used in antivenom purification to digest Fc fragment of immunoglobulins to obtain F(ab´)2. Materials and Methods: A new method was developed for the production of F(ab´)2 antivenom whereby whole equine antiserum was mixed with equal amount of 0.15 M HCl and pH 3.4 with pepsin 660 mg/L of diluted antivenom and incubated for 4 h at 37°C. After digestion the pH were brought to 4.8 with sodium hydroxide solution (1.5 M) and then 1.5% caprylic acid and 10% ammonium sulfate was added and mixed for 60 minutes and passed through filter paper. Results: Caprylic acid caused precipitation of albumin, and ammonium sulfate reduced turbidity of solution, resulting in a reduced protein load presented to the digestion enzymes culminating in substantial reductions in processing time. Conclusions: The equine F(ab´)2 obtained using these novel caprylic acid methods were comparable in terms of yield, purity and specific activity to those obtained by multi-step and time consuming conventional salt fractionation with ammonium sulfate.

Hydrogen molecules can modulate enzymatic activity and structural properties of pepsin in vitro

Hydrogen gas therapy has been recognized as the promising application merit. However, the underlying mechanism on the biological effects remains far from being understood. In this work, pepsin used as a research model, the effects of hydrogen-rich water on the protein activities and structural properties were investigated by enzymatic assay, atomic force microscopy-based peakforce quantitative nanomechanical mapping (PF-QNM) and terahertz time-domain spectroscopy (THz-TDS). We found that hydrogen-rich water can increase the protein activity and its apparent height while alter the mechanical properties (Young's modulus) and the terahertz dynamics. These results suggest a possible mode of hydrogen molecules acting with pepsin through the local changes of hydrophobic interfaces in the protein molecules, thus provide the first evidence for the direct interaction between hydrogen with proteins and a biophysical insight into the mechanism of hydrogen as well as other gases on the biological effects.

Histological, histochemical and biochemical characterization of larval digestive system ontogeny in black tetra Gymnocorymbus ternetzi to inform aquaculture weaning protocols

Identifying an appropriate larval diet is a significant challenge in aquaculture. To overcome this bottleneck, characterization of larval fish digestive system ontogeny is critical for optimizing nutritional protocols. Intensive culture of characids for ornamental trade is highly dependent on live Artemia nauplii through the larval stage. Live feeds have unique challenges relative to prepared microparticulate diets (MDs), specifically pertaining to availability, labor, and cost. In this study, a feeding trial was conducted to compare three commercially available MDs in larval Black Tetra Gymnocorymbus ternetzi. Larvae exhibited poor survival and growth for MDs relative to Artemia. Digestive system ontogeny was characterized using histology, histochemistry, and digestive enzyme activity assays from the onset of exogenous feeding through the larval stage. G. terntzi exhibited an agastric larval stage, as well as low digestive enzyme activity at the onset of exogenous feeding, followed by increases in trypsin, lipase, and pepsin activity. Histological differentiation of the stomach, including gastric gland formation and production of periodic acid Schiff positive neutral mucopolysaccharides, as well as the onset of pepsin activity, occurred at 22 days post hatch (dph). This shift from agastric to gastric digestion indicated elevated digestive capacity and increased ability to consume a diversity of prey.Based on these data, experiments were conducted to evaluate different weaning times from Artemia to a MD. Larvae were fed Artemia exclusively from hatch through the end of the trial, a MD exclusively, or were transitioned from Artemia to a MD at three different time points. For G. ternetzi fed until 33 dph, weaning beginning at 13 dph exhibited higher survival (20.6 ± 1.8%) than live Artemia and weaning beginning at 18 dph and 23 dph (mean: 13.6 ± 1.5%), while the MD resulted in the lowest survival (0.8 ± 0.2%). These results indicate that weaning is possible prior to gastric differentiation, suggesting that it is possible to rely less on Artemia as a larval feed in this characid species.

Comparison of digestive, immune and antioxidant enzyme activities of mackerel tuna (Euthynnus affinis) in the wild and under captivity

ABSTRACT To explore the feasibility of land-based farming on mackerel tuna (Euthynnus affinis), the changes of antioxidant, immune and digestive enzyme activities of wild caught and indoor cultured juvenile mackerel tuna were evaluated. The activities of total superoxide dismutase, catalase, peroxidase and alkaline phosphatase in the liver of the wild group were significantly higher than those cultured under captivity. The total superoxide dismutase and peroxidase activities in the indoor cultured group were significantly increased in the head kidney, and the MDA content was significantly increased. The activities of lysozyme in the liver and head kidney, the activities of amylase, lipase and trypsin activities in the intestine were significantly increased in the indoor cultured group. The pepsin activity in the stomach was significantly reduced. The land-based farming increased the digestive enzyme activity compared to the wild caught group. The mode of land-based farming is suitable for growth and fattening of young fish, but it leads to inhibition of antioxidant capacity and increase in immune enzyme activity. This study adds knowledge of our understanding on the physiological response of mackerel tuna and improves future farming practice of this species in an indoor condition.

Citral as food additive for common snook - zootechnical parameters and digestive enzymes

ABSTRACT: Essential oils of plants whose main compound is citral showed beneficial effects when added to fish feed. The objective of the present study was to evaluate the dietary effect of the addition of citral on zootechnical parameters and digestive enzyme activities of Centropomus undecimalis. Juveniles were fed for 45 days with diets containing different amounts of citral (0.0 - control, 0.5, 1.0, and 2.0 mL per kg of diet). The water quality parameters were kept stable during the experiment and no mortality was observed. At the end of the experimental period, the treatment 0.5 mL citral per kg of diet had the lowest weight gain and specific growth rate, and the highest feed conversion, while the same parameters did not differ between the other treatments. Pepsin activity was higher in the stomach of fish fed with 0.5 mL citral per kg of diet and amylase activity was higher in the intestine of fish fed with 0.5 and 2.0 mL citral per kg of diet compared to the control group. Intestinal lipase activity was higher in all groups that were fed with citral compared to the control group. Chymotrypsin and trypsin activities showed no difference between groups. Consequently, dietary addition of citral at any of the levels tested is not recommended for common snook.

New Use of Non-natural Catalytic Activity of Pepsin: Highly Efficient Synthesis of Tetrahydroquinazolines

New Use of Non-natural Catalytic Activity of Pepsin: Highly Efficient Synthesis of Tetrahydroquinazolines

ANTIULCER ACTIVITY OF ELECTROHOMEOPATHIC DRUG (SPAGYRIC ESSENCE) CANCEROUS 15 - 3rd DILUTION AGAINST INDOMETHACIN-INDUCED GASTRIC ULCER IN RATS

Objective: To evaluate the curative protects activity of electrohomeopathic drug Cancerous15 (C15) against Indomethacin (IND)-induced gastric ulcer in albino rats. None of the data have been reported on antiulcer activity of C15 drug. Hence, the present study focuses on the scientific investigation of antiulcer activity of C15 against Indomethacin-induced peptic ulceration of rat models.&#x0D; Methods: A total number of 35 albino rats were divided into five groups equally. Pure water was given to Group 1 (normal). IND was given orally to Group 2 (ulcerated control). Cancerous15 third dilution was given to Group 3. IND was given to Group 4 and Group 5 animals, thereafter pretreatment with Esomeprazole (ESM) and C15-third dilution, respectively. The pretreatments were treated in each day’s interval for 21 days before IND administration. There was provision of food and water throughout the experimental period. On the 23rd day, the stomachs of the sacrificed rats were removed, and (1) ulcer index, (2) percentage of ulcer inhibition, (3) gastric volume, (4) pH, (5) pepsin activity, (6) mucin content, (7) malondialdehyde (MDA) concentration, and (8) stearidonic acid activity were studied.&#x0D; Results: The pre-treated animals with ESM and C15 shown as significantly decreases in ulcer index, gastric volume, pepsin activity and MDA concentration, and significant increase in the percentage ulcer inhibition, pH value, mucin content, and superoxide dismutase activity concentration in comparison to ulcer-induced rats.&#x0D; Conclusion: The study shows the significant ulcer protective activity of the C15 drug against IND-induced peptic ulceration in rats.

Effects of Pseudoalteromonas espejiana W115 on Enzyme Activity and Disease Resistance of the Yesso Scallop Patinopecten yessoensis

A 49-day rearing trial was conducted to investigate the effects of Pseudoalteromonas espejiana W115 on digestive and immune–related enzyme activities in larval and juvenile Yesso scallop Patinopecten yessoensis. Larvae (day 2 post-fertilization) (day 2 PF) were distributed into three groups and provided with a diet only of microalgae (control group) and supplemented with W115 at final concentrations of 1 × 104 and 1 × 106 cells/mL, respectively. Results showed that umbo larvae (day 13 PF) and/or crawling-pediveliger larvae (day 28 PF) and/or juveniles (day 51 PF) fed with W115 supplementation diet at 104 cells/mL showed a significant increase in pepsin, amylase and cellulase activities compared to those fed the diet of microalgae, however, no significant difference was observed between the 106 cells/mL supplementation and control groups at days 13, 28 and 51 PF. The lysozyme and superoxide dismutase activities of umbo larvae and/or crawling-pediveliger larvae and juveniles treated with 104 and/or 106 cells/mL W115 supplementations were significantly higher than those in the control group, while there were no significant differences in enzyme activities in larvae and juveniles treated with two doses of W115 at days 13, 28 and 51 PF. No significant difference in catalase activity was observed among the three groups at days 13, 28 and 51 PF. In addition, there was increased survival in W115-amended treatments after juveniles were challenged with Vibrio splendidus. The results suggest that a 104 cells/mL supplementation of W115 can be useful in Yesso scallop larviculture.

Antiangiogenic potentials of ahpatinins obtained from a Streptomyces species

While exploring new angiogenesis inhibitors from microbial metabolites, we recently isolated ahpatininsC, E, and G from a soil‑derived Streptomycessp. 15JA150. AhpatininsC, EandG are known to have pepsin and renin inhibitory activities; however, their antiangiogenic activities and underlying molecular mechanisms have not been fully elucidated. In the present study, the antiangiogenic properties of ahpatininsC, EandG were investigated. The results revealed that the natural compounds significantly inhibited the vascular endothelial growth factor (VEGF)‑induced proliferation, invasion, adhesion, and tube formation of human umbilical vein endothelial cells (HUVECs) without exhibiting any cytotoxicity. It was also revealed that ahpatininE effectively suppressed the neovascularization of the chorioallantoic membranes in growing chick embryos. Notably, ahpatininsC, E, and G led to the downregulation of VEGF‑induced activation of VEGF receptor2 (VEGFR2) and its downstream signaling mediators, including AKT, ERK1/2, JNK, p38, and NF‑κB, in HUVECs. Moreover, they reduced the expression of matrix metalloproteinase (MMP)‑2 and MMP‑9 in the HUVECs following stimulation with VEGF. Furthermore, ahpatininsC, E, and G reduced the tumor cell‑induced invasion and tube forming abilities of HUVECs, as well as the expression of VEGF, by suppressing hypoxia‑inducible factor‑1α (HIF‑1α) activity in U87MG glioblastoma cells. Collectively, the present findings indicated that ahpatininsC, E, and G may be used in anticancer therapy by targeting tumor angiogenesis through the inhibition of both VEGFR2 and HIF‑1α pathways.

Pepsin activity from gastric of milkfish and catfish in Indonesian Waters

Pepsin is the main protease enzyme in the gastric including the gastric of the fish. The enzyme has many benefits for both food and non-food. The purpose of this study was to determined the pepsin activity of the catfish and milkfish. The study was divided into 2 stages, namely pepsin extraction stage and pepsin characterization. The results showed that pepsin from milkfish and catfish can be extracted properly using tris-Cl buffer with the activity of 526.5 U/mL (catfish) and 397 U/mL (milkfish). The pepsin isolated from the gastric of catfish and milkfish can work well at a temperature range of 20-40°C and pH 1-3. Pepsin from the gastric of catfish had Km = 0.3333 mM and Vmax = 1666.6667 mmol/s. Pepsin from milkfish has Km = 0.091 and Vmax = 909,091 mmol/s.