Background: Somatic cell nuclear transfer (SCNT) is a prominent technology that can preserve superior genetic traits of animals and expand the population in a short time. Hematological characters and endocrine profiles are important elements that demonstrate the stability of the physiological state of cloned animals. To date, several studies regarding cloned camels with superior genes have been conducted. However, detailed hemato-physiological assessments to prove that cloned camels are physiologically normal are limited. In this study, We evaluated the hemato-physiological characteristics of cloned male and female dromedary camels (Camelus dromedaries). Methods: Therefore, we analyzed variations in hematological characteristics and endocrine profiles between cloned and non-cloned age-matched male and female dromedary camels (Camelus dromedaries ). Two groups each of male and female cloned and non-cloned camels were monitored to investigate the differences in hemato-physiological characteristics. Results: All the animals were evaluated by performing complete blood count (CBC), serum chemistry, and endocrinological tests. We found no significant difference between the cloned and non-cloned camels. Furthermore, the blood chemistry and endocrine profiles in male and female camels before maturity were similar. Conclusions: These results suggest that cloned and non-cloned camels have similar hematological characteristics and endocrine parameters.

Background: The small intestine plays a crucial role in animals in maintaining homeostasis as well as a series of physiological events such as nutrient uptake and immune function to improve productivity. Research on intestinal organoids has recently garnered interest, aiming to study various functions of the intestinal epithelium as a potential alternative to an in vivo system. These technologies have created new possibilities and opportunities for substituting animals for testing with an in vitro model. Methods: Here, we report the establishment and characterisation of intestinal organoids derived from jejunum tissues of adult pigs. Intestinal crypts, including intestinal stem cells from the jejunum tissue of adult pigs (10 months old), were sequentially isolated and cultivated over several passages without losing their proliferation and differentiation using the scaffold-based and three-dimensional method, which indicated the recapitulating capacity. Results: Porcine jejunum-derived intestinal organoids showed the specific expression of several genes related to intestinal stem cells and the epithelium. Furthermore, they showed high permeability when exposed to FITC-dextran 4 kDa, representing a barrier function similar to that of in vivo tissues. Collectively, these results demonstrate the efficient cultivation and characteristics of porcine jejunum-derived intestinal organoids. Conclusions: In this study, using a 3D culture system, we successfully established porcine jejunum-derived intestinal organoids. They show potential for various applications, such as for nutrient absorption as an in vitro model of the intestinal epithelium fused with organ-on-a-chip technology to improve productivity in animal biotechnology in future studies.

The optimization of deacetylation process parameters for producing chitosan from isolated chitin shrimp shell waste was investigated using response surface methodology with central composite design (RSM-CCD). Three independent variables viz, NaOH concentration (X1), radiation power (X2), and reaction time (X3) were examined to determine their respective effects on the degree of deacetylation (DD). The DD of chitosan was also calculated using the baseline approach of the Fourier Transform Infrared (FTIR) spectra of the yields. RSM-CCD analysis showed that the optimal chitosan DD value of 96.45 % was obtained at an optimized condition of 63.41 % (w/v) NaOH concentration, 227.28 W radiation power, and 3.34 min deacetylation reaction. The DD was strongly controlled by NaOH concentration, irradiation power, and reaction duration. The coefficients of correlation were 0.257, 0.680, and 0.390, respectively. Because the procedure used microwave radiation absorption, radiation power had a substantial correlation of 0.600~0.800 compared to the two low variables, which were 0.200~0.400. This independently predicted robust quadratic model interaction has been validated for predicting the DD of chitin.

Black phosphorus (BP) is incorporated in the electrochemical detection of uric acid (UA) to form few layers of BP nanosheets (BPNS)-modified glassy carbon electrodes (BPNS/GCE), investigated by means of ultrasound-assisted liquid-phase exfoliation. We find a significant increase in the peak current magnitude and positive potential shift in the electrochemical response of BPNS/GCE, which may be attributed to the larger specific surface area and good charge transfer ability of BPNS. Further, the electrochemical response of BPNS/GCE is evaluated under different conditions to achieve the optimal conditions. UA detection using differential pulse voltammetry (DPV) shows linear response within the range of 1–1000 μM with a detection limit of 0.33 μM. This work reveals new applications of BP nanomaterials in the electrochemical sensing, thereby promoting further advancement in terms of practical applications of two-dimensional nanomaterials.

Hypertension is characterized by excessive renin-angiotensin system activity, leading to blood vessel constriction. Several synthetic compounds have been developed to inhibit renin and angiotensin-converting enzyme (ACE). These drugs often have adverse side effects, driving the exploration of plant protein-derived peptides as alternative or supplementary treatments. This study assessed the phenolic compound and amino acid content and the antioxidant and antihypertensive activity of 5 South Korean staple crops. Sorghum had the highest phenolic compound content and exhibited the highest antioxidant activity. Millet grains, particularly finger millet (38.86%), showed higher antihypertensive activity than red beans (14.42%) and sorghum (17.16%). Finger millet was found to contain a large proportion of branched-chain, aromatic, and sulfur-containing amino acids, which are associated with ACE inhibition. In particular, cysteine content was positively correlated with ACE inhibition in the crops tested (r=0.696, p<0.01). This study confirmed that the amino acid composition was more correlated with the antihypertensive activity of grains than the phenolic compound content. Finger millet mainly contained amino acids, which have higher ACE inhibitory activity, resulting in the strongest antihypertensive activity. These findings underscore the antihypertensive potential of select crops as plant-based food ingredients, offering insight into their biological functions.

Background: Largemouth bass (Micropterus salmoides) is introduced species that has caused aquatic ecology activity both in vitro and in vivo were investigated for the possibility of application of the bass extract as an alternative feed ingredient. Methods: The bass oil was extracted using a 1-L supercritical extractor, while the protein was extracted from 250 g of bass dry matter, which was dissolved in 1 mL of H2O at 50℃. Both oil and protein extracts were evaluated antioxidant activities and the level of DPPH radical scavenging assay and nitric oxide (NO) production assay with lipopolysaccharide response. Oral administration of 6.6 μL/g bass protein and 5.38 μL/g bass oil conducted for investigating serological and physiological effect. Results: DPPH radical scavenging showed similar radical scavenging ability of 50 μM of ascorbic acid at 200 μg of protein and 10% of oil treatment. NO concentration was diminished by the treatment of bass oil. Oral administration of both bass oil and proteins to mice showed that the body weight increase rate of the bass oil treated group was significantly reduced by 1.55 g compared to the other groups. The number of white blood cells (WBC) was increased by 4.52 k/μL in the bass protein-treated group and 4.44 k/μL in the bass oil-treated group compared to the control group. However, the serum IgG level did not show a significant difference between the bass extract-treated groups and the control group. Conclusions: These studies demonstrate that both bass oil and proteins extracted from the bass not only provide excellent effects of antioxidant and immune activity but can also be used as functional food supplements.

Pasture formation and management are crucial to avoid yield reduction. This experiment aimed to examine the effects of tall fescue-centered mixed-seeding combinations on yield and vegetation changes in perennial pastures in the central region for two years, from September 2020 to October 2022. The treatments were arranged in three replications in a randomized block design: control (C), tall fescue-based mixture-1 (T-1), and tall fescue-based mixture-2 (T-2). The tall fescue (TF), orchard grass (OG), perennial ryegrass (PRG), Kentucky bluegrass (KBG), and white clover (WC) were used. The emergency rate of grasses (70.0 to 73.3%) did not differ among mixed seeding combinations. Overwintering rates (81.7 to 83.3%) were similar among treatments. The plant height of grasses was similar at each harvest date, with the highest height (86.2 cm) recorded in the second harvest of the first year, followed by that (58.4 cm) in the third harvest of the first year; it was least (38.9 cm) in the fourth harvest of the second year. There was no significant difference in the dry matter yield of grasses among the mixed seeding combination treatments in the first, third, or fourth harvests of the first year (p>0.05). For second-year grasses, dry matter yield was not significantly different in harvest date among the treatments (p>0.05). Based on mixed seeding ratio, orchard grass showed the highest yield at 70% in the C treatment, followed by tall fescue at 80% and 60% in the T-1 and T-2 treatments, respectively, in the first harvest after seeding. There was no significant difference in feed value between treatments (p>0.05), but a significant difference was observed between the third and fourth harvest (p<0.05). Therefore, it indicated that it is important to create perennial pastures in the central region through mixed seeding combinations centered on tall fescue.

Canine hyperadrenocorticism, a prevalent endocrine disorder characterized by excessive cortisol production. Notably, hypercoagulability leading to pulmonary thromboembolism (PTE) poses a substantial concern. PTE may be underestimated because of the rapid dissolution of canine thrombi postmortem. However, traditional coagulation assays face challenges in early detection of hypercoagulability. Therefore, this study explored the use of thromboelastography (TEG) as a diagnostic tool for identifying hypercoagulability in dogs with hyperadrenocorticism. A total of 31 dogs visited the Gyeongsang Animal Medical Center between 2018 and 2022, comprising 21 dogs with hyperadrenocorticism and 10 controls who underwent clinical and coagulation analyses. Hyperadrenocorticism was diagnosed using a low-dose dexamethasone stimulation test or adrenocorticotropin hormone stimulation test, and conventional laboratory parameters and coagulation parameters, such as the prothrombin time, activated partial thromboplastic time, fibrinogen, and TEG results, were compared between the groups. Clinical data revealed significantly elevated monocyte, platelet, alanine aminotransferase, alkaline phosphatase, triglyceride, and cholesterol concentrations in dogs with hyperadrenocorticism, which were attributed to excess cortisol secretion (p<0.05). TEG analysis demonstrated significantly decreased K values and increased α and MA values in hyperadrenocorticism dogs (p<0.05), indicating a shortened clotting time and enhanced clot strength, suggestive of hypercoagulability. TEG effectively highlights hypercoagulability in dogs with hyperadrenocorticism and provides valuable insights in predicting blood clot formation. Although predicting clot formation in dogs remains complex owing to multifactorial influences, this study underscores the potential utility of TEG in enhancing such predictions for dogs with hyperadrenocorticism.

The spherical mesophases are the main precursors for the high tap density of carbonaceous anode batteries. However, it is challenging to control mesophase size without coalescence and no deformation since it quickly coalesces into a regular large sphere. Here, we propose a feasible extraction method to refine the spherical size of mesophase using benzene. Thermogravimetric and differential scanning calorimetry analysis of untreated pitch revealed that the maximum temperature for mesophase nucleation should not exceed 410 °C to provoke the nucleation of mesophase spheres while maintaining a high pyrolysis yield. The extraction results showed that the extraction weight tends to decrease with an increase in the solvent ratio. There is an exponential relationship between the influence of solvent ratio and the ability for extraction. The solubility of the spherical mesophase in benzene is size-dependent and can dissolve selectively spherical mesophases smaller than 5 μm. Consequently, a monodisperse spherical mesophase was obtained. The reason for forming uniform mesophase spheres can be explained by their thermodynamic state, as described by the “two-step” classical nucleation theory. Benzene effectively improves the size distribution of spherical mesophase by dissolving small sizes while retaining large ones.

Insect cuticle is an extracellular matrix formed primarily from two different biopolymers, chitin and protein. During each molt cycle, a new cuticle is deposited simultaneously with degradation of the old cuticle by molting fluid cuticle degrading-enzymes, including epidermal chitinases (CHTs). Insect CHTs, belonging to family 18 glycosylhydrolase (GH18), have been classified into at least eleven subgroups based on phylogenetic analyses, and group I (CHT5) and group II (CHT10) epidermal CHTs present in molting fluid. In this study we report the physiological function of MaCHT5 and MaCHT10 in the Japanese pine sawyer, Monochamus alternatus. RNAi for either MaCHT5 or MaCHT10 resulted in larval-pupal and pupal-adult molting defects, in which the insects were unable to shed completely their old cuticle and died entrapped in their exuviae. Furthermore, TEM analysis revealed a failure of degradation of the old cuticle in both MaCHT5- and MaCHT10-deficient pharate adults. In the old pupal cuticle, the chitinous horizontal laminar and vertical pore canal essentially remained intact in the endocuticular layer. These results indicate that both CHTs are required for turnover of the chitinous old cuticle, which is critical for completion of insect molting. We also discuss the possible function of two spliced variants of MaCHT10, MaCHT10a and MaCHT10b.

Structural cuticular proteins (CPs) and the liner polysaccharide, chitin, are the primary components of insect cuticle or exoskeleton. A large number of insect CP family proteins are divided into several distinct subfamilies defined by the presence of specific amino acid sequence motifs. One of these subfamilies is composed of Cuticular Protein Analogous to Peritrophins (CPAPs), containing one (CPAP1s) or three (CPAP3s) type-2 chitin-binding domains. In this study, we report a novel function of TcCPAP1-C from Tribolium castaneum in movement of legs. RNAi for TcCPAP1-C at larval stage has no effect on insect molting, growth and development. However, the resulting adults exhibit impaired leg movement, in which internal tendon cuticles are ruptured near the femur-tibia joint. The exoskeletal cuticle, hemiadherens junctions, microtubule array, myotendinous junctions and muscle fibers exhibit normal morphology before the tendon breakage. These results indicate functional specialization of TcCPAP1-C in structural integrity of the internal tendon cuticle, and loss of function of TcCPAP1-C caused breakage of the tendon cuticle, resulting in defective limb movement and locomotion.

Lytic polysaccharide monooxygenases (LPMOs) catalyze the oxidative cleavage of glycosidic bonds in crystalline polysaccharides including chitin and cellulose. The recent discovery of LPMO family proteins in many insect species suggests that they presumably play a role in chitin degradation in the cuticle/exoskeleton, tracheae and peritrophic matrix during insect development. Insect LPMOs belong to auxiliary activity family 15 (AA15/LPMO15) and have been classified into at least four groups based on phylogenetic analysis. In this study, we identified, characterized and investigated the physiological functions of group I LPMO15 (MaLPMO15-1 and PhLPMO15-1) in two longhorn beetle species, Monochamus alternatus and Psacothea hilaris. In both species, depletion of LPMO15-1 transcripts in last instar larvae by RNAi had no effect on subsequent larval-pupal molting and the resulting pupae developed normally. However, adverse effects on their development were observed during the pupal-adult molting period. The pharate adults were unable to shed their old pupal cuticle and died entrapped in their exuviae probably due to a failure of degradation of the chitin in their old cuticle, which is critical for completion of the insect molting and continuous growth.

Insect eggshell and cuticle/exoskeleton play vital roles in protecting them from natural environmental stresses. However, these chitinous cuticular extracellular matrices must be degraded at least in part during embryo hatching and molting/ecdysis periods to accommodate continuous growth all the way to the adult stage. In this study, we investigated the functional importance of groups I and II chitinases, TcCHT5 and TcCHT10, in the turnover of chitinous cuticle during both embryonic and post-embryonic development in Tribolium castaneum. RNAi and TEM analyses revealed that TcCHT10 is required for digestion of chitin in the serosal cuticle for embryo hatching as well as in the old cuticle during post-embryonic molts including larval-pupal and pupal-adult metamorphosis. TcCHT10 appears to be able to substitute for TcCHT5 in all these vital physiological events except for the pupal-adult molting in which TcCHT5 is indispensable for complete digestion of chitin in the old pupal cuticle.

Many insects form mutualistic relationships with microbial symbionts, crucial for their physiological processes. The bean bug, Riptortus pedestris, establishes a unique gut symbiosis with the genus Caballeronia and consistently acquires these symbionts from surrounding soil with each generation. As a result, the bean bug unavoidably consumes a variety of environmental microbes, including potential pathogens. To address this, the bean bug has developed a specialized organ in its midgut that selectively filters out non-symbiotic microbes, thereby preventing contamination of its symbiotic organ. In this study, we identified a pathogenic strain from the genus Burkholderia that lethally affects the bean bug post-invasion of the symbiotic organ. This pathogen employs a strategy of mimicking the motility of native symbionts to infiltrate the symbiont sorting organ and displays a pronounced resistance against antimicrobial agents produced by the host. Upon establishing itself in the symbiotic organ, the pathogen breaches the midgut cells, leading to host mortality, and subsequently disperses into the external environment. Our findings unveil a cunning pathogenic tactic that exploits the mimicry of native symbionts within an insect's symbiotic framework.