Pig Liver Research Articles

Mesenchymal Stromal Cell on Liver Decellularised Extracellular Matrix for Tissue Engineering

In end-stage chronic liver disease, transplantation represents the only curative option. However, the shortage of donors results in the death of many patients. To overcome this gap, it is mandatory to develop new therapeutic options. In the present study, we decellularised pig livers and reseeded them with allogeneic porcine mesenchymal stromal cells (pMSCs) to understand whether extracellular matrix (ECM) can influence and/or promote differentiation into hepatocyte-like cells (HLCs). After decellularisation with SDS, the integrity of ECM-scaffolds was examined by histological staining, immunofluorescence and scanning electron microscope. DNA quantification was used to assess decellularisation. pMSCs were plated on scaffolds by static seeding and maintained in in vitro culture for 21 days. At 3, 7, 14 and 21 days, seeded ECM scaffolds were evaluated for cellular adhesion and growth. Moreover, the expression of specific hepatic genes was performed by RT-PCR. The applied decellularisation/recellularisation protocol was effective. The number of seeded pMSCs increased over the culture time points. Gene expression analysis of seeded pMSCs displayed a weak induction due to ECM towards HLCs. These results suggest that ECM may address pMSCs to differentiate in hepatocyte-like cells. However, only contact with liver-ECM is not enough to induce complete differentiation.

Serological Searching for Hepatitis E Virus Infection Among Pig Liver Transudate from Bulgaria: An Alternative Method for Seroepidemiological Survey.

Background: The aim of this research was to perform a seroepidemiological survey of hepatitis E virus (HEV) by an alternative method-serological establishment of the virus from pig liver transudate. This is the first such research in Bulgaria. Moreover, no studies have been conducted on HEV in pig liver in our country. Materials and Methods: A total of 90 liver samples from healthy pigs were collected from slaughterhouses in three parts of the country (Sofia, Lovech, and Stara Zagora districts). An equal number of samples (n = 30) were obtained from each district. Results: The overall HEV seropositivity was 67.7% of all 90 tested pig liver transudate samples; in Sofia district 20.0%, in Lovech district 90.0%, and in Stara Zagora district 93.3%. Conclusion: These data are similar to our previous serological studies for HEV in serum samples from industrial pigs, wild boars, and East Balkan swine. In this regard, testing the liver transudate could be a fine alternative method for seroepidemiological survey on HEV in swine.

Nitrogen-Doped Carbon Dots with Free Radical Scavenging Abilities as Fluorescence Probes for Cr(VI) Sensing in Bio-Systems

Carbon dots (CDs) are a new class of fluorescent nanomaterials that can be used in optical devices, fluorescence sensing and bio-imaging. Recently, the preparation of CDs from biomass gained great interest because the source is plentiful and they are friendly to environment. However, there still exist some problems including low quantum yield (QY) and limited application. Although CDs have been successfully applied in detection of various ions, the sensing of Cr(VI) and the corresponding mechanism is rarely proposed. Here, a facile and effective preparation of N-doped carbon dots (N-CDs) from sweet potatoes was developed through hydrothermal synthesis, using urea as passivator. The characterization was performed to demonstrate the morphology and chemical composition of the N-CDs. In particular, it is confirmed that N element exists in NH2 group on the surface of the N-CDs. Because of doing with N element, the QY of these CDs significantly increases to 31%. In addition, the N-CDs exhibit favorable free radical scavenging activities toward DPPH, hydroxyl and superoxide anion free radicals. This antioxidant property, combined with inner filter effect, endows the N-CDs with florescence response to Cr(VI). The further study showed that linear range of Cr(VI) concentration is from 1[Formula: see text][Formula: see text]M to 80[Formula: see text][Formula: see text]M with a detection limit of 0.61[Formula: see text][Formula: see text]M. Subsequently, the N-CDs were competent to detect trace Cr(VI) both in human serum and pig liver. Based on the fact that the N-CDs are biocompatible by MTT experiment, the N-CDs were excellent in fluorescent sensing of Cr(VI) in living cells. In a word, the N-CDs are suitable to be applied for Cr(VI) detection in bio-systems, as well as in antioxidant fields.

Detection of Hepatitis E Virus (HEV) in Pigs and in the Wild Boar (Sus scrofa) Population of Chieti Province, Abruzzo Region, Italy

Hepatitis E virus (HEV) is a zoonotic pathogen, causing infectious hepatitis in man. Pigs and wild boars are the natural asymptomatic reservoirs, while the disease in humans could be either asymptomatic or evolve in hepatitis. In Europe, an increasing number of human infections from HEV have been reported over the last few years. The main route of transmission is through contaminated food, by direct or indirect consumption of raw or undercooked pork and wild boar meat and meat products. Up to now, HEV prevalence in Italian northern regions has been extensively determined in wild boars and pigs, while less data have been collected from the southern ones. There is a need to report more data about HEV prevalence from wild boars and pigs in southern Italy in consideration of the potential risk posed by some specific traditional food products manufactured in these areas and produced from pig and wild boar livers (e.g., sausages and salami). The aim of this study was to demonstrate the circulation of the Hepatitis E virus (HEV) in pigs and in the wild boar population of the province of Chieti, Abruzzo Region, Central Italy. Moreover, potential HEV seroprevalence in hunters from that area was also assessed. The overall prevalence of HEV RNA in wild boars was 9.5% (CI 5.4–16.2%), but no HEV RNA was detected in samples from pigs.

Manganese content in some organs and tissues of Landrace pigs

In the article, the authors presented the results of assessing the level of manganese in the skeletal muscles, liver, kidneys, and bristles of Landrace pigs. The studies were performed on clinically healthy six-monthold animals from a large pig farm in the Altai Territory. The conditions of keeping the animals corresponded to the standard ones, the feeding was typical for meat fattening. Elemental analysis of samples of parenchymal organs and muscle tissue was performed using atomic absorption spectrometry with flame and electrothermal atomization, bristles - by atomic emission spectrometry with inductively coupled plasma. Data processing was performed using Microsoft Office Excel and the R programming language in the data analysis environment RStudio version 1.3.1093 (2009–2020 RStudio, PBC). The authors found that the distribution in all cases was normal, and the dispersions are not homogeneous. Based on the average value and median, an increasing ranked series of manganese content in organs and tissues were established: skeletal muscles < kidneys < liver < bristles, in numerical terms: 1: 12.5: 15.7: 207. Manganese levels in skeletal muscles, kidneys, liver, and bristles were 0.13; 1.62; 2.04, and 26.91 mg/kg, respectively. Greater homogeneity is characteristic of the accumulation of microelements in the muscles and kidneys. Based on the Kruskal-Wallis test, it was established that the accumulation of manganese differs in the examined organs and tissues of pigs (H = 88.4, df = 3, p < 0.0001). Pairwise comparison showed significant differences for all pairs, except for the liver-kidney pair. Based on the cluster analysis, the closest results are typical for the liver and kidneys; the bristle is a separate opposite cluster, characterized by a significant level of manganese accumulation. The results obtained can serve as a preliminary corresponding physiological norm for the concentration of manganese in individual organs, tissues, and skin derivatives of Landrace pigs under the conditions of Western Siberia.

Validation of a LC-MS/MS method for the quantitative analysis of four antibiotics in pig tissues and plasma to assess the risk of transfer of residues to edible matrices after exposure to cross-contaminated feed

Cross-contamination between medicated and non-medicated feed can occur during production, processing, transport or storage of animal feed. This may lead to the presence of low concentrations of antibiotics in supposedly drug-free feed for food production animals, which potentially could also harm consumers due to residues. In addition, consumption of sub-therapeutic concentrations of antibiotics may increase the risk of emergence of resistant bacteria. In this study, LC-MS/MS methods were developed to quantify four antibiotics (sulfadimethoxine, oxytetracycline, trimethoprim and amoxicillin) in several pig matrices, i.e. plasma, muscle, liver, kidneys and faeces. All methods were validated using the accuracy profile, except for amoxicillin in faeces, for which extraction could not be optimised for low concentrations. These methods were then applied as part of an animal study during which several pigs received contaminated feed at a concentration corresponding to 2% of therapeutic dose, in order to evaluate the risk of the presence of residues in animal faeces and tissues. The results showed that sulfadimethoxine is well absorbed and accumulates in the muscle, kidneys and liver, where concentrations were higher than the maximum residue limits (MRLs) authorised in EU legislation. Conversely, oxytetracycline was mostly found in faeces as its oral absorption is very low. Trimethoprim concentrations were slightly higher than the tolerated MRL in the kidneys, but they were below this level in the other tissues. Finally, amoxicillin concentrations remained below the lower limit of quantification of the methods in all matrices.

Synthesis of Cyclic Carbonates Catalyzed by CaI2–Et3N and Studies on Their Biocatalytic Kinetic Resolution

Herein, we report a simple catalytic system consisting of CaI2 in combination with Et3N as a ligand for the efficient cycloaddition of CO2 to a variety of epoxides. Terminal (bio-based) carbonates were obtained under mild conditions (23 °C, 1 bar CO2 pressure) in excellent yields up to 97%. Under elevated temperature (70 °C) and higher CO2 pressure (10 bar), even challenging internal epoxides were converted, leading to the desired carbonates in yields up to 90%. 1H NMR experiments indicated the in situ coordination of the amine to the Lewis acidic calcium center as well as the activation of the epoxide. Furthermore, the enzymatic kinetic resolution of seven cyclic carbonates using readily available pig liver esterase was investigated. Styrene carbonate proved to be the most suitable substrate for the kinetic resolution, leading to (R)-styrene carbonate in 99% ee.

Deciphering the species differences in CES1A-mediated hydrolytic metabolism by using a bioluminescence substrate

Carboxylesterases 1A (CES1A) is a key enzyme responsible for the hydrolytic metabolism of a great deal of endogenous and exogenous substrates bearing ester- or amide-bond(s). This study aimed to decipher the species difference in CES1A-mediated hydrolytic metabolism by using a newly developed bioluminescence CES1A sensor (termed NLMe) as the probe substrate, while the liver microsomes from six different mammalian species (human, cynomolgus monkey, dog, minipig, rat and mouse) were used as the enzyme sources. Metabolite profiling demonstrated that all tested liver microsomes from various species could catalyze NLMe hydrolysis, but significant difference in hydrolytic rate was observed. Kinetic plots of NLMe hydrolysis in liver microsomes from different species showed that the inherent clearance rates (Clint) of NLMe in human liver microsomes (HLM), cynomolgus monkey liver microsomes (CyLM), and pig liver microsome (PLM) were comparable, while the Clint values of NLMe in dog liver microsomes (DLM), mouse liver microsomes (MLM), and rat liver microsomes (RLM) were relatively small. Moreover, chemical inhibition assays showed that NLMe hydrolysis in all tested liver microsomes could be competently inhibited by BNPP (a potent broad-spectrum inhibitor of CES), but CUA (a selective inhibitor of human CES1A) only inhibited NLMe hydrolysis in human liver microsomes and dog liver microsomes. In summary, the species differences in CES1A-catalyzed NLMe hydrolysis were carefully investigated from the views of the similarities in metabolite profile, hydrolytic kinetics and inhibitor response. All these findings provide new insights into the species differences in CES1A-mediated hydrolytic metabolism and suggest that it is necessary for the pharmacologists to choose appropriate animal models to replace humans for evaluating the in vivo effects of CES1A inhibitors.

An Efficacious Transgenic Strategy for Triple Knockout of Xeno-Reactive Antigen Genes GGTA1, CMAH, and B4GALNT2 from Jeju Native Pigs.

Pigs are promising donors of biological materials for xenotransplantation; however, cell surface carbohydrate antigens, including galactose-alpha-1,3-galactose (α-Gal), N-glycolylneuraminic acid (Neu5Gc), and Sd blood group antigens, play a significant role in porcine xenograft rejection. Inactivating swine endogenous genes, including GGTA1, CMAH, and B4GALNT2, decreases the binding ratio of human IgG/IgM in peripheral blood mononuclear cells and erythrocytes and impedes the effectiveness of α-Gal, Neu5Gc, and Sd, thereby successfully preventing hyperacute rejection. Therefore, in this study, an effective transgenic system was developed to target GGTA1, CMAH, and B4GALNT2 using CRISPR-CAS9 and develop triple-knockout pigs. The findings revealed that all three antigens (α-Gal, Neu5Gc, and Sd) were not expressed in the heart, lungs, or liver of the triple-knockout Jeju Native Pigs (JNPs), and poor expression of α-Gal and Neu5G was confirmed in the kidneys. Compared with the kidney, heart, and lung tissues from wild-type JNPs, those from GGTA1/CMAH/ B4GALNT2 knockout-recipient JNPs exhibited reduced human IgM and IgG binding and expression of each immunological rejection component. Hence, reducing the expression of swine xenogeneic antigens identifiable by human immunoglobulins can lessen the immunological rejection against xenotransplantation. The findings support the possibility of employing knockout JNP organs for xenogeneic transplantation to minimize or completely eradicate rejection using multiple gene-editing methods.

P2.21: Protection of Normal Metabolic Function of Healthy Pig Livers Following Prolonged Normothermic Ex-situ Perfusion

P2.21: Protection of Normal Metabolic Function of Healthy Pig Livers Following Prolonged Normothermic Ex-situ Perfusion

Identification and characterization of MicroRNAs in pig liver after the LPS challenge using RNA-seq

ABSTRACT This study was conducted to test the expression profiles of miRNAs in the liver in piglets after the LPS challenge using sequencing technology. Twelve barrows were assigned to two groups, including the saline group and the LPS group. Six small RNA (sRNA) libraries were constructed, and novel miRNAs were identified using mirevo and mirdeep2 software and validated by quantitative real-time PCR (qRT-PCR). The results showed the mRNA expression of IL-1β, IL-6 and TNF-α in the liver increased after the LPS challenge (P < .05). A total of 29 differentially expressed miRNAs were identified in the liver after the LPS challenge. And 11 miRNAs were validated by qRT-PCR. In addition, the results of statistics of pathway enrichment showed that these miRNAs could be related to lipid metabolism response. This study provides the first miRNA expression profiles of LPS-mediated changes in the liver, which might provide potential insights into miRNAs involved in regulating lipid metabolism in pigs challenged by LPS.

Mild-temperature photothermal treatment method and system based on photoacoustic temperature measurement and control

As a non-invasive technology for tumor treatment, photothermal therapy (PTT) is increasingly applied during clinical treatment. However, due to the lack of non-invasive temperature measurement methods, excessive therapeutic light dose will damage the healthy tissue around the lesion. Monitoring and control the therapy temperature during PTT application is necessary to ensure effective therapy while minimizing thermal damage to adjacent tissue. In order to ensure that the therapy temperature in PTT is stable in the mild range (below 43 °C), it is necessary to design a mild-temperature PTT method and system that can detect and control the temperature in real-time. A mild-temperature photothermal treatment method and system based on photoacoustic temperature measurement and control is proposed in this paper. On the basis of reference temperature, the relative temperature change is measured based on the photoacoustic information, and the treatment temperature of the target area is precisely controlled in real time using the closed-loop temperature control algorithm. Based on the experimental results of agar-based phantoms and pig liver tissue, the proposed system can achieve temperature imaging in real time, and the control steady-state error and overshoot are 0.62 °C and 0.3 % respectively. The results show that our method has the potential for accurate temperature monitoring and closed-loop temperature control during PTT.

Occurrence of Toxic Metals and Metalloids in Muscle and Liver of Italian Heavy Pigs and Potential Health Risk Associated with Dietary Exposure.

Muscle and liver tissues from Italian heavy pigs were analyzed to investigate whether the chronic consumption of these products by local consumers could represent a health risk in relation to the contamination by some toxic metals and metalloids (TMMs). The concentrations of Al, As, Cd, Cr, Cu, Fe, Ni, Pb, Sn, U, and Zn were measured with an inductively coupled plasma–mass spectrometer, while Hg analysis was performed by using a mercury analyzer. Fe, Zn, and Cu were the most abundant elements in both tissues, while U was detected only at ultra-trace levels. As, Cd, Cu, Fe, Hg, Pb, U, and Zn showed significantly higher concentrations in livers compared to muscles (p ≤ 0.01), with Cd and Cu being 60- and 9-fold more concentrated in the hepatic tissue. Despite this, concentrations of all TMMs were found to be very low in all the samples to the point that the resulting estimated dietary intakes did not suggest any food safety concern. Indeed, intakes were all below the toxicological health-based guidance values or resulted in low margins of exposure. Nevertheless, in the calculation of the worst-case exposure scenario, the children’s estimated intake of Cd, Fe, and Zn through the sole consumption of pig liver contributed to more than 23, 38, and 39% of the tolerable weekly intakes of these elements, while the combined consumption of pig liver and pig muscle to more than 24, 46, and 76%. These findings alert about the probability of exceeding the toxicological guidance values of Cd, Fe, and Zn though the whole diet, suggesting long-term negative health effects for the younger population.

Variations in Fibrinogen-like 1 (FGL1) Gene Locus as a Genetic Marker Related to Fat Deposition Based on Pig Model and Liver RNA-Seq Data.

The goal of this study was to evaluate the effects of mutations in the FGL1 gene associated with pig productive traits to enrich the genetic marker pool for further selection and to support the studies on FGL1 in the context of the fat deposition (FD) process. The variant calling and χ2 analyses of liver RNA-seq data were used to indicate genetic markers. FGL1 mutations were genotyped in the Złotnicka White (n = 72), Polish Large White (n = 208), Duroc (n = 72), Polish Landrace (PL) (n = 292), and Puławska (n = 178) pig breeds. An association study was performed using a general linear model (GLM) implemented in SAS® software. More than 50 crucial mutations were identified in the FGL1 gene. The association study showed a significant effect of the FGL1 on intramuscular fat (IMF), loin eye area, backfat thickness at the lumbar, ham mass (p = 0.0374), meat percentage (p = 0.0205), and loin fat (p = 0.0003). Alternate homozygotes and heterozygotes were found in the PL and Duroc, confirming the selective potential for these populations. Our study supports the theory that liver FGL1 is involved in the FD process. Moreover, since fat is the major determinant of flavor development in meat, the FGL1 rs340465447_A allele can be used as a target in pig selection focused on elevated fat levels.

Adhesive polyethylene glycol-based hydrogel patch for tissue repair

Polyethylene glycol (PEG)-based tissue glue has been widely used in surgery, but the sealants were weak in strength and could not be in place of sutures. This paper prepared a PEG-based tissue patch made of F127-DA hydrogel and PEG-DA as adhesive. For the patch adhesion on tissues, photoinitiator of lithium phenyl-2,4,6-trimethylbenzoylphosphinate (LAP, 0.1 % w/v) and UV flashlight at 395 nm for 30 s irradiation was used. The adhesion energy on pig skin, lung, heart, liver, kidney and stomach was 85.33, 56.40, 75.56, 70.22, 66.67 and 36.89 N/m, indicating the well stitching to different tissues. The hydrogel patch could also seal the punched tissues to prevent the leakage of liquid or air. Finally, the patch showed good tissue compatibility after 2 days adhesion on pig skin. In this regard, the hydrogel patch is expected to repair the wounded tissues without suture, being a biocompatible adhesive for wound healing of various tissues.

RNA-Seq identification of lipid metabolism genes associated with intramuscular fat by transcriptome analyses of Longissimus dorsi and liver in pigs feed with avocado meal supplement

El objetivo de esta investigación fue identificar los genes involucrados en el metabolismo de lípidos y grasa intramuscular (GIM), por medio del análisis de transcriptoma en Longissimus dorsi e hígado, en cerdos de engorde con dieta suplementada con harina de aguacate (HA), así como su efecto sobre el peso final, pH45 y GIM. Se alimentaron ocho cerdos castrados en cada dieta, una con HA al 0 % (HA0) y otra con 10 % (HA10). El análisis de transcriptoma fue realizado con secuenciación masiva (ARN-Seq), con el método DESeq2 se identificaron genes a partir de 12 muestras, tres por dieta y tejido, para obtener diferencialmente la expresión génica (DEG) de los Log2 Fold Change (Log2FC) de HA0 vs HA10, considerando valores de P&lt;0,10 a P&lt;0,01-E10. Suplementar con HA10 no afectó el peso final (107,6 kg), pero si aumentó el pH45 (6,0) y disminuyó la GIM (5,3 %). Comparando las dietas HA0 vs HA10, se identificaron en L. dorsi más genes con alta DEG Log2FC (UP) en HA0, que se han relacionado con mayor influencia en el aumento de GIM. Con valores más altos de Log2FC y P&lt;0,001 se identificaron a genes UP en HA10 más asociados al metabolismo de los ácidos grasos; once genes para L. dorsi (ABCG1, ADORA1, BMPR1B, FABP3, FRZB, ITGB6, MYLIP, RGN, RORC, RXRG, SPOCK3) y trece genes para hígado (CA3, CDKN1A, FADS1, FADS2, GABRB2, PCK2, PLIN4, RETSAT, ROBO2, RORC, SLC27A6, SOCS2, SPOCK3). Las correlaciones entre los valores Log2 de expresión de los genes con GIM, identifican en L. dorsi a 19 genes con correlación positiva (0,80-0,97) y 13 con correlación negativa (0,80-0,98), y en hígado a seis genes con correlación positiva (0,80-0,92) y nueve con correlación negativa (0,80-0,94). La HA afecta la GIM y ocasiona cambios en la expresión de genes asociados al metabolismo de lípidos.

Mechanisms of regenerative preconditioning in pigs with subtotal hepatectomies.

Portal vein occlusion shortly before extended hepatic resections has hepatoprotective properties, but its molecular effects have not been elucidated. We characterized the impact of regenerative preconditioning by portal vein embolization (PVE) on hepatic energy metabolism and cytokine expression. About 90% hepatectomies were performed in normal pigs (Control) and in pigs that underwent a PVE 24 h before the surgery (n = 10/group). Blood biochemistry and coagulation, liver damage, liver function (ICG), hepatic content of adenine nucleotides, and hepatic expression of inflammatory mediators (RT-PCR and WB) were determined before the hepatectomy, 15 min, and 24 h later. All PVE and hepatectomies were successfully accomplished. The 90% hepatectomy resulted in: Immediate reduction of ATP, leading to persistent decreases of energy load and ATP/ADP ratio up to the 24-h time-point; and pro-inflammatory expression profile of cytokines in the remnant liver. Prior performance of PVE attenuated the bioenergetic alterations and prevented many of the changes in hepatic cytokine expression. Regenerative preconditioning by PVE improved hepatic energy metabolism and modulated inflammatory mediators in the remnant liver in pigs undergoing major hepatectomies, potentially contributing to its hepatoprotective effects.

Co-Circulation of Different Hepatitis E Virus Genotype 3 Subtypes in Pigs and Wild Boar in North-East Germany, 2019.

Hepatitis E is a major cause of acute liver disease in humans worldwide. The infection is caused by hepatitis E virus (HEV) which is transmitted in Europe to humans primarily through zoonotic foodborne transmission from domestic pigs, wild boar, rabbits, and deer. HEV belongs to the family Hepeviridae, and possesses a positive-sense, single stranded RNA genome. This agent usually causes an acute self-limited infection in humans, but in people with low immunity, e.g., immunosuppressive therapy or underlying liver diseases, the infection can evolve to chronicity and is able to induce a variety of extrahepatic manifestations. Pig and wild boar have been identified as the primary animal reservoir in Europe, and consumption of raw and undercooked pork is known to pose a potential risk of foodborne HEV infection. In this study, we analysed pig and wild boar liver, faeces, and muscle samples collected in 2019 in Mecklenburg-Western Pomerania, north-east Germany. A total of 393 animals of both species were investigated using quantitative real-time reverse transcription polymerase chain reaction (RT-qPCR), conventional nested RT-PCR and sequence analysis of amplification products. In 33 animals, HEV RNA was detected in liver and/or faeces. In one individual, viral RNA was detected in muscle tissue. Sequence analysis of a partial open reading frame 1 region demonstrated a broad variety of genotype 3 (HEV-3) subtypes. In conclusion, the study demonstrates a high, but varying prevalence of HEV RNA in swine populations in Mecklenburg-Western Pomerania. The associated risk of foodborne HEV infection needs the establishment of sustainable surveillance and treatment strategies at the interface between humans, animals, and the environment within a One Health framework.

In Vivo Evaluation of a Novel Radiofrequency Ablation Electrode in Pig Livers

(1) Background: Endoscopic ultrasound-guided radiofrequency ablation (EUS-RFA) is an alternative treatment for pancreatic tumors. Currently, EUS-RFA has been trialled to treat hepatic tumors. However, little has been reported about optimal settings for EUS-RFA in patients with hepatic tumors. We evaluated the ablation effect after in vivo RFA using a new EUS-RFA electrode in a pig model. (2) Methods Four pigs were used for the in vivo test. The in vivo testing was divided into two tests based on the length of the RFA electrode (0.5, 0.7, 1 or 1.5 cm), the ablation power (30 W or 50 W), and the ablation time (10 or 15 s). In test one, ablation effect was evaluated based on the electrode length and power. In test two, ablation effect was assessed based on power and time. (3) Results: In test one, the ablation width and depth correlated with the length of the electrode and power (0.5 cm, 10 W, 10 s: width 0.46 cm, depth 0.65 cm vs. 1.5 cm, 75 W, 10 s: width 0.77 cm, depth 1.80 cm). In test two, ablation width and depth were similar when RFA was set at 1.5cm, 50 W, and 10 s or 1 cm, 30 W, and 15 s (0.65 cm, 1.14 cm vs. 0.65cm, 1.26 cm). (4) Conclusions: The relationship between electrode length, ablation power, and ablation time, and the resulting ablation effect in pig livers suggest that EUS-RFA produces effective ablation while minimizing thermal injury.

Identification of ASPDH as a novel NAADP-binding protein

Nicotinic acid adenine dinucleotide phosphate (NAADP) is a signaling molecule that can induce calcium release from intracellular acidic stores. However, proteins that bind to NAADP are understudied. Here, we identify aspartate dehydrogenase domain-containing protein (ASPDH) as an NAADP-binding protein through biochemical purification from pig livers. Isothermal titration calorimetry (ITC) experiment using the recombinantly expressed protein shows a 1:1 binding stoichiometry and a Kd of 455 nM between NAADP and mouse ASPDH. In contrast, recombinantly expressed Jupiter microtubule-associated homolog 2 (JPT2) and SM-like protein LSM12, two proteins previously identified as NAADP-receptors, show no binding in ITC experiments.