Selecting serum-free hepatocyte cryopreservation stage and storage temperature for the application of an "off-the-shelf" bioartificial liver system.

The bioartificial liver (BAL) system can potentially rescue acute liver failure (ALF) patients by providing partial liver function until a suitable donor liver can be found or the native liver has self-regenerated. In this study, we established a suitable cryopreservation process for the development of an off-the-shelf BAL system. The viability of hepatocyte spheroids cryopreserved in liquid nitrogen was comparable to that of fresh primary hepatocyte spheroids. When hepatocyte spheroids were subjected to cryopreservation in a deep freezer, no statistically significant differences were observed in ammonia removal rate or urea secretion rate based on the cryopreservation period. However, the functional activity of the liver post-cryopreservation in a deep freezer was significantly lower than that observed following liquid nitrogen cryopreservation. Moreover, cryopreserving spheroid hydrogel beads in a deep freezer resulted in a significant decrease (approximately 30%) in both ammonia removal and urea secretion rates compared to the group cryopreserved in liquid nitrogen. The viabilities of spheroid hydrogel beads filled into the bioreactor of a BAL system were similar across all four groups. However, upon operating the BAL system for 24 h, the liver function activity was significantly higher in the group comprising hydrogel beads generated after thawing hepatocyte spheroids cryopreserved in liquid nitrogen. Consequently, the manufacturing of beads after the cryopreservation of hepatocyte spheroids is deemed the most suitable method, considering efficiency, economic feasibility, and liver function activity, for producing a BAL system.

Cytotoxic Activities on Human Ovarian Cancer Cell Lines by Neolignans and Diarylnonanoids from the Seed of Myristica fragrans Houtt

Ovarian cancer is a primary global health concern, often diagnosed at advanced stages with limited treatment options and poor prognosis. Natural products have emerged as potential sources of safe and effective therapies. From the seeds of Myristica fragrans Houtt. (nutmeg), 24 compounds, including neolignans and diarylnonanoid derivatives, were isolated and structurally elucidated. The cytotoxic activities of these isolated metabolites against cisplatin-sensitive and resistant human ovarian cancer cell lines were evaluated. In particular, myrifragranone C (23) exhibited cytotoxicity against all test cancer cell lines A2780, TOV-112D, and SK-OV3 with IC50 values of 14.1, 16.9, and 33.4 μM, respectively. Furthermore, compound 23 induced the death of A2780 and SK-OV3 cancer cells via apoptosis. Western blotting revealed that compound 23 significantly increased the expression of cleaved caspase-3 and poly-ADP ribose polymerase and promoted apoptosis via the mitogenactivated protein kinase signaling pathway. Our findings may provide a preliminary understanding of the antiovarian cancer effect of the active compound myrifragranone C as a potential treatment using natural products.

Electronic alert outpatient protocol improves the quality of care for the risk of postcontrast acute kidney injury following computed tomography

Prevention and diagnosis of postcontrast acute kidney injury (AKI) after contrast-enhanced computed tomography is burdensome in outpatient department. We investigated whether an electronic alert system could improve prevention and diagnosis of postcontrast AKI. Methods: In March 2018, we launched an electronic alert system that automatically identifies patients with a baseline estimated glomerular filtration rate of <45 mL/min/1.73 m2, provides a prescription of fluid regimen, and recommends a follow-up for serum creatinine measurement. Participants prescribed contrast-enhanced computed tomography at outpatient department before and after the launch of the system were categorized as historical and alert group, respectively. Propensity for the surveillance of postcontrast AKI was compared using logistic regression. Risks of AKI, admission, mortality, and renal replacement therapy were analyzed. Results: The historical and alert groups included 289 and 309 participants, respectively. The alert group was more likely to be men and take diuretics. The most frequent volume of prophylactic fluid in historical and alert group was 1,000 and 750 mL, respectively. Follow-up for AKI was more common in the alert group (adjusted odds ratio, 6.00; p < 0.001). Among them, incidence of postcontrast AKI was not statistically different. The two groups did not differ in risks of admission, mortality, or renal replacement therapy. Conclusion: The electronic alert system could assist in the detection of high-risk patients, prevention with reduced fluid volume, and proper diagnosis of postcontrast AKI, while limiting the prescribing clinicians’ burden. Whether the system can improve long-term outcomes remains unclear.

The predictive value of procalcitonin for septic shock in sepsis patients

Objective@#This study examined the difference in procalcitonin between sepsis and septic shock. @*Methods@#The single-center retrospective cohort study was conducted from July 2017 to June 2018 at an emergency department (ED) of a university hospital. The inclusion criteria were patients over 18 years old who visited the ED with an infection. The exclusion criteria were the patients without organ failure by sepsis-3 definition, those with missing serum lactate data, and those discharged without workup. The sepsis patients were divided into those with and without septic shock, and the two groups were compared with biomarkers, including procalcitonin. @*Results@#Of the 406 patients who visited the ED with an infection, 36 were excluded because they did not have sepsis or an unknown infection. Finally, 369 patients were enrolled, and 61.5% fitted the septic shock definition. A comparison of the septic shock and non-shock sepsis groups showed that a history of chronic liver disease, malignancy, pulse rate, prothrombin time, blood urea nitrogen, aspartate and alanine transaminase, troponin-I, Sequential Organ Failure Assessment score and procalcitonin levels were significantly higher in the septic shock group. In multivariate analysis, however, procalcitonin was an independent predictor for septic shock (adjusted odd ratio, 1.05; 95% confidential interval, 1.01-1.09). The area under the receiver operating characteristic curve was 0.729, and the cutoff value was 4.0 ng/mL. @*Conclusion@#The procalcitonin levels were higher in the septic shock group than in the non-shock sepsis group. This could help predict septic shock independently. Further prospective multicenter research is needed to determine if procalcitonin can predict the severity of sepsis.

Textural and Volumetric Changes of the Temporal Lobes in Semantic Variant Primary Progressive Aphasia and Alzheimer’s Disease

Background@#Texture analysis may capture subtle changes in the gray matter more sensitively than volumetric analysis. We aimed to investigate the patterns of neurodegeneration in semantic variant primary progressive aphasia (svPPA) and Alzheimer’s disease (AD) by comparing the temporal gray matter texture and volume between cognitively normal controls and older adults with svPPA and AD. @*Methods@#We enrolled all participants from three university hospitals in Korea. We obtained T1-weighted magnetic resonance images and compared the gray matter texture and volume of regions of interest (ROIs) between the groups using analysis of variance with Bonferroni posthoc comparisons. We also developed models for classifying svPPA, AD and control groups using logistic regression analyses, and validated the models using receiver operator characteristics analysis. @*Results@#Compared to the AD group, the svPPA group showed lower volumes in five ROIs (bilateral temporal poles, and the left inferior, middle, and superior temporal cortices) and higher texture in these five ROIs and two additional ROIs (right inferior temporal and left entorhinal cortices). The performances of both texture- and volume-based models were good and comparable in classifying svPPA from normal cognition (mean area under the curve [AUC] = 0.914 for texture; mean AUC = 0.894 for volume). However, only the texture-based model achieved a good level of performance in classifying svPPA and AD (mean AUC = 0.775 for texture; mean AUC = 0.658 for volume). @*Conclusion@#Texture may be a useful neuroimaging marker for early detection of svPPA in older adults and its differentiation from AD.

The Experiences of Overcoming Turnover Intention among Experienced Nurses / 간호행정학회지

Purpose@#The purpose of this study was to explore the experiences through which nurses overcame turnover intention when first joining the field. @*Methods@#Study subjects included 10 nurses, each of whom had four to six years of work experience at a general hospital. The experiences of overcoming turnover intentions were analyzed from the perspectives of beginner and experienced nurses, using qualitative contents analysis as a method of deduction. @*Results@#The subjects’ turnover intention was low, with an average score of 2.25±0.35. Five themes and 15 sub-themes were extracted from their experiences. The five themes included: “gaining an understanding of the nursing scene”, “experiencing a support system”, “being satisfied with my job”, “developing into a skilled nurse”, and “overcoming and growing”. @*Conclusion@#Subjects maintained low levels of turnover intention, and they were able handle difficulty and grow into their careers as nurses. These findings may inform the development of programs to overcome turnover intention, specifically in terms of factors affecting norvice nurses’ positive achievement factor.

Establishment of a Serum-Free Hepatocyte Cryopreservation Process for the Development of an "Off-the-Shelf" Bioartificial Liver System.

To use hepatocytes immediately when necessary for hepatocyte transplantation and bioartificial liver (BAL) systems, a serum-free cryopreservation protocol ensuring the high survival of hepatocytes and maintenance of their functions should be developed. We established a serum-free protocol for the cryopreservation of primary hepatocytes, hepatocyte spheroids, and hepatocyte spheroid beads in liquid nitrogen. The serum-free cryopreservation solutions showed a significantly higher performance in maintaining enhanced viability and ammonia removal, urea secretion, and the albumin synthesis of hepatocyte spheroids and spheroid beads. The serum-free thawing medium, containing human serum albumin (HSA) and N-acetylcysteine (NAC), was compared with a fetal bovine serum-containing thawing medium for the development of a serum-free thawing medium. Our results show that hepatocyte spheroids and spheroid beads thawed using a serum-free thawing medium containing HSA and NAC exhibited increased hepatocyte viability, ammonia removal, urea secretion, and albumin synthesis compared to those thawed using the serum-containing medium. Finally, we evaluated the liver functions of the cryopreserved BAL system-applied serum-free cryopreservation process compared to the fresh BAL system. The ammonia removal efficiency of the cryopreserved hepatocyte spheroids BAL was lower than or similar to that of the fresh BAL system. Additionally, the urea concentrations in the media of all three BAL systems were not significantly different during BAL system operation. This cryopreserved spheroid-based BAL system using a serum-free process will be a good candidate for the treatment of patients.

Anti-inflammatory activity and cytotoxicity against ovarian cancer cell lines by amide alkaloids and piperic esters isolated from Piper longum fruits: In vitro assessments and molecular docking simulation.

Three new amide alkaloids, piperlongumamides D-F (14, 19, and 32); a new piperic ester, piperlongumester A (45); and two new natural compounds, methyl (2E,4Z)-5-(1,3-benzodioxol-5-yl)penta-2,4-dienoate (46) and trans-piperolein B ester (47), along with 41 known compounds were isolated from the fruits of Piper longum L. Their structures were identified by analyzing spectroscopic data, including mass spectrometry, 1D, and 2D NMR data. The anti-inflammatory and cytotoxic activities of all isolated compounds (1-47) were evaluated. Compounds 3, 6, and 19 inhibited nitric oxide production with IC50 values of 16.1 ± 0.94, 14.5 ± 0.57, and 27.3 ± 1.11 µM, respectively, whereas compound 1 exhibited strong cytotoxic activity toward three ovarian cancer cell lines A2780, TOV-112D, and SK-OV3, with IC50 values of 6.7 ± 0.77, 5.8 ± 0.29, and 48.3 ± 0.40 µM, respectively. Molecular docking simulations were performed to identify the interaction and binding mechanisms of these active metabolites with proteins related to inflammation and cancer.

TAZ links exercise to mitochondrial biogenesis via mitochondrial transcription factor A.

Mitochondria are energy-generating organelles and mitochondrial biogenesis is stimulated to meet energy requirements in response to extracellular stimuli, including exercise. However, the mechanisms underlying mitochondrial biogenesis remain unknown. Here, we demonstrate that transcriptional coactivator with PDZ-binding motif (TAZ) stimulates mitochondrial biogenesis in skeletal muscle. In muscle-specific TAZ-knockout (mKO) mice, mitochondrial biogenesis, respiratory metabolism, and exercise ability were decreased compared to wild-type mice. Mechanistically, TAZ stimulates the translation of mitochondrial transcription factor A via Ras homolog enriched in brain (Rheb)/Rheb like 1 (Rhebl1)-mTOR axis. TAZ stimulates Rhebl1 expression via TEA domain family transcription factor. Rhebl1 introduction by adeno-associated virus or mTOR activation recovered mitochondrial biogenesis in mKO muscle. Physiologically, mKO mice did not stimulate exercise-induced mitochondrial biogenesis. Collectively, our results suggested that TAZ is a novel stimulator for mitochondrial biogenesis and exercise-induced muscle adaptation.

The pioneer round of translation ensures proper targeting of ER and mitochondrial proteins.

The pioneer (or first) round of translation of newly synthesized mRNAs is largely mediated by a nuclear cap-binding complex (CBC). In a transcriptome-wide analysis of polysome-associated and CBC-bound transcripts, we identify RN7SL1, a noncoding RNA component of a signal recognition particle (SRP), as an interaction partner of the CBC. The direct CBC-SRP interaction safeguards against abnormal expression of polypeptides from a ribosome-nascent chain complex (RNC)-SRP complex until the latter is properly delivered to the endoplasmic reticulum. Failure of this surveillance causes abnormal expression of misfolded proteins at inappropriate intracellular locations, leading to a cytosolic stress response. This surveillance pathway also blocks protein synthesis through RNC-SRP misassembled on an mRNA encoding a mitochondrial protein. Thus, our results reveal a surveillance pathway in which pioneer translation ensures proper targeting of endoplasmic reticulum and mitochondrial proteins.

Translation mediated by the nuclear cap-binding complex is confined to the perinuclear region via a CTIF-DDX19B interaction.

Newly synthesized mRNA is translated during its export through the nuclear pore complex, when its 5'-cap structure is still bound by the nuclear cap-binding complex (CBC), a heterodimer of cap-binding protein (CBP) 80 and CBP20. Despite its critical role in mRNA surveillance, the mechanism by which CBC-dependent translation (CT) is regulated remains unknown. Here, we demonstrate that the CT initiation factor (CTIF) is tethered in a translationally incompetent manner to the perinuclear region by the DEAD-box helicase 19B (DDX19B). DDX19B hands over CTIF to CBP80, which is associated with the 5'-cap of a newly exported mRNA. The resulting CBP80-CTIF complex then initiates CT in the perinuclear region. We also show that impeding the interaction between CTIF and DDX19B leads to uncontrolled CT throughout the cytosol, consequently dysregulating nonsense-mediated mRNA decay. Altogether, our data provide molecular evidence supporting the importance of tight control of local translation in the perinuclear region.

Increased ribosomal protein levels and protein synthesis in the striatal synaptosome of Shank3-overexpressing transgenic mice.

The SH3 and multiple ankyrin repeat domains 3 (Shank3) protein is a core organizer of the macromolecular complex in excitatory postsynapses, and its defects cause numerous synaptopathies, including autism spectrum disorders. Although the function of Shank3 as a postsynaptic scaffold is adequately established, other potential mechanisms through which Shank3 broadly modulates the postsynaptic proteome remain relatively unexplored. In our previous quantitative proteomic analysis, six up-regulated ribosomal proteins were identified in the striatal synaptosome of Shank3-overexpressing transgenic (TG) mice. In the present study, we validated the increased levels of RPLP1 and RPL36A in synaptosome, but not in whole lysate, of the TG striatum. Moreover, protein synthesis and extracellular signaling-regulated kinase (ERK) activity were enhanced in the TG striatal synaptosome. To understand the potential contribution of increased protein synthesis to the proteomic change in the TG striatal synaptosome, we performed RNA-sequencing analyses on both whole synaptosomal and synaptic polysome-enriched fractions. Comparative analyses showed a positive correlation only between the polysome-associated transcriptome and up-regulated proteome in the TG striatal synaptosome. Our findings suggest a novel mechanism through which Shank3 may remodel the postsynaptic proteome by regulating synaptic protein synthesis, whose dysfunction can be implicated in SHANK3-associated synaptopathies.

Umbilical trocar port site keloid management using a transposition flap after laparoscopic surgery

Background@#Keloids can occur anywhere in the human body. They are difficult to remove and can cause distress in patients. Although many options are available to treat keloids, no single method is considered the optimal treatment of choice. The authors encountered cases where an umbilical keloid developed at the trocar site after laparoscopic surgery and managed the keloid using a transposition flap. @*Methods@#A total of 10 umbilical keloid patients treated from 2013 to 2020 were included in this study. All patients developed a keloid due to the placement of a laparoscopic trocar incision port, and their major complaints varied from an asymptomatic nodule to pruritus or pain. All excisions were performed under local anesthesia, and transposition flaps were planned afterward. The surrounding tissue was rearranged so that the shape of the umbilicus was deformed to the minimum extent possible. The keloid scars were examined both preoperatively and 6 months postoperatively using the Patient and Observer Scar Assessment Scale (POSAS). @*Results@#All surgical wounds healed well without complications. The average time interval from laparoscopic surgery to keloid scar revision was 4.3 years. The mean postoperative follow-up period was 10.9 months, and no patient underwent reoperation. Four patients were treated with triamcinolone after surgery due to mild hypertrophy or pruritus. The POSAS observer scale showed significantly decreasing scores over time in all patients (P=0.002). @*Conclusions@#Cosmetically unfavorable keloids that form in the umbilicus following laparoscopic surgery can be improved with a simple procedure using excision and transposition flaps.

Nonsense-mediated mRNA decay factor UPF1 promotes aggresome formation.

Nonsense-mediated mRNA decay (NMD) typifies an mRNA surveillance pathway. Because NMD necessitates a translation event to recognize a premature termination codon on mRNAs, truncated misfolded polypeptides (NMD-polypeptides) could potentially be generated from NMD substrates as byproducts. Here, we show that when the ubiquitin-proteasome system is overwhelmed, various misfolded polypeptides including NMD-polypeptides accumulate in the aggresome: a perinuclear nonmembranous compartment eventually cleared by autophagy. Hyperphosphorylation of the key NMD factor UPF1 is required for selective targeting of the misfolded polypeptide aggregates toward the aggresome via the CTIF-eEF1A1-DCTN1 complex: the aggresome-targeting cellular machinery. Visualization at a single-particle level reveals that UPF1 increases the frequency and fidelity of movement of CTIF aggregates toward the aggresome. Furthermore, the apoptosis induced by proteotoxic stresses is suppressed by UPF1 hyperphosphorylation. Altogether, our data provide evidence that UPF1 functions in the regulation of a protein surveillance as well as an mRNA quality control.

A radiation shielding sensitivity analysis based on metal hydrides multilayers.

In order to shield neutron and gamma rays efficiently, a multilayer model is designed with metal hydrides and heavy metals and is analysed based on Monte Carlo simulations. In terms of shielding performance, the hydrogen in metal hydrides acts as a moderator to slow down the neutron energy and heavy metals are good for absorbing gamma rays. A simulation and calculational analysis are carried out with various parameters such as spectrum change, shield thickness, and number of multilayers. In addition, the rate of DPA (displacement per atom) is analysed to estimate both the lifetime and radiation resistance with the MCNP code. From lots of simulations, ZrH2 and W couples are the best candidate especially for shielding gamma rays, while TiH2 with W is good for neutron shielding. The concept of multilayer metal hydride such as TiH2 and ZrH2 coupled with W could be one of best combinations to shield both neutron and gamma-rays in many nuclear facilities such as nuclear reactor, fusion reactor and other applications.

Diabetic Muscle Infarction in a Young Patient with Prader-Willi Syndrome and Type 2 Diabetes Mellitus / 임상당뇨병

Diabetic muscle infarction (DMI), also known as diabetic myonecrosis, is a rare complication of diabetes mellitus (DM); hence, it is often underdiagnosed. Thus, timely diagnosis and treatment are essential for a better prognosis. We describe a 24-year-old woman with Prader-Willi syndrome and an 8-year history of uncontrolled type 2 DM, who presented with a sudden onset of fever and subacute painful swelling of her left thigh. She was finally diagnosed with DMI based on magnetic resonance imaging and muscle biopsy after excluding other infectious and inflammatory diseases of proximal muscles. The patient was treated with bed rest, strict glycemic control, and analgesics, and her symptoms gradually resolved. DMI should be considered in the differential diagnosis of patients with poorly controlled DM, who present with subacute pain and swelling of lower extremity muscles, without a history of trauma.

Diabetic Muscle Infarction in a Young Patient with Prader-Willi Syndrome and Type 2 Diabetes Mellitus / 임상당뇨병

Diabetic muscle infarction (DMI), also known as diabetic myonecrosis, is a rare complication of diabetes mellitus (DM); hence, it is often underdiagnosed. Thus, timely diagnosis and treatment are essential for a better prognosis. We describe a 24-year-old woman with Prader-Willi syndrome and an 8-year history of uncontrolled type 2 DM, who presented with a sudden onset of fever and subacute painful swelling of her left thigh. She was finally diagnosed with DMI based on magnetic resonance imaging and muscle biopsy after excluding other infectious and inflammatory diseases of proximal muscles. The patient was treated with bed rest, strict glycemic control, and analgesics, and her symptoms gradually resolved. DMI should be considered in the differential diagnosis of patients with poorly controlled DM, who present with subacute pain and swelling of lower extremity muscles, without a history of trauma.

Quisqualis indica extract ameliorates low urinary tract symptoms in testosterone propionate-induced benign prostatic hyperplasia rats / 한국실험동물학회지

Benign prostate hyperplasia (BPH) is a common disease in old-age males, accounting for approximately 77% of morbidity within the age range of 40 to 70 years. It has been shown that morbidity increases with social graying. Quisqualis indica linn (QI) has been used to treat inflammation, stomach pain, and digestion problems. In this study, we evaluated the symptom-regulating effects of QI extract on a testosterone-induced BPH rat model. After inducing BPH in rats using testosterone propionate (TP) injection, we assessed basal intraurethral pressure (IUP) and increments of IUP elicited by electrical field stimulation (5 V, 5, 10, or 20 Hz) or phenylephrine (Phe) (0.01, 0.03, 0.1 mg/kg IV). To induce BPH, 8-week-old rats were subjected to a daily subcutaneous TP (3 mg/kg) injection for 4 weeks. Finasteride (Fina) (10 mg/kg PO) was administered to the rats in the first treatment, while QI (150 mg/kg PO) was administered to those in the second group. Blood pressure was measured together with IUP, after which low urinary tract (LUT), ventral prostate (VP), testicle, and corpus spongiosum were isolated and weighed. Basal IUPs for the Fina- and QI-treated groups were 87.6 and 86.8%, respectively.LUT and VP organ weights in the QI group were lower than those in the Fina group. However, the QI group showed significantly reduced electrical stimulated or Phe-induced IUP increment compared to the Fina and BPH groups. These results proved that QI can be beneficial for BPH symptoms by inhibiting 5α-reductase and consequently decreasing prostate and releasing urinary pressure.

Nursing Salary Guideline for Korean Hospitals / 임상간호연구

Purpose@#The purpose of this study is to develop a nursing salary guideline for Korean hospitals. @*Methods@#Literature review and a mobile survey were conducted regarding staff nurses’ salary. Regression analysis and simulation model were applied to develop the nurses’ salary guideline. @*Results@#The United Kingdom, Australia, and Germany have standard salary guidelines which demonstrated the standard salary of nurses. These were determined mainly by nursing experience and expertise. The results of the mobile survey indicated that the maximum to minimum ratio of the salary was as high as 4.5 among staff nurses working in Korean hospitals. Two models (exponential and linear) for a standard nursing salary guideline were developed and the simulation results demonstrated an improved salary structure for staff nurses. @*Conclusion@#This developed salary guideline for staff nurses is recommended to be applied in Korean hospitals which provide total nursing care services.

One-stage nipple and breast reconstruction using a deep inferior epigastric perforator flap after a skin-sparing mastectomy

Background@#Nipple reconstruction is usually performed as a delayed procedure in patients with breast cancer who undergo skin-sparing mastectomy and breast reconstruction surgery using a deep inferior epigastric perforator (DIEP) flap. The authors designed this study to evaluate the utility of breast reconstruction based on a DIEP flap and immediate nipple reconstruction. @*Methods@#A retrospective review was conducted of all patients who underwent breast reconstruction performed by a single plastic surgeon from October 2016 to June 2018. Through a questionnaire and chart review, we compared surgical results and complications in cases of single-stage nipple reconstruction after skin-sparing mastectomy (n=17) with patients who underwent delayed nipple reconstruction after skin-sparing mastectomy, modified radical mastectomy, or simple mastectomy (n=7). @*Results@#In a subjective analysis using clinical photos, the immediate nipple reconstruction group had higher scores than their counterparts in an evaluation of the nipple-areolar complex (NAC) (NAC placement, 3.34 vs. 3.04; nipple projection, 3.05 vs. 3.03; nipple size, 3.30 vs. 3.29). No significant differences between the groups were found in terms of complications. @*Conclusions@#Simultaneous nipple reconstruction is a reliable surgical method with economic advantages. No differences were found in terms of outcomes and complications in comparison to delayed reconstruction. Therefore, surgeons can consider simultaneous nipple reconstruction without particular concerns about asymmetry or necrosis.