Characteristics of Purified Horse Oil by Supercritical Fluid Extraction with Different Deodorants Agents.

This study investigated the impact of activated carbon, palm activated carbon, and zeolite on horse oil (HO) extracted from horse neck fat using supercritical fluid extraction with deodorant-untreated HO (CON) as a comparison. The yield and lipid oxidation of deodorant untreated HO (CON) were not significantly affected by the three deodorants. However, deodorant-treated HOs exhibited significantly elevated levels of α-linolenic acid (C18:3n3) and eicosenoic acid (C20:1n9) compared to CON (p<0.05), while other fatty acids remained consistent. Zeolite-purified HO demonstrated significantly lower levels of volatile organic compounds (VOCs) than other treatments (p<0.05). Remarkably, zeolite decreased the concentration of pentane, 2,3-dimethyl (gasoline odor), by over 90%, from 177.17 A.U. ×106 in CON to 15.91 A.U. ×106. Zeolite also effectively eliminates sec-butylamine (ammonia and fishy odor) as compared to other deodorant-treated HOs (p<0.05). Additionally, zeolite reduced VOCs associated with the fruity citrus flavor, such as nonanal, octanal, and D-limonene in HO (p<0.05). This study suggests that integrating zeolite in supercritical fluid extraction enhances HO purification by effectively eliminating undesirable VOCs, presenting a valuable approach for producing high-quality HO production in the cosmetic and functional food industries.

The Mst1/2-BNIP3 axis is required for mitophagy induction and neuronal viability under mitochondrial stress.

Mitophagy induction upon mitochondrial stress is critical for maintaining mitochondrial homeostasis and cellular function. Here, we found that Mst1/2 (Stk3/4), key regulators of the Hippo pathway, are required for the induction of mitophagy under various mitochondrial stress conditions. Knockdown of Mst1/2 or pharmacological inhibition by XMU-MP-1 treatment led to impaired mitophagy induction upon CCCP and DFP treatment. Mechanistically, Mst1/2 induces mitophagy independently of the PINK1-Parkin pathway and the canonical Hippo pathway. Moreover, our results suggest the essential involvement of BNIP3 in Mst1/2-mediated mitophagy induction upon mitochondrial stress. Notably, Mst1/2 knockdown diminishes mitophagy induction, exacerbates mitochondrial dysfunction, and reduces cellular survival upon neurotoxic stress in both SH-SY5Y cells and Drosophila models. Conversely, Mst1 and Mst2 expression enhances mitophagy induction and cell survival. In addition, AAV-mediated Mst1 expression reduced the loss of TH-positive neurons, ameliorated behavioral deficits, and improved mitochondrial function in an MPTP-induced Parkinson's disease mouse model. Our findings reveal the Mst1/2-BNIP3 regulatory axis as a novel mediator of mitophagy induction under conditions of mitochondrial stress and suggest that Mst1/2 play a pivotal role in maintaining mitochondrial function and neuronal viability in response to neurotoxic treatment.

Selective induction of Rab9-dependent alternative mitophagy using a synthetic derivative of isoquinoline alleviates mitochondrial dysfunction and cognitive deficits in Alzheimer's disease models.

Rationale: Promotion of mitophagy is considered a promising strategy for the treatment of neurodegenerative diseases including Alzheimer's disease (AD). The development of mitophagy-specific inducers with low toxicity and defined molecular mechanisms is essential for the clinical application of mitophagy-based therapy. The aim of this study was to investigate the potential of a novel small-molecule mitophagy inducer, ALT001, as a treatment for AD. Methods: ALT001 was developed through chemical optimization of an isoquinolium scaffold, which was identified from a chemical library screening using a mitophagy reporter system. In vitro and in vivo experiments were conducted to evaluate the potential of ALT001 as a mitophagy-targeting therapeutic agent and to investigate the molecular mechanisms underlying ALT001-induced mitophagy. The therapeutic effect of ALT001 was assessed in SH-SY5Y cells expressing mutant APP and mouse models of AD (5×FAD and PS2APP) by analyzing mitochondrial dysfunction and cognitive defects. Results: ALT001 specifically induces mitophagy both in vitro and in vivo but is nontoxic to mitochondria. Interestingly, we found that ALT001 induces mitophagy through the ULK1-Rab9-dependent alternative mitophagy pathway independent of canonical mitophagy pathway regulators such as ATG7 and PINK1. Importantly, ALT001 reverses mitochondrial dysfunction in SH-SY5Y cells expressing mutant APP in a mitophagy-dependent manner. ALT001 induces alternative mitophagy in mice and restores the decreased mitophagy level in a 5×FAD AD model mouse. In addition, ALT001 reverses mitochondrial dysfunction and cognitive defects in the PS2APP and 5×FAD AD mouse models. AAV-mediated silencing of Rab9 in the hippocampus further confirmed that ALT001 exerts its therapeutic effect through alternative mitophagy. Conclusion: Our results highlight the therapeutic potential of ALT001 for AD via alleviation of mitochondrial dysfunction and indicate the usefulness of the ULK1-Rab9 alternative mitophagy pathway as a therapeutic target.

A Study on Breast Augmentation Using Fat Grafting With Stromal Vascular Fraction.

BACKGROUND: Stromal vascular fraction (SVF) and fat grafting are emerging alternatives to implants for breast augmentation. However, the lack of controlled clinical data has led to conflicting results regarding the effectiveness of surgical treatments. This study aimed to identify the key factors affecting the outcomes of fat grafting with SVF and to recognize novel methods to improve the retention rate. PATIENTS AND METHODS: In total, 384 women underwent breast augmentation using fat grafting with SVF. The patients were preoperatively and postoperatively managed and recalled for follow-up at 3, 6, and 18 months. RESULTS: The average volume of the injection in the left breast was 162.35 mL (range, 50-260 mL). The postoperative retention rates were 78.65% in 384 patients at 3 months, 77.17% in 273 patients at 6 months, and 77.48% in 102 patients at 18 months. The retention rates were compared based on the number of SVF cells; patients with more than 60 million cells had a retention rate of 70.77%, and those with less than 60 million cells had a retention rate of 85.60% at 18 months. The retention rates at the 18-month follow-up were 65.62% and 85.09% in stiff and soft breasts, respectively. A higher number of cells in the SVF was associated with a greater retention volume, and the retention volume was greater in patients with soft breasts.Given the higher use of the right arm, after 18 months of the surgery, the retention rate of the right breast (60.35%) was lower than that of the left breast (77.48%) ( P < 0.05; t = -13.199). CONCLUSIONS: Limiting arm movement, increasing the number of cells in the SVF, and improving the skin tension might enhance the retention rate in patients undergoing breast augmentation.

Plasma Circulating Tumor Necrosis Factor Receptors at Multiple Time Points Can Predict the Outcome of Severe Acute Kidney Injury.

INTRODUCTION: This prospective cohort study investigated the clinical role of circulating tumor necrosis factor receptor (cTNFR) levels as prognostic biomarkers in severe acute kidney injury (AKI) patients requiring continuous renal replacement therapy (CRRT). METHODS: We enrolled 136 patients from 7 hospitals participating in the VENUS (VolumE maNagement Under body composition monitoring in critically ill patientS on CRRT) trial from July 2017 to October 2019. The levels of cTNFR1 and cTNFR2 were measured using plasma samples collected on days 0 (D0), 2 (D2), and 7 (D7). Patients were divided into high- and low-cTNFR groups based on their receptor concentrations. RESULTS: D0 concentrations of cTNFR1 and cTNFR2 were positively correlated with one another (R2 = 0.37, p < 0.001). The high-cTNFR1 group displayed a higher in-hospital mortality rate than the low-TNFR1 group (p = 0.002). Moreover, the mortality rate was significantly higher in the high-TNFR1 group than in the low-TNFR1 group after adjusting for age, sex, and acute physiology, and chronic health evaluation II scores (hazard ratio 1.82, 95% confidence interval 1.09-3.03, p = 0.025). D2 and D7 cTNFR1 levels were also associated with in-hospital mortality; contrastingly, cTNFR2 levels were not associated with this outcome. Additionally, patients were divided into three groups according to the change in cTNFR levels from D0 to D2 (ΔcTNFR). Those in the highest ΔcTNFR tertile had a higher mortality rate than the remaining patients (p = 0.033 for ΔcTNFR1; p = 0.025 for ΔcTNFR2). Patients who underwent AKI-to-chronic kidney disease transition had higher concentrations of cTNFR1 (p = 0.014). DISCUSSION/CONCLUSION: Plasma cTNFR1 concentrations at CRRT initiation and changes in cTNFR1 and 2 levels immediately following CRRT initiation are significant biomarkers for predicting the outcomes of patients with severe AKI.

Effect of cutting time and cooking temperature on physicochemical properties of chicken breast meat emulsion sausage with olive oil.

This study aimed to optimize the emulsification of olive oil in chicken sausage production at varying cutting times (30, 45, and 60 s) and cooking temperatures (63, 73, and 83°C). Pork backfat sausages were prepared as controls, using the same variables. The quality attributes of the sausages were analyzed, and the distribution of lipid droplets was identified using confocal laser scanning microscopy. The combinations of cutting time and cooking temperature in olive oil sausages showed different emulsifying characteristics. Meat emulsion with olive oil at a cutting time of 60 s and cooking temperature of 73°C showed the highest emulsion stability with lowest water and lipid loss (2.49%, P < 0.05). The pH values were lower for olive oil samples than for the controls (5.9 vs. 6.2, P < 0.05). Cutting time of 60 s and cooking temperature of 73°C generated higher hardness, gumminess, and chewiness in olive oil sausages (P < 0.05). The replacement of pork backfat with olive oil resulted in a higher b*, C*, and h values, as well as lower lipid oxidation (P < 0.05). In addition, microstructural images exhibited a finer distribution of lipid droplets in olive oil sausages with a cutting time of 60 s. In conclusion, chicken sausage at a cooking temperature of 73°C and cutting time of 60 s was optimal for producing sausages with olive oil. Given the condition, the sausages produced from olive oil had better emulsion and oxidative stability than sausages produced from pork backfat.

p53-mediated regulation of mitochondrial dynamics plays a pivotal role in the senescence of various normal cells as well as cancer cells.

The tumor suppressor p53 is known as a critical mediator of many cellular processes, including cellular senescence, but its role in mitochondrial dynamics is not fully understood. We have previously shown that p53 regulates mitochondrial dynamics via the PKA-Drp1 pathway to induce cellular senescence. In this study, to further understand the role of p53-dependent regulation of mitochondrial dynamics, the effect of p53 expression on mitochondrial morphology was examined in various cancer cell lines and normal human cells. We found that p53 induced remarkable mitochondrial elongation and cellular senescence in various cancer cells regardless of their p53 status. p53 also induced mitochondrial elongation in various human primary normal cells, suggesting that p53-mediated mitochondrial elongation is a general phenomenon. Moreover, we found that p53 plays an essential role in mitochondrial elongation in H-Ras-induced cellular senescence and in the replicative senescence of normal human cells. Treatment with the MDM-2 antagonist Nutlin-3a also induced mitochondrial elongation through the PKA-Drp1 pathway in IMR90 normal human cells. Furthermore, the inhibition of PKA activity in late-passage normal cells significantly reduced both mitochondrial elongation and cellular senescence, suggesting that the p53-PKA pathway is essential for maintaining the senescence phenotype in normal cells. Together, these results further confirm the direct regulation of mitochondrial dynamics by p53 and the important role of p53-mediated mitochondrial elongation in cellular senescence.

PINK1 alleviates thermal hypersensitivity in a paclitaxel-induced Drosophila model of peripheral neuropathy.

Paclitaxel is a representative anticancer drug that induces chemotherapy-induced peripheral neuropathy (CIPN), a common side effect that limits many anticancer chemotherapies. Although PINK1, a key mediator of mitochondrial quality control, has been shown to protect neuronal cells from various toxic treatments, the role of PINK1 in CIPN has not been investigated. Here, we examined the effect of PINK1 expression on CIPN using a recently established paclitaxel-induced peripheral neuropathy model in Drosophila larvae. We found that the class IV dendritic arborization (C4da) sensory neuron-specific expression of PINK1 significantly ameliorated the paclitaxel-induced thermal hyperalgesia phenotype. In contrast, knockdown of PINK1 resulted in an increase in thermal hypersensitivity, suggesting a critical role for PINK1 in sensory neuron-mediated thermal nociceptive sensitivity. Interestingly, analysis of the C4da neuron morphology suggests that PINK1 expression alleviates paclitaxel-induced thermal hypersensitivity by means other than preventing alterations in sensory dendrites in C4da neurons. We found that paclitaxel induces mitochondrial dysfunction in C4da neurons and that PINK1 expression suppressed the paclitaxel-induced increase in mitophagy in C4da neurons. These results suggest that PINK1 mitigates paclitaxel-induced sensory dendrite alterations and restores mitochondrial homeostasis in C4da neurons and that improvement in mitochondrial quality control could be a promising strategy for the treatment of CIPN.

Breast Augmentation by Fat Transplantation With Adipose-Derived Stem/Stromal Cells.

BACKGROUND: Surgical methods using implants were broadly selected for breast augmentation surgery until recently; however, owing to several associated problems, fat transplantation using adipose-derived stem/stromal cells (ADSCs) has been suggested as an alternative. OBJECTIVES: This study evaluated the clinical benefits of fat transplantation using ADSCs for breast augmentation. METHODS: The clinical effects were investigated in 105 patients who underwent breast augmentation with ADSCs and fat transplantation. Liposuction was performed in the abdominal and/or thigh regions; ADSCs were isolated from the fat, mixed with refined fat, and transplanted into each breast; and changes in the breast volume were measured. RESULTS: The average increase in breast volume was approximately 185 mL at 2 weeks after operation. Fat engraftment rates were 85.1, 75.1, and 73.7% of augmented volumes after 1, 3, and 6 months, respectively. A total of 39 patients who received >60 million ADSCs exhibited a transplanted fat engraftment rate of 90.5% (average increase, 162 mL), whereas this rate was 68.9% (average increase, 115 mL) in 31 patients who received <60 million ADSCs. CONCLUSIONS: This study demonstrates that breast augmentation with ADSCs and fat transplantation is effective. Surgical outcomes substantially improved with increased numbers of implanted ADSCs.

Prevalence of radiologic acetabular dysplasia in asymptomatic Asian volunteers.

The purpose of this study was to evaluate the prevalence of acetabular dysplasia in an asymptomatic Asian population as one of the most important risk factors of hip osteoarthritis. From December 2014 to March 2015, we investigated the data of 200 asymptomatic volunteers (400 hips) aged 18-50 years recruited from our institution. Pelvic radiographs were taken and reviewed by two experienced orthopaedic surgeons. Lateral centre-edge (LCE) angle, Sharp angle, Tonnis angle and acetabular depth-to-width ratio (AD/WR) were measured. We investigated the mean values and identified the statistical differences between the sexes and evaluated the prevalence and bilaterality of acetabular dysplasia defined by each parameter. Mean LCE angle, Sharp angle, Tonnis angle and AD/WR were 26.2°, 41.3°, 8.5° and 0.28, respectively. All parameters showed more dysplastic results in females than in males and were statistically significantly different, except for AD/WR. When defined acetabular dysplasia as LCE angle <20°, Sharp angle >45°, Tonnis angle >14° or AD/WR <0.25, the prevalence of acetabular dysplasia by each parameter was 15.0%, 12.8%, 13.3% and 12.8%, respectively. There was a higher prevalence in females than in males; however, only Sharp and Tonnis angles showed significant differences. The bilaterality of acetabular dysplasia was 18.6-39.5% for all subjects. There is high prevalence of asymptomatic dysplastic hips in the Asian population.

The Surgical Choice of Incidental Periampullary Carcinoma Management in Emergency Laparotomy.

BACKGROUND Laparotomy patients are occasionally diagnosed as having incidental periampullary cancers, making emergency pancreaticoduodenectomy (PD) inevitable. In this situation is difficult to decide whether to perform an emergency PD or a two-stage PD. MATERIAL AND METHODS A total of 27 patients who underwent emergency abdominal laparotomy were diagnosed with periampullary or pancreatic cancer during the operation without enough preoperative preparation. Ten patients underwent emergency one-stage PD and 17 patients underwent two-stage PD. Data of 137 patients with elective PD were selected as the control group. The preoperative, operative, and postoperative parameters, including hospital stay, medical cost, blood loss, and postoperative complications between elective PD and emergency PD (one-stage and two-stage) and between one-stage PD and two-stage PD were analyzed by chi-square test, Fisher test, or t test. RESULTS Patients undergoing emergency two-stage PD had less blood loss (P=0.014), while patients with one-stage PD had shorter hospital stay (P=0.004), shorter operation time (P=0.047), and lower treatment costs (P=0.003). Additionally, the complications rates between one-stage and two-stage PD had no significant difference (P=0.365). Elective PD was the optimal method due to shorter hospital stay (P<0.001), less hemorrhage (P<0.001), shorter operative time (P<0.001), and lower cost (P<0.001) compared with emergency PD. CONCLUSIONS Based on our experience, one-stage PD had advantages of shorter hospital stay, shorter operation time, and lower treatment costs, while two-stage PD had less blood loss. The emergency two-stage PD may be more suitable for patients with unstable vital signs if emergency PD is inevitable in an emergency laparotomy.

A sample-to-answer droplet magnetofluidic assay platform for quantitative methylation-specific PCR.

Dysregulation of DNA methylation has been identified as an epigenetic biomarker for numerous cancer types. Gene-specific identification techniques relying on methylation-specific PCR (MSP) require a lengthy manual benchtop process that is susceptible to human-error and contamination. This MSP assay requires a series of discrete sample processing steps including genomic DNA extraction, bisulfite conversion and readout via PCR. In this work, we present a streamlined assay platform utilizing droplet magnetofluidic principles for integration of all sample processing steps required to obtain quantitative MSP signal from raw biological samples. We present a streamlined protocol for solid-phase extraction and bisulfite conversion of genomic DNA, which minimizes reagent use and simplifies the sample preparation protocol for implementation on a compact assay platform. Furthermore, we present a thermally robust assay chip that enables DNA extraction, bisulfite conversion and quantitative PCR from biological samples on a single device. Technical improvements to facilitate DNA extraction and PCR on a single chip in addition to chip performance characterization data are presented.

Lotus (Nelumbo nucifera) Rhizome as an Antioxidant Dietary Fiber in Cooked Sausage: Effects on Physicochemical and Sensory Characteristics.

The objective of this study was to determine the physicochemical and sensory properties of cooked emulsion sausages containing different levels of lotus rhizome powder (0, 1, 2, and 3%, based on total weight). Lotus rhizome powder had no significant (p>0.05) impact on pH, moisture, protein, or ash content of sausage. However, fat content was slightly but significantly (p<0.05) decreased when the level of lotus rhizome powder was increased in the sausages. The addition of lotus rhizome powder to sausages at over 1% resulted in significantly (p<0.05) darker and less red color of cooked sausage compared to control. Increase in lotus rhizome level slightly improved the emulsion stability and apparent viscosity. Significant (p<0.05) reduction in cooking loss was observed when more than 1% of lotus rhizome powder was added to sausages. The textural properties of sausages were unaffected by the inclusion of lotus rhizome except for springiness and chewiness. On the manufacture day, control sausage had significantly (p<0.05) higher TBARS value than treatments. Regarding sensory characteristics, increased levels of lotus rhizome powder decreased (p<0.05) color and juiciness scores. However, cooked sausages exhibited similar overall acceptability regardless of the level of lotus rhizome powder added to sausages. Therefore, lotus rhizome powder, an antioxidant dietary fiber, could be used as an effective natural ingredient in meat products for the development of healthier and functional food.

A parallelized microfluidic DNA bisulfite conversion module for streamlined methylation analysis.

Aberrant methylation of DNA has been identified as an epigenetic biomarker for numerous cancer types. The vast majority of techniques aimed at detecting methylation require bisulfite conversion of the DNA sample prior to analysis, which until now has been a benchtop process. Although microfluidics has potential benefits of simplified operation, sample and reagent economy, and scalability, bisulfite conversion has yet to be implemented in this format. Here, we present a novel droplet microfluidic design that facilitates rapid bisulfite conversion by reducing the necessary processing steps while retaining comparable performance to existing methods. This new format has a reduced overall processing time and is readily scalable for use in high throughput DNA methylation analysis.

Droplet Array Platform for High-Resolution Melt Analysis of DNA Methylation Density.

High-resolution melting (HRM) has garnered significant interest as an analytical technique for a number of applications, including DNA methylation detection, due to its inherent sensitivity and robustness. In this study, we describe a miniaturized assay platform for quantitative methylation density analysis using a microfluidic droplet array cartridge. We demonstrate that the DNA methylation level of the RASSF1A promoter can be directly analyzed using HRM. PCR products were generated by amplifying bisulfite-treated DNA with varying CpG densities using CpG island-flanking primer sets. Subsequent HRM analysis on the miniaturized droplet platform shows distinct melting curve profiles associated with methylation levels, which was verified using a conventional benchtop PCR-HRM system. The characteristic melting temperature (Tm) of the PCR products was used to directly quantify the respective levels of DNA methylation density. Our approach provides a key advantage over current gold standard methods such as methylation-specific PCR (MSP), which are incapable of providing specific information regarding the overall methylation density of the target genes. The miniaturized platform establishes a practical approach to methylation density profiling from multiple DNA samples with a potential application in point-of-care diagnostics.

Topography-assisted electromagnetic platform for blood-to-PCR in a droplet.

This paper presents an electromagnetically actuated platform for automated sample preparation and detection of nucleic acids. The proposed platform integrates nucleic acid extraction using silica-coated magnetic particles with real-time polymerase chain reaction (PCR) on a single cartridge. Extraction of genomic material was automated by manipulating magnetic particles in droplets using a series of planar coil electromagnets assisted by topographical features, enabling efficient fluidic processing over a variety of buffers and reagents. The functionality of the platform was demonstrated by performing nucleic acid extraction from whole blood, followed by real-time PCR detection of KRAS oncogene. Automated sample processing from whole blood to PCR-ready droplet was performed in 15 min. We took a modular approach of decoupling the modules of magnetic manipulation and optical detection from the device itself, enabling a low-complexity cartridge that operates in tandem with simple external instruments.

Evolution of abnormal eye movements in Wernicke's encephalopathy: correlation with serial MRI findings.

A 33-year-old woman with Wernicke's encephalopathy (WE) due to poor oral intake after allogeneic stem cell transplantation for acute myeloid leukemia showed a sequential development of bilateral gaze-evoked nystagmus (GEN), rightward gaze palsy, and upbeat nystagmus. Initial MRIs obtained when she had GEN only showed a lesion involving the medullary tegmentum, and follow-up MRIs revealed additional lesions in the pontine and midbrain tegmentum along with development of rightward gaze palsy, and finally bilateral medial thalamus lesions in association with upbeat nystagmus. The evolution of abnormal ocular motor findings and serial MRI changes in our patient with WE provide imaging evidence on relative vulnerability of the neural structures, and on the progression of lesions and ocular motor findings in thiamine deficiency.

A Case of Action-Induced Clonus that Mimicked Action Tremors and was Associated with Cervical Schwannoma.

Clonus is the rhythmic muscle contraction which usually occurs in patients with lesions involving descending motor pathways. Sometimes, rhythmic oscillation of action induced clonus could be confused to action tremor. We report a case of action induced clonus associated with cervical schwannoma which was misdiagnosed as essential tremor. The patient had spasticity in all limbs with exaggerated tendon reflexes, and passive stretch-induced clonus. Imaging and histological examinations revealed a schwannoma extending from C2 to C7. The lesion was partially removed by surgery. Even though essential tremor is a common disease, clinician have to do sufficient neurologic examination considering differential diagnosis.

Ophthalmoplegic migraine with reversible thalamic ischemia shown by brain SPECT.

Two patients who presented with ophthalmoplegic migraine underwent electroencephalography, brain magnetic resonance imaging, cerebral angiography, and technetium Tc 99m ethyl cysteinate dimer single photon emission computed tomography (SPECT) during an attack. Follow-up SPECT was performed after neurologic symptoms resolved. In both patients, SPECT during an attack of ophthalmoplegia and headache demonstrated significantly decreased regional cerebral blood flow (rCBF) in the thalamus on the side of the ophthalmoplegia; rCBF reverted to normal on a follow-up SPECT during the symptom-free period. These findings suggest reversible ischemia in the territories of perforating branches of the posterior cerebral artery may accompany ophthalmoplegic migraine and possibly bear some relationship to the clinical features.