Changes of antioxidant enzymes in the kidney after cardiac arrest in the rat model

Globally, cardiac arrest (CA) is a leading cause of death and disability. Asphyxial CA (ACA)-induced kidney damage is a crucial factor in reducing the survival rate. The purpose of this study was to investigate the role of antioxidant enzymes in histopathological renal damage in an ACA rat model at different time points. A total of 88 rats were divided into five groups and exposed to ACA except for the sham group. To evaluate glomerular function and oxidative stress, serum levels of blood urea nitrogen (BUN) and creatinine (Crtn) and malondialdehyde (MDA) levels in renal tissues were measured. To determine histopathological damage, hematoxylin and eosin staining, periodic acid-Schiff staining, and Masson's trichrome staining were performed. Expression levels of antioxidant enzymes including superoxide dismutase-1 (SOD-1), superoxide dismutase-2 (SOD-2), catalase (CAT), and glutathione peroxidase (GPx) were measured by immunohistochemistry (IHC). Survival rate of the experimental rats was reduced to 80% at 6 h, 55% at 12 h, 42.9% at 1 day, and 33% at 2 days after return of spontaneous circulation. Levels of BUN, Crtn, and MDA started to increase significantly in the early period of CA induction. Renal histopathological damage increased markedly from 6 h until two days post-CA. Additionally, expression levels of antioxidant enzymes were significantly decreased at 6 h, 12 h, 1 day, and 2 days after CA. CA-induced oxidative stress and decreased levels of antioxidant enzymes (SOD-1, SOD-2, CAT, GPx) from 6 h to two days could be possible mediators of severe renal tissue damage and increased mortality rate.

Germline Cancer Testing in Unselected Patients with Gastric and Esophageal Cancers: A Multi-center Prospective Study.

BACKGROUND AND AIMS: To determine prevalence and clinical utility of pathogenic germline variants (PGV) in gastric and esophageal cancer patients using universal genetic testing approach. METHODS: We undertook a prospective study of germline sequencing using an > 80 gene next-generation sequencing platform among patients with gastric and esophageal cancers receiving care at Mayo Clinic Cancer Center between April 1, 2018, and March 31, 2020. Patients were not selected based on cancer stage, family history of cancer, ethnicity, or age. Family cascade testing was offered at no cost. RESULTS: A total of 96 patients were evaluated. Median age was 66 years, 80.2% were male, 89.6% were white. Nearly 39% of the cohort had esophageal cancer, 35.4% gastric cancer and 26% gastroesophageal junction cancers. Approximately half (52%) of the patients had metastatic disease. Pathogenic germline variants (PGV) were detected in 15.6% (n = 15) patients. The prevalence of PGV was 10.8% in esophageal cancer, 17.6% in gastric cancer and 20% in gastroesophageal cancer. Eighty percent of patients with a positive result would not have been detected by screening with standard guidelines for genetic testing. Most PGV detected included genes with high and moderate penetrance related to DNA damage response including BRCA1, BRCA2, PALB2 and ATM. CONCLUSIONS: Universal multi-gene panel testing in gastric and esophageal cancers was associated with detection of heritable mutations in 15% of patients. The majority of PGV would not be detected with current screening guidelines and are related to DNA damage response.

Increased salivary syndecan-1 level is associated with salivary gland function and inflammation in patients with Sjögren's syndrome.

OBJECTIVE: Syndecan-1 (SDC-1), a transmembrane heparin sulphate proteoglycan predominantly expressed on epithelial cells, also exists in a soluble form through ectodomain shedding. SDC-1 expression and shedding may be modulated in the inflammatory milieu of primary Sjögren's syndrome (SS). We investigated SDC-1 expression in minor salivary glands (MSGs) and analysed the association between salivary or plasma levels of SDC-1 and clinical parameters in SS. METHOD: We measured salivary and plasma SDC-1 levels via an enzyme-linked immunosorbent assay and assessed the salivary flow rates (SFRs) in 70 patients with SS and 35 healthy subjects. Disease activity indices, serological markers, salivary gland scintigraphy, and MSG biopsy were evaluated in patients with SS. RESULTS: SDC-1 expression was upregulated on ductal epithelial cells in inflamed salivary glands. Salivary SDC-1 levels in patients significantly exceeded those in healthy subjects [median (interquartile range) 49.0 (20.7-79.1) vs 3.7 (1.7-6.3) ng/mL, p < 0.001] and inversely correlated with SFRs (r = -0.358, p = 0.032) and ejection fractions of the parotid (r = -0.363, p = 0.027) and submandibular (r = -0.485, p = 0.002) glands in salivary gland scintigraphy. Plasma SDC-1 levels were significantly correlated with the EULAR Sjögren's Syndrome Disease Activity Index (r = 0.507, p < 0.001) and EULAR Sjögren's Syndrome Patient Reported Index (r = 0.267, p = 0.033). Focus scores were correlated with salivary SDC-1 levels (r = 0.551, p = 0.004). CONCLUSIONS: Salivary and plasma SDC-1 levels may constitute potential biomarkers for salivary gland function and disease activity, respectively, in SS.

Recurrence following hemithyroidectomy in patients with low- and intermediate-risk papillary thyroid carcinoma.

BACKGROUND: This study evaluated the incidence, patterns and risk factors for recurrence after hemithyroidectomy in patients with low- and intermediate-risk papillary thyroid carcinoma (PTC), and verified the predictive role of the risk staging systems in current use. METHODS: The clinicopathological characteristics and risk categories were analysed according to recurrence in patients who underwent hemithyroidectomy for low- and intermediate-risk conventional PTC, and were followed up for at least 24 months. Five risk staging systems were used to stratify risk: the 2015 American Thyroid Association (ATA) system; Age, Metastases, Extent and Size (AMES) system; Metastases, Age, Complete resection, Invasion and Size (MACIS) system; Grade, Age, Metastases, Extent and Size (GAMES) system; and the eighth AJCC system. RESULTS: The study included 561 patients; 93·9 per cent of the study population (527 of 561) had a papillary thyroid microcarcinoma 1 cm or smaller in size. At a mean follow-up of 83 months, 25 patients (4·5 per cent) had recurrence; among these patients, 23 (92%) presented with a remaining thyroid lobe. Multifocality was significantly associated with recurrence in univariable and multivariable analyses (adjusted hazard ratio 3·16, 95 per cent c.i. 1·25 to 7·98; P = 0·015). Disease-free survival (DFS) varied according to multifocality (P = 0·010). The five risk staging systems were not associated with recurrence, and their Harrell's C-index ranged from 0·500 to 0·531. DFS rates did not differ between the risk categories in each system. CONCLUSION: Although the recurrence rate after hemithyroidectomy in patients with low- and intermediate-risk PTC was low, meticulous follow-up focusing on the remaining thyroid lobe is needed for early detection and timely management of recurrence. The risk scoring systems in current use have no predictive role in these patients.

ANTECEDENTES: Este estudio evaluó la incidencia, patrones y factores de riesgo de recidiva tras hemitiroidectomía en pacientes con carcinoma papilar de tiroides (papillary thyroid carcinoma, PTC) de riesgo bajo e intermedio y verificó el papel predictivo de los sistemas de estadificación del riesgo utilizados en la actualidad (risk staging systems, RSSs). MÉTODOS: Se analizaron las características clinicopatológicas y las categorías de riesgo en base a la recidiva en 561 pacientes que fueron sometidos a hemitiroidectomía por PTC convencional de riesgo bajo e intermedio y seguidos durante ≥ 24 meses. Para estratificar el riesgo se utilizaron cinco RSSs, incluyendo el sistema de la American Thyroid Association (ATA) de 2015; la edad, las metástasis, la extensión y el tamaño del sistema AMES; las metástasis, la edad, la resección completa, la invasión y el tamaño del sistema GAMES; y la octava edición de la American Joint Committee on Cancer system (AJCC). RESULTADOS: La proporción de la población de estudio con microcarcinoma papilar de tiroides de tamaño ≤ 1 cm fue 93,9% (527/561). A los 83 meses de seguimiento, 25 pacientes (4,5%) presentaron recidiva y entre estos pacientes, 23 (92%) no habían sido sometidos a tiroidectomía total. La multifocalidad se asoció significativamente con la recidiva en los análisis univariado y multivariable con un cociente de riesgos instantáneos (hazard ratio, HR) ajustado de 3,163; i.c. del 95% 1,253-7,983; P = 0,015. La supervivencia libre de enfermedad (disease-free survival, DFS) varió según la multifocalidad (P = 0,010). Los cinco RSSs no se asociaron con la recidiva, y su índice C de Harrell fue 0,500-0,531. Las DFSs no fueron diferentes entre las categorías de riesgo de cada RSS. CONCLUSIÓN: La tasa de recidiva tras hemitiroidectomía en pacientes con PTC de riesgo bajo e intermedio fue baja. Sin embargo, es necesario efectuar un seguimiento meticuloso, centrándose en el lóbulo tiroideo restante, para la detección precoz y el tratamiento oportuno de la recidiva. Los RSSs que se utilizan en la actualidad no tienen valor predictivo en estos pacientes.

In vivo investigation of temporomandibular joint regeneration: development of a mouse model.

Temporomandibular joint (TMJ) reconstruction is traditionally invasive. Several investigators have developed animal models, including mouse models, to study the TMJ. However, there are no detailed descriptions of a mouse model to be followed for additional research. The goal of this project was to study minimally invasive TMJ regeneration using tissue engineering in mice. As part of the project, a detailed mouse model was developed, which is described in this article. Eight carcasses were used to study the anatomy of the TMJ of the mouse and 36 mice were used to describe the surgical approach and perioperative management. The study showed similarities and differences when compared to humans. One mouse died suddenly 10 days postoperatively, while 35 mice survived the operation. Keratitis and wound dehiscence were the most common complications. Investigators reviewing this paper should be able to use this mouse model to further study TMJ regeneration in mice.

Phase 1b investigation of the MEK inhibitor binimetinib in patients with advanced or metastatic biliary tract cancer.

Background The MAPK pathway plays a central role in regulation of several cellular processes, and its dysregulation is a hallmark of biliary tract cancer (BTC). Binimetinib (MEK162), a potent, selective oral MEK1/2 inhibitor, was assessed in patients with advanced BTC. Patients and Methods An expansion cohort study in patients who received ≤1 line of therapy for advanced BTC was conducted after determination of the maximum tolerated dose in this Phase 1 trial. Patients received binimetinib 60 mg twice daily. The primary objectives were to characterize the safety profile and pharmacokinetics of binimetinib in advanced BTC. Secondary objectives included assessment of clinical efficacy, changes in weight and lean body mass, and pharmacodynamic effects. Tumor samples were assessed for mutations in relevant genes. Results Twenty-eight patients received binimetinib. Common adverse events (AEs) were mild, with rash (82%) and nausea (54%) being most common. Two patients experienced grade 4 AEs, one generalized edema and the other pulmonary embolism. The pharmacokinetics in this patient population were consistent with those previously reported (Bendell JC et al., Br J Cancer 2017;116:575-583). Twelve patients (43%) experienced stable disease and two had objective responses (1 complete response, 1 partial response) per Response Evaluation Criteria in Solid Tumors and stable metabolic disease by positron emission tomography/computed tomography. Most patients (18/25; 72%) did not have KRAS, BRAF, NRAS, PI3KCA, or PTEN mutations, nor was there correlation between mutation status and response. The average non-fluid weight gain was 1.3% for lean muscle and 4.7% for adipose tissue. Conclusion Binimetinib was well tolerated and showed promising evidence of activity in patients with BTC. Correlative studies suggested the potential for binimetinib to promote muscle gain in patients with BTC.

Feasibility of a new V-shaped incision for parotidectomy: a preliminary report.

We report the design of a new V-shaped incision for parotidectomy that involves only preauricular and postauricular incisions and no hairline or upper cervical incision. It can be used to approach almost all the superficial parotid region, including the superior and anterior divisions, with minimal scarring. To evaluate its technical feasibility, safety, and cosmetic results, we prospectively enrolled 15 patients (between September 2015 and September 2016) who had partial parotidectomy as the primary treatment for benign parotid tumours. Operations were successfully completed through this approach alone in 14 (mean (range) operating time: 120 (105-142) minutes; drainage volume: 51 (23-70) ml; and duration of drainage: 2.6 (2-4) days). There were no serious complications such as paralysis of the facial nerve or necrosis of the wound. The mean (range) visual analogue scale (VAS) and Vancouver Scar Scale scores for the scars were 9 (8-10) and 0.9 (0-3), respectively. A V-shaped incision for partial parotidectomy is technically feasible and safe, and can produce good cosmetic results in selected patients with benign parotid tumours. Our results need to be confirmed in larger studies and case-control trials.

Antioxidant, angiotensin-converting enzyme inhibitory activity and other functional properties of egg white proteins and their derived peptides - A review.

Egg white contains many functionally important proteins: ovalbumin (54%), ovotransferrin (12%), ovomucoid (11%), ovoglobulin (G2 and G3, 8%), ovomucin (3.5%), and lysozyme (3.5%) are major proteins, while ovoinhibitors, ovomacroglobulin, ovoglycoprotein, ovoflavoprotein, thiamine-binding proteins, and avidin are minor proteins present in egg white. These proteins, as well as the peptides derived from the proteins, have been recognized for their functional importance as antioxidant, antimicrobial, metal-chelating, anti-viral, anti-tumour, and angiotensin-converting enzyme (ACE)-inhibitory activities. Among the functional properties of the peptides, antioxidant and antimicrobial activities are important characteristics for food processing while other properties such as ACE-inhibitory activity of the peptides can have important health-related functionalities. Bioactive peptides can be produced from egg white proteins by enzyme hydrolysis, chemical treatments, or thermal treatments at different pH conditions. The effective functional peptides produced from egg white proteins are usually smaller than 2 kDa in molecular size. However, these peptides are known for their beneficial activities in vitro only, and little work has been done to prove their beneficial effects in vivo. Therefore, further studies are needed to see if the bioactive peptides derived from egg white proteins are helpful for humans in the future.

Effect of oregano oil and tannic acid combinations on the quality and sensory characteristics of cooked chicken meat.

The antioxidant effects of oregano essential oil and tannic acid combinations on ground chicken breast and thigh meats were studied. Six treatments, including: 1) control (none added), 2) 100 ppm oregano essential oil + 5 ppm tannic acid, 3) 100 ppm oregano essential oil + 10 ppm tannic acid, 4) 200 ppm oregano essential oil + 5 ppm tannic acid, 5) 200 ppm oregano essential oil + 10 ppm tannic acid, and 6) 5 ppm butylated hydroxyanisole (BHA) for breast or 14 ppm for thigh meat, were prepared. Cooked meat samples were individually vacuum-packaged in oxygen-impermeable vacuum bags and then cooked in-bag to an internal temperature of 75°C. After cooling to room temperature, the cooked meat was re-packaged in new oxygen-permeable bags and stored at 4°C for 7 days. Cooked ground chicken meats were analyzed for lipid and protein oxidation and volatiles at 0, 3, and 7 d of storage. The significant differences among the treatments were very clear in cooked meat samples: Thigh meat patties showed higher 2-thiobarbituric acid reactive substances (TBARS), total carbonyl, and volatiles content compared to the breast meat during storage. A combination of 200 ppm oregano oil with 10 ppm tannic acid showed the most significant effects (P < 0.05) on TBARS, total carbonyl, and off-odor volatile formation for both breast and thigh meats. Oregano oil (200 ppm) and 10 ppm tannic acid combination also showed positive effects on the sensory scores of chicken thigh meat. In conclusion, the combination of 200 ppm oregano oil and 10 ppm tannic acid could be a good replacement for the synthetic antioxidants in ground cooked chicken meat.

Immune-enhancing activity of phosvitin by stimulating the production of pro-inflammatory mediator.

Egg yolk phosvitin is one of the most phosphorylated proteins in nature, and the extraordinarily high concentration of phosphate groups in its structure provides a strong metal-binding ability. Phosvitin is known to possess various functional activities, including metal-chelating, antioxidant, emulsifying, antimicrobial, and cytotoxic activities. However, little is known about the immune-enhancing activity of phosvitin. The objective of this study was to evaluate the immune-enhancing activity of phosvitin in murine RAW 264.7 macrophages. Griess reagents and quantitative real-time PCR were used to determine the effect of phosvitin (at 12.5, 25, 50, and 100 µg/mL) on the levels of pro-inflammatory mediators NO and inducible nitric oxide synthase (iNOS), cytokines TNF-α and IL-1ß in RAW 264.7 macrophages. The effect of phosvitin on the phagocytic activity of RAW 264.7 macrophages was also measured using the Neutral-Red Uptake method. Lipopolysaccharides was used as a positive control. Phosvitin significantly (P < 0.05) increased the production of NO in RAW 264.7 macrophages in a dose-dependent manner, but did not show any cytotoxicity. The amounts of NO produced were 3.47, 7.12, 10.23, and 14.57 µM in 12.5 to 100 µg/mL range of phosvitin (control: 0.46 µM). Compared with the untreated group, phosvitin treatment at a 100 µg/mL level increased the production of NO by 31.67 times. Phosvitin also significantly increased the mRNA expression of the RAW 264.7 macrophages: 100 µg/mL of phosvitin treatment increased the expression of mRNA for iNOS, TNF-α, and IL-1ß by 46.25, 9.09, and 85.18 times of the control, respectively. The phagocytic activity of RAW 264.7 macrophages was also increased significantly by phosvitin treatment. These results demonstrated that phosvitin dramatically improved the immune functions RAW 264.7 macrophages by enhancing the production of immune mediators and increasing phagocytic activity. Therefore, phosvitin has a potential to be used as an immune-enhancing agent by food or nutraceutical industries.

In vitro cytotoxic and ACE-inhibitory activities of promod 278P hydrolysate of ovotransferrin from chicken egg white.

Peptides released from egg proteins via enzymatic hydrolysis show various bioactivities such as antimicrobial, antioxidant, antihypertensive, and immunomodulatory properties. The objective of this study was to investigate the cytotoxic and Angiotensin Converting Enzyme (ACE)-inhibitory activities of ovotransferrin and its promod 278P enzyme hydrolysate. Ovotransferrin from egg white was hydrolyzed using promod 278P at 45°C for 3 hours. Using the MTT assay, the cytotoxicity of ovotransferrin and promod 278P hydrolysate of ovotransferrin were evaluated in human cancer cell lines of various tissue origins. The ACE-inhibitory activity was determined using the cleavage of a chromogenic substrate -Hip-His-Leu. The promod 278P hydrolysate of ovotransferrin showed a potent cytotoxicity (>90%) at 20 mg/mL in all cancer cell lines tested, but ovotransferrin did not. The IC50 value of the promod 278P hydrolysate of ovotransferrin against 5 different cancer cells were 10.05 ± 1.55, 3.45 ± 0.94, 4.43 ± 1.87, 4.92 ± 0.63, and 10.43 ± 3.91 mg/mL for MCF-7, HeLa, HepG2, HT-29, and LoVo cells, respectively. The promod 278P hydrolysate of ovotransferrin showed a strong ACE-inhibiting activity: at 10 mg/mL level, the hydrolysate showed 76.82 ± 1.28% inhibition to ACE-inhibitory activity, and 73.33 ± 2.56%, 56.85 ± 1.84%, 50.32 ± 3.71%, 17.30 ± 0.13%, and 4.52 ± 6.83% inhibitory activity at 5, 2.5, 1.25, 0.625, and 0.3125 mg/mL level, respectively. The IC50 value of the promod 278P hydrolysate of ovotransferrin was 1.53 ± 0.20 mg/mL. However, ovotransferrin did not show any inhibitory effect to angiotensin-converting enzyme activity. This result indicated that the promod 278P hydrolysate of ovotransferrin has a great potential as an anticancer and antihypertension agent for humans, but the information on the peptides responsible for the functional activities is not available yet.

Antinociceptive Effects of Botulinum Toxin Type A on Trigeminal Neuropathic Pain.

Previous studies have demonstrated that botulinum toxin type A (BoNT-A) attenuates orofacial nociception. However, there has been no evidence of the participation of the voltage-gated sodium channels (Navs) in the antinociceptive mechanisms of BoNT-A. This study investigated the cellular mechanisms underlying the antinociceptive effects of BoNT-A in a male Sprague-Dawley rat model of trigeminal neuropathic pain produced by malpositioned dental implants. The left mandibular second molar was extracted under anesthesia, followed by a miniature dental implant placement to induce injury to the inferior alveolar nerve. Mechanical allodynia was monitored after subcutaneous injection of BoNT-A at 3, 7, or 12 d after malpositioned dental implant surgery. Subcutaneous injections of 1 or 3 U/kg of BoNT-A on postoperative day 3 significantly attenuated mechanical allodynia, although 0.3 U/kg of BoNT-A did not affect the air-puff threshold. A single injection of 3 U/kg of BoNT-A produced prolonged antiallodynic effects over the entire experimental period. Treatment with BoNT-A on postoperative days 7 and 12, when pain had already been established, also produced prolonged antiallodynic effects. Double treatments with 1 U/kg of BoNT-A produced prolonged, more antiallodynic effects as compared with single treatments. Subcutaneous administration of 3 U/kg of BoNT-A significantly inhibited the upregulation of Nav isoform 1.7 (Nav1.7) expression in the trigeminal ganglion in the nerve-injured animals. These results suggest that antinociceptive effects of BoNT-A are mediated by an inhibition of upregulated Nav1.7 expression in the trigeminal ganglion. BoNT-A is therefore a potential new therapeutic agent for chronic pain control, including neuropathic pain.

Modified irinotecan and infusional 5-fluorouracil (mFOLFIRI) in patients with refractory advanced pancreas cancer (APC): a single-institution experience.

Pancreatic adenocarcinoma is the fourth leading cause of cancer death. Recently, MM-398 (nanoliposomal irinotecan) was shown to be associated with significant improvement in outcome measures with acceptable toxicities when combined with 5-fluorouracil (5-FU)/leucovorin (LV) compared to 5-FU/LV alone in patients failing one line of gemcitabine-based therapy. There is a paucity of data evaluating the role of irinotecan in combination with 5FU in advanced pancreas cancer (APC). We performed a retrospective analysis of all patients who received mFOLFIRI (minus bolus 5FU and LV). All patients with metastatic disease who had failed at least one line of gemcitabine-based therapy prior to receiving mFOLFIRI were included in this study. Descriptive statistics were used to assess the continuous variables and adverse events (AEs), and Kaplan-Meier methods were used to calculate the median progression-free survival (PFS) and overall survival (OS). Forty patients were included in this analysis. Patients received 1-5 lines of prior therapy (25 % with more than 3 lines of prior therapy). The mean age at diagnosis was 60, and 98 % had ECOG of 1. The mean CA 19-9 at the start of therapy was 33,169 U/ml. The median PFS was 2.59 months [95 % confidence interval (CI) (1.90, 3.54)], and OS was 4.75 months [95 % CI (3.14, 8.98)]. The most common AEs included fatigue (98 %), neuropathy (83 %), anorexia (68 %), nausea (60 %) and constipation (55 %). Grade 3 toxicities included fatigue (13 %) and rash (3 %). There were no observed grade 4 toxicities. In this single-institution retrospective analysis, mFOLFIRI was found to be both tolerable and relatively effective in a heavily pretreated patient population with APC. Future prospective studies should consider evaluating the role of mFOLFIRI in refractory APC.

The NAD(+) salvage pathway modulates cancer cell viability via p73.

The involvement of the nicotinamide adenine dinucleotide (NAD(+)) salvage pathway in cancer cell survival is poorly understood. Here we show that the NAD(+) salvage pathway modulates cancer cell survival through the rarely mutated tumour suppressor p73. Our data show that pharmacological inhibition or knockdown of nicotinamide phosphoribosyltransferase (NAMPT), a rate-limiting enzyme in the NAD(+) salvage pathway, enhances autophagy and decreases survival of cancer cells in a p53-independent manner. Such NAMPT inhibition stabilizes p73 independently of p53 through increased acetylation and decreased ubiquitination, resulting in enhanced autophagy and cell death. These effects of NAMPT inhibition can be effectively reversed using nicotinamide mononucleotide (NMN), the enzymatic product of NAMPT. Similarly, knockdown of p73 also decreases NAMPT inhibition-induced autophagy and cell death, whereas overexpression of p73 alone enhances these effects. We show that the breast cancer cell lines (MCF-7, MDA-MB-231 and MDA-MB-468) harbour significantly higher levels of NAMPT and lower levels of p73 than does the normal cell line (MCF-10A), and that NAMPT inhibition is cytotoxic exclusively to the cancer cells. Furthermore, data from 176 breast cancer patients demonstrate that higher levels of NAMPT and lower levels of p73 correlate with poorer patient survival, and that high-grade tumours have significantly higher NAMPT/p73 mRNA ratios. Therefore, the inverse relationship between NAMPT and p73 demonstrable in vitro is also reflected from the clinical data. Taken together, our studies reveal a new NAMPT-p73 nexus that likely has important implications for cancer diagnosis, prognosis and treatment.

Enzymatic hydrolysis of ovomucin and the functional and structural characteristics of peptides in the hydrolysates.

Ovomucin was hydrolyzed using enzymes or by heating under alkaline conditions (pH 12.0), and the functional, structural and compositional characteristics of the peptides in the hydrolysates were determined. Among the treatments, heating at 100 °C for 15 min under alkaline conditions (OM) produced peptides with the highest iron-binding and antioxidant capacities. Ovomucin hydrolyzed with papain (OMPa) or alcalase (OMAl) produced peptides with high ACE-inhibitory activity. The mass spectrometry analysis indicated that most of the peptides from OMPa were <2 kDa, but peptides from OMTr and OM were >2 kDa. OMAl hydrolyzed ovomucin almost completely and no peptides within 700-5000 Da were found in the hydrolasate. The results indicated that the number and size of peptides were closely related to the functionality of the hydrolysates. Considering the time, cost and activities of the hydrolysates, OM was the best treatment for hydrolyzing ovomucin to produce functional peptides.

Oncolytic reovirus induces intracellular redistribution of Ras to promote apoptosis and progeny virus release.

Reovirus is a naturally oncolytic virus that preferentially replicates in Ras-transformed cells and is currently undergoing clinical trials as a cancer therapeutic. Ras transformation promotes reovirus oncolysis by enhancing virion disassembly during entry, viral progeny production, and virus release through apoptosis; however, the mechanism behind the latter is not well understood. Here, we show that reovirus alters the intracellular location of oncogenic Ras to induce apoptosis of H-RasV12-transformed fibroblasts. Reovirus infection decreases Ras palmitoylation levels and causes accumulation of Ras in the Golgi through Golgi fragmentation. With the Golgi being the site of Ras palmitoylation, treatment of target cells with the palmitoylation inhibitor, 2-bromopalmitate (2BP), prompts a greater accumulation of H-RasV12 in the Golgi, and a dose-dependent increase in progeny virus release and subsequent spread. Conversely, tethering H-RasV12 to the plasma membrane (thereby preventing its movement to the Golgi) allows for efficient virus production, but results in basal levels of reovirus-induced cell death. Analysis of Ras downstream signaling reveals that cells expressing cycling H-RasV12 have elevated levels of phosphorylated JNK (c-Jun N-terminal kinase), and that Ras retained at the Golgi body by 2BP increases activation of the MEKK1/MKK4/JNK signaling pathway to promote cell death. Collectively, our data suggest that reovirus induces Golgi fragmentation of target cells, and the subsequent accumulation of oncogenic Ras in the Golgi body initiates apoptotic signaling events required for virus release and spread.

Influence of soy oil source and dietary supplementation of vitamins E and C on the oxidation status of serum and egg yolk, and the lipid profile of egg yolk.

An experiment was conducted to determine the effects of adding vitamins E and C to diets containing 3.5% refined soy oil (SO), recycled soy oil (RSO), or acidulated soy oil soapstocks (ASS) on 1) fatty acid (FA) profile, and cholesterol, triglyceride (TG) and α-tocopherol (α-T) concentrations of yolk, and 2) the oxidation status of serum and yolk. Twelve dietary treatments, using 3 oil sources, 2 levels of vitamin E (0 vs. 250 mg/kg), and 2 levels of vitamin C (0 vs. 250 mg/kg), were prepared. A total of 300 W36 Hy-line laying hens, from 44 to 56 weeks of age, were placed in 60 cages (5 birds/cage) and 5 cages were randomly assigned to one of the 12 diets. Blood samples and eggs were collected after 84 d on trial. No interactions among main effects were found for any of the traits studied. Oil sources had little effects on the FA profile of the yolk, except for C18:3 that was higher (P-value of < 0.01) in the hens fed SO than those fed RSO or ASS. Vitamin E supplementation significantly (P-value of < 0.05) increased the concentration of C16:0, C18:0, and C16:1 but decreased that of C18:2 and C22:6n3 in the yolk. Vitamin C supplementation significantly (P-value of < 0.05) increased C18:0 and C18:3 concentrations in the yolk but decreased the n6 to n3 FA ratio. The concentrations of cholesterol and triglyceride in serum and yolk were not affected by dietary treatment but α-tocopherol concentration increased (P-value of < 0.01) by the dietary vitamin E. Compared with the hens fed the SO diets, malondialdehyde (MDA) concentration in serum was higher with RSO diet but lower with ASS diet. Vitamin E and vitamin C supplementation decreased (P-value of < 0.05) serum MDA. Yolk FA profile was affected not only by the FA profile of the oil source used in diet, but also by the supplementation of vitamin E and C. The results showed that triglyceride profile, but not cholesterol content, of egg was affected by fatty acid profile of the supplemental oil and the vitamin C and E supplementations.

Enzymatic hydrolysis of ovomucoid and the functional properties of its hydrolysates.

Ovomucoid is well known as a "trypsin inhibitor" and is considered to be the main food allergen in egg. However, the negative functions of ovomucoid can be eliminated if the protein is cut into small peptides. The objectives of this study were to hydrolyze ovomucoid using various enzyme combinations, and compare the functional properties of the hydrolysates. Purified ovomucoid was dissolved in distilled water (20 mg/mL) and treated with 1% of pepsin, α-chymotrypsin, papain, and alcalase, singly or in combinations. Sodium sodium dodecyl sulfate-polyacrylamide (SDS-PAGE) results of the hydrolysates indicated that pepsin (OMP), alcalase (OMAl), alcalase+trypsin (OMAlTr), and alcalase+papain (OMAlPa) treatments best hydrolyzed the ovomucoid, and the 4 treatments were selected to determine their functional characteristics. Among the 4 enzyme treatments, hydrolysate from OMAlTr showed the highest iron-chelating and antioxidant activities, while OMP showed higher ACE-inhibitory activity, but lower Fe-chelating activity than the other treatments. However, no difference in the copper-chelating activity among the treatments was found. MS/MS analysis identified numerous peptides from the hydrolysates of OMAlPa and OMAlTr, and majority of the peptides produced were <2 kDa. Pepsin treatment (OMP), however, hydrolyzed ovomucoid almost completely and produced only amino acid monomers, di- and tri-peptides. The ACE-inhibitory, antioxidant and iron-chelating activities of the enzyme hydrolysates were not consistent with the number and size of peptides in the hydrolysates, but we do not have information about the quantity of each peptide present in the hydrolysates at this point.

Enzymatic hydrolysis of ovalbumin and the functional properties of the hydrolysates.

Ovalbumin is the predominant protein in egg white and is widely used in cell culture. However, it also can be used to produce peptides with various functional properties. The objectives of this study were to hydrolyze ovalbumin using various enzyme, incubation time, and temperature combinations, and to compare the functional properties of the hydrolysates. Ovalbumin (20 mg/mL) was hydrolyzed with 1% of pepsin, trypsin, α-chymotrypsin, papain, and alcalase, singly or in combination at 37°C, and then the enzymes were inactivated at 100°C for 15 min. Hydrolyzing ovalbumin with pepsin (OAPe), pepsin + papain (OAPePa), pepsin + alcalase (OAPeAl), alcalase + trypsin (OAAlTr), and α-chymotrypsin (OACh) was also effective in producing peptides from ovalbumin, and the peptides produced had strong iron- and copper-binding capacities and antioxidant capability. However, the best treatment of all was the OAAlTr treatment, which showed the highest iron-chelating and antioxidant activities among the enzyme treatments (P < 0.05). Electrospray-ionization mass spectrometry (MS/MS) analysis identified numerous peptides (<5 kDa) from the OAPe, OAPeAl, OACh, OAAlTr, and OAPePa hydrolysates of ovalbumin, but the number and size of peptides varied widely depending on the treatments. The enzymatic hydrolysis significantly increased the functionality of ovalbumin, and the improvement depended upon the composition of peptides produced rather than the number of the peptides produced.