Tumor hypoxia represses γδ T cell-mediated antitumor immunity against brain tumors.

The anatomic location and immunologic characteristics of brain tumors result in strong lymphocyte suppression. Consequently, conventional immunotherapies targeting CD8 T cells are ineffective against brain tumors. Tumor cells escape immunosurveillance by various mechanisms and tumor cell metabolism can affect the metabolic states and functions of tumor-infiltrating lymphocytes. Here, we discovered that brain tumor cells had a particularly high demand for oxygen, which affected γδ T cell-mediated antitumor immune responses but not those of conventional T cells. Specifically, tumor hypoxia activated the γδ T cell protein kinase A pathway at a transcriptional level, resulting in repression of the activatory receptor NKG2D. Alleviating tumor hypoxia reinvigorated NKG2D expression and the antitumor function of γδ T cells. These results reveal a hypoxia-mediated mechanism through which brain tumors and γδ T cells interact and emphasize the importance of γδ T cells for antitumor immunity against brain tumors.

Exosomal miR-205-5p Improves Endometrial Receptivity by Upregulating E-Cadherin Expression through ZEB1 Inhibition.

Endometrial receptivity is a complex process that prepares the uterine endometrium for embryo implantation; insufficient endometrial receptivity is one of the causes of implantation failure. Here, we analyzed the microRNA expression profiles of exosomes derived from both receptive (RL95-2) and non-receptive (AN3-CA) endometrial epithelial cell (EEC) lines to identify exosomal miRNAs closely linked to endometrial receptivity. Among the 466 differentially expressed miRNAs, miR-205-5p was the most highly expressed in exosomes secreted from receptive RL95-2 cells. miR-205-5p, enriched at the adhesive junction, was closely related to endometrial receptivity. ZEB1, a transcriptional repressor of E-cadherin associated with endometrial receptivity, was identified as a direct target of miR-205-5p. miR-205-5p expression was significantly lower in the endometrial tissues of infertile women than in that of non-infertile women. In vivo, miR-205-5p expression was upregulated in the post-ovulatory phase, and its inhibitor reduced embryo implantation. Furthermore, administration of genetically modified exosomes overexpressing miR-205-5p mimics upregulated E-cadherin expression by targeting ZEB1 and improved spheroid attachment of non-receptive AN3-CA cells. These results suggest that the miR-205-5p/ZEB1/E-cadherin axis plays an important role in regulating endometrial receptivity. Thus, the use of exosomes harboring miR-205-5p mimics can be considered a potential therapeutic approach for improving embryo implantation.

Ceria-Zirconia nanoparticles reduce intracellular globotriaosylceramide accumulation and attenuate kidney injury by enhancing the autophagy flux in cellular and animal models of Fabry disease.

BACKGROUND: Fabry disease (FD) is a lysosome storage disease (LSD) characterized by significantly reduced intracellular autophagy function. This contributes to the progression of intracellular pathologic signaling and can lead to organ injury. Phospholipid-polyethyleneglycol-capped Ceria-Zirconia antioxidant nanoparticles (PEG-CZNPs) have been reported to enhance autophagy flux. We analyzed whether they suppress globotriaosylceramide (Gb3) accumulation by enhancing autophagy flux and thereby attenuate kidney injury in both cellular and animal models of FD. RESULTS: Gb3 was significantly increased in cultured human renal proximal tubular epithelial cells (HK-2) and human podocytes following the siRNA silencing of α galactosidase A (α-GLA). PEG-CZNPs effectively reduced the intracellular accumulation of Gb3 in both cell models of FD and improved both intracellular inflammation and apoptosis in the HK-2 cell model of FD. Moreover these particles attenuated pro fibrotic cytokines in the human podocyte model of FD. This effect was revealed through an improvement of the intracellular autophagy flux function and a reduction in reactive oxygen species (ROS). An FD animal model was generated in which 4-week-old male B6;129-Glatm1Kul/J mice were treated for 8 weeks with 10 mg/kg of PEG-CZNPs (twice weekly via intraperitoneal injection). Gb3 levels were reduced in the kidney tissues of these animals, and their podocyte characteristics and autophagy flux functions were preserved. CONCLUSIONS: PEG-CZNPs alleviate FD associated kidney injury by enhancing autophagy function and thus provide a foundation for the development of new drugs to treat of storage disease.

The miR-182-5p/NDRG1 Axis Controls Endometrial Receptivity through the NF-κB/ZEB1/E-Cadherin Pathway.

Endometrial receptivity is essential for successful pregnancy, and its impairment is a major cause of embryo-implantation failure. MicroRNAs (miRNAs) that regulate epigenetic modifications have been associated with endometrial receptivity. However, the molecular mechanisms whereby miRNAs regulate endometrial receptivity remain unclear. Therefore, we investigated whether miR-182 and its potential targets influence trophoblast cell attachment. miR-182 was expressed at lower levels in the secretory phase than in the proliferative phase of endometrium tissues from fertile donors. However, miR-182 expression was upregulated during the secretory phase in infertile women. Transfecting a synthetic miR-182-5p mimic decreased spheroid attachment of human JAr choriocarcinoma cells and E-cadherin expression (which is important for endometrial receptivity). miR-182-5p also downregulated N-Myc downstream regulated 1 (NDRG1), which was studied further. NDRG1 was upregulated in the secretory phase of the endometrium tissues and induced E-cadherin expression through the nuclear factor-κΒ (NF-κΒ)/zinc finger E-box binding homeobox 1 (ZEB1) signaling pathway. NDRG1-overexpressing or -depleted cells showed altered attachment rates of JAr spheroids. Collectively, our findings indicate that miR-182-5p-mediated NDRG1 downregulation impaired embryo implantation by upregulating the NF-κΒ/ZEB1/E-cadherin pathway. Hence, miR-182-5p is a potential biomarker for negative selection in endometrial receptivity and a therapeutic target for successful embryo implantation.

Aconitate Decarboxylase 1 Deficiency Exacerbates Mouse Colitis Induced by Dextran Sodium Sulfate.

Ulcerative colitis is a complex inflammatory bowel disorder disease that can induce rectal and colonic dysfunction. Although the prevalence of IBD in Western countries is almost 0.5% of the general population, genetic causes are still not fully understood. In a recent discovery, itaconate was found to function as an immune-modulating metabolite in mammalian immune cells, wherein it is synthesized as an antimicrobial compound from the citric acid cycle intermediate cis-aconitic acid. However, the association between the Acod1 (Aconitate decarboxylase 1)-itaconate axis and ulcerative colitis has rarely been studied. To elucidate this, we established a DSS-induced colitis model with Acod1-deficient mice and then measured the mouse body weights, colon lengths, histological changes, and cytokines/chemokines in the colon. We first confirmed the upregulation of Acod1 RNA and protein expression levels in DSS-induced colitis. Then, we found that colitis symptoms, including weight loss, the disease activity index, and colon shortening, were worsened by the depletion of Acod1. In addition, the extent of intestinal epithelial barrier breakdown, the extent of immune cell infiltration, and the expression of proinflammatory cytokines and chemokines in Acod1-deficient mice were higher than those in wild-type mice. Finally, we confirmed that 4-octyl itaconate (4-OI) alleviated DSS-induced colitis in Acod1-deficient mice and decreased the expression of inflammatory cytokines and chemokines. To our knowledge, this study is the first to elucidate the role of the Acod1-itaconate axis in colitis. Our data clearly showed that Acod1 deletion resulted in severe DSS-induced colitis and substantial increases in inflammatory cytokine and chemokine levels. Our results suggest that Acod1 may normally play an important regulatory role in the pathogenesis of colitis, demonstrating the potential for novel therapies using 4-OI.

Comparison of Macrophage Immune Responses and Metabolic Reprogramming in Smooth and Rough Variant Infections of Mycobacterium mucogenicum.

Mycobacterium mucogenicum (Mmuc), a rapidly growing nontuberculous mycobacterium (NTM), can infect humans (posttraumatic wound infections and catheter-related sepsis). Similar to other NTM species, Mmuc exhibits colony morphologies of rough (Mmuc-R) and smooth (Mmuc-S) types. Although there are several case reports on Mmuc infection, no experimental evidence supports that the R-type is more virulent. In addition, the immune response and metabolic reprogramming of Mmuc have not been studied on the basis of morphological characteristics. Thus, a standard ATCC Mmuc strain and two clinical strains were analyzed, and macrophages were generated from mouse bone marrow. Cytokines and cell death were measured by ELISA and FACS, respectively. Mitochondrial respiration and glycolytic changes were measured by XF seahorse. Higher numbers of intracellular bacteria were found in Mmuc-R-infected macrophages than in Mmuc-S-infected macrophages. Additionally, Mmuc-R induced higher levels of the cytokines TNF-α, IL-6, IL-12p40, and IL-10 and induced more BMDM necrotic death. Furthermore, our metabolic data showed marked glycolytic and respiratory differences between the control and each type of Mmuc infection, and changes in these parameters significantly promoted glucose metabolism, extracellular acidification, and oxygen consumption in BMDMs. In conclusion, at least in the strains we tested, Mmuc-R is more virulent, induces a stronger immune response, and shifts bioenergetic metabolism more extensively than the S-type. This study is the first to report differential immune responses and metabolic reprogramming after Mmuc infection and might provide a fundamental basis for additional studies on Mmuc pathogenesis.

Quantitative proteomic profiling of Cervicovaginal fluid from pregnant women with term and preterm birth.

BACKGROUND: Preterm birth (PTB) is one of major causes of perinatal mortality and neonatal morbidity, but knowledge of its complex etiology is still limited. Here we present cervicovaginal fluid (CVF) protein profiles of pregnant women who subsequently delivered at spontaneous preterm or term, aiming to identify differentially expressed CVF proteins in PTB and term birth. METHODS: The CVF proteome of women who sequentially delivered at preterm and term was analyzed using isobaric tags for relative and absolute quantitation (iTRAQ) coupled with two-dimensional nanoflow liquid chromatography-tandem mass spectrometry (2D-nLC-MS/MS). We compared the CVF proteome of PTB (n = 5) and control subjects (term birth, n = 7) using pooled control CVF (term birth, n = 20) as spike-in standard. RESULTS: We identified 1294 CVF proteins, of which 605 were newly identified proteins. Of 990 proteins quantified in both PTB and term birth, 52 proteins were significantly up/down-regulated in PTB compared to term birth. The differentially expressed proteins were functionally associated to immune response, endopeptidase inhibitors and structural constituent of cytoskeleton. Finally, we confirm the down-regulation of SERPINB7 (a serine-type protease inhibitor) in PTB compared to control by Western blot. CONCLUSIONS: Taken together, our study provide quantitative CVF proteome profiles of pregnant women who ultimately delivered at preterm and term. These promising results could help to improve the understanding of PTB etiology and to discover biomarkers for asymptomatic PTB.

Tendril extract of Cucurbita moschata suppresses NLRP3 inflammasome activation in murine macrophages and human trophoblast cells.

Inflammation is the root cause of many diseases that pose a serious threat to human health. Excessive inflammation can also result in preterm birth or miscarriage in pregnant women. Pumpkin (Cucurbita moschata Duchesne, CMD) is a well-known traditional health food and medicinal herb used in many countries to treat diabetes, obesity, osteoporosis, cancer and other diseases. In this study, we investigated the effects of hot water extract derived from the tendrils of C. moschata Duchesne (TCMD) on NLRP3 inflammasome activation in murine macrophages and human trophoblast cells. The TCMD treatment of LPS-primed bone marrow-derived macrophages (BMDMs) and human trophoblast cells attenuated NLRP3 inflammasome activation induced by inflammasome activators such as ATP, nigericin, and monosodium urate (MSU). TCMD treatment suppressed IL-1ß secretion in a dose-dependent manner, without affecting IL-6 secretion. In addition, TCMD inhibited NLRP3-dependent pyroptosis in BMDMs. TCMD also suppressed the release of mature IL-1ß and activation of cleaved-caspase-1 via limited ASC oligomerization. Furthermore, TCMD significantly inhibited IL-1ß secretion and pyroptotic cell death in human trophoblast cells. These results suggest that TCMD exhibits anti-inflammatory effects mediated via inhibition of NLRP3 inflammasome activation suggesting therapeutic potential against inflammatory diseases, preterm birth, and miscarriage.

Verbascoside-Rich Abeliophyllum distichum Nakai Leaf Extracts Prevent LPS-Induced Preterm Birth Through Inhibiting the Expression of Proinflammatory Cytokines from Macrophages and the Cell Death of Trophoblasts Induced by TNF-α.

Background: Preterm birth is a known leading cause of neonatal mortality and morbidity. The underlying causes of pregnancy-associated complications are numerous, but infection and inflammation are the essential high-risk factors. However, there are no safe and effective preventive drugs that can be applied to pregnant women. Objective: The objectives of the study were to investigate a natural product, Abeliophyllum distichum leaf (ADL) extract, to examine the possibility of preventing preterm birth caused by inflammation. Methods: We used a mouse preterm birth model by intraperitoneally injecting lipopolysaccharides (LPS). ELISA, Western blot, real-time PCR and immunofluorescence staining analyses were performed to confirm the anti-inflammatory efficacy and related mechanisms of the ADL extracts. Cytotoxicity and cell death were measured using Cell Counting Kit-8 (CCK-8) analysis and flow cytometer. Results: A daily administration of ADL extract significantly reduced preterm birth, fetal loss, and fetal growth restriction after an intraperitoneal injection of LPS in mice. The ADL extract prevented the LPS-induced expression of TNF-α in maternal serum and amniotic fluid and attenuated the LPS-induced upregulation of placental proinflammatory genes, including IL-1ß, IL-6, IL-12p40, and TNF-α and the chemokine gene CXCL-1, CCL-2, CCL3, and CCL-4. LPS-treated THP-1 cell-conditioned medium accelerated trophoblast cell death, and TNF-α played an essential role in this effect. The ADL extract reduced LPS-treated THP-1 cell-conditioned medium-induced trophoblast cell death by inhibiting MAPKs and the NF-κB pathway in macrophages. ADL extract prevented exogenous TNF-α-induced increased trophoblast cell death and decreased cell viability. Conclusions: We have demonstrated that the inhibition of LPS-induced inflammation by ADL extract can prevent preterm birth, fetal loss, and fetal growth restriction.

Quality comparison of retorted Samgyetang made from white semi-broilers, commercial broilers, Korean native chickens, and old laying hens.

OBJECTIVE: The aim of this study was to compare the quality characteristics of retorted Samgyetang (Korean ginseng chicken soup) made from white semi-broilers (WSB; Ross× Hyline white, 3 weeks old), commercial broilers (CB; Ross, 4 weeks old), Korean native chickens (KNC; Hanhyup-3-ho, 12 weeks old) and old laying hens (OLH; Hyline white, 72 weeks old) and to explore the possibility of using the carcasses of KNCs and OLHs as raw material for product diversification. METHODS: Raw and cooked meat quality, fatty acid composition and consumer acceptance were analyzed. RESULTS: Among the chicken breeds, OLH and KNC showed a higher shear force value than WSB and CB due to high insoluble collagen contents. However, the meat of KNC was more tender than that of OLH. The meat of OLH was characterized by the lowest moisture content and highest crude fat content. The meat of KNC was characterized by a higher proportion of saturated fatty acids, α-linolenic acid and arachidonic acid than that of OLH. The meat of OLH showed the highest content of unsaturated fatty acid, particularly linoleic acid, in its thigh meat. Electronic nose readings revealed that the meat aroma pattern was clearly different across breeds. OLH had the lowest overall acceptance score, while no differences were found in flavor, texture, juiciness and appearance among WSB, CB, and KNC. CONCLUSION: KNC shows potential as raw material for Samgyetang, while additional preprocessing methods, such as tenderization and fat removal, are required for the utilization of OLH as raw material for retorted Samgyetang.

The effect of finishing diet supplemented with methionine/lysine and methionine/α-tocopherol on performance, carcass traits and meat quality of Hanwoo steers.

OBJECTIVE: This study aimed to compare the effects of diets with and without supplements of methionine/lysine (met/lys) and methionine/α-tocopherol (met/α-tocopherol) on the performance, carcass traits and meat quality of Hanwoo steers. METHODS: A total of 21 Hanwoo steers were divided into three different groups. Each group consisted of 7 animals that received different dietary treatments for 120 days as follows: a control (C) diet composed of a basal diet with 74% total digestible nutrient and 12% crude protein; treatment 1 (T1) composed of a basal diet enriched with methionine 69%+lysine 31%; and treatment (T2) composed of a basal diet enriched with methionine 84.65%+α-tocopherol 15.35%. Daily supplementation was given using the top-dressing method (20 g/animal). RESULTS: The met/lys groups yielded a longissimus lumborum with increased water holding capacity (p<0.01) and lower shear force value (p<0.05). Dietary met/lys did not have an adverse effect on the animal performance and carcass traits. Instead, these supplements contributed significantly to the higher protein content compared to the control diet (p<0.05). Myristic acid (C14:0) was the only fatty acid affected by the supplementation. While the met/α-tocopherol group led to significantly higher protein and oxymyoglobin contents during storage (p<0.05). It also contributed significantly to the lower shear force value and 2-thiobarbituric acid reactive substances score during storage (p<0.05) compared to the control and treatment 1 since the initial storage day. The met/α-tocopherol diet also yielded meat with a redder color (p<0.05) after 3 days of storage. However, it did not significantly contribute to the fatty acid profiles of Hanwoo steers. CONCLUSION: Met/lys supplementation resulted in higher protein scores, water holding capacity and lower shear force scores. While met/α-tocopherol supplementation contributes to the production of redder meat, reduces lipid oxidation, production of more tender meat and increases the content of protein and oxymyoglobin percentage.

Comparison of meat quality, fatty acid composition and aroma volatiles of dry-aged beef from Hanwoo cows slaughtered at 60 or 80 months old.

OBJECTIVE: The objective of this study was to compare the quality of dry-aged beef from cull Hanwoo cows slaughtered at 60 or 80 months old. METHODS: A total of eight cull Hanwoo carcasses with a quality grade of 3 (low-grade) were selected and divided into two age groups: 63.5±2.5 months old (n = 4) and 87.8±4.5 months old (n = 4). Whole longissimus thoracis et lumborum from the 11th rib to the last lumbar vertebrae, including the back fat, was removed from the carcass at 24 h postmortem and aged for 50 days in darkness at a temperature of 2°C±1°C, a relative humidity of 85% and an air flow of 2 m/s. The sampling was performed aseptically after 0, 20, 24, 40, and 50 days of aging. RESULTS: Regardless of the aging period, aging increased the lightness (p<0.05), redness (p<0.05) and yellowness (p<0.05) at initial blooming (90 min after slicing) and the overall acceptance (p<0.05). No further tenderization effect was found after 20 days of aging, but aging for 50 days significantly increased the lipid oxidation (p<0.05). The generation of aroma volatiles in the roast steak from aged samples was higher (p<0.05) than that of non-aged samples. No significant effect of age at slaughter was found on the color, pH, water-holding capacity, cooking loss, shear force value, bacterial counts, volatile basic nitrogen, consumer acceptance, lipid oxidation, fatty acid composition or aroma volatiles. CONCLUSION: The quality of dry-aged beef obtained from cull Hanwoo cows slaughtered at either 60 or 80 months old with similar quality grade was comparable and extending dry aging for more than 40 days is not recommended considering the costs and further lipid oxidation.

Deuterium Oxide Labeling for Global Omics Relative Quantification: Application to Lipidomics.

A novel quantitative mass spectrometric method based on partial metabolic deuterium oxide (D2O) labeling, named "Deuterium Oxide Labeling for Global Omics Relative Quantification (DOLGOReQ)", was developed for relative quantification of lipids on a global scale. To assess the precision and robustness of DOLGOReQ, labeled and unlabeled lipids from HeLa cells were mixed in various ratios based on their cell numbers. Using in-house software developed for automated high-throughput data analysis of DOLGOReQ, the number of detectable mass isotopomers and the degree of deuterium labeling were exploited to filter out low quality quantification results. Quantification of an equimolar mixture of HeLa cell lipids exhibited high reproducibility and accuracy across multiple biological and technical replicates. Two orders of magnitude of effective dynamic range for reasonable relative quantification could be established with HeLa cells mixed from 10:1 to 1:10 ratios between labeled and unlabeled samples. The quantification precision of DOLGOReQ was also illustrated with lipids commonly detected in both positive and negative ion modes. Finally, quantification performance of DOLGOReQ was demonstrated in a biological sample by measuring the relative change in the lipidome of HeLa cells under normal and hypoxia conditions.

Prolactin receptor gene polymorphism and the risk of recurrent pregnancy loss: a case-control study.

Since the first study was published reporting the candidate association between the prolactin receptor gene intron C/T polymorphism (rs37389) and recurrent miscarriage, no replication study has been performed. In this study, we investigated the role of the prolactin receptor gene C/T polymorphism in 311 Korean women with recurrent pregnancy loss and 314 controls. Genotyping for prolactin receptor gene intron C/T polymorphism was performed using a TaqMan assay. The significance of difference in the genotype distribution was assessed using a chi-square test, and continuous variables were compared using a Student's t-test. The genotype distribution of the prolactin receptor gene C/T polymorphism in the recurrent pregnancy loss group did not differ from that in the control group (CC/CT/TT rates were 49.8%/41.5%/8.7% and 52.5%/37.6%/9.9% for the recurrent pregnancy loss patient and control groups, respectively, p = .587). When the analysis was restricted to patients with three or more consecutive spontaneous miscarriages or patients without prior live birth, there were also no differences in the genotype distribution between these subgroups and controls. In conclusion, the findings of the current study suggest that the prolactin receptor gene intron C/T polymorphism is not a major determinant of the development of recurrent pregnancy loss. Impact statement What is already known: Many studies have investigated whether there is a genetic component for the risk of recurrent pregnancy loss. Recently, one study investigated whether genetic polymorphisms involved in the regulation of the hypothalamic-pituitary-ovarian axis would be associated with recurrent miscarriage. Among 35 polymorphisms in 20 candidate genes, genotype distribution with regard to the prolactin receptor gene intron C/T polymorphism (rs37389) differed between the recurrent miscarriage and the control groups. Since this study reporting the candidate association between the prolactin receptor gene and recurrent miscarriage, no replication study has been performed. What the results of this study add: The genotype distribution of the prolactin receptor gene C/T polymorphism in the recurrent miscarriage group did not differ from that in the control group. What the implications are of these findings: Our study may be useful in that it is the first replication study since the initial report of the association of prolactin receptor gene polymorphism with recurrent miscarriage. Although no association was found, the potential role of prolactin in pregnancy loss needs to be further investigated because prolactin and its receptor have been postulated to play an important role in the maintenance of normal pregnancy.

Comparison of meat quality, fatty acid composition and aroma volatiles of Chikso and Hanwoo beef.

OBJECTIVE: Although Hanwoo has been selected as the superior commercial beef cattle breed in Korea, Chikso (Korean brindle cattle) is still recognized as a valuable breed for beef production. The aim of this study was to compare the meat quality, fatty acid composition and aroma volatiles of beef from Chikso and Hanwoo steers maintained under identical feed management, as information regarding these characteristics is still limited. METHODS: A total of 19 carcasses with a quality grade of 1 were selected, and strip loin (longissimus lumborum) cuts were collected from 11 Hanwoo carcasses and 8 Chikso carcasses. Meat quality and aroma analyses were performed at day four postmortem. RESULTS: Though Hanwoo strip loin tended to have higher fat content (15.37%) than Chikso (12.01%), no significant differences were observed. Meat pH, water-holding capacity, cooking loss, shear force value, instrumental surface color (Commission International De L'eclairage L*, a*, b*, chroma, and hue angle) and fatty acid composition were not significantly different. Roasted Chikso beef released more intense aroma than roasted Hanwoo beef based on the total area units of identified volatiles. Among identified volatiles, the amounts of toluene, heptanal, octanal, and nonanal were higher in roasted Chikso beef than in roasted Hanwoo beef. In addition, the aroma pattern of the roasted beef from these breeds was well-discriminated by electronic nose. CONCLUSION: No distinct differences were found in terms of meat quality between Hanwoo and Chikso beef in this study. However, the aroma pattern and volatiles of roasted Hanwoo and Chikso beef were different according to instrumental analysis.

Effects of high-pressure processing on taste-related ATP breakdown compounds and aroma volatiles in grass-fed beef during vacuum aging.

OBJECTIVE: This study aimed to observe whether high-pressure processing (HPP) affected aroma development and the degradation rate of umami taste-related ATP breakdown products, specifically inosinic acid in grass-fed beef during vacuum aging. METHODS: Strip loin (longissimus lumborum) cuts obtained from six grass-fed Friesian Holstein steers (32 months old) on day 4 post slaughter were vacuum-packed and subjected to pressurization at 300 and 500 MPa for 180 s at 15°C±2°C. The samples were then stored for 4 weeks at 5°C±0.5°C under vacuum and compared with the control (0.1 MPa). RESULTS: HPP increased the shear force value, promoted moisture loss and lipid oxidation, induced surface paleness, stabilized pH during aging, and reduced bacterial load and growth. The shear force value of 500 MPa-treated samples remained higher than the control after aging, while no significant differences were found between the control and 300 MPa-treated samples. Degradation of inosinic acid and inosine occurred during pressurization, resulting in an increase in hypoxanthine content. However, the degradation rate in HPP-treated samples during aging was slower; therefore, inosinic acid and inosine content remained higher than in control samples. No significant differences were found in hypoxanthine content at the end of aging. HPP intensified the levels of hexanal, octanal, 2-methylbutanal, 3-methylbutanal, benzaldehyde, and 2,5-dimethylpyrazine in cooked-aged beef samples. CONCLUSION: HPP induced aroma development and delayed the degradation of inosinic acid. However, it also reduced the postmortem tenderization rate.

Effects of cooking method and final core-temperature on cooking loss, lipid oxidation, nucleotide-related compounds and aroma volatiles of Hanwoo brisket.

OBJECTIVE: This study observed the effects of cooking method and final core temperature on cooking loss, lipid oxidation, aroma volatiles, nucleotide-related compounds and aroma volatiles of Hanwoo brisket (deep pectoralis). METHODS: Deep pectoralis muscles (8.65% of crude fat) were obtained from three Hanwoo steer carcasses with 1+ quality grade. Samples were either oven-roasted at 180°C (dry heat) or cooked in boiling water (moist heat) to final core temperature of 70°C (medium) or 77°C (well-done). RESULTS: Boiling method reduced more fat but retained more moisture than did the oven roasting method (p<0.001), thus no significant differences were found on cooking loss. However, samples lost more weight as final core temperature increased (p<0.01). Further, total saturated fatty acid increased (p = 0.02) while total monounsaturated fatty acid decreased (p = 0.03) as final core temperature increased. Regardless the method used for cooking, malondialdehyde (p<0.01) and free iron contents (p<0.001) were observed higher in samples cooked to 77°C. Oven roasting retained more inosinic acid, inosine and hypoxanthine in samples than did the boiling method (p<0.001), of which the concentration decreased as final core temperature increased except for hypoxanthine. Samples cooked to 77°C using oven roasting method released more intense aroma than did the others and the aroma pattern was discriminated based on the intensity. Most of aldehydes and pyrazines were more abundant in oven-roasted samples than in boiled samples. Among identified volatiles, hexanal had the highest area unit in both boiled and oven-roasted samples, of which the abundance increased as the final core temperature increased. CONCLUSION: The boiling method extracted inosinic acid and rendered fat from beef brisket, whereas oven roasting intensified aroma derived from aldehydes and pyrazines and prevented the extreme loss of inosinic acid.

Trends in Fetal and Perinatal Mortality in Korea (2009-2014): Comparison with Japan and the United States.

Fetal death is an important indicator of national health care. In Korea, the fetal mortality rate is likely to increase due to advanced maternal age and multiple births, but there is limited research in this field. The authors investigated the characteristics of fetal deaths, the annual changes in the fetal mortality rate and the perinatal mortality rate in Korea, and compared them with those in Japan and the United States. Fetal deaths were restricted to those that occurred at 20 weeks of gestation or more. From 2009 to 2014, the overall mean fetal mortality rate was 8.5 per 1,000 live births and fetal deaths in Korea, 7.1 in Japan and 6.0 in the United States. While the birth rate in Korea declined by 2.1% between 2009 and 2014, the decrease in the number of fetal deaths was 34.5%. The fetal mortality rate in Korea declined by 32.9%, from 11.0 in 2009 to 7.4 in 2014, the largest decline among the 3 countries. In addition, rates for receiving prenatal care increased from 53.9% in 2009 to 75.0% in 2014. Perinatal mortality rate I and II were the lowest in Japan, followed by Korea and the United States, and Korea showed the greatest decrease in rate of perinatal mortality rate II. In this study, we identified that the indices of fetal deaths in Korea are improving rapidly. In order to maintain this trend, improvement of perinatal care level and stronger national medical support policies should be maintained continuously.

Methylenetetrahydrofolate Reductase Polymorphisms and Risk of Recurrent Pregnancy Loss: a Case-Control Study.

The balance between coagulation and fibrinolysis is an essential part in early pregnancy. Mutations in methylenetetrahydrofolate reductase (MTHFR) gene lead to decreased activity of the enzyme and hyperhomocysteinemia, which then induces platelet aggregation by promoting endothelial oxidative damage, possibly resulting in adverse effect on maintenance of pregnancy. We investigated the role of MTHFR single nucleotide polymorphisms (SNPs), C677T and A1298C, in Korean patients with recurrent pregnancy loss (RPL). We conducted a prospective case-control study in the Korean population. Subjects included 302 women with 2 or more consecutive, unexplained, spontaneous miscarriages before 20 weeks of gestation and 315 control women without a history of recurrent miscarriages. The genotyping for C677T and A1298C polymorphisms was performed using the TaqMan assay. Continuous variables were compared using Student's t-test, and χ² test was used to evaluate differences in the genotype distributions between the RPL and the controls. The genotype distribution of both polymorphisms in the RPL group did not differ from those of the controls. For further analysis, if RPL patients were divided according to the numbers of pregnancy losses (≥ 2 and ≥ 3) neither group was significantly different compared with controls. MTHFR gene C677T and A1298C polymorphisms are not associated with idiopathic RPL in Korean women, suggesting that those may not be susceptible allelic variants or be deficient to cause RPL.

High pressure processing for dark-firm-dry beef: effect on physical properties and oxidative deterioration during refrigerated storage.

OBJECTIVE: Study on the application of high pressure processing (HPP) for dark-firm-dry (DFD) beef was conducted to observe whether HPP has any impact on physical properties and to evaluate oxidative deterioration during refrigerated storage under vacuum. METHODS: The longissimus lumborum muscles obtained from Friesian Holstein steers (33±0.5 months old) with 24-h postmortem pH higher than 6.0 were vacuum-packed and subjected to pressurization at 200, 400, and 600 MPa for 180 s at 15°C±2°C; the samples were then stored for 9 days at 4°C±1°C and compared with control (0.1 MPa). RESULTS: HPP increased meat pH by 0.1 to 0.2 units and the tenderness of cooked DFD beef significantly with no significant effects on meat texture profile. The stability of meat pH was well maintained during refrigerated storage under vacuum. No clear effects were found on the activity of catalase and superoxide dismutase, however, glutathione peroxidase activity was significantly reduced by high pressure. HPP and storage time resulted in aroma changes and the increasing amount of malondialdehyde and metmyoglobin relative composition. CONCLUSION: Although the increasing amount of malondialdehyde content, metmyoglobin formation and aroma changes in HPP-treated samples could not be avoided, HPP at 200 MPa increased L* and a* values with less discoloration and oxidative deterioration during storage.