The Association of Freezer Storage Time With Vitamin K and Vitamin D Concentrations in Human Brain Tissue

Vitamins K and D are present in the human brain and have been implicated in Alzheimer’s disease and related dementias (ADRD). Because the use of banked brain tissue in ADRD research is increasing, we evaluated the stability of vitamin K and vitamin D in human brain tissue over long-term freezer storage using samples obtained from the Rush Memory and Aging Project (n=500, mean age=91, 29% male). Specimens were stored at -80□C until analyzed. Vitamin K (menaquinone-4, MK4) and vitamin D (25(OH)D) were measured in four regions (mid-temporal and mid-frontal cortexes, cerebellum, anterior watershed white matter) and averaged across regions. Storage time was categorized into two-year increments. Differences in MK4 and 25(OH)D concentrations according to storage time were evaluated using general linear models. MK4 concentrations did not differ in brains stored ≤8 years (geometric mean±SEM MK4 pmol/g: storage ≤2.0 years=1.2±0.1, 2.1-4.0 years=1.2±0.1, 4.1-6.0 years=1.4±0.1, 6.1-8.0 years=1.4±0.2; p≥0.21). MK4 in brains stored >8.0 years (0.8±0.1 pmol/g) was 33% lower than the concentration in brains stored ≤2.0 years (p=0.005). The 25(OH)D concentrations did not differ in brains stored ≤6 years (geometric mean±SEM 25(OH)D pmol/g: storage ≤2.0 years=1.2±0.1, 2.1-4.0 years=1.1±0.1, 4.1-6.0 years=1.2±0.1; p≥0.37). The 25(OH)D concentration in brains stored >6.0 years was 31-37% lower than that in brains stored ≤2.0 years (6.1-8.0 years=0.8±0.06, >8.0 years=0.7±0.04; p<0.001). MK4 and 25(OH)D appeared to be stable in human brain tissue stored at -80oC for up to 8 and 6 years, respectively. Storage time merits consideration when designing and interpreting studies that relate brain nutrient concentrations to ADRD.

Vitamin D and Vitamin K Concentrations in Human Brain Tissue Are Influenced by Freezer Storage Time: The Memory and Aging Project

ABSTRACT Background Vitamins D and K, which are present in human brain, may have a role in neurodegenerative disease. Objectives Given the interest in measuring nutrient concentrations in archived brain samples, it is important to evaluate whether freezer storage time affects these concentrations. Therefore, we evaluated differences in vitamin D and vitamin K concentrations in human brain samples stored for various lengths of time. Methods Postmortem brain samples were obtained from 499 participants in the Rush Memory and Aging Project (mean age 92 y, 72% female). Concentrations of vitamins D and K and their metabolites were measured in 4 regions (midtemporal cortex, midfrontal cortex, cerebellum, anterior watershed white matter) using LC-MS/MS and HPLC, respectively. The predominant forms were 25-hydroxycholecalciferol [25(OH)D3] and menaquinone-4 (MK4). ANOVA was used to determine if concentrations differed according to storage time. Results The geometric mean of the mean 25(OH)D3 concentration (across 4 regions) in brains stored for 1.1 to 6.0 y did not differ from that in brains stored ≤1.0 y (all P ≥ 0.37), whereas 25(OH)D3 in brains stored >6.0 y was 31–40% lower (P ≤ 0.003). MK4 had similar results, with the geometric mean MK4 concentration in the brains stored ≥9.0 y being 48–52% lower than those in brains stored ≤1.0 y (P ≤ 0.012). The 25(OH)D3 and MK4 concentrations were positively correlated across all 4 regions (all Spearman ρ ≥ 0.79, P < 0.001). Conclusions 25(OH)D3 and MK4 appear to be stable in brain tissue from older adults stored at -80°C for up to 6 and 9 y, respectively, but not longer. Freezer storage time should be considered in the design and interpretation of studies using archived brain tissue.

Onboard Evaluation of Variable Water Flow and Recirculation Effects on Bleeding of Atlantic Cod (Gadus morhua)

The aim of the study was to explore the effects of different design variables in the onboard bleeding process of cod on bleeding efficiency and the resulting product quality. A time- and flow-controlled process was used to create variable bleeding conditions for whole gutted cod onboard a wet-fish trawler. Two main design variables influencing the bleeding process are the pump flow recirculation (PFR) and the water replacement ratio (WRR); they were studied in five different combinations (groups). The effects of different bleeding conditions were evaluated by measurements of free fatty acids (FFAs), phospholipids (PLs), and total heme iron (HI) content during freezer storage for up to four months. The results for PL content and the regression model indicate that the enzyme activity in the fish muscle is lower in cases where PFR exerts greater influence in the bleeding process than WRR. The effects of successful blood removal also seem to be most noticeable after one month of freezer storage, rather than in fresh cod after seven days or after four months of simulated frozen food-chain storage. The study indicates that, with the bleeding medium to fish ratio of around 3:1 and enough WRR (over 100% replacement in 20 min), the PFR becomes the limiting design parameter regarding efficient blood removal and should be at least 10% of the tank volume per minute to ensure enough recirculation and flow of water in the bleed-out tanks.

Bacterial stability with freezer storage of human milk

ObjectivesHuman milk supports the development of a beneficial newborn intestinal microflora. We have shown previously that human milk had reduced bacteria but unchanged nutrient composition when stored at −20 °C for up to nine months. We suspected declining bacterial colony counts were manifestations of bacterial dormancy and not failure of survival. We investigated differences in selected bacterial colony counts (lactobacillus, bifidobacteria, staphylococcus, streptococcus and enterococcus) in human milk stored for 2 and 12 weeks at −20 °C in either manual or automatic defrost freezers and whether reduced bacterial counts at 12 weeks were the result of dormancy or failure of survival.MethodsFreshly expressed milk was obtained from mothers in the NICU, divided into aliquots and stored for 2 and 12 weeks at −20 °C in either automatic or manual defrost freezers. Subsequently, duplicate aliquots, one thawed and the other thawed and maintained at room temperature for 4 h, were plated to assess bacterial colony counts.ResultsSignificant declines in bacterial colony counts were seen from 2 to 12 weeks freezer storage for all bacteria. There were no differences in colony counts between freezer types. Once thawed, no further bacterial growth occurred.ConclusionsShort-term freezer storage for 12 weeks resulted bacterial killing. Type of freezer used for storage did not have an impact on bacterial survival. It is unknown whether the paucity of important probiotic bacteria in stored human milk has adverse effects on infants.

An Assessment of Storability of Norway Spruce Container Seedlings in Freezer Storage as Affected by Short-Day Treatment

Determination of safe times at which to transfer seedlings to freezer storage is problematic in forest tree nurseries. The present study aimed to determine the relationship between pre-storage frost hardiness (FH) of different plant parts, dry matter content (DMC), chilling hours (the sum of hours when temperature was between −5 °C and +5 °C), and post-storage vitality, and the impact of short-day (SD) treatment on these relationships. One and a half year old control seedlings and SD-treated seedlings of Norway spruce were transferred to freezer storage (−3 °C) on five occasions during autumn. On each occasion, the FH of buds, needles, stem, and roots, as well as DMC, were determined, and chilling hours were calculated. The vitality of the freezer-stored seedlings was determined through their root growth capacity in the subsequent spring, and through the field performance of the seedlings (shoot growth and seedling damage) at the end of the following two growing seasons. Seedlings were considered to be storable when the FH of the needles was at least −25 °C, and the FH of the roots was about −10 °C in both treatments. Early storage reduced the vitality of the seedlings. SD treatment did not advance the storability of the seedlings, although it alleviated some of the negative effects of early storage by improving the FH of needles and stem, but not that of the roots. The DMC value, indicating storability, was higher for SD-treated seedlings than for control seedlings. When data from five experiments conducted in Suonenjoki were combined, it was found that the relationship between accumulation of chilling hours and needle FH was dependent on nursery treatment and assessment year, which reduces the reliability of using chilling hours in predicting the storability of Norway spruce seedlings. The predicted climate change may complicate the fall acclimation of seedlings. New, user-friendly methods for determining storability of seedlings are urgently needed.

The Stability of Vitamins D and K of the Human Brain During Freezer Storage: The Memory and Aging Project (MAP)

Objectives Vitamins D and K are present in human brain tissue, and evidence is emerging that these nutrients may have a role in neurodegenerative disease. Given the increasing interest in conducting analyses of archived brain samples, it is important to evaluate the stability of these nutrients in brain tissue over long-term storage. Therefore, we evaluated the influence of freezer storage time on vitamin D and vitamin K concentrations in human brain samples. Methods Post-mortem brain samples were obtained from 500 participants in the Rush Memory and Aging Project (mean age 91 yrs, 71% female). At autopsy and brain dissection, the tissues were immediately frozen on brass plates atop dry ice then stored at −80o C until analysis. Storage time was categorized in 1-year increments. Vitamin D [25(OH)D] and vitamin K [menaquinone-4 (MK4)] concentrations were measured in four regions (mid-temporal cortex, mid-frontal cortex, cerebellum, anterior watershed white matter) using LC/MS/MS and HPLC, respectively. We calculated the mean 25(OH)D and MK4 concentrations across the four regions and then applied a natural-log transformation to the means to improve normality. Analysis of variance was used to determine if the geometric mean ± SEM 25(OH)D and MK4 concentrations differed according to storage time. The samples stored ≤1.0 year served as the reference group. Results The mean 25(OH)D concentration in brains stored >6.0 years (n = 203; 0.8 ± 0.05 pmol/g), was significantly lower than the concentration in brains stored ≤1.0 year (n = 79; 1.2 ± 0.09 pmol/g, P ≤ 0.004). The 25(OH)D in the brains stored ≤1.0 year did not differ from the concentration in brains stored for 1.1–6.0 years (n = 212; 1.2 ± 0.05 pmol/g, P ≥ 0.37). The mean MK4 concentration in the brains stored ≥9.0 years (n = 81; 0.7 ± 0.6 pmol/g) was significantly lower than that in the brains stored ≤1.0 year (1.2 ± 1.3 pmol/g; P ≤ 0.01). The MK4 in brains stored ≤1.0 year did not differ from the concentration in brains stored for 1.1–9.0 years (n = 339; 1.3 ± 0.8 pmol/g; P ≥ 0.11). Conclusions 25(OH)D and MK4 appear to be stable in brain tissue stored at −80 °C for up to 6 and 9 years, respectively, but not longer. This should be considered in the design and interpretation of studies linking brain concentrations of these nutrients to neurodegenerative diseases. Funding Sources Supported by the National Institute of Aging.

Main Health-Promoting Compounds Response to Long-Term Freezer Storage and Different Thawing Methods in Frozen Broccoli Florets

The effects of long-term freezer storage and different defrosting methods on the retention of glucosinolates, vitamin C, and total phenols in frozen broccoli florets were investigated in the present study. Frozen broccoli florets were stored in a freezer at −20 °C for 165 days or subjected to defrosting by three different house-hold thawing methods (water, air, and refrigerator defrosting). Results showed that all glucosinolates were well preserved, while vitamin C and total phenols were reduced by less than 12% and 19% of the control, respectively, during the storage. Besides, refrigerator and air defrosting were better than water defrosting in glucosinolates retention, and refrigerator defrosting was the best in vitamin C preservation. No difference was observed in reserving phenolic compounds among the three methods. In conclusion, long-term freezer storage is an excellent way to preserve broccoli florets, and refrigerator defrosting is the best way to maintain the nutritional compounds in frozen broccoli florets.

IDENTIFIKASI POTENSI BAHAYA DAN PEMANTAUAN CRITICAL POINT, (HACCP) PRODUK MAKANAN PENERBANGAN

ABSTRACTHACCP has been known as a system that uses a systematic and preventive approach that is shown to biological, chemical and physical hazards through anticipatory and preventive measures by no relying on inspection and testing on the final product. The application of HACCP is not only for the food industry but can be applied to the catering industry, catering services and food at hotels and restaurants. For this reason, modern food companies really need to determine quality standards for the consumers they serve. The purpose of this study was to analyze the application of HACCP on aviation food products (Aerofood ACS SUB) by identifying potential hazards and applying Critical Control Points (CCP). The methods are 1) observation of Critical Control Points (CCP) performed at receiving (CCP 1), chiller and freezer (Storage) (CCP 2), cooking (CCP 3), blast chilling (CCP 4), portioning (CCP 5). 2) Microbiological sampling consisting of random raw material samples at each arrival, hand swab samples randomly in the production and operational areas, swab equipment samples were also taken according to random, Dry good samples were taken randomly in storage, random ice cube samples , water tab samples are taken according to the sample. The company has HACCP planning as a guide for all processes that will take place within the company. All are based on the principles of HACCP for the whole process. The implementation of critical control points is in 5 places, namely receiving, storage, cooking, chilling and portioning blast. At each of these critical points, the standard critical temperature is different. Materials that do not meet the standards are rejected for further processing. In terms of microbiological hazards, it is checked by testing samples on foodstuffs, ready to eat food, dry good, air test, hand swab, production equipment, water and ice cube. The implementation of each sample test has been determined by PT. Aerofood ACS Surabaya based on standard procedures. If the results of checking is not the standards, repairs are handled or changes in the flow of handling procedures. Keywords: HACCP, CCP, Critical Limits, management system ABSTRAKHACCP telah dikenal luas diseluruh dunia sebagai suatu sistem yang menggunakan pendekatan sistimatis dan preventif yang ditunjukan kepada bahaya biologis, kimia dan fisik melalui langkah-langkah antisipatif dan pencegahan dengan tidak lagi mengandalkan pada pemeriksaan dan pengujian pada produk akhir. Penerapan HACCP tidak hanya untuk industri pangan melainkan dapat diterapkan pada industri catering dan jasa boga serta makanan di hotel dan restauran. Untuk itu perusahaan pangan modern sangat perlu untuk menentukan standart mutu untuk konsumen yang dilayaninya. Tujuan penelitian ini adalah menganalisis penerapan HACCP pada produk makanan penerbangan (Aerofood ACS SUB) dengan identifikasi potensi bahaya dan penerapan Critical Control Point (CCP). Metode yang digunakan dalam penelitian ini adalah 1) Pengamatan Critical Control Point (CCP) yang dilakukan di receiving (CCP 1), chiller dan freezer (Storage) (CCP 2), cooking (CCP 3), blast chilling (CCP 4), portioning (CCP 5). 2) Pengambilan sampel Mikrobiologi yang terdiri dari sampel raw material secara random disetiap kedatangan, sampel hand swab secara random di area produksi dan operasional, sampel equipment swab juga diambil sesuai random, sampel Dry good diambil secara random di storage, sampel ice cube secara random, sampel Water tab diambil sesuai sampel. Perusahan telah membuat perencanaan HACCP sebagai panduan untuk semua proses yang akan berlangsung didalam perusahaan. Semua disusun berdasarkan prinsip-prinsip HACCP untuk keseluruhan proses. Penerapan critical control point terdapat di 5 tempat yaitu receiving, storage, cooking, blast chilling dan portioning. Disetiap titik kritis ini, suhu kritis standart adalah berbeda beda. Bahan yang tidak memenuhi standard ditolak untuk diproses selanjutnya. Dalam hal bahaya mikrobiologi dilakukan pengecekkan melalui uji sampel pada bahan makanan, makanan ready to eat, dry good, uji udara, hand swab, peralatan produksi, air dan ice cube. Pelaksanaan masing-masing pengujian sampel sudah ditetapkan oleh PT. Aerofood ACS Surabaya berdasarkan prosedur standar. Jika hasil pengecekkan tidak memenuhi standart maka dilakukan perbaikan penanganan atau pengubahan alur prosedur handling. Kata kunci: HACCP, CCP, Batas Kritis, management system