Journal articles on the topic 'Frozen foods – Evaluation – Case studies'

Nowadays antimicrobial drugs of different pharmacologic classes are widely used in aquaculture all over the world. Therefore, in accordance with EU Regulation 37/2010 “On pharmacologically active substances and their classification in regard to maximum allowable amounts in nourishing food of animal origin”, there are certain amount of antimicrobial drugs acceptable in fish. The aim of the work was to determine toxico-biological assessment of flesh of frozen fish where there are residues of antimicrobial drugs on the culture of infusorias Tetrachymena pyriformis. The availability of residual amounts of antimicrobial drugs was carried out by screening method for antibiotics in products of animal origin РХ/МС/МС and microbiological method. Toxico-biological assessment of flesh of frozen fish was determined in accordance with “Methodological recommendations on toxico-biological assessment of flesh, flesh products and milk with infusoria Tetrachymena pyriformis”. It was found that samples of frozen fish containing residues of antimicrobial drugs of aminoglycosides, penicillic and fluoroquinolonic class were not toxic for Tetrachymena pyriformis cultures. Since infusorias were active, natural movement – progressive and circular, inhibition of growth or change of body shape were not showed compared to infusorias within control group. It was shown that in the fish flesh with residues of tetracycline antibiotics, there was observed the inhibition of growth and decreased activity and mobility of infusorias Tetrachymena pyriformis up to 20% compared to control without antibiotics. However, unnatural movements (affected or oscillatory) and pathological changes in the shape of infusoria cultures were not detected. This flesh fish was characterized as moderate toxic. It was stated that relative biological value of frozen fish flesh with antibiotic residues of aminoglycosides, and fluoroquinolonic class was lower by 0,27–0,73% compared to relative biological value of control fish flesh without antibiotics. At the same time relative biological value of fish flesh with the presence of residues of tetracycline antibiotics was by 4.12% lower compared to control fish flesh. Consequently, conducted studies indicate the need to control the residues of antimicrobial drugs in frozen fish which is imported to Ukraine. In case of detection the exceeding permissible amount of antibiotic residues in frozen fish it should be discarded, as it exhibits a moderately toxic effect.

The antioxidant activity (AOA) of plant foods is recognized as an index of the potential health benefits resulting from their consumption. Due to their high perishability and seasonality, plant foods are largely consumed or used as processed products and freezing is one of the technologies used for the production of high-quality foods. However, cell breakages occurring during freezing and frozen storage can lead to the release of antioxidant compounds and their degradation due to chemical and enzymatic oxidation reactions, and thus, they could present a lower antioxidant activity compared to the corresponding fresh product. In this context, process conditions, freezing pre-treatments and the use of cryoprotectants can limit the extent of freeze-induced damages and preserve the antioxidant activity of plant foods. This review collects and discusses the state-of-the-art knowledge on the single and combined effect of freezing and frozen storage conditions on the antioxidant activity of fruits and vegetables as well as the role of cryoprotectants. Classes of compounds responsible for the antioxidant activity of plant foods and the most common methods used for the evaluation of the antioxidant activity in vitro are also presented. The freezing principles and the effects of ice nucleation and crystallization on fruits, vegetables and their main derivatives (juices, pulps) have been addressed to highlight their impact on the AOA of plant foods. The effect of freezing and frozen storage on the AOA of plant foods resulted dependant on a series of intrinsic factors (e.g., composition and structure), while the role of extrinsic processing-related factors, such as freezing and storage temperatures, is ambiguous. In particular, many conflicting results are reported in the literature with a high variability depending on the method of analysis used for the AOA evaluation and data expression (fresh or dry weight). Other intrinsic raw material properties (e.g., cultivar, ripening degree), post-harvest conditions, as well as defrosting methods that in the majority of the studies are scarcely reported, contribute to the aforementioned discrepancies. Finally, due to the limited number of studies reported in the literature and the high variability in product processing, the effect of cryoprotectants on the AOA of plant foods remains unclear.

The history and applications of food irradiation are reviewed. The term wholesomeness when applied to food irradiation, embodies the concepts of microbiological and toxicological safety, and nutritional adequacy. The status of these areas of concern is reviewed. Nutritional studies have addressed the effects of irradiation on nutrient content and bioavailability, and evaluation of potential consequences of changes in either. Results of rat studies are presented in which we tested for the presence of anti-thiamin and anti-pyridoxine activity in radappertized chicken and beef. Test meats were analyzed for thiamin and pyridoxine to establish a basis for incorporation into repletion diets. Thiamin levels in gamma- and electron-irradiated, and thermally processed (commercial canning) chicken were 74, 34 and 78%, respectively, of the vitamin level in a frozen meat reference; the levels in beef were 77, 56 and 79%, respectively. Pyridoxine levels in chicken were 50, 38 and 17%, respectively, of the reference level. Rats were depleted in each vitamin, then repleted at two vitamin levels with diets containing test meats. Activities of transketolase, aspartate aminotransferase and alanine aminotransferase in erythrocytes from these rats provided no consistent evidence of antivitamin presence. It was concluded that these irradiated meats pose no problem regarding vitamins B1 and B6 if part of a complete diet.

A thaumatin-like protein gene from Basrai banana was cloned and expressed in Escherichia coli. Amplified gene product was cloned into pTZ57R/T vector and subcloned into expression vector pET22b(+) and resulting pET22b-basrai TLP construct was introduced into E. coli BL21. Maximum protein expression was obtained at 0.7 mM IPTG concentration after 6 hours at 37°C. Western blot analysis showed the presence of approximately 20 kDa protein in induced cells. Basrai antifungal TLP was tried as pharmacological agent against fungal disease. Independently Basrai antifungal protein and amphotericin B exhibited their antifungal activity against A. fumigatus; however combined effect of both agents maximized activity against the pathogen. Docking studies were performed to evaluate the antimicrobial potential of TLP against A. fumigatus by probing binding pattern of antifungal protein with plasma membrane ergosterol of targeted fungal strain. Ice crystallization primarily damages frozen food items; however addition of antifreeze proteins limits the growth of ice crystal in frozen foods. The potential of Basrai TLP protein, as an antifreezing agent, in controlling the ice crystal formation in frozen yogurt was also studied. The scope of this study ranges from cost effective production of pharmaceutics to antifreezing and food preserving agent as well as other real life applications.

ABSTRACT: This research presents the case of a patient with a frozen shoulder and assesses the impact of osteopathic techniques and physical exercises for this condition. The article describes the epidemiology, etiology, pathogenesis, clinical symptoms and treatment options for frozen shoulder. It also contains information on how to conduct clinical tests and diagnosis of the adhesive capsulitis. We assessed the range of flexion, external rotation, internal rotation and the intensity of pain of the patients frozen shoulder. In this case study frozen shoulder developed typically, in three stages “freezing”, “frozen”, “thawing”. The treatment significantly increased the range of motion of the shoulder joint and reduced sensation of pain. The analysis shows that the osteopathic techniques and physical exercises are an effective form of rehabilitation and pain management. Further studies are needed in this area.

Egg is a basic foodstuff in many cultures and has a special place, also represent a rich source of nutrients such as, high quality proteins and a large variety of minerals and vitamins. The main derived products which are processed nowadays are dehydrated products (egg powder) and the liquid ones (refrigerated or frozen) as well as pasteurized egg m�lange which was the studied food in the current research. A number of 20 samples of egg m�lange were gathered from two different processing units. Samples were subjected to chemical analysis being determined content in water, dry matter, proteins, lipids, minerals, non-nitrogenous extractive substances, fatty acids and amino acids. Based on the obtained values was realised a nutritional evaluation for the analysed product by calculating the chemical indexes of essential amino acids (CI), essential amino acids index (EAAI), biological value (BV) and nutritional index (NI) for three categories of persons (children, young people and adults). The results of the current study showed the fact that chemically speaking, the analysed pasteurized egg m�lange presented values which were into the limits from literature; the analysed samples contented 75.59% water, 24.41% dry matter, 12.22% protein, 9.71% lipids, 0.99% total minerals, 1.48% non-nitrogenous extractive substances, 12.34 g/100 g content in amino acids (5.5 g/100 g essential amino acids) and 22.86 g/100 g lipids for polyunsaturated fatty acids. After nutritional evaluation of proteins from the studied food we observed that those ones had a very close value (EAA=50.77 g/16 g N) to the one of hen egg albumen (51.2 g/16 g N), value which is considered etalon. Calculus of some foresight indicators for nutritional value of proteins from studied egg m�lange showed the fact that this one could represent a good source of proteins and implicitly of essential amino acids for humans even if in children�s case the values were a little bit lower and two amino acids (tryptophan and leucine) were slightly limiting.

Abstract Damaging effects of low temperatures on the functionality of sperm is well known. Stallion sperm cryoresistance has a high individual variability. The genetic basis of cryoresistance and its heritability are little studied. The aim of the study was to search for genome-wide associations with sperm motility after freezing. 96 sperm samples were collected. The collected sperm was diluted to a final concentration 100 mln/ml and frozen in 0.5 ml straws or 18–20 ml tubes. The evaluation of semen was carried out no earlier than 24 hours after freezing. Sperm motility was assessed by computer-assisted semen analysis (CASA). Genomic DNA was purified form semen samples and genotyped using Affymetrix Equine HD array. Genotypes quality control and association studies were performed in Plink 1.9 and EMMAX software respectively. Evaluation of sperm motility showed high individual variability in both total and progressive motility after freezing. Total motility varied from 0.5 to 92.5%, and progressive motility from 0.5 to 70.8%. Sufficient associations of PTF with SNP’s were detected on chromosome (Chr) 1 (P < 1.33e-09) and 4 (P < 2.00e-09). Found SNP’s were located in genes PAS domain-containing protein 3 (Chr1) and UBAP1-MVB12 (Chr4). Expression of these genes in human body was found in cerebrum and male genitals. Suggestive SNPs were found lying nearby to genes responsible for formation of cell wall and proteins affecting mitochondrial activity. Performed studies are presenting first step of understanding genetic background of cryoresistancy of semen in horses. Found markers could be used for selection of stallions based on cryopreservation ability. Authors acknowledge financial support from Russian Science Foundation, Grant No: 18-16-00071.

Abstract Context.—Frozen section (FS) evaluation of central nervous system (CNS) lesions provides an assessment of specimen adequacy and facilitates triage for ancillary studies. Frozen section also provides an accurate preliminary diagnosis; however, certain lesions are recognized to cause diagnostic challenges at FS. Objective.—To identify cases in which there was a discrepancy between the FS diagnosis and final diagnosis in the case to heighten awareness of common diagnostic pitfalls in surgical neuropathology. Design.—All CNS FS cases involving a tumor diagnosis at FS or permanent section (N = 2156) from September 1997 until June 2005 were retrospectively reviewed. Discrepancies between the FS and final diagnoses were identified. Results.—Of the 2156 cases identified, 57 (2.7%) discrepant diagnoses were found. Twelve (21.1%) of 57 discrepancies involved errors in classification of spindle cell lesions, most commonly confusing schwannomas or meningiomas with other lesions. Twelve (21.1%) of 57 cases involved errors in differentiating oligodendrogliomas from astrocytomas. Nine (15.8%) of 57 discrepancies involved errors in the diagnosis of CNS lymphoma. Eight (14.0%) of 57 cases involved errors in differentiating reactive from neoplastic processes, most frequently gliosis versus glioma. Four (7.0%) of 57 discrepancies involved errors in the overgrading of tumors. The remaining 12 (21.1%) of 57 cases included an assortment of other discrepancies. Conclusions.—Frozen section of CNS neoplastic processes can be highly accurate. Less than 3% of FS diagnoses in 1 institution's experience were discrepant with the final diagnoses. Approximately 80% of the discrepant cases were classified into 5 categories: spindle cell lesions, astrocytoma versus oligodendroglioma, differential diagnosis of CNS lymphoma, reactive versus neoplastic process, and tumor overgrading. Awareness of these pitfalls may help in further increasing diagnostic accuracy.

Abstract The VIDAS® Listeria monocytogenes Xpress (LMX) test is an enzyme-linked fluorescent immunoassay designed for use with the automated VIDAS or mini-VIDAS instruments for the specific detection of L. monocytogenes using a 26 h proprietary enrichment broth. The VIDAS LMX method was validated according to harmonized AOAC Research Institute (RI) and Official Methods of Analysis guidelines in both the AOAC Performance Tested MethodSM (PTM) and GovVal programs. In the PTM comparison studies, the VIDAS LMX method was compared to the U. S. Department of Agriculture-Food Safety and Inspection Service Microbiology Laboratory Guidebook, the U. S. Food and Drug Administration Bacteriological Analytical Manual, and AOAC Official Methods. The comparative food studies consisted of two main parts: internal testing and AOAC independent laboratory testing, which included seven food matrixes (deli ham, processed cheese, vanilla ice cream, cooked shrimp, smoked white fish, frozen spinach, and peanut butter). As part of the AOAC RI GovVal program, the VIDAS LMX method was compared to the Health Canada MFHPB-30 method for the detection of L. monocytogenes in five ready-to-eat (RTE) meats (hot dogs, deli turkey, deli ham, fermented sausage, and liver paté). Twenty replicates of each inoculation level and five uninoculated controls were evaluated in each study. The LMX method also included the use of chromogenic media, chromID™ Ottaviani Agosti agar and chromID™ L. mono. agar, for confirmation of LMX presumptive results. In both the PTM and GovVal evaluations, there were no significant differences in the Chi-square values for the LMX method when compared to reference methods. The additional parameters tested in the PTM evaluation (inclusivity, exclusivity, ruggedness, stability, and lot-to-lot) satisfied the AOAC RI performance requirements. In both the PTM and GovVal validation studies, the VIDAS LMX method demonstrated reliability as a rapid qualitative method for next-day detection of L. monocytogenes in a variety of foods, including RTE meats.

AbstractGauge-calibrated radar estimates of daily precipitation are compared with daily observed values of precipitation from National Weather Service (NWS) Cooperative Observer Network (COOP) stations to evaluate the multisensor precipitation estimate (MPE) product that is gridded by the National Centers for Environmental Prediction (NCEP) for the eastern United States (defined as locations east of the Mississippi River). This study focuses on a broad evaluation of MPE across the study domain by season and intensity. In addition, the aspect of precipitation type is considered through case studies of winter and summer precipitation events across the domain. Results of this study indicate a north–south gradient in the error of MPE and a seasonal pattern with the highest error in summer and autumn and the lowest error in winter. Two case studies of precipitation are also considered in this study. These case studies include instances of intense precipitation and frozen precipitation. These results suggest that MPE is less able to estimate convective-scale precipitation as compared with precipitation variations at larger spatial scales. In addition, the results suggest that MPE is subject to errors related both to the measurement gauges and to the radar estimates used. Two case studies are also included to discuss the differences with regard to precipitation type. The results from these case studies suggest that MPE may have higher error associated with estimating the liquid equivalent of frozen precipitation when compared with NWS COOP network data. The results also suggest the need for more analysis of MPE error for frozen precipitation in diverse topographic regimes.

In North American cold regions, terrestrial spill-response tactics have evolved through clean-up experience with crude oil and refined petroleum products. Alaska has developed response tactics as guidelines for clean-up of petroleum-based spills. Generic application of any response tactic without regard for season, site-specific conditions, and equipment limitations can further damage an ecosystem. For example, the practice of igniting and burning petroleum product spilled onto frozen tundra without consideration of the anthropogenic effect on the surface energy balance may actually increase the vertical migration of the spilled product. Prior to application of any mitigation strategy to a release of petroleum product, the movement of the product through freezing soil needs to be better understood. Case studies are presented, and lessons learned from them are discussed.

Smell is the second-most used sense in marketing strategies in the food industry. Sensory marketing appeals to the senses with the aim of creating sensory experiences and converting them into specific emotions associated with a specific product. There is a strong relationship between sensory marketing, aromachology, and neuroscience. In this review, studies were searched on the use of scents in food experiences such as restaurants and food establishments, and a critical evaluation was performed on their aims, target population, place of the study, scents tested, foods tested, and measured parameters, and the main findings were reviewed. Case studies carried out by private companies are also presented. A small number of scientific studies on aromachology related to food are available, and most of them are conducted in artificial laboratory conditions. Methodological procedures largely diverge among studies, making them very difficult to compare and extrapolate results. There is a clear need for research on aromachology related to food in the fields of sensory marketing and appetite modulation. After a brief presentation of the state of the art, we briefly mention future improvements and ideas for future research.

Abstract A validation study was conducted to extend the matrix claim for the Nissui Compact Dry Total Count (TC), Performance Tested Method(s)SM (PTM) Certification No. 010404, to cooked chicken, lettuce, frozen fish, milk powder, and pasteurized whole milk. The method was originally certified by the AOAC Research Institute Performance Tested Method(s)SM Program for raw meat products. The Compact Dry TC is a ready-to-use dry media sheet that is rehydrated by adding 1 mL of diluted sample. A total aerobic colony count can be determined in the sample following 48 h of incubation. Matrix extension studies were conducted by Campden BRI (formerly Campden and Chorleywood Food Research Association Technology Limited), Chipping Campden, UK. Single-laboratory data were collected for cooked chicken, lettuce, frozen fish, and milk powder, whereas a multilaboratory study was conducted on pasteurized milk. Fourteen laboratories participated in the collaborative study. The Compact Dry TC was tested at two time points, 48 ± 3 h and 72 ± 3 h and compared with the current International Organization for Standardization (ISO) method at the time of the study, ISO 4833:2003 (this standard is withdrawn and has been replaced by: ISO 4833-1:2013 and ISO 4833-2:2013), Microbiology of food and animal feeding stuffs–Horizontal method for the enumeration of microorganisms–Colony-count technique at 30°C. The data were logarithmically transformed and evaluated for repeatability (plus reproducibility for pasteurized milk), RSD of repeatability (plus RSD of reproducibility for milk), r2, and mean difference between methods with 95% confidence interval (CI). A CI outside of (−0.5 to 0.5) on the log10 mean difference was used as the criterion to establish significant statistical difference between methods. No significant differences were found between the Compact Dry TC 48 and 72 h time points, with the exception of one contamination level of cooked chicken and one contamination level of dry milk powder. Mean differences were small at these levels (<0.5 log10), but the upper CIs were above 0.5. Statistical differences were indicated between the Compact Dry TC and ISO 4833 in two of five contamination levels tested each for lettuce and frozen fish. In each case, mean differences were >0.5 log10, and the total aerobic colony count was higher for the ISO method. In most cases, mean differences between the Compact Dry and ISO methods were small (<0.5 log10) with CIs within the acceptance criterion. Repeatability, reproducibility, and RSD were similar for both methods, and r2 values were >0.97 for all matrixes. The Compact Dry TC, at 48 h, offers the advantage of a shorter time to results than ISO 4833 in an easy-to-use format.

Aldehyde fixatives are the reagents of choice to preserve human tissues for diagnostic electron microscopic examination. Since the need for ultrastructural studies is often unanticipated at the time of frozen sections, tissues are not routinely fixed in aldehydes. Alternatively, even if properly fixed, the sample may not contain representative lesions. This may require retrieval of tissue for electron microscopy from paraffin-embedded blocks or slides in which preservation of fine structure is markedly inadequate.The present study was undertaken to evaluate the use of noncryoprotected frozen tissue for diagnostic electron microscopic evaluation. The tissues samples from the following lesions were available for study: bronchial carcinoid, duodenal somatostatinoma, granular cell tumor of breast, interstitial myocarditis (from an autopsy). The effect of the rapidity of freezing on ultrastructural preservation was studied in a normal rat liver. Each case had its own control in companion samples that had been routinely fixed by immersion in 2.5% glutaraldehyde and 2% paraformaldehyde in 0.1M sodium cacodylate, pH = 7.4, (Karnovsky’s fixative).

With growing interest in food system solutions to address poor health outcomes related to preventable chronic diseases, organizations and researchers are examining the value of community gardens as interventions to promote individual and community health. Research suggests that participation in community gardens improves access to fresh, healthy foods and increases fruit and vegetable consumption. In addition to these physical benefits, research also documents a variety of social and communal benefits, by expanding social capital, stabilizing neighborhoods, and cultivating relationships. Unfortunately, most of these studies focus on a specific case, cross case, or intervention studies within a geographically specific locale. Learning lessons from successful community garden programs can be difficult because community gardens often rely on the synergy of a complex network of support agencies that assist in various technical and educational capacities. The purpose of the study was to demonstrate the use of a framework for program development and evaluation that stakeholders, including extension, can adopt to show program outcomes. The framework used a Delphi approach with a diverse panel of community garden stakeholders to reach consensus about program outcomes. The study demonstrated that the panel could reach consensus on a variety of short-, medium-, and long-term outcomes.

Abstract An enzymatic method for the determination of sulfite in foods was collaboratively studied in Nordic industry and government laboratories. The sulfite in liquid foods or extracts of solid foods is analyzed according to the following principle: Sulfite ions are oxidized to sulfate ions by oxygen in the presence of sulfite oxidase, thereby forming hydrogen peroxide. Hydrogen peroxide is transformed to water by reduced nicotinamide adenine dinucleotide (NADH) in the presence of NADH peroxidase. In this reaction, NAD+ is formed (and NADH is consumed) in amounts proportional to the sulfite concentration. Consumption of NADH can be measured spectro photo metrically at 340 nm. The method was collaboratively tested in 2 separate studies with high and low levels of sulfite tested. Results of both studies are reported here. The study samples consisted of potato flakes, wine, juice, and dried apples containing between 0 and about 960 mg SCVkg. Eleven laboratories participated in the full study and analyzed 12 samples. Six laboratories analyzed 8 samples in the complementary study. Before statistical evaluation of the collaborative study data, results were adjusted for the time-dependent decrease of sulfite in the case of materials with high sulfite content (dried apples and wine). For 2 blind duplicate samples of wine containing 75 mg S02/kg, the relative standard deviation for repeatability (RSDR, within-laboratory variation) was 3.9%. Relative standard deviation for reproducibility (RSDR, between-laboratory variation) was 7.6%. For 2 samples of dried apples containing 800 and 960 mg S02/kg, an RSDr value of 13.3% and an RSDR value of 13.9% were calculated. The corresponding parameters for 2 juice samples containing about 270 mg S02/kg were 4.8 and 10.4% for repeatability and reproducibility, respectively.

Purpose This paper is a systemic review on enset plant’s role in Ethiopian people’s life as the source of food. This paper aims to summarize the traditional processing and preparation methods of enset-based foods and their nutritional composition. Design/methodology/approach Available scientific articles were collected and reviewed for enset plant evaluation, description, enset plant’s role in Ethiopian people’s food security, post harvesting and traditional processing of enset plants, microbiology of the fermented enset foods, different foods reported from enset, nutritional profile of the three food from enset base (kocho, bulla and amicho) and other non-food applications of enset plant. Findings Enset plant has a predominant role in people living in the southern part of Ethiopia. This plant is drought-tolerant and provides many non-food applications. Harvesting of the enset plant, preparing for fermentation and food preparations follow the traditional route by using the indigenous knowledge and practices. Limited studies have been reported on the microbiology of the enset fermentation, but various types of microbes have been reported. In case of nutritional composition, foods from enset are reported to contain high carbohydrate and minerals content, such as calcium, potassium and zinc, but limited protein content; they are also the best source of the essential amino acids such as lysine and leucine. Limited data are available on vitamins, anti-oxidant and fatty acids profiles of enset-based foods. The existing data indicate variations, and the reasons for variability are discussed in this paper. Originality/value Scientific reviews on enset food nutrition profile and related issues are scarce; this paper will compile information about enset plant-based foods for researchers for their future research.

Background There are conflicting reports of allograft performance, immune response, tissue incorporation, and rerupture rates when used for anterior cruciate ligament reconstruction. Purpose To evaluate the clinical outcome of a fresh-frozen, nonirradiated, patellar tendon allograft for primary anterior cruciate ligament reconstruction surgery. Study Design Case series; Level of evidence, 4. Methods Patients who underwent endoscopic primary anterior cruciate ligament reconstruction with allograft tissue a minimum of 2 years ago were evaluated with physical examinations, the KT-1000 arthrometer, functional testing, radiographic evaluation, subjective assessment, and outcomes tools. Results Fifty-nine patients (60 knees) were evaluated at an average of 51 months after surgery. Ninety-four percent of patients were mostly or completely satisfied. A negative pivot shift test result was noted in 90% of subjects. The KT-1000 arthrometer side-to-side differences were = 3 mm in 95% of patients, and no patient exceeded 5 mm. The mean International Knee Documentation Committee score was 78 (SD = 19), and the mean Lysholm score was 82 (SD = 17). There were no clinical symptoms consistent with graft rejection or infection. Radiographic evaluation demonstrated infrequent significant tunnel widening. Conclusions Use of a fresh-frozen, nonirradiated allograft for primary reconstruction of the anterior cruciate ligament is a successful procedure both subjectively and functionally for restoring stability in patients selected for allograft reconstruction. In the patients selected for this surgical procedure, clinical, arthrometric stability testing, and subjective satisfaction were comparable to our previously published cohort studies using patellar tendon autograft at similar postoperative follow-up.

Introduction. To date, one of the priority tasks of the agro-industrial complex of Ukraine is to provide the population with benign and safe environmental and veterinary-sanitary products of animal products. Aquaculture is one of the fastest growing industries in the rapidly developing agro-industrial complex. The mentioned industry is able to provide the population with high quality nutritious and dietary fishery products in a relatively short period. Fish products replenish the human diet with essential amino acids, polyunsaturated fatty acids, trace elements and other nutrients to meet the needs of the body. But important indicators are the quality and safety of fish and fish products. The use of hazardous fish products by humans can lead to human diseases. Therefore, research on determining the safety and quality of fish remains an important link. The goal of the work. The purpose of our research was to investigate commercial fish sold in the Sumy trade network and to carry out its veterinary and sanitary evaluation. Materials and methods of research. These studies were conducted under the conditions of the Department of Veterinary Expertise, Microbiology, Zohygiene and Safety and Quality of Livestock Products of Sumy National Agrarian University and in Sumy Regional State Laboratory of the State Consumer Service. Conducted research on commercial fish, which came to the sale in the trade network of the city of Sumy, namely in supermarkets, specialized stores and agri-food markets. Fresh, dried and frozen fish were studied.. Results of research and discussion. As a result of the study of the selected samples was found in live fish, namely the fathead, characteristic signs of postodiplastomosis. This disease manifested itself as small black spots on the body of a fish. It was also revealed in the study of carp in two specimens that the chronic course of aeromonosis was manifested by the presence of fissured ulcers on the body of the fish. Since the detection of the disease did not spoil the product indicators of fish, then all living fish are allowed to sell. When examining dried fish, no deviations from the requirements for dried fish were found in the submitted samples. All fish are identified as benign and admitted for sale. Studies of fresh frozen fish (hake) revealed the presence in the muscle tissue of the nematode Contracoecum squalii Affected fish are characterized by an increased content of microorganisms in the deep and superficial muscle layers, a doubtful or negative reaction to peroxidase, a doubtful or substandard reaction with a Nesler reagent, a doubtful or substandard reaction with sulfuric acid, acidification. Conclusions and prospects for further research: 1. In a study of 323 samples of live, dried, frozen fish, which entered the trade network in Sumy, it was found that 8 (2.47%) samples did not correspond to good-quality fish.2. Signs of diseases characteristic of aeromonosis (2 specimens), postodiplastomosis (1 specimen) and contracenosis (5 specimens) were established in the selected fish samples.3. A batch of hake (170 kg) in which the pathogen Contracoecum squalii was detected is not allowed for sale and is intended for technical disposal. In the future, it is planned to develop and implement a permanent monitoring system for the quality and safety of fish and fishery products that are commercially available.

2540 Background: There are different requirements of biopsies for diagnosis vs. pharmacologic evaluation of drug mechanism biomarkers. Evaluation of core needle biopsy pairs collected pre-dose and at a defined timepoint post-dose provides insight into the pharmacodynamics of agents in early development. Adequate biopsies are key for quantifying response of the tumor cell population to molecular drug action. Tumor heterogeneity and variable tumor content make many biopsy pairs unsuitable for biomarker evaluation with any assay platform (microscopy, immunoassay, etc.). We analyzed biopsies obtained from the Developmental Therapeutics Clinic (DTC) for suitability for PD assays. Methods: Specimens obtained from 2010-2016 across 4 trials were analyzed. For microscopy measurements, biopsy pairs collected using image guidance are snap-frozen, thawed under fixative, and embedded in paraffin with control tissues. The likelihood of finding optimal regions for analysis is maximized by preparing a series of sections with H&E stained flanking slides, and annotation by an anatomic pathologist. Results: Of 112 biopsies evaluated, 26% were found to contain < 5% tumor, making them inadequate for quantitative microscopy due to a mixture of factors, with the primary factor being predominantly non-neoplastic tissue, followed by extensive mucin content, necrosis, fibrosis, inadequate size and tissue artifact post-collection. Another 20% contained ≤25% tumor, and in this group, tumor segmentation methodology during image analysis increased the rate of evaluable biopsies. 56% of tissues had > 25% tumor and were suitable for analysis. Conclusions: Improved communication between oncologists, radiologists and pathologists is key to a better understanding of factors that affect suitability of biopsies for robust PD biomarker analyses. NCI’s DTC has implemented protocol modifications including increased tissue collection and frequent case reviews by the Phase 1 team, interventional radiologists and PD team aimed at understanding features during image guidance that relate to suitability. Implementation of a scoring system has allowed the assessment of suitability of biopsies for analysis. Funded by NCI Contract No HHSN261200800001E.

Abstract BACKGROUND In Europe, the number of frozen embryo transfer (FET) cycles is steadily increasing, now accounting for more than 190 000 cycles per year. It is standard clinical practice to postpone FET for at least one menstrual cycle following a failed fresh transfer or after a freeze-all cycle. The purpose of this practice is to minimise the possible residual negative effect of ovarian stimulation on the resumption of a normal ovulatory cycle and receptivity of the endometrium. Although elective deferral of FET may unnecessarily delay time to pregnancy, immediate FET may be inefficient in a clinical setting, following an increased risk of irregular ovulatory cycles and the presence of functional cysts, increasing the risk of cycle cancellation. OBJECTIVE AND RATIONALE This review explores the impact of timing of FET in the first cycle (immediate FET) versus the second or subsequent cycle (postponed FET) following a failed fresh transfer or a freeze-all cycle on live birth rate (LBR). Secondary endpoints were implantation, pregnancy and clinical pregnancy rates (CPR) as well as miscarriage rate (MR). SEARCH METHODS We searched PubMed (MEDLINE) and EMBASE databases for MeSH and Emtree terms, as well as text words related to timing of FET, up to March 2020, in English language. There were no limitations regarding year of publication or duration of follow-up. Inclusion criteria were subfertile women aged 18-46 years with any indication for treatment with IVF/ICSI. Studies on oocyte donation were excluded. All original studies were included, except for case reports, study protocols and abstracts only. Covidence, a Cochrane-tool, was used for sorting and screening of literature. Risk of bias was assessed using the Robins-I tool and the quality of evidence using the Grading of Recommendations, Assessment, Development and Evaluation framework. OUTCOMES Out of 4124 search results, 15 studies were included in the review. Studies reporting adjusted odds ratios (aOR) for LBR, CPR and MR were included in meta-analyses. All studies (n = 15) were retrospective cohort studies involving a total of 6,304 immediate FET cycles and 13,851 postponed FET cycles including 8,019 matched controls. Twelve studies of very low to moderate quality reported no difference in LBR with immediate versus postponed FET. Two studies of moderate quality reported a statistically significant increase in LBR with immediate FET and one small study of very low quality reported better LBR with postponed FET. Trends in rates of secondary outcomes followed trends in LBR regarding timing of FET. The meta-analyses showed a significant advantage of immediate FET (n =2,076) compared to postponed FET (n =3,833), with a pooled aOR of 1.20 (95% CI 1.01–1.44) for LBR and a pooled aOR of 1.22 (95% CI 1.07–1.39) for CPR. WIDER IMPLICATIONS The results of this review indicate a slightly higher LBR and CPR in immediate versus postponed FET. Thus, the standard clinical practice of postponing FET for at least one menstrual cycle following a failed fresh transfer or a freeze-all cycle may not be best clinical practice. However, as only retrospective cohort studies were assessed, the presence of selection bias is apparent, and the quality of evidence thus seems low. Randomised controlled trials including data on cancellation rates and reasons for cancellation are highly needed to provide high-grade evidence regarding clinical practice and patient counselling.

Microorganisms can contaminate food, thus causing food spoilage and health risks when the food is consumed. Foods are not sterile; they have a natural flora and a transient flora reflecting their environment. To ensure food is safe, we must destroy these microorganisms or prevent their growth. Recurring hazards due to lapses in the handling, processing, and distribution of foods cannot be solved by obsolete methods and inadequate proposals. They require positive approach and resolution through the pooling of accumulated knowledge. As the industrial domain evolves rapidly and we are faced with pressures to continually improve both products and processes, a considerable competitive advantage can be gained by the introduction of predictive modeling in the food industry. Research and development capital concerns of the industry have been preserved by investigating the plethora of factors able to react on the final product. The presence of microorganisms in foods is critical for the quality of the food. However, microbial behavior is closely related to the properties of food itself such as water activity, pH, storage conditions, temperature, and relative humidity. The effect of these factors together contributing to permitting growth of microorganisms in foods can be predicted by mathematical modeling issued from quantitative studies on microbial populations. The use of predictive models permits us to evaluate shifts in microbial numbers in foods from harvesting to production, thus having a permanent and objective evaluation of the involving parameters. In this vein, predictive microbiology is the study of the microbial behavior in relation to certain environmental conditions, which assure food quality and safety. Microbial responses are evaluated through developed mathematical models, which must be validated for the specific case. As a result, predictive microbiology modeling is a useful tool to be applied for quantitative risk assessment. Herein, we review the predictive models that have been adapted for improvement of the food industry chain through a built virtual prototype of the final product or a process reflecting real-world conditions. It is then expected that predictive models are, nowadays, a useful and valuable tool in research as well as in industrial food conservation processes.

AbstractThe impact of ingesting various foods on psychological and behavioural functions is a topic of both interest and concern to the general public. In this article, the scientific literature concerning demonstrated cause-and-effect relationships is reviewed, beginning with methodological considerations specific to the quantification of particular behaviours and psychological events. The essential function of food is to satisfy hunger and the need for essential nutrients. The contributions of macronutrients to appetite and satiety are described, as well as their impact on metabolism and energy balance. Functional properties of macronutrient substitutes (high intensity sweeteners, fat replacers) and flavour enhancers are examined in relation to their contribution to hunger, satiety, and energy balance. The effects of foods and individual nutrients on the performance of diverse psychomotor tasks are studied with consideration given to the various validated quantitative tools used to assess behaviour. The effects of food components on activation, sedation, and affective states such as dysphoria are also reviewed, with special attention given to brain function and neuroactive substances such as serotonin and the endorphins. The case of hyperactivity in children is given special emphasis with reference to the potential influence of sugar and food additives. Safety issues related to food constituents and additives are discussed. Finally, a set of criteria is proposed for the evaluation and elaboration of studies in the behavioural and psychological fields, along with suggestions for future research.

Abstract The purpose of this study was to perform an updated reclassification of all definite prion disease cases with available fresh-frozen samples referred to the Danish Reference Center over the past 40 years, putting a special emphasis on the molecular characterization of novel disease subtypes. Investigation of the Danish prion diseases cohort revealed rare sporadic Creutzfeldt-Jakob disease cases with mixed subtypes and subtypes with previously uncharacterized white matter plaques, a new case of sporadic fatal insomnia, and 3 novel mutations, including 2 large octapeptide repeat insertions, and a point mutation in the prion protein gene. The evaluation of methionine and valine distribution at codon 129 among the prion disease patients in the cohort revealed the increased prevalence of methionine homozygotes compared to the general population. This observation was in line with the prevalence reported in other Caucasian prion disease cohort studies. Reclassification of the old prion diseases cohort revealed unique cases, the molecular characterization of which improves prion diseases classification, diagnostic accuracy, genetic counseling of affected families, and the understanding of disease biology.

Introduction: Meningiomas in childhood are infrequently seen. Furthermore, they are extremely rare at a pineal location with few reported cases above 5-years of age. We present a very young child with a large pineal region meningioma which mimicked the usually occurring pathologies at this site. Case Presentation: A 2.5-year-old child presented with symptoms of raised intracranial pressure. MRI demonstrated a homogenously enhancing pineal lesion with small cystic areas. After an initial cerebrospinal fluid evaluation for germ cell tumors, the child underwent excision of the lesion by the Krause approach. The tumor showed no definitive dural attachment, had well-defined arachnoid interface, and was completely excised. The final histopathology was meningioma. Conclusion: Although unusual, we highlight the importance of considering meningiomas among the childhood pineal region lesions, given their good outcome with total resection. Also, pertinent brief literature of the pediatric pineal region meningiomas has been provided. An assessment of preoperative and intraoperative features (clear arachnoid plane) along with adjuncts such as frozen studies can help discern various entities of this region, and decide the extent of excision.

The aim of this study was to investigate the intratumor heterogeneity of gene expression profiles in epithelial ovarian cancer (EOC). This was done to evaluate whether sampling of a single macrodissected tissue sample from each EOC case would bias the data and result in, eg, prognostic studies based on gene expression microarray experiments. From nine EOCs removed at Odense University Hospital, Denmark, three tumor samples of 200–300 mg each were taken with greatest possible mutual distance. The samples were immediately flash frozen. A parallel section was taken for histopathologic comparison. RNA was extracted from the tissue samples. Five micrograms of each RNA sample was used for labeling. The fragmented biotin-labeled complementary RNA was hybridized to Affymetrix GeneChip Human Genome U133 plus 2.0 arrays, and scanning was performed on the GeneArray scanner 3000 (Affymetrix, Santa Clara, CA). Data were evaluated using hierarchical clustering and intraclass correlation coefficient (ICC) from reliability analysis. All evaluation methods revealed low intratumor heterogeneity. Intratumor ICCs ranged from 0.888 to 0.978. In contrast, “between-tumor” ICC was 0.549 indicating much lower intra- than intertumor heterogeneity. Due to a low degree of intratumor variation, we conclude that it is sufficiently accurate in a clinical setup to use single, macrodissected tumor samples in the evaluation of gene expression in EOCs.

The health benefits attributed to pomegranate have been associated with its high content in polyphenols, particularly ellagitannins. This is also the case for other ellagitannin-containing fruits and nuts including strawberry, raspberry, blackberry, walnuts, and muscadine grapes. The bioavailability of ellagitannins and ellagic acid is however very low. These molecules suffer extensive metabolism by the gut microbiota to produce urolithins that are much better absorbed. Urolithins circulate in plasma as glucuronide and sulfate conjugates at concentrations in the range of 0.2–20 μM. It is therefore conceivable that the health effects of ellagitannin-containing products can be associated with these gut-produced urolithins, and thus the evaluation of the biological effects of these metabolites is essential. Recent research, mostly based onin vitrotesting, has shown preliminary evidence of the anti-inflammatory, anticarcinogenic, antiglycative, antioxidant, and antimicrobial effects of urolithins, supporting their potential contribution to the health effects attributed to pomegranate and ellagitannin-rich foods. The number ofin vivostudies is still limited, but they show preventive effects of urolithins on gut and systemic inflammation that encourage further research. Bothin vivoand mechanistic studies are necessary to clarify the health effects of these metabolites. Attention should be paid when designing these mechanistic studies in order to use the physiologically relevant metabolites (urolithins in gut models and their conjugated derivatives in systemic models) at concentrations that can be reachedin vivo.

Aims This pilot study tested the performance of a rapid assay for diagnosing prosthetic joint infection (PJI), which measures synovial fluid calprotectin from total hip and knee revision patients. Methods A convenience series of 69 synovial fluid samples from revision patients at the Norfolk and Norwich University Hospital were collected intraoperatively (52 hips, 17 knees) and frozen. Synovial fluid calprotectin was measured retrospectively using a new commercially available lateral flow assay for PJI diagnosis (Lyfstone AS) and compared to International Consensus Meeting (ICM) 2018 criteria and clinical case review (ICM-CR) gold standards. Results According to ICM, 24 patients were defined as PJI positive and the remaining 45 were negative. The overall accuracy of the lateral flow test compared to ICM was 75.36% (52/69, 95% CI 63.51% to 84.95%), sensitivity and specificity were 75.00% (18/24, 95% CI 53.29% to 90.23%) and 75.56% (34/45, 95% CI 60.46% to 87.12%), respectively, positive predictive value (PPV) was 62.07% (18/29, 95% CI 48.23% to 74.19%) and negative predictive value (NPV) was 85.00% (34/40, 95% CI 73.54% to 92.04%), and area under the receiver operating characteristic (ROC) curve (AUC) was 0.78 (95% CI 0.66 to 0.87). Patient data from discordant cases were reviewed by the clinical team to develop the ICM-CR gold standard. The lateral flow test performance improved significantly when compared to ICM-CR, with accuracy increasing to 82.61% (57/69, 95% CI 71.59% to 90.68%), sensitivity increasing to 94.74% (18/19, 95% CI 73.97% to 99.87%), NPV increasing to 97.50% (39/40, 95% CI 85.20% to 99.62%), and AUC increasing to 0.91 (95% CI 0.81 to 0.96). Test performance was better in knees (100.00% accurate (17/17, 95% CI 80.49% to 100.00%)) compared to hips (76.92% accurate (40/52, 95% CI 63.16% to 87.47%)). Conclusion This study demonstrates that the calprotectin lateral flow assay could be an effective diagnostic test for PJI, however additional prospective studies testing fresh samples are required. Cite this article: Bone Joint Res. 2020;9(5):202–210.

Background: Meniscal allograft transplant (MAT) is considered an effective procedure for reducing pain and improving knee function. Nevertheless, the current knowledge regarding the results of MAT is limited to short- to mid-term follow-up studies, especially for arthroscopic techniques. Purpose: To evaluate the long-term clinical outcomes, reoperations, and failures with a minimum follow-up of 10 years after fresh-frozen MAT performed arthroscopically with soft tissue fixation. Study Design: Case series; Level of evidence, 4. Methods: A total of 46 patients (age, 36.6 ± 10.6 years; 36 male, 10 female) who underwent medial MAT (n = 27) or lateral MAT (n = 19) with a minimum 10-year follow-up were evaluated with the Lysholm score, Knee injury and Osteoarthritis Outcome Score (KOOS), visual analog scale for pain, and Tegner score. Surgical failure was defined as the need for partial or total graft removal (meniscectomy or knee replacement), and clinical failure was defined as the need for partial or total graft removal in addition to a poor Lysholm score (<65 points) at final follow-up. Survival analysis was performed with Kaplan-Meier curve, and clinical scores were analyzed based on the Patient Acceptable Symptom State (PASS) for MAT. Results: 10-year survival and clinical data were available for 38 patients. Because 6 meniscectomies were required, the rate of survival free from surgical failure was 91% at 5 years and 86% at 10 years. Lower survival was reported in lateral MAT (73%) compared with medial MAT (96%). Because a further 4 patients had poor Lysholm scores, the rate of survival free from surgical and clinical failure was 87% at 5 years and 70% at 10 years. The average Lysholm score at final follow-up was 82 ± 20, and 60% to 82% of patients achieved PASS of the various KOOS subscales. The Tegner score and the KOOS Sport score significantly decreased from mid-term to long-term evaluation ( P < .001 and P < .05, respectively). Other KOOS subscales and the Lysholm score remained stable at long-term evaluation. No significant differences were found between isolated MAT or combined MAT regarding subjective scores, surgical failures, or clinical failures. Conclusion: Up to 10 years after surgery, 86% of fresh-frozen MATs with soft tissue fixation were still in situ, and satisfactory clinical results were present for about 70% of patients. Decline of clinical outcomes from midterm to long term was noted only in sports-related scores. A higher number of overall reoperations was noted in female patients, whereas a higher risk of failure was present in the patients with lateral MAT.

Abstract. Warm conveyor belts (WCBs) are important cyclone-related airstreams that are responsible for most of the cloud and precipitation formation in the extratropics. They can also substantially influence the dynamics of cyclones and the upper-level flow. So far, most of the knowledge about WCBs is based on model data from analyses, reanalyses and forecast data with only a few observational studies available. The aim of this work is to gain a detailed observational perspective on the vertical cloud and precipitation structure of WCBs during their inflow, ascent and outflow and to evaluate their representation in the new ERA5 reanalysis dataset. To this end, satellite observations from the CloudSat radar and the Cloud-Aerosol Lidar and Infrared Pathfinder Satellite Observation (CALIPSO) lidar are combined with an ERA5-based WCB climatology for nine Northern Hemisphere winters. Based on a case study and a composite analysis, the main findings can be summarized as follows. (i) WCB air masses are part of deep, strongly precipitating clouds, with cloud-top heights at 9–10 km during their ascent and an about 2–3 km deep layer with supercooled liquid water co-existing with ice above the melting layer. The maximum surface precipitation occurs when the WCB is at about 2–4 km height. (ii) Convective clouds can be observed above the inflow and during the ascent. (iii) At upper levels, the WCB outflow is typically located near the top of a 3 km deep cirrus layer. (iv) There is a large variability between WCBs in terms of cloud structure, peak reflectivity and associated surface precipitation. (v) The WCB trajectories with the highest radar reflectivities are mainly located over the North Atlantic and North Pacific, and – apart from the inflow – they occur at relatively low latitudes. They are associated with particularly deep and strongly precipitating clouds that occur not only during the ascent but also in the inflow and outflow regions. (vi) ERA5 represents the WCB clouds remarkably well in terms of position, thermodynamic phase and frozen hydrometeor distribution, although it underestimates the high ice and snow values in the mixed-phase clouds near the melting layer. (vii) In the lower troposphere, high potential vorticity is diabatically produced along the WCB in areas with high reflectivities and hydrometeor contents, and at upper levels, low potential vorticity prevails in the cirrus layer in the WCB outflow. The study provides important observational insight into the internal cloud structure of WCBs and emphasizes the ability of ERA5 to essentially capture the observed pattern but also reveals many small- and mesoscale structures observed by the remote sensing instruments but not captured by ERA5.

On 16 October 1996, a malfunction at the Swan Hills Special Waste Treatment Center (SHSWTC) in Alberta, Canada, released an undetermined quantity of persistent organic pollutants (POPs) into the atmosphere, including polychlorinated biphenyls, dioxins, and furans. The circumstances of exposure are detailed in Part 1, Background and Policy Issues. An ecologically based, staged health risk assessment was conducted in two parts with two levels of government as sponsors. The first, called the Swan Hills Study, is described in Part 2. A subsequent evaluation, described here in Part 3, was undertaken by Health Canada and focused exclusively on Aboriginal residents in three communities living near the lake, downwind, and downstream of the SHSWTC of the area. It was designed to isolate effects on members living a more traditional Aboriginal lifestyle. Aboriginal communities place great cultural emphasis on access to traditional lands and derive both cultural and health benefits from “country foods” such as venison (deer meat) and local fish. The suspicion of contamination of traditional lands and the food supply made risk management exceptionally difficult in this situation. The conclusion of both the Swan Hills and Lesser Slave Lake studies was that although POPs had entered the ecosystem, no effect could be demonstrated on human exposure or health outcome attributable to the incident. However, the value of this case study is in the detail of the process, not the ultimate dimensions of risk. The findings of the Lesser Slave Lake Study have not been published previously and are incomplete.

15071 Background: Cancer cells have been shown to exhibit many defects in gap junctions, which contribute to the loss of growth control and tissue homeostasis. In ovarian tissues, it is known that gap junction communication is predominantly mediated by connexin 43 (cx43) proteins. Recent studies have demonstrated that while human ovarian surface epithelial cells exhibit extensive expression of cx43, this protein is nearly absent in the vast majority of ovarian cancers. Differences in cx43 expression between normal and malignant ovarian tissue at the mRNA level has heretofore not been reported. Methods: In the present study, specimens of normal ovaries and serous adenocarcinomas were obtained at the time of laparotomy and flash frozen in liquid nitrogen. Results: Evaluation of cx43 mRNA levels by semiquantitative RT-PCR indicated significantly reduced expression in tissues obtained from ovarian adenocarcinomas compared to that from normal ovaries; the average relative levels of cx43 mRNA in tumors were less than 30% of that found in normal tissue. In one case in which tumor and adjacent normal tissue was obtained from the same patient, cx43 mRNA expression was reduced in the former to less than 10% of that found in the normal section. This data represents, to our knowledge, the first demonstration showing reduced cx43 mRNA in ovarian carcinoma. Conclusion: These findings suggest that the defect in cx43 expression in this tumor type is at the level of transcription and support the contention that its downregulation is involved in neoplastic transformation of ovarian epithelial cells. (This study was supported in part by the Vesa W. and William J. Hardman, Jr. Charitable Foundation Inc.) No significant financial relationships to disclose.

Object. Endolymphatic sac (ELS) tumors are low-grade malignancies of the temporal bone that are associated with von Hippel—Lindau (VHL) disease but can also occur sporadically. The VHL gene product VHL protein is important in the regulation of hypoxia inducible factor (HIF)-1α, which controls expression of molecules that are important in angiogenesis and cell metabolism. In this study the authors examine the role of VHL and HIF-1 in ELS tumors. Methods. The ELS tumors from three patients were examined using the following method: DNA from tumor tissue was isolated, amplified by polymerase chain reaction and the VHL gene sequence was compared with the known wild-type sequence. Loss of heterozygosity (LOH) studies were performed to confirm the sequencing data. Immunohistochemical evaluation for VHL, HIF-1α, vascular endothelial growth factor (VEGF), and carbonic anhydrase IX (CA IX) was performed. Snap-frozen tumor tissue was examined using Western blot and HIF-1 immunoassays for HIF-1α and VHL expression. Two patients had sporadic ELS tumors and the other one suffered from VHL disease. Results of VHL gene sequencing were normal in the tissue derived from the sporadic ELS tumors. The ELS tumor, pheochromocytoma, and spinal hemangioblastoma were heterozygous for the same C-to-A transversion found in the germline carried by the patient with VHL disease. No LOH was detected in the tumor tissue obtained in the patient with VHL disease. Expression of HIF-1α, VEGF, and CA IX evaluated using immunohistochemical studies was elevated in the VHL-associated tumors. Nevertheless, Western blots and immunoassays for HIF-1α did not show elevated expression in these tumors. Conclusions. The sporadic and VHL disease—associated ELS tumors in this study had normal VHL-mediated HIF-1 regulation. This is a result of normal VHL gene expression in the case of the sporadic ELS tumor. In the VHL-associated ELS tumor, this is due to one normal copy of the VHL gene and adequate VHL gene expression.

Background Frozen shoulder causes pain and stiffness. It affects around 10% of people in their fifties and is slightly more common in women. Costly and invasive surgical interventions are used, without high-quality evidence that these are effective. Objectives To compare the clinical effectiveness and cost-effectiveness of three treatments in secondary care for adults with frozen shoulder; to qualitatively explore the acceptability of these treatments to patients and health-care professionals; and to update a systematic review to explore the trial findings in the context of existing evidence for the three treatments. Design This was a pragmatic, parallel-group, multicentre, open-label, three-arm, randomised superiority trial with unequal allocation (2 : 2 : 1). An economic evaluation and a nested qualitative study were also carried out. Setting The orthopaedic departments of 35 hospitals across the UK were recruited from April 2015, with final follow-up in December 2018. Participants Participants were adults (aged ≥ 18 years) with unilateral frozen shoulder, characterised by restriction of passive external rotation in the affected shoulder to < 50% of the opposite shoulder, and with plain radiographs excluding other pathology. Interventions The inventions were early structured physiotherapy with a steroid injection, manipulation under anaesthesia with a steroid injection and arthroscopic capsular release followed by manipulation. Both of the surgical interventions were followed with post-procedural physiotherapy. Main outcome measures The primary outcome and end point was the Oxford Shoulder Score at 12 months post randomisation. A difference of 5 points between early structured physiotherapy and manipulation under anaesthesia or arthroscopic capsular release or of 4 points between manipulation under anaesthesia and arthroscopic capsular release was judged clinically important. Results The mean age of the 503 participants was 54 years; 319 were female (63%) and 150 had diabetes (30%). The primary analyses comprised 473 participants (94%). At the primary end point of 12 months, participants randomised to arthroscopic capsular release had, on average, a statistically significantly higher (better) Oxford Shoulder Score than those randomised to manipulation under anaesthesia (2.01 points, 95% confidence interval 0.10 to 3.91 points; p = 0.04) or early structured physiotherapy (3.06 points, 95% confidence interval 0.71 to 5.41 points; p = 0.01). Manipulation under anaesthesia did not result in statistically significantly better Oxford Shoulder Score than early structured physiotherapy (1.05 points, 95% confidence interval –1.28 to 3.39 points; p = 0.38). No differences were deemed of clinical importance. Serious adverse events were rare but occurred in participants randomised to surgery (arthroscopic capsular release, n = 8; manipulation under anaesthesia, n = 2). There was, however, one serious adverse event in a participant who received non-trial physiotherapy. The base-case economic analysis showed that manipulation under anaesthesia was more expensive than early structured physiotherapy, with slightly better utilities. The incremental cost-effectiveness ratio for manipulation under anaesthesia was £6984 per additional quality-adjusted life-year, and this intervention was probably 86% cost-effective at the threshold of £20,000 per quality-adjusted life-year. Arthroscopic capsular release was more costly than early structured physiotherapy and manipulation under anaesthesia, with no statistically significant benefit in utilities. Participants in the qualitative study wanted early medical help and a quicker pathway to resolve their shoulder problem. Nine studies were identified from the updated systematic review, including UK FROST, of which only two could be pooled, and found that arthroscopic capsular release was more effective than physiotherapy in the long-term shoulder functioning of patients, but not to the clinically important magnitude used in UK FROST. Limitations Implementing physiotherapy to the trial standard in clinical practice might prove challenging but could avoid theatre use and post-procedural physiotherapy. There are potential confounding effects of waiting times in the trial. Conclusions None of the three interventions was clearly superior. Early structured physiotherapy with a steroid injection is an accessible and low-cost option. Manipulation under anaesthesia is the most cost-effective option. Arthroscopic capsular release carries higher risks and higher costs. Future work Evaluation in a randomised controlled trial is recommended to address the increasing popularity of hydrodilatation despite the paucity of high-quality evidence. Trial registration Current Controlled Trials ISRCTN48804508. Funding This project was funded by the National Institute for Health Research (NIHR) Health Technology Assessment programme and will be published in full in Health Technology Assessment; Vol. 24, No. 71. See the NIHR Journals Library website for further project information.

Background Acquired coagulopathies are a common problem in Hematology, and are most often due either to medication effect, liver disease, or consumption. Among the uncommon causes of acquired coagulopathy, inhibitory auto-antibodies may develop, either in the setting of autoimmune diseases, in the setting of lymphoproliferative disorders, or as isolated inhibitory immunoglobulins. Uncommonly, the adsorption of coagulation factors from the circulation into the tissues by extracellular deposition of pathologic amyloid results in an acquired factor deficiency, due to clearance of factor from the circulation that exceeds the body's ability to produce factor. When amyoidosis does cause a coagulapathy, it is most often the result of the adsorption of Factor X by the amyloid protein, resulting in an acquired Factor X deficiency. However, there are rare reports of amyloidosis being associated with other factor deficiencies. We report a case of amyloidosis that was associated with a severe bleeding diathesis, with the etiology of the bleeding disorder being due to both acquired Factor V deficiency and concomitant acquired von Willebrand Disease. Case Report A previously healthy 51-year-old gentleman presented to an outside medical center for evaluation and management of recurrent bleeding episodes. The patient had a prior medical history significant only for right ankle trauma in the year 2005, following which he underwent a total of 4 surgical procedures; there was no excessive bleeding complicating the patient's surgeries. He was then in his usual state of health until September, 2012 when he developed onset of severe abdominal pain and was admitted to the outside facility. Following hospitalization for several months at the outside facility, he was admitted to our institution. Physical examination was remarkable for extensive ecchymoses, and for splenomegaly to 18 cm. span by exam, confirmed by imaging. CT scan showed multiple peri-caval and periaortic nodes present up to 1.7 cm in size, with shotty inguinal lymph nodes. A complete blood count showed White blood count 21,600, hemoglobin 8.0 g/dL, hematocrit 24%, platelet count 370,000, Hepatic function studies and renal function studies, as well as electrolytes, were normal on admission. Coagulation studies revealed Prothrombin Time prolonged at 16.8 seconds (normal < 12.7), aPTT prolonged at 44. Mixing patient plasma with equal volume normal plasma corrected both the PT and aPTT. Detailed factor assays showed markedly decreased Factor V activity of 27%; Ristocetin Cofactor activity was markedly decreased at 49%, but von Willebrand antigen was elevated at 213%. Multimer analysis was consistent with Type II vWD (see figure 1). The patient received fresh frozen plasma and Humate P, with transient correction of the bleeding diathesis. This permitted inguinal lymph node biopsy, which documented AL amyloidosis. Extraction of the protein from the lymph node documented AL lambda light chain amyloid (see figure 2). Marrow biopsy documented IgG lambda multiple myeloma. The patient was treated using Bortezumib plus Dexamethasone, and achieved a complete remission, with normalization of the coagulation parameters and factor levels over the following several months. His bleeding diathesis has fully resolved, and Karnofsky performance status improved to 100%. Conclusion Although there are several case reports of acquired von Willebrand disease on the basis of amyloidosis, and several case reports of acquired Factor V deficiency on the basis of amyloidosis, this appears to be the first reported case of both acquired vWD and acquired Factor V deficiency on the basis of amyloidosis. Disclosures: No relevant conflicts of interest to declare.

Abstract Case: A Caucasian male infant, born at 39 weeks and 2 days was assessed at 7 hours of life for new onset purpuric changes in toes 1-2, and 5 of the right foot. The course of the pregnancy was complicated by a fetal cardiac mass seen on ultrasound, thus requiring a C-section. Initial course of delivery was complicated by chorioamnionitis due to maternal fever. The patient's mother was treated appropriately with antibiotics prior to delivery. After delivery, APGAR scores were 9/9/9. At day 6 of life, there was progressive discoloration of the R. foot digits 1,2,3, and 5, disproportionate to the general acrocyanosis of the rest of the body. NICU was consulted and he was promptly transferred. Initial examination showed good pulses and oxygenation of the involved limb. No other areas of disproportionate discoloration were seen. Initial evaluation included arterial and venous duplex studies. Each exam showed no evidence of decreased blood flow. Pediatric hematology and vascular surgery were consulted. Per vascular surgery recommendations an aortic duplex was obtained, showing no evidence of thrombosis. Due to clinical picture of purpuric digits, Neonatal Purpura Fulminans was strongly suspected. Initial lab work obtained showed a WBC of 20.5, Hgb/Hct 18.2/55.4, and a platelet count of 196,000. Initial fibrinogen was undetectable. PTT and PT/INR were unable to be obtained. Protein S and Protein C Levels were drawn prior to any intervention. Initial Protein S level was 35%, and initial Protein C level was <20%. After intervention, Protein C levels were redrawn on day 4 of life, and shown to be 48% Due to the initial lab work obtained, the patient was initially started on Fresh Frozen Plasma 20 ml/kg, and Low Molecular Weight Heparin 1.5 mg/kg q 12 hrs. Nitroglycerin paste was applied to the R. foot and toes. After Protein C levels returned undetectable, the patient was switched from FFP to protein C concentrate, (100 U/kg q12h). Warfarin was started on day 3 of life, for transition to long term anticoagulation, with a titration to goal INR of 2.5-3.0. During hospitalization, CT head w/o contrast, and a kidney ultrasound were ordered and seen to be normal. An ophthalmologic examination was also done on day 4 of life, and showed no retinal or vitreous hemorrhage. Following normal screening lab work, and proper transition to Warfarin therapy for outpatient management, patient was discharged home on day 14 of life, with close follow with hematology and genetics. Patient was discharged home on Warfarin 2 mg daily, with a goal INR >2.2. The patient was followed by pediatric hematology after discharge. A second opinion was sought at an outside hospital, at which time patient was trialed off of Warfarin. Genetic studies showed a heterozygous mutation for Factor V Leiden R506Q polymorphism, with normal protein C levels after a trial off of Warfarin, making inherited Protein C deficiency less likely. Discussion: Neonatal Purpura Fulminans is a rare disorder that can be either acquired or inherited. The differential diagnosis for acquired protein C deficiency is vast, ranging from infectious causes with and without DIC, hypoxia, severe hepatic dysfunction, and severe congenital heart disease. Congenital forms of protein C deficiency are generally homozygous. However, the clinician should be aware of the possibility of Factor V Leiden heterozygous mutation causing activated protein C resistance and acquired protein C deficiency in infancy. The concern in this specific case was that the infant may have otherwise been considered to have inherited protein C deficiency until genetic testing revealed otherwise. While waiting for genetic testing results, the infant was carefully monitored on oral anticoagulation. While these measures were taken and helped to prevent thrombosis, no official guidelines are in place for surveillance, treatment, or testing frequency of Protein C levels in infancy. To date, only one other case of Neonatal Purpura Fulminans in association with Factor V R506Q mutation has been reported. In this case, the authors concluded that activated protein C resistance should be included in the evaluation of Neonatal Purpura Fulminans. We hope to build on the existing management guidelines in order to more promptly identify infants with Neonatal Purpura Fulminans and transient protein C deficiency. Disclosures No relevant conflicts of interest to declare.

Abstract Abstract 2259 Introduction: In Congenital Bleeding Disorders (CBD), efficacy of Replacement Therapy (RT) can be indirectly measured on the basis of pharmacokinetic (PK) methods. Therapeutic concentrations of clotting factors in the blood-stream at a given time depend on the dose and frequency of RT and the fall off patterns specific for each clotting factor (i.e. PK properties). The issue of the frequency of RT administration is particularly relevant in Factor VII (FVII) deficiency, a CBD characterized by the lack of a rare protein with a very short half-life. Recombinant FVIIa (rFVIIa), plasma-derived FVII (pdFVII) concentrates and Fresh Frozen Plasma (FFP) are used for on-demand or prophylaxis replacement despite reported half-lives in the range of 2–5 hours. Very limited information is available with reference to the pharmacokinetic parameters of infused FVII in FVII deficiency. The same holds for the In Vivo Recovery (IVR). In this study, we evaluated the PK of rFVIIa in 11 severe (FVIIc <2%) FVII deficient patients. Further, evaluation of “incremental” IVR was performed in 116 patients with a FVII deficiency (90 for rFVIIa, 19 for pdFVII concentrates and 7 for FFP). Methods & Results: Eleven severe FVII-deficient individuals in the non-bleeding state were given rFVIIa doses ranging from 16 to 24 μg/Kg/bw (mean 19.7, median 20). Pharmacokinetic parameters of rFVIIa were analyzed with reference to FVIIc post-infusion levels. Analyses of the PK parameters are detailed in Tab. 1. In Tab. 2 results regarding the IVR analyses of rFVIIa, pdFVII and FFP are reported, on the basis of FVIIc 15' after replacement. IVR data were also evaluated with reference to the baseline FVIIc, age and FVII mutation zygosity. Conclusions: PK data are characterized by an optimal correspondence between FVIIc levels and time; data variability is ample considering that all individuals were adults or teen-agers. Terminal half-life appeared longer than the Mean Residence Time (MRT) or T1/2, a data that can be interpreted as a reduction of the factor flow from the plasma pool to the extravascular space at the end of the fall off curve; this is confirmed by the Apparent volume of distribution based on the terminal phase (Vz) that is higher than the Apparent Volume of distribution at equilibrium (Vss). MRT and T1/2 are in keeping with previous studies and appear lower than those already reported for the pdFVII concentrates. Quite variable is also the Clearance (CL) showing that metabolic degradation and/or FVIIa vascular uptake greatly differ among patients. The latter aspect points towards the need for a CL individualized evaluation for patients who are eligible for continuous infusion protocols. Incremental IVRs of rFVIIa and FFP were lower (p< 0.001) than those calculated for the pdFVII concentrates. The difference between the recoveries of rFVIIa and pdFVII can either be ascribed to the FVIIc assay (only FVIIa assayed in the case of rFVIIa administration, FVII zymogen and FVIIa assayed in the case of pdFVII concentrates), to a more rapid disappearance rate of FVIIa from the vascular compartment or, finally, to a more rapid uptake by the FVIIa receptors (TF on the pericytes and the PC receptor on the endothelial cells). No difference was found between IVRs in children and adults nor between individuals with baseline FVIIc <2% or ≥ 2%. Also, no differences were found between patients homozygous or compound heterozygous for FVII gene lesions. Disclosures: No relevant conflicts of interest to declare.

Congenital plasminogen deficiency (C-PLGD) is a rare autosomal recessive multisystem disorder of the fibrinolytic system that is often underdiagnosed. It is caused by mutations in the PLG gene and is characterized by extravascular fibrinous deposits on mucus membranes resulting in tissue injury or organ dysfunction. The abnormal accumulation or growth of fibrin-rich pseudomembranous lesions on mucous membranes, such as the conjunctiva, gingiva, linings of airways and genitourinary tract have been termed ligneous for their "woody" appearance. Ligneous conjunctivitis is the most common manifestation of C-PLGD occurring in greater than 80% of affected patients. Ligneous lesions affecting the female reproductive system have been reported in one large case series in 9% of patients (Shuster et. al. JTH 2007). An extensive review of the literature (Baithun et. al. Haemophilia 2018) identified 30 cases of C-PLGD patients with gynecologic involvement. A diagnosis of infertility, when performed, was reported in 8 (26.7%) of these females of child-bearing age. Due to the rarity of the disease, infertility may be under diagnosed and under reported in the literature as no natural history studies of this disease have been conducted. We report a case of a patient with C-PLGD with a documented diagnosis of infertility who received intravenous plasminogen therapy. Although the protocol stated that adequate contraceptive measures be taken to avoid becoming pregnant during the study, she became pregnant, went to term and had a successful delivery of a healthy child. She was a 33-year-old woman with a diagnosis of C-PLGD in childhood, after developing ligneous conjunctivitis before the age of 1, developed uterine ligneous lesions and dysmenorrhea as a teenager. At age 27, a hysteroscopy was attempted and laparoscopy was performed which revealed fibrosis and web-like fibrin-rich adhesions in the uterus and fallopian tubes. She was referred to the Oslo University Hospital, Norway, for evaluation and management of her infertility and eye lesions at the age of 28. She was treated with fresh frozen plasma (FFP) infusions with improvement in eye symptoms and dysmenorrhea. At the age of 29, a vaginoscopy and hysteroscopy were performed which again demonstrated fibrin membranes in the uterus. A repeat evaluation at the age of 30 revealed fibrin membranes in the cervix, fibrin accumulation in the uterus, and tube ostia obstructed. At age 33 she was enrolled in a clinical trial, an open-label phase 2/3 study of human Glu-plasminogen administered intravenously at 6.6 mg/kg every 2-4 days in 15 patients with plasminogen deficiency, registered at www.clinicaltrials.gov as #NCT02690714. She received intravenous plasminogen replacement therapy, starting in October 2016. At baseline, she had ligneous conjunctivitis and gynecologic lesions with regular, but atypical menstrual bleeding, discharging very small amounts of thick, mucus-like material over 1-2 days. Her eye lesions had resolved by week 12 of treatment. At the end of the first menstrual cycle on plasminogen treatment, she discharged a large mucus-like plug from the vagina, and had regular and normal menstrual bleeds after that. Approximately 20 weeks into treatment, she reported being pregnant to the investigator. After discussion with the clinical study sponsor, regulatory agencies and the regional ethical committee, the patient and her physician, it was agreed to continue plasminogen replacement therapy through the pregnancy, delivery and recovery period. After approximately 55 weeks into the study treatment, the patient had an uncomplicated pregnancy and delivered a healthy child with APGAR score of 9. The patient remained on PLG treatment post-partum, and her PLG activity trough levels were maintained until she came off the study when her levels went back to baseline. (Figure 1) This is the first reported case of intravenous plasminogen used in a patient with C-PLGD and documented infertility who subsequently became pregnant and delivered a healthy child. Figure Disclosures Parker: Liminal BioSciences: Current Employment, Current equity holder in publicly-traded company. Trybul:Liminal BioSciences: Current Employment, Current equity holder in publicly-traded company. Sandset:Liminal BioSciences: Other: Investigator Clinical Trial.

Abstract Introduction: JAK2 V617Fmutation is present in majority of polycythemia vera (PV) and in 50-60% of patients with primary myelofibrosis (MF). JAK1/2 inhibitor ruxolitinib (RUX) is approved for treatment of both PV and intermediate/high risk MF. Two independent randomized studies showed the benefit of RUX in MF with significant reduction in spleen volume and improvement in constitutional symptoms compared to best supportive care. Clonal evolution may occur in PV and MF upon acquisition of new mutations and transformation to acute myeloid leukemia. Rarely, patients with JAK2V617Fpositive PV or MF have also concomitant BCR-ABL1 fusion gene with phenotype of chronic myelogenous leukemia (CML). Imatinib, a BCR-ABL1 inhibitor, induces long-term cytogenetic and molecular remissions in CML. Single agent imatinib has shown no clinical benefit in MF patients despite in vitro efficacy (Gaikwad et al Exp. Hematol. 2007). We describe here a patient with JAK2V617F positive post-PV MF who later developed concomitant BCR-ABL+ CML. Splenomegaly was initially resolved by RUX but returned coincidently with the diagnosis of CML and was again normalized by imatinib alone. Massive splenomegaly reappeared along with elevated hematocrit on imatinib and was resolved by the addition of low dose RUX. Case report: A 71-year old woman initially presented in September 2011 with isolated erythrocytosis and splenomegaly. A diagnosis of PV was established based on the presence of the JAK2V617Fmutation and hypercellular marrow. Bone marrow karyotype was normal (Table 1). The patient was randomized to pegylated interferon arm in the MPD Consortium study. A complete hematologic response was achieved after nearly 2 years on maximal dose 180 mcg weekly. In early 2015, she developed constitutional symptoms, and progressive splenomegaly. Progression to post-PV MF was confirmed on bone marrow evaluation. The patient was taken off study and started on RUX 20 mg BID. She developed severe anemia 4 months later requiring dose reduction to 10 mg BID. The anemia markedly improved, while JAK2V617F allele burden remained high at 88% with normal cytogenetics. In December 2017, the patient developed anemia, fatigue and rapid regrowth of spleen. Marrow karyotype showed Philadelphia (Ph) chromosome in 25% of the cells analyzed, PCR was positive for BCR-ABL1 p210, and JAK2V617F allele burden was reduced to 56.4%. In January 2018, she started imatinib 400 mg daily and RUX was discontinued due to toxicity concerns. In April 2018, her hematocrit increased and massive splenomegaly returned. Restart of initial RUX 20 mg BID was followed by severe pancytopenia which normalized when RUX was reduced to 5 mg BID with continuing imatinib regimen. After 8 weeks, splenomegaly completely resolved (Figure 1). A complete cytogenetic response and major molecular response were achieved after 3 months and 6 months of combination therapy respectively. Methods: Mononuclear cells from peripheral blood collected in June 2018 were plated in methylcellulose without erythropoietin or other cytokines. After 14 days, colonies were plucked, with ½ of each colony used for JAK2V617F allele-specific PCR and ½ prepared as cytospins for BCR-ABL fluorescent in situ hybridization (FISH) with the Vysis LSI BCR/ABL Dual Color, Dual Fusion Translocation Probe from Abbott Laboratories. Results: Analysis of 3-6 EPO-independent colonies of each CFU-E, G, M, GM, GEMM, BFU-E showed no BCR-ABL1 positive cells, while a single CFU-GEMM colony was positive and also homozygous for JAK2V617F. Frozen marrow cells from the time of the original diagnosis are being analyzed. Conclusion: The recurrence of splenomegaly in this patient following single agent RUX indicates emergence of novel BCR-ABL1+ clone. While single agent imatinib temporarily normalized the splenomegaly and leukocytosis for several months, MF phenotype reoccurred. Splenomegaly resolved completely in 8 weeks with the addition of very low dose RUX to standard dose imatinib, providing the first clinical observation of in vivo synergism of RUX and imatinib. We are investigating serial samples during her course, which will be presented at the meeting. The analysis of a single clone demonstrate that BCR-ABL1 mutation did not occur in previously normal dormant stem cell but in JAK2V617F positive progenitors and represent in this pts a novel subclone of previous PV/MF stem cell. Disclosures Deininger: Pfizer: Consultancy, Membership on an entity's Board of Directors or advisory committees; Blueprint: Consultancy.

Introduction: Nesidioblastosis is a hyperplasia of the beta cells that causes persistent hyperinsulinemic hypoglycemia, unfortunately, it is difficult to diagnose through imaging studies. Case presentation: A 68-year-old woman with palpitations, diaphoresis, dizziness and alterations in the level of consciousness that improve with food intake. Laboratories: glucose 35.0 mg/dl, insulin 12.5 mUI/ml, proinsulin 14.1 pmol/L and peptide C 2.55 ng/ml. Octreoscan reveals an abnormal epigastric uptake area and the tomography shows a focal hypervascular lesion in pancreatic body of 12×11 mm. A distal pancreatectomy was performed without palpating the tumor in transoperative period, and a diffuse nesiodioblastosis was reported in the histopathological study. The patient persists with hypoglycemia and an additional pancreatic resection was performed, resecting 90% of the organ. Although an insulinoma was not located in the piece, an area surrounded by connective tissue was observed where islet hyperplasia was more accentuated. Discussion: The pathophysiology of nesidioblastosis is unknown, its main differential diagnosis is insulinoma and this should be suspected when the tumor can not be identified. Most agree that a pancreatectomy that involves 60-80% of the total organ can control glucose levels with low risk of diabetes and pancreatic insufficiency. Conclusion: The nesidioblastosis should be suspected when a hyperinsulinemic hypoglycemia is difficult to control and when a tumor can not be identified. The extension of the pancreatectomy should be individualized and if an insulinoma is not localized and a nesidioblastosis is suspected, an intraoperative histopathological examination with frozen section evaluation for the margins could determine the extent of pancreatectomy.

Abstract Background Norovirus is the etiology for about 60% of foodborne outbreaks identified in Minnesota. Contamination of food during preparation by food handlers is by far the most common cause of these outbreaks. Norovirus outbreaks due to commercially distributed foods are rarely reported in the United States, and only 2 have been previously identified in Minnesota, both due to oysters. Methods In August 2016, we investigated an outbreak of norovirus gastroenteritis in Minnesota that was linked to consumption of commercially distributed ice cream at multiple venues. Sanitarians from local public health agencies visited the facilities involved for follow-up, and case-control studies were conducted. The outbreak was identified by linking multiple independent illness reports to a centralized foodborne illness complaint system and subsequently confirmed though genotyping of stool specimens. Results A total of 15 cases from 4 venues were reported. Raspberry chocolate chip ice cream was statistically associated with illness in 2 analytic studies (6 of 7 cases vs 0 of 7 controls; odds ratio, undefined; P = .005). Norovirus GII.17[P17] (GII.17 Kawasaki) strains from case stool specimens matched norovirus found in frozen raspberries imported from China that were used to make the implicated ice cream. Conclusions To our knowledge, this is the first norovirus outbreak due to commercially distributed frozen berries identified in the United States. To detect norovirus outbreaks associated with commercially distributed food vehicles, investigators should thoroughly investigate all norovirus outbreaks (including stool testing and genotyping), coordinate complaint and response activities across agencies and jurisdictions, and consider testing food for norovirus when appropriate.

During evaluation of physical and chemical properties of sandy soils and theirfertility in Southern part of Republic of Chad it has been revealed that some soilshave very high content of strontium. Its content varies from 10 to 270 mg/kg ofsoil depending on type of soil, depth of soil layers, clay and organic content.Strontium content negatively correlates with total content of calcium andphosphorus in layers of soil. Low CEC (CEC - Cation-exchange capacity) of soilmay be a reason of possible translocation of strontium from higher to lower layersof soils. Strontium content in soils do not relates with level of radioactivity of soilmeasured. The highest content of strontium has been found in soils developed onsome eolian and colluvio-alluvium deposits. Some researchers hypothesize thatsome endemic and chronic diseases such as Kashin-Beck disease, `Dysostosisenchondralis endemic`, endemic hoiter, osteoarthritis might be caused by highcontent of strontium in water and plant foods contaminated with it. Absence ofconsensus on etiological factors of these diseases confirms that it is worthconsidering necessity of further studies of different affects of high content ofstrontium in water and foods on human health directly or indirectly throughcausing misbalance in mineral nutrition.

Introduction: Adhesive capsulitis, also called frozen shoulder, affects about 3-5% of the population, especially in the age group 40 to 60 years, affecting more women and having an insidious onset. Objective: Analyze the isolated effect of joint manipulation by Maitland method in treating patients with adhesive capsulitis of the shoulder, the level of pain, range of motions of the shoulder and functional capacity. Method: It is about a cases studies in which four volunteers underwent protocol Maitland to shoulder for 15 sessions. The evaluation instruments used were the Visual Analog Scale, Goniometer and Shoulder Pain and Disability Index to measure the level of disability of the shoulder. Results: There was decreased pain from 55.5 to 80.0%, increased all range of motions, especially flexion of the shoulder, with more than 100% gains in the four cases investigated and improved functional capacity between 27.9 – 91.9%. Conclusion: These results suggest that the therapy had improved outcomes assessed, as clinically relevant contribution, but indicate the need to develop more studies to prove the effectiveness of the method isolated.

Abstract Milk and its derivatives are fundamental foods in the human diet. However, milk is highly susceptible to microbial contamination, among which is Aflatoxin M1 (AFM1), presenting a risk to public health. Aflatoxin M1 is the main hydroxylated metabolite of aflatoxin B1 present in the milk of animals that have ingested feed contaminated with B1. These toxins have carcinogenic and genotoxic potential and similar toxicities. According to Resolution RDC 07/2011, published by the National Health Surveillance Agency (ANVISA), the level of tolerance for AFM1 is 0.5 µg / Kg in Brazil. The present study aimed to evaluate qualitatively and quantitatively Aflatoxin M1 in raw milk samples, being 2 samples of bovine milk and one sample of goat milk. The raw, bovine and goat milk samples were purchased from local stores and were kept at approximately 5 ºC for 8 hours and subsequently frozen at 6 ºC for conservation purposes. The tests were carried out at the premises of Hexis Científica, Av. Antonieta Piva Barranqueiros, 385 - Distrito Industial - Jundiaí-SP, which provided training and use of the laboratory. The Scientific Charm Kits, Test SLAFMQ-EZ-20K were used. It is a quantitative test that uses ROSA immunoreceptors (Rapid Assay Step One) lateral flow technology. The milk sample interacts with colored spheres and the intensity of the color in the test area and is read as ppt (parts per trillion) by the reader. According to the analyzes, all the samples studied presented Aflatoxin M1, with a concentration of 2,507 ppb; 2,410 ppb; 2,627 ppb, respectively for bovine and caprine milk samples, which characterizes a concentration five times above the allowed by the current legislation, according to RDC 07/2011, published by the National Health Surveillance Agency (ANVISA). Key messages According to studies, most of the negative results obtained on the incidence of aflatoxin M1 can be attributed to the low sensitivity of the analytical methods employed. The presence of Aflatoxins M1, represents a public health issue, since milk is among the products most consumed by man and, constituting the basis of infant feeding.

AbstractThe role of dietary phyto-oestrogens in health has been of continued interest and debate, but data available on the distribution of intake in the Australian diet are scarce. Therefore, we aimed to estimate phyto-oestrogen consumption in Australian women, describe the pattern of intake and identify correlates of high phyto-oestrogen intake. Study participants were 2078 control women (18–79 years) from two population-based case–control studies on gynaecological cancers (2002–2007). Dietary information was obtained using a 135-item FFQ, and the intakes of isoflavones, lignans, enterolignans and coumestans, including their individual components, were estimated using a database of phyto-oestrogen content in food developed in the UK. Median total intake (energy-adjusted) of phyto-oestrogens was 1·29 mg/d, including 611 µg/d isoflavones, 639 µg/d lignans, 21 µg/d enterolignans and 8 µg/d coumestrol. Both isoflavone and lignan intakes were strongly skewed towards higher values and positively correlated with age. Women consumed on average two servings of soyabean foods/week. Compared to lower phyto-oestrogen consumers (≤1·29 mg/d, median split), higher phyto-oestrogen consumers (>1·29 mg/d) were slightly older, less likely to be smokers, had a higher educational and physical activity level, lower BMI, lower intake of dietary fat, and higher intake of fibre, selected micronutrients and soyabean foods (all P < 0·03). The daily intake of phyto-oestrogens in Australian women with predominantly Caucasian ethnicity is approximately 1 mg; this is similar to other Western populations, but considerably lower than that among Asian women. However, those with a relatively high phyto-oestrogen diet seem to have a healthier lifestyle and a more favourable dietary profile compared to others.

Abstract Background Despite adherence to WHO guidelines, inpatient mortality among sick children admitted to hospital with complicated severe acute malnutrition (SAM) remains unacceptably high. Several studies have examined risk factors present at admission for mortality. However, risks may evolve during admission with medical and nutritional treatment or deterioration. Currently, no specific guidance exists for assessing daily treatment response. This study aimed to determine the prognostic value of monitoring clinical signs on a daily basis for assessing mortality risk during hospitalization in children with SAM. Methods This is a secondary analysis of data from a randomized trial (NCT02246296) among 843 hospitalized children with SAM. Daily clinical signs were prospectively collected during ward rounds. Multivariable extended Cox regression using backward feature selection was performed to identify daily clinical warning signs (CWS) associated with time to death within the first 21 days of hospitalization. Predictive models were subsequently developed, and their prognostic performance evaluated using Harrell’s concordance index (C-index) and time-dependent area under the curve (tAUC). Results Inpatient case fatality ratio was 16.3% (n=127). The presence of the following CWS during daily assessment were found to be independent predictors of inpatient mortality: symptomatic hypoglycemia, reduced consciousness, chest indrawing, not able to complete feeds, nutritional edema, diarrhea, and fever. Daily risk scores computed using these 7 CWS together with MUAC<10.5cm at admission as additional CWS predict survival outcome of children with SAM with a C-index of 0.81 (95% CI 0.77–0.86). Moreover, counting signs among the top 5 CWS (reduced consciousness, symptomatic hypoglycemia, chest indrawing, not able to complete foods, and MUAC<10.5cm) provided a simpler tool with similar prognostic performance (C-index of 0.79; 95% CI 0.74–0.84). Having 1 or 2 of these CWS on any day during hospitalization was associated with a 3 or 11-fold increased mortality risk compared with no signs, respectively. Conclusions This study provides evidence for structured monitoring of daily CWS as recommended clinical practice as it improves prediction of inpatient mortality among sick children with complicated SAM. We propose a simple counting-tool to guide healthcare workers to assess treatment response for these children. Trial registration NCT02246296

Background “The sense of taste,” write Nelson and colleagues in a 2002 issue of Nature, “provides animals with valuable information about the nature and quality of food. Mammals can recognize and respond to a diverse repertoire of chemical entities, including sugars, salts, acids and a wide range of toxic substances” (199). The authors go on to argue that several amino acids—the building blocks of proteins—taste delicious to humans and that “having a taste pathway dedicated to their detection probably had significant evolutionary implications”. They imply, but do not specify, that the evolutionary implications are positive. This may be the case with some amino acids, but contemporary tastes, and changes in them, are far from universally beneficial. Indeed, this article argues that modern food production shapes and distorts human taste with significant implications for health and wellbeing. Take the western taste for fried chipped potatoes, for example. According to Schlosser in Fast Food Nation, “In 1960, the typical American ate eighty-one pounds of fresh potatoes and about four pounds of frozen french fries. Today [2002] the typical American eats about forty-nine pounds of fresh potatoes every year—and more than thirty pounds of frozen french fries” (115). Nine-tenths of these chips are consumed in fast food restaurants which use mass-manufactured potato-based frozen products to provide this major “foodservice item” more quickly and cheaply than the equivalent dish prepared from raw ingredients. These choices, informed by human taste buds, have negative evolutionary implications, as does the apparently long-lasting consumer preference for fried goods cooked in trans-fats. “Numerous foods acquire their elastic properties (i.e., snap, mouth-feel, and hardness) from the colloidal fat crystal network comprised primarily of trans- and saturated fats. These hardstock fats contribute, along with numerous other factors, to the global epidemics related to metabolic syndrome and cardiovascular disease,” argues Michael A. Rogers (747). Policy makers and public health organisations continue to compare notes internationally about the best ways in which to persuade manufacturers and fast food purveyors to reduce the use of these trans-fats in their products (L’Abbé et al.), however, most manufacturers resist. Hank Cardello, a former fast food executive, argues that “many products are designed for ‘high hedonic value’, with carefully balanced combinations of salt, sugar and fat that, experience has shown, induce people to eat more” (quoted, Trivedi 41). Fortunately for the manufactured food industry, salt and sugar also help to preserve food, effectively prolonging the shelf life of pre-prepared and packaged goods. Physiological Factors As Glanz et al. discovered when surveying 2,967 adult Americans, “taste is the most important influence on their food choices, followed by cost” (1118). A person’s taste is to some extent an individual response to food stimuli, but the tongue’s taste buds respond to five basic categories of food: salty, sweet, sour, bitter, and umami. ‘Umami’ is a Japanese word indicating “delicious savoury taste” (Coughlan 11) and it is triggered by the amino acid glutamate. Japanese professor Kikunae Ikeda identified glutamate while investigating the taste of a particular seaweed which he believed was neither sweet, sour, bitter, or salty. When Ikeda combined the glutamate taste essence with sodium he formed the food additive sodium glutamate, which was patented in 1908 and subsequently went into commercial production (Japan Patent Office). Although individual, a person’s taste preferences are by no means fixed. There is ample evidence that people’s tastes are being distorted by modern food marketing practices that process foods to make them increasingly appealing to the average palate. In particular, this industrialisation of food promotes the growth of a snack market driven by salty and sugary foods, popularly constructed as posing a threat to health and wellbeing. “[E]xpanding waistlines [are] fuelled by a boom in fast food and a decline in physical activity” writes Stark, who reports upon the 2008 launch of a study into Australia’s future ‘fat bomb’. As Deborah Lupton notes, such reports were a particular feature of the mid 2000s when: intense concern about the ‘obesity epidemic’ intensified and peaked. Time magazine named 2004 ‘The Year of Obesity’. That year the World Health Organization’s Global Strategy on Diet, Physical Activity and Health was released and the [US] Centers for Disease Control predicted that a poor diet and lack of exercise would soon claim more lives than tobacco-related disease in the United States. (4) The American Heart Association recommends eating no more than 1500mg of salt per day (Hamzelou 11) but salt consumption in the USA averages more than twice this quantity, at 3500mg per day (Bernstein and Willett 1178). In the UK, a sustained campaign and public health-driven engagement with food manufacturers by CASH—Consensus Action on Salt and Health—resulted in a reduction of between 30 and 40 percent of added salt in processed foods between 2001 and 2011, with a knock-on 15 percent decline in the UK population’s salt intake overall. This is the largest reduction achieved by any developed nation (Brinsden et al.). “According to the [UK’s] National Institute for Health and Care Excellence (NICE), this will have reduced [UK] stroke and heart attack deaths by a minimum of 9,000 per year, with a saving in health care costs of at least £1.5bn a year” (MacGregor and Pombo). Whereas there has been some success over the past decade in reducing the amount of salt consumed, in the Western world the consumption of sugar continues to rise, as a graph cited in the New Scientist indicates (O’Callaghan). Regular warnings that sugar is associated with a range of health threats and delivers empty calories devoid of nutrition have failed to halt the increase in sugar consumption. Further, although some sugar is a natural product, processed foods tend to use a form invented in 1957: high-fructose corn syrup (HFCS). “HFCS is a gloopy solution of glucose and fructose” writes O’Callaghan, adding that it is “as sweet as table sugar but has typically been about 30% cheaper”. She cites Serge Ahmed, a French neuroscientist, as arguing that in a world of food sufficiency people do not need to consume more, so they need to be enticed to overeat by making food more pleasurable. Ahmed was part of a team that ran an experiment with cocaine-addicted rats, offering them a mutually exclusive choice between highly-sweetened water and cocaine: Our findings clearly indicate that intense sweetness can surpass cocaine reward, even in drug-sensitized and -addicted individuals. We speculate that the addictive potential of intense sweetness results from an inborn hypersensitivity to sweet tastants. In most mammals, including rats and humans, sweet receptors evolved in ancestral environments poor in sugars and are thus not adapted to high concentrations of sweet tastants. The supranormal stimulation of these receptors by sugar-rich diets, such as those now widely available in modern societies, would generate a supranormal reward signal in the brain, with the potential to override self-control mechanisms and thus lead to addiction. (Lenoir et al.) The Tongue and the Brain One of the implications of this research about the mammalian desire for sugar is that our taste for food is about more than how these foods actually taste in the mouth on our tongues. It is also about the neural response to the food we eat. The taste of French fries thus also includes that “snap, mouth-feel, and hardness” and the “colloidal fat crystal network” (Rogers, “Novel Structuring” 747). While there is no taste receptor for fats, these nutrients have important effects upon the brain. Wang et al. offered rats a highly fatty, but palatable, diet and allowed them to eat freely. 33 percent of the calories in the food were delivered via fat, compared with 21 percent in a normal diet. The animals almost doubled their usual calorific intake, both because the food had a 37 percent increased calorific content and also because the rats ate 47 percent more than was standard (2786). The research team discovered that in as little as three days the rats “had already lost almost all of their ability to respond to leptin” (Martindale 27). Leptin is a hormone that acts on the brain to communicate feelings of fullness, and is thus important in assisting animals to maintain a healthy body weight. The rats had also become insulin resistant. “Severe resistance to the metabolic effects of both leptin and insulin ensued after just 3 days of overfeeding” (Wang et al. 2786). Fast food restaurants typically offer highly palatable, high fat, high sugar, high salt, calorific foods which can deliver 130 percent of a day’s recommended fat intake, and almost a day’s worth of an adult man’s calories, in one meal. The impacts of maintaining such a diet over a comparatively short time-frame have been recorded in documentaries such as Super Size Me (Spurlock). The after effects of what we widely call “junk food” are also evident in rat studies. Neuroscientist Paul Kenny, who like Ahmed was investigating possible similarities between food- and cocaine-addicted rats, allowed his animals unlimited access to both rat ‘junk food’ and healthy food for rats. He then changed their diets. “The rats with unlimited access to junk food essentially went on a hunger strike. ‘It was as if they had become averse to healthy food’, says Kenny. It took two weeks before the animals began eating as much [healthy food] as those in the control group” (quoted, Trivedi 40). Developing a taste for certain food is consequently about much more than how they taste in the mouth; it constitutes an individual’s response to a mixture of taste, hormonal reactions and physiological changes. Choosing Health Glanz et al. conclude their study by commenting that “campaigns attempting to change people’s perception of the importance of nutrition will be interpreted in terms of existing values and beliefs. A more promising strategy might be to stress the good taste of healthful foods” (1126). Interestingly, this is the strategy already adopted by some health-focused cookbooks. I have 66 cookery books in my kitchen. None of ten books sampled from the five spaces in which these books are kept had ‘taste’ as an index entry, but three books had ‘taste’ in their titles: The Higher Taste, Taste of Life, and The Taste of Health. All three books seek to promote healthy eating, and they all date from the mid-1980s. It might be that taste is not mentioned in cookbook indexes because it is a sine qua non: a focus upon taste is so necessary and fundamental to a cookbook that it goes without saying. Yet, as the physiological evidence makes clear, what we find palatable is highly mutable, varying between people, and capable of changing significantly in comparatively short periods of time. The good news from the research studies is that the changes wrought by high salt, high sugar, high fat diets need not be permanent. Luciano Rossetti, one of the authors on Wang et al’s paper, told Martindale that the physiological changes are reversible, but added a note of caution: “the fatter a person becomes the more resistant they will be to the effects of leptin and the harder it is to reverse those effects” (27). Morgan Spurlock’s experience also indicates this. In his case it took the actor/director 14 months to lose the 11.1 kg (13 percent of his body mass) that he gained in the 30 days of his fast-food-only experiment. Trivedi was more fortunate, stating that, “After two weeks of going cold turkey, I can report I have successfully kicked my ice cream habit” (41). A reader’s letter in response to Trivedi’s article echoes this observation. She writes that “the best way to stop the craving was to switch to a diet of vegetables, seeds, nuts and fruits with a small amount of fish”, adding that “cravings stopped in just a week or two, and the diet was so effective that I no longer crave junk food even when it is in front of me” (Mackeown). Popular culture indicates a range of alternative ways to resist food manufacturers. In the West, there is a growing emphasis on organic farming methods and produce (Guthman), on sl called Urban Agriculture in the inner cities (Mason and Knowd), on farmers’ markets, where consumers can meet the producers of the food they eat (Guthrie et al.), and on the work of advocates of ‘real’ food, such as Jamie Oliver (Warrin). Food and wine festivals promote gourmet tourism along with an emphasis upon the quality of the food consumed, and consumption as a peak experience (Hall and Sharples), while environmental perspectives prompt awareness of ‘food miles’ (Weber and Matthews), fair trade (Getz and Shreck) and of land degradation, animal suffering, and the inequitable use of resources in the creation of the everyday Western diet (Dare, Costello and Green). The burgeoning of these different approaches has helped to stimulate a commensurate growth in relevant disciplinary fields such as Food Studies (Wessell and Brien). One thing that all these new ways of looking at food and taste have in common is that they are options for people who feel they have the right to choose what and when to eat; and to consume the tastes they prefer. This is not true of all groups of people in all countries. Hiding behind the public health campaigns that encourage people to exercise and eat fresh fruit and vegetables are the hidden “social determinants of health: The conditions in which people are born, grow, live, work and age, including the health system” (WHO 45). As the definitions explain, it is the “social determinants of health [that] are mostly responsible for health iniquities” with evidence from all countries around the world demonstrating that “in general, the lower an individual’s socioeconomic position, the worse his or her health” (WHO 45). For the comparatively disadvantaged, it may not be the taste of fast food that attracts them but the combination of price and convenience. If there is no ready access to cooking facilities, or safe food storage, or if a caregiver is simply too time-poor to plan and prepare meals for a family, junk food becomes a sensible choice and its palatability an added bonus. For those with the education, desire, and opportunity to break free of the taste for salty and sugary fats, however, there are a range of strategies to achieve this. There is a persuasive array of evidence that embracing a plant-based diet confers a multitude of health benefits for the individual, for the planet and for the animals whose lives and welfare would otherwise be sacrificed to feed us (Green, Costello and Dare). Such a choice does involve losing the taste for foods which make up the lion’s share of the Western diet, but any sense of deprivation only lasts for a short time. The fact is that our sense of taste responds to the stimuli offered. It may be that, notwithstanding the desires of Jamie Oliver and the like, a particular child never will never get to like broccoli, but it is also the case that broccoli tastes differently to me, seven years after becoming a vegan, than it ever did in the years in which I was omnivorous. When people tell me that they would love to adopt a plant-based diet but could not possibly give up cheese, it is difficult to reassure them that the pleasure they get now from that specific cocktail of salty fats will be more than compensated for by the sheer exhilaration of eating crisp, fresh fruits and vegetables in the future. Conclusion For decades, the mass market food industry has tweaked their products to make them hyper-palatable and difficult to resist. They do this through marketing experiments and consumer behaviour research, schooling taste buds and brains to anticipate and relish specific cocktails of sweet fats (cakes, biscuits, chocolate, ice cream) and salty fats (chips, hamburgers, cheese, salted nuts). They add ingredients to make these products stimulate taste buds more effectively, while also producing cheaper items with longer life on the shelves, reducing spoilage and the complexity of storage for retailers. Consumers are trained to like the tastes of these foods. Bitter, sour, and umami receptors are comparatively under-stimulated, with sweet, salty, and fat-based tastes favoured in their place. Western societies pay the price for this learned preference in high blood pressure, high cholesterol, diabetes, and obesity. Public health advocate Bruce Neal and colleagues, working to reduce added salt in processed foods, note that the food and manufacturing industries can now provide most of the calories that the world needs to survive. “The challenge now”, they argue, “is to have these same industries provide foods that support long and healthy adult lives. And in this regard there remains a very considerable way to go”. If the public were to believe that their sense of taste is mutable and has been distorted for corporate and industrial gain, and if they were to demand greater access to natural foods in their unprocessed state, then that journey towards a healthier future might be far less protracted than these and many other researchers seem to believe. References Bernstein, Adam, and Walter Willett. “Trends in 24-Hr Sodium Excretion in the United States, 1957–2003: A Systematic Review.” American Journal of Clinical Nutrition 92 (2010): 1172–1180. Bhaktivedanta Book Trust. 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MacGregor, Graham, and Sonia Pombo. “The Amount of Hidden Sugar in Your Diet Might Shock You.” The Conversation 9 January (2014). 2 Feb. 2014 ‹http://theconversation.com/the-amount-of-hidden-sugar-in-your-diet-might-shock-you-21867›. Mackeown, Elizabeth. “Cold Turkey?” [Letter]. New Scientist 2787 (2010): 31. Martindale, Diane. “Burgers on the Brain.” New Scientist 2380 (2003): 26–29. Mason, David, and Ian Knowd. “The Emergence of Urban Agriculture: Sydney, Australia.” The International Journal of Agricultural Sustainability 8.1–2 (2010): 62–71. Neal, Bruce, Jacqui Webster, and Sebastien Czernichow. “Sanguine About Salt Reduction.” European Journal of Preventative Cardiology 19.6 (2011): 1324–1325. Nelson, Greg, Jayaram Chandrashekar, Mark A. Hoon, Luxin Feng, Grace Zhao, Nicholas J. P. Ryba, & Charles S. Zuker. “An Amino-Acid Taste Receptor.” Nature 416 (2002): 199–202. O’Callaghan, Tiffany. “Sugar on Trial: What You Really Need to Know.” New Scientist 2954 (2011): 34–39. 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The Norwegian Scientific Committee for Food Safety (Vitenskapskomiteen for mattrygghet, VKM) has, at the request of the Norwegian Food Safety Authority (Mattilsynet; NFSA), assessed the risk of "other substances" in food supplements and energy drinks sold in Norway. VKM has assessed the risk of doses given by NFSA. These risk assessments will provide NFSA with the scientific basis while regulating "other substances" in food supplements. "Other substances" are described in the food supplement directive 2002/46/EC as substances other than vitamins or minerals that have a nutritional and/ or physiological effect. It is added mainly to food supplements, but also to energy drinks and other foods. In this series of risk assessments of "other substances" the VKM has not evaluated any claimed beneficial effects from these substances, only possible adverse effects. The present report is limited to the use of L-lysine in food supplements. Risks related to lysine added to food and drinks, protein hydrolysates or high dietary protein intake are outside the scope of the opinion. The report is based on previous risk assessments of lysine and scientific papers retrieved from a comprehensive literature search. According to information from the NFSA, L-lysine is an ingredient in food supplements sold in Norway. NSFA has requested a risk assessment of 1000, 2000, 2500, 2750 and 3000 mg/day of L-lysine from food supplements. Foods rich in L-lysine are generally protein rich foods such as meat, dairy products, eggs, legumes, and some fish. Based on NHANES III (19881994), the overall mean intake of L-lysine from food and food supplements in the United States was 5.3 g/day (IOM, 2005). L-Lysine, an indispensable amino acid, is present in all proteins in the human body. Its catabolisation takes place mainly in the liver. The two nitrogen groups are transferred to alpha-ketoglutarate to form glutamate. The remaining carbon skeleton is broken down to acetoacetyl-CoA. Lysine is exclusively ketogenic i.e. does not enter gluconeogenesis for the production of glucose. In the first phase of the present evaluation of L-lysine, previous reports evaluating the safety of L-lysine supplementation in humans were identified. In the second phase, two systematic literature searches have been performed to retrieve scientific papers published before 11 May 2016 (human studies literature search) and before 28 September 2016 (animal studies literature search). Based on these searches, we identified two human studies and one study in rats that could be used for risk assessment of L-lysine in food supplement. According to a report from the Institute of Medicine in the USA (IOM, 2005), several clinical trials of lysine with intakes ranging from 0.6 to 3.0 g/day for 3 to 6 months have not reported any adverse effects. The same was the case for the two human randomised controlled trials (RCTs) included in this report providing 6 g/day L-lysine orally for 8 weeks to schizophrenic patients. One 90-days subchronic toxicity study with rats was identified, showing a no observed adverse effect level (NOAEL) of 3357 mg/kg bw per day (the highest dose tested), with no functional, biochemical or histological changes in renal function. In the present report, the value of comparison is set to 86 mg/kg bw per day, corresponding to 6000 mg per day in a 70 kg adult; the daily dose provided in the two human RCTs. The calculated margins of exposures (MOE-values) range from 1.2 to 6.0 for the specified supplement doses with 1000-3000 mg/day of L-lysine. MOE-values below 10 (for interindividual differences in humans) is regarded as acceptable since L-lysine is a nutrient that does not cause any known adverse effects. In addition, the overall mean lysine intake according to NHANES III (5.3 g/day) is close to the the doses considered in the present risk assessment. The requirement for lysine, 30 mg/kg bw per day, corresponding to 2.1 g/day in a 70 kg adult, is close to the doses considered in the present risk assessment. The NOAEL suggested in the 90 days subchronic rat study supports the suggestion that these doses are well tolerated in humans even with somewhat low MOE-values. VKM concludes that: In adults (≥18 years), the specified doses 1000, 2000, 2500, 2750 and 3000 mg/day L-lysine in food supplements are unlikely to cause adverse health effects. In adolescents (14 to <18 years), the specified doses 1000, 2000, 2500, 2750 and 3000 mg/day L-lysine in food supplements are unlikely to cause adverse health effects. In children (10 to <14 years), the specified doses 1000, 2000, 2500, 2750 and 3000 mg/day L-lysine in food supplements are unlikely to cause adverse health effects. Children younger than 10 years were not within the scope of the present risk assessment.

The Norwegian Scientific Committee for Food Safety (Vitenskapskomiteen for mattrygghet, VKM) has, at the request of the Norwegian Food Safety Authority (Mattilsynet; NFSA), assessed the risk of "other substances" in food supplements and energy drinks sold in Norway. VKM has assessed the risk of doses given by NFSA. These risk assessments will provide NFSA with the scientific basis while regulating "other substances" in food supplements. "Other substances" are described in the food supplement directive 2002/46/EC as substances other than vitamins or minerals that have a nutritional and/ or physiological e ffect. It is added mainly to food supplements, but also to energy drinks and other foods. In this series of risk assessments of "other substances" the VKM has not evaluated any claimed beneficial effects from these substances, only possible adverse effects. The present report is a risk assessment of specified doses of L-threonine in food supplements, and it is based on previous risk assessments and articles retrieved from literature searches. According to information from NFSA, L-threonine is an ingredient in food supplements sold in Norway. NSFA has requested a risk assessment of 1000, 1200, 1500, 2000 and 2400 mg/day of L-threonine from food supplements. L-threonine is an essential amino acid not known to cause any adverse health effects. Previous reports do not indicate a tolerable upper intake level, apart from an approval of a dose of 1150 mg/day by the Scientific Committee of the Spanish Agency for Food Safety and Nutrition (AESAN). Long-term studies in humans were not found. The only available human studies were: a small uncontrolled one-year pilot study with doses ranging from 0.5 to 2.5 g/day, one eight-week randomised controlled trial (RCT) using a dose of 7.5 g/day, and two 2-week RCTs using doses of 6 and 4.5 g/day. No adverse effects (diary method of registration of adverse effects) were reported in the eight-week clinical trial, and the only adverse effects observed in the two-week trials were one case of indigestion and one case of diarrhoea. A four-week rodent toxicity study indicated a no observed adverse effect level (NOAEL) of 854.3 mg/kg bw per day (only dose tested, no adverse effects observed). The value used for comparison with the estimated exposure in the risk characterisation is the NOAEL defined in an 8-week randomised placebo controlled study in humans, 7500 mg/day. For a 70-kg individual, this corresponds to 107 mg/kg bw per day. Two human two-week studies and a small one-year pilot study support the notion that this dose will be well tolerated. The overall mean threonine intake according to NHANES III (3 g/day) is slightly larger than the doses requested for evaluation in the present risk assessment. No studies in children (10 to <14 years) and adolescents (14 to <18 years) were identified. Based on the included literature there was no evidence indicating that age affects the tolerance for relevant doses of threonine. Therefore, in this risk characterisation a tolerance as for adults, based on body weight, was assumed for these age groups. VKM concludes that: In adults (≥18 years), the specified doses 1000, 1200, 1500, 2000 and 2400 mg/day L-threonine in food supplements are unlikely to cause adverse health effects. In adolescents (14 to <18 years), the specified doses 1000, 1200, 1500, 2000 and 2400 mg/day L-threonine in food supplements are unlikely to cause adverse health effects. In children (10 to <14 years), the specified doses 1000, 1200, 1500, 2000 and 2400 mg/day L-threonine in food supplements are unlikely to cause adverse health effects. Children younger than 10 years were not within the scope of the present risk assessment.