Evaluating the Impact of the Standardized Letter of Recommendation on a Successful Match in Urology Residency.

OBJECTIVES: To evaluate the contribution of the SLOR to the likelihood of matching in urology residency by applying a novel scoring system and characterize utilization in the 2022 application cycle. METHODS: We conducted an investigation of all applicants to our urology residency program during the 2022 cycle. We developed a scoring system to assess SLOR strength across all templates. Match outcomes were verified with the Society of Academic Urologists listing. Statistical analysis was performed to assess for factors predictive of a successful match. RESULTS: Out of 386 total applicants, 239 (61.9%) had at least one SLOR in their application. SLOR utilization was more prevalent in MD applicants, in those with higher Step 2 scores, and in those who matched (p < 0.01). The majority of SLOR scores (66.5%) were above a 3.5/5 in our cohort. Step 1 score, number of research entries, and presence of a SLOR were predictive of successful match. However, a SLOR score of < 3 was strongly associated with not matching (OR 0.021, p < 0.01). CONCLUSIONS: The presence of a SLOR in our cohort overall was associated with a successful match. A poor SLOR score was highly deleterious to an applicant's chance of matching in urology. Our SLOR scoring system can be used across all letter templates and demonstrates that strength of SLOR can significantly impact an applicant's chance of matching.

Perinephric fat stranding is associated with increased age and stone size but not with serum creatinine in patients with obstructing ureterolithiasis.

Previous reports show increased severity of perinephric fat stranding (PFS) with elevated serum creatinine in obstructing ureterolithiasis. We sought to investigate this association with our institution's patient population.We reviewed charts of patients diagnosed with obstructive ureterolithiasis or nephrolithiasis in our emergency department between January and October 2018. Patient demographics, lab results, and computed tomography (CT) imaging were reviewed. A blinded radiologist reviewed all CTs and graded hydronephrosis and PFS. Subjects were stratified by degree of PFS and compared via paired t-test, chi-squared test, univariate analysis, and multivariate analysis.We identified 141 patients; 114 had no-mild (Group 1) PFS, while 27 had moderate-severe (Group 2) PFS. Group 1 had a mean age of 56 (SD = 16.1) and mean stone size of 7.3 mm (SD = 4.22); 77% of the cohort had symptoms under 24 h. Group 2 was older with a mean age of 65 (SD = 16.2, p = 0.01) and mean stone size of 10.1 mm (SD = 6.07, p < 0.01); 50% had symptoms less than 24 h (p = 0.01). PFS did not correlate with change in serum creatinine. Univariate and multivariate analysis showed increasing age increased the odds of moderate-severe PFS by 3.5% (OR = 1.035, p < 0.05) while increased stone size increased the odds of moderate-severe PFS by 13.7% (OR = 1.137, p = 0.01).Although increased PFS correlated with increased age and stone size, no correlation was found with presenting creatinine or change in creatinine. Degree of PFS is likely a poor predictor of renal disease severity in acute ureterolithiasis.

Ex-vivo ureteroscopy, laser lithotripsy, and stone basketing extraction of deceased donor kidney stones during machine perfusion preservation.

The incidence of nephrolithiasis in kidney donors is rare. The timing and treatment of nephrolithiasis in deceased donor kidneys are not well established. While some programs have proposed ex-situ rigid or flexible ureteroscopy treatment before transplantation, we report on two cases of kidney stones in the same deceased donor that we treated by flexible ureteroscopy and laser lithotripsy performed during the storage time on a hypothermic perfusion machine. Two deceased donor kidneys were found to have multiple kidney stones discovered on preprocurement CT imaging. The right kidney had less than five 2-3 mm stones, whereas the left had five to ten 1 mm stones with a single 7 mm stone. Both organs were placed on a hypothermic perfusion machine and maintained at a temperature of 4°C. An ex-vivo flexible ureteroscopy with laser lithotripsy and basket extraction was performed while the kidneys were maintained on Lifeport* perfusion machine. The cold ischemia time was 16.9 and 23.1 h. After 12 months of observational follow-up, neither recipient had nephrolithiasis, UTI, or other urologic complications. The creatinine values now are 1.17 and 2.44 mg/dL (103.4 and 215.7 µmol/L), respectively. Ex-vivo flexible ureteroscopy with laser lithotripsy and stone removal on machine-perfused kidneys appears to be safe and offers a good option to treat graft nephrolithiasis and prevent posttransplant complications. Ureteroscopy serves as a minimally invasive treatment option with direct stone removal. Performing this while on machine perfusion minimizes the ischemic time of the kidney and resultant complications or delays in graft function.

Postpartum women's experiences in a randomized controlled trial of a web-based lifestyle intervention following Gestational Diabetes: a qualitative study.

INTRODUCTION: Gestational diabetes mellitus (GDM) is associated with an increased maternal risk for the development of type 2 diabetes (T2DM). We previously demonstrated in a randomized trial that a web-based postpartum lifestyle intervention program, Balance After Baby, increased weight loss among postpartum women with recent pregnancies complicated by GDM. The aim of this analysis is to identify the impact of the intervention on study participants as assessed by exit interviews after completion of the 12 month study. METHODS: We conducted structured exit interviews created with a concurrent-contextual design with subjects randomized to the intervention group at the conclusion of their participation (∼12 months) in the Balance After Baby study, with the objectives of 1) understanding the impact of the intervention on participants and their family members, 2) identifying which program components were most and least helpful, and 3) identifying the perceived best timing for diabetes prevention interventions in postpartum women with recent GDM. RESULTS: Seventy-nine percent (26/33) of eligible intervention participants participated in interviews. Participants noted changes in diet and physical activity as a result of the intervention. Several components of the intervention, particularly the online modules and support from the lifestyle coach, were perceived by intervention participants to have had a positive effect on personal and familial lifestyle change, while other components were less utilized, including the community forum, YMCA memberships, and pedometers. Nearly all participants felt that the timing in the intervention study, beginning about 6 weeks postpartum, was ideal. DISCUSSION: Results of this study identify the importance of individualized coaching, impact on family members, and demonstrate that postpartum women feel ready to make changes by 6 weeks postpartum. Findings from this study will help inform the development of future technologically-based lifestyle interventions for postpartum women with recent GDM.

Development and Usability of a Text Messaging Program for Women With Gestational Diabetes: Mixed Methods Study.

BACKGROUND: Gestational diabetes mellitus (GDM) affects 5%-10% of pregnancies and can lead to serious fetal and maternal complications. SMS text messaging is an effective way to improve diabetes management outside of pregnancy, but has not been well studied in GDM. OBJECTIVE: This study aimed to perform user experience testing and assess usability and acceptability of an SMS text messaging program (Text 4 Success) for women with GDM. METHODS: An automated 2-way texting program was developed. It included (1) reminders to check blood glucose levels, (2) positive feedback to user-reported glucose levels, (3) weekly educational messages, and (4) weekly motivational messages. For the user experience testing, women received simulated messages. For the usability study, women were enrolled in the program and received messages for 2 weeks. All women participated in semistructured interviews. For women in the usability study, data from glucose measuring devices were downloaded to assess adherence to self-monitoring of blood glucose (SMBG), measured as the percentage of recommended SMBG checks performed (a secondary outcome). RESULTS: Ten women participated in user experience testing. Suggestions for optimization included further customization of message timing and minimization of jargon, which were incorporated. Ten women participated in the usability study. All 10 would recommend the program to other women with GDM. Participants liked the immediate feedback to glucose values. Suggestions included further flexibility of messages related to mealtimes and the ability to aggregate blood glucose data into a table or graph. Overall, adherence to SMBG testing was high at baseline (222/238 recommended checks, 93%). In comparing the week prior to the trial with the 2 weeks during the trial, there was a small but statistically insignificant difference (P=.48) in the percentage of recommended SMBG performed (median 93% [25th-75th IQR 89%-100%] vs median 97% [25th-75th IQR 92%-100%]). CONCLUSIONS: Overall, women with GDM would recommend the Text 4 Success in GDM program and think it is helpful for GDM self-management. The program was usable and acceptable. The program may be better suited to those who have low levels of adherence to SMBG at baseline or to women at time of their diagnosis of GDM. Adaptations to the program will be made based on user suggestions. Further study of SMS text messaging to improve SMBG in GDM is needed.

Breastfeeding Associated with Lower Prevalence of Metabolic Syndrome in Women with Gestational Diabetes in the Very Early Postpartum Period.

OBJECTIVE: The aim of this study is to examine the association of breastfeeding with metabolic syndrome (MetS) in women with recent gestational diabetes mellitus (GDM) in the very early postpartum (PP) period. STUDY DESIGN: We performed a secondary analysis of the Balance After Baby Intervention (BABI) study which enrolled women with recent GDM. Data collected during an early (∼8 weeks) PP visit were used in this analysis. At this visit, weight, height, waist circumference (WC), blood pressure (BP), fasting plasma glucose (FPG), and lipids were obtained. MetS was classified per National Cholesterol Education Program Adult Treatment Program III (NCEP-ATP III) criteria. We defined breastfeeding as currently breastfeeding or not currently breastfeeding for the main analysis. RESULTS: Of 181 women enrolled in BABI, 178 were included in this analysis (3 excluded for missing lipids). Thirty-four percent were Hispanic. Of non-Hispanics, 31.5% were White, 18.5% Asian, and 12.9% Black/African American. The prevalence of MetS was 42.9% in women not breastfeeding versus 17.1% in women breastfeeding (p < 0.001; adjusted odds ratio [aOR] = 0.16 [95% confidence interval (CI): 0.06-0.41]). Breastfeeding women had significantly lower odds of FPG ≥100 mg/dL (aOR = 0.36 [95% CI: 0.14-0.95], p = 0.039), HDL < 50 mg/dL (aOR = 0.19 [95% CI: 0.08-0.46], p < 0.001), and triglycerides (TG) ≥ 150 mg/dL (aOR = 0.26 [95% CI: 0.10-0.66], p = 0.005). When evaluated as continuous variables, WC, FPG, and TG were significantly lower and HDL significantly higher in women breastfeeding in the very early PP period (vs. not breastfeeding). CONCLUSION: In a diverse population of women with recent GDM, there was lower prevalence of MetS in women breastfeeding compared with those not breastfeeding in the very early PP period. This study extends the findings of an association of breastfeeding with MetS previously reported at time points more remote from pregnancy to the very early PP period and to an ethnically and racially diverse population. KEY POINTS: · MetS prevalence in women with recent GDM was lower in breastfeeding than not breastfeeding women.. · FPG, HDL, WC, and TG were improved in the breastfeeding group.. · This study extends prior findings to the very early PP period and to a diverse population..

Safety evaluation of the food enzyme isoamylase from a Dyella sp. strain.

The food enzyme isoamylase (glycogen α-1,6-glucanohydrolase, EC 3.2.1.68) is produced with Dyella sp. by Hayashibara Co. Ltd. Whole genome sequence analysis of the production strain identified a sequence with high homology with a gene conferring resistance to an antimicrobial, which may confer cross-resistance to a critically important antimicrobial, as defined by the World Health Organisation. This is a concern, since DNA from the production strain was detected in the food enzyme. The isoamylase food enzyme is intended to be used in starch processing for the production of various starch hydrolysates. Since residual amounts of total organic solids are removed by the purification steps applied during the production of saccharides from starch, dietary exposure was not calculated. The batch used for toxicological testing was not sufficiently characterised; therefore, the toxicological data provided were not considered. Similarity of the amino acid sequence to those of known allergens was searched and no match was found. The Panel considered that, under the intended conditions of use, the risk of allergic sensitisation and elicitation reactions by dietary exposure cannot be excluded, but the likelihood for this to occur is considered to be low. Overall, the Panel cannot conclude on the safety of the food enzyme isoamylase produced with Dyella sp.

Extraction of Essential Oils from Medicinal Plants and their Utilization as Food Antioxidants.

BACKGROUND: The use of essential oils is receiving increasing attention worldwide, as these oils are good sources of several bioactive compounds. Nowadays essential oils are preferred over synthetic preservatives thanks to their antioxidant and antimicrobial properties. Several studies highlight the beneficial effect of essential oils extracted from medicinal plants to cure human diseases such as hypertension, diabetes, or obesity. However, to preserve their bioactivity, the use of appropriate extraction technologies is required. METHODS: The present review aims to describe the studies published so far on the essential oils focusing on their sources and chemical composition, the technologies used for their recovery and their application as antioxidants in food products. RESULTS: The review has been structured in three parts. In the first part, the main compounds present in essential oils extracted from medicinal plants have been listed and described. In the second part, the most important technologies used for extraction and distillation, have been presented. In detail, conventional methods have been described and compared with innovative and green technologies. Finally, in the last part, the studies related to the application of essential oils as antioxidants in food products have been reviewed and the main findings discussed in detail. CONCLUSION: In summary, an overview of the aforementioned subjects is presented by discussing the results of the most recent published studies.

Urinary tract infections following radical cystectomy with enhanced recovery protocol: A prospective study.

OBJECTIVES: Urinary tract infection (UTI) following radical cystectomy (RC) is a common complication associated with significant morbidity and risk of readmission. Recent literature has assessed the effect of perioperative antibiotic regimens on the rate of postoperative infections but not yet yielded with significant changes in UTI rates. Our study focused on the effect of postoperative suppressive regimens on the rate of UTI following radical cystectomy with Enhanced Recovery After Surgery (ERAS) protocol. METHODS: We retrospectively reviewed 427 patients who underwent RC with ERAS protocol between May 2012 and January 2017 at our institution. The ERAS protocol infection prevention measures included 24-hr perioperative antibiotic followed by suppressive antibiotic until removal of catheter/stents. A patient was found to have a UTI if they had a positive urine culture and documented symptoms, positive urine culture with treatment per practitioner discretion, or negative or unavailable urine culture but the clinical presumption of UTI that got treatment. Urosepsis was defined if any of UTI episodes were associated with positive blood culture. Patients' characteristics, UTI events, and urine culture sensitivities were reviewed for analysis. RESULTS: The incidence of UTI and urosepsis was 36.1% and 7.13% within 90-days following RC, respectively. The median time to the first UTI was 13 days (IQR 8-35). Candida (25.57%) and Escherichia coli (22.16%) were the most commonly identified pathogens. UTI and urosepsis were significantly lower in patients who received suppressive fluoroquinolones compared to other antibiotic regimens (32.72% vs. 45.24%, P = 0.04 for UTI and 5.25% vs. 11.90%, P = 0.04 for urosepsis). In multivariable analysis, orthotopic neobladder and perioperative transfusion were significantly associated with increased UTI rate (OR = 2.3 and 1.71, p < 0.05, respectively). CONCLUSIONS: UTI is common following RC and urinary diversion with ERAS protocol. The most common isolated pathogens are candida and Escherichia coli. Orthotopic neobladder and perioperative transfusion are independent risk factors for postoperative UTI. The use of suppressive fluoroquinolones is associated with a significant decrease in UTI rate.

Safety evaluation of the food enzyme α-amylase from a genetically modified strain of Bacillus licheniformis (DP-Dzb25).

The food enzyme α-amylase (4-α-d-glucan glucanhydrolase; EC 3.2.1.1) is produced with the genetically modified strain Bacillus licheniformis DP-Dzb25 by Danisco US Inc. It is intended to be used in distilled alcohol production, starch processing for the production of glucose syrups, and in brewing processes. Since residual amounts of the food enzyme are removed by distillation and during starch processing, no dietary exposure was calculated for these food processes. Based on the maximum use levels recommended for brewing processes and individual data from the EFSA Comprehensive European Food Database, dietary exposure to the food enzyme-total organic solids (TOS) was estimated to be up to 0.138 mg TOS/kg body weight (bw) per day. The production strain of the food enzyme contains multiple copies of a known antimicrobial resistance gene and consequently, it does not fulfil the requirements for the Qualified Presumption of Safety (QPS) approach to safety assessment. However, considering the absence of viable cells and DNA from the production organism in the food enzyme, this is not considered to be a risk. As no other concerns arising from the microbial source and its subsequent genetic modification or from the manufacturing process have been identified, the Panel considers that toxicological tests are not needed for the assessment of this food enzyme. Similarity of the amino acid sequence to those of known allergens was searched for and no match was found. The Panel notes that the food enzyme may contain a known allergen. Therefore, allergenicity cannot be excluded for uses other than distilled alcohol production. Apart from potential allergenicity, the Panel concluded that the food enzyme 4-α-d-glucan glucanhydrolase produced with the genetically modified B. licheniformis strain DP-Dzb25 does not give rise to safety concerns under the intended conditions of use.

Safety evaluation of the food enzyme glucose oxidase from Aspergillus niger (strain ZGL).

The food enzyme glucose oxidase (ß-d-glucose:oxygen 1-oxidoreductase; EC 1.1.3.4) is produced with a genetically modified Aspergillus niger strain ZGL by DSM Food Specialties B.V.. The genetic modifications do not give rise to safety concerns. The food enzyme is free from viable cells of the production organism and recombinant DNA. The glucose oxidase is intended to be used in baking processes. Based on the maximum use levels, dietary exposure to the food enzyme-total organic solids (TOS) was estimated to be up to 0.004 mg TOS/kg body weight (bw) per day. The toxicity studies were carried out with an asparaginase from A. niger (strain ASP). The Panel considered this enzyme as a suitable substitute to be used in the toxicological studies, because they derive from the same recipient strain, the location of the inserts are comparable, no partial inserts were present and the production methods are essentially the same. Genotoxicity tests did not raise a safety concern. The systemic toxicity was assessed by means of a repeated dose 90-day oral toxicity study in rats. The Panel identified a no observed adverse effect level (NOAEL) at the highest dose of 1,038 and 1,194 mg TOS/kg bw per day (for males and females, respectively) that, compared with the estimated dietary exposure, results in a sufficiently high margin of exposure (MoE) (of at least 260,000). Similarity of the amino acid sequence to those of known allergens was searched and one match was found. The Panel considered that, under the intended conditions of use, the risk of allergic sensitisation and elicitation reactions by dietary exposure cannot be excluded, but the likelihood to occur is considered to be low. Based on the data provided, the Panel concluded that this food enzyme does not give rise to safety concerns under the intended conditions of use.

Safety evaluation of the food enzyme triacylglycerol lipase from Aspergillus niger (strain LFS).

The food enzyme triacylglycerol lipase (triacylglycerol acylhydrolase, EC 3.1.1.3) is produced with a genetically modified Aspergillus niger strain LFS by DSM Food Specialties B.V.. The genetic modifications do not give rise to safety concerns. The food enzyme is free from viable cells of the production organism and recombinant DNA. The triacylglycerol lipase food enzyme is intended to be used in baking processes. Based on the maximum use levels, dietary exposure to the food enzyme-total organic solids (TOS) was estimated to be up to 0.020 mg TOS/kg body weight (bw) per day. The toxicity studies were carried out with an asparaginase from A. niger (strain ASP). The Panel considered this enzyme as a suitable substitute to be used in the toxicological studies, because they derive from the same recipient strain, the location of the inserts are comparable, no partial inserts were present and the production methods are essentially the same. Genotoxicity tests did not raise a safety concern. The systemic toxicity was assessed by means of a repeated dose 90-day oral toxicity study in rats. The Panel identified a no observed adverse effect level (NOAEL) at the highest dose of 1,038 and 1,194 mg TOS/kg bw per day (for males and females, respectively) that, compared with the estimated dietary exposure, results in a sufficiently high margin of exposure (MoE) (of at least 51,900). Similarity of the amino acid sequence to those of known allergens was searched and no match was found. The Panel considered that, under the intended conditions of use, the risk of allergic sensitisation and elicitation reactions by dietary exposure cannot be excluded, but the likelihood to occur is considered to be low. Based on the data provided, the Panel concluded that this food enzyme does not give rise to safety concerns under the intended conditions of use.

Safety evaluation of the food enzyme α-amylase and 1,4-α-glucan 6-α-glucosyltransferase from Paenibacillus alginolyticus.

The food enzyme α-amylase (4-α-d-glucan glucanohydrolase, EC 3.2.1.1) and 1,4-α-glucan 6-α-glucosyltransferase ((1→4)-α-d-glucan:(1→4)-α-d-glucan(d-glucose) 6-α-d-glucosyltransferase, EC 2.4.1.24) is produced with a Paenibacillus alginolyticus by Hayashibara Co., Ltd. The food enzyme is free from viable cells of the production organism. The α-amylase and 1,4-α-glucan 6-α-glucosyltransferase is intended to be used in starch processing for the production of isomaltodextrins. Residual amounts of total organic solids are removed by the purification steps applied during the production of isomaltodextrins and consequently dietary exposure is considered negligible. Similarity of the amino acid sequences to those of known allergens was searched and no matches were found. The Panel considered that, under the intended conditions of use, the risk of allergic sensitisation and elicitation reactions by dietary exposure to the food enzyme cannot be excluded, but the likelihood is considered to be low. Based on the data provided, the Panel concluded that this food enzyme does not give rise to safety concerns under the intended conditions of use.

Availability of long-acting reversible contraceptives in Los Angeles County clinics through a Medicaid state plan amendment program.

OBJECTIVE: To assess the availability of long acting reversible contraceptive (LARC) methods in Los Angeles County through providers participating in a California State Medicaid State Plan Amendment Program called Family Planning, Access, Care and Treatment (Family PACT). STUDY DESIGN: This was a cross-sectional telephone survey utilizing "secret shopper" methodology. From 855 Family PACT providers in Los Angeles County in 2015, a representative sample of 400 providers was selected for study. Young female researchers posing as potential patients called each sampled clinic to ask a scripted series of questions about LARC availability for Family PACT patients in that practice. RESULTS: All but one eligible practice (99.7%) responded to our questions. Among the 336 responding practices, 284 said they accepted Family PACT. Of those accepting Family PACT, staff answering the telephone call at 61% said they did not provide any LARC method onsite, 2% provided all currently available LARC methods, and 6% provided same-day placement of any LARC. CONCLUSION: The majority of Family PACT practices surveyed said that they did not provide any LARC onsite, and very few provided same-day LARC placement despite easy patient enrollment procedures, relatively reasonable reimbursement and concerted efforts to increase LARC use. Substantial barriers to greater uptake may rest at the provider level with either actual unavailability of LARC or staff perception of unavailability. IMPLICATIONS: Only a minority of Family PACT practices said that LARC methods were available onsite, which imposes substantial restriction to access for women who are entitled to have access without cost. Other states developing programs should be aware of this challenge.

Urinary tract infections following radical cystectomy and urinary diversion: a review of 1133 patients.

OBJECTIVE: To investigate the incidence and microbiology of urinary tract infection (UTI) within 90 days following radical cystectomy (RC) and urinary diversion. METHODS: We reviewed 1133 patients who underwent RC for bladder cancer at our institution between 2003 and 2013; 815 patients (72%) underwent orthotopic diversion, 274 (24%) ileal conduit, and 44 (4%) continent cutaneous diversion. 90-day postoperative UTI incidence, culture results, antibiotic sensitivity/resistance and treatment were recorded through retrospective review. Fisher's exact test, Kruskal-Wallis test, and multivariable analysis were performed. RESULTS: A total of 151 urinary tract infections were recorded in 123 patients (11%) during the first 90 days postoperatively. 21/123 (17%) had multiple infections and 25 (20%) had urosepsis in this time span. Gram-negative rods were the most common etiology (54% of positive cultures). 52% of UTI episodes led to readmission. There was no significant difference in UTI rate, etiologic microbiology (Gram-negative rods, Gram-positive cocci, fungi), or antibiotic sensitivity and resistance patterns between diversion groups. Resistance to quinolones was evident in 87.5% of Gram-positive and 35% of Gram-negative bacteria. In multivariable analysis, Charlson Comorbidity Index > 2 was associated with higher 90-day UTI rate (OR = 1.8, 95% CI 1.1-2.9, p = 0.05) and Candida UTI (OR 5.6, 95% CI 1.6-26.5, p = 0.04). CONCLUSIONS: UTI is a common complication and cause of readmission following radical cystectomy and urinary diversion. These infections are commonly caused by Gram-negative rods. High comorbidity index is an independent risk factor for postoperative UTI, but diversion type is not.

Safety evaluation of the food enzyme acetolactate decarboxylase from a genetically modified Bacillus licheniformis (strain NZYM-JB).

The food enzyme acetolactate decarboxylase (α-acetolactate decarboxylase; EC 4.1.1.5) is produced with a genetically modified Bacillus licheniformis strain NZYM-JB by Novozymes A/S. The genetic modifications do not give rise to safety concerns. The food enzyme is free from viable cells of the production organism and recombinant DNA. This acetolactate decarboxylase is intended to be used in distilled alcohol production and brewing processes. Residual amounts of total organic solids (TOS) are removed by distillation; consequently, dietary exposure was not calculated for distilled alcohol products. For other brewery products, based on the maximum use level recommended for the brewing processes and individual data from the EFSA Comprehensive European Food Consumption Database, dietary exposure to the food enzyme-TOS was estimated to be up to 0.003 mg TOS/kg body weight (bw) per day in European populations. Genotoxicity tests with the food enzyme did not indicate a genotoxic concern. The systemic toxicity was assessed by means of a repeated dose 90-day oral toxicity study in rats. The Panel identified a no observed adverse effect level at the highest dose of 1,018 mg TOS/kg bw per day. When compared with the dietary exposure, this results in a sufficiently high margin of exposure (at least 300,000). The amino acid sequence of the food enzyme did not match to those of known allergens. The Panel considered that, under the intended condition of use, the risk of allergic sensitisation and elicitation reactions upon dietary exposure to this food enzyme cannot be excluded, but the likelihood is considered low. Based on the data provided, the Panel concluded that this food enzyme does not raise safety concerns under the intended conditions of use.

Safety evaluation of the food enzyme endo-1,4-ß-xylanase from a genetically modified Aspergillus oryzae (strain NZYM-FA).

The food enzyme is an endo-1,4-ß-xylanase (EC 3.2.1.8) produced with a genetically modified strain of Aspergillus oryzae by Novozymes A/S. The genetic modifications do not give rise to safety concerns. The food enzyme is free from viable cells of the production organism and recombinant DNA. This xylanase is intended to be used in baking and cereal-based processes. Based on the proposed maximum use levels, dietary exposure to the food enzyme-total organic solids (TOS) was estimated to be up to 0.027 mg TOS/kg body weight (bw) per day in European populations. Genotoxicity tests did not raise a safety concern. Subchronic toxicity was assessed by means of a repeated dose 90-day oral toxicity study in rats. In this study, effects were seen in the control group on reproductive parameters (particularly the seminiferous epithelium atrophy in the testes) at incidences that far exceeded the background range. Consequently, a combined repeated dose toxicity study with the reproduction/developmental toxicity screening test, including haematological parameters and the immunology cohort, was conducted. The Panel identified a no observed adverse effect level at the highest dose tested of 1,101.3 mg TOS/kg bw per day. When compared with the dietary exposure, resulted in a sufficiently high margin of exposure (at least 40,000). Similarity of the amino acid sequence to those of known allergens was searched and no match was found. The Panel considered that under the intended conditions of use the risk for allergic sensitisation and elicitation reactions by dietary exposure cannot be excluded, but the likelihood is considered low. Based on the data provided, the Panel concluded that this food enzyme does not raise safety concerns under the intended conditions of use.

Age-dependent hepatic lymphoid organization directs successful immunity to hepatitis B.

Hepatitis B virus (HBV) is a major human pathogen that causes immune-mediated hepatitis. Successful immunity to HBV is age dependent: viral clearance occurs in most adults, whereas neonates and young children usually develop chronic infection. Using a mouse model of HBV infection, we sought mechanisms underpinning the age-dependent outcome of HBV and demonstrated that hepatic macrophages facilitate lymphoid organization and immune priming within the adult liver and promote successful immunity. In contrast, lymphoid organization and immune priming was greatly diminished in the livers of young mice, and of macrophage-depleted adult mice, leading to abrogated HBV immunity. Furthermore, we found that CXCL13, which is involved in B lymphocyte trafficking and lymphoid architecture and development, is expressed in an age-dependent manner in both adult mouse and human hepatic macrophages and plays an integral role in facilitating an effective immune response against HBV. Taken together, these results identify some of the immunological mechanisms necessary for effective control of HBV.

Measuring energy metabolism in cultured cells, including human pluripotent stem cells and differentiated cells.

Measurements of glycolysis and mitochondrial function are required to quantify energy metabolism in a wide variety of cellular contexts. In human pluripotent stem cells (hPSCs) and their differentiated progeny, this analysis can be challenging because of the unique cell properties, growth conditions and expense required to maintain these cell types. Here we provide protocols for analyzing energy metabolism in hPSCs and their early differentiated progenies that are generally applicable to mature cell types as well. Our approach has revealed distinct energy metabolism profiles used by hPSCs, differentiated cells, a variety of cancer cells and Rho-null cells. The protocols measure or estimate glycolysis on the basis of the extracellular acidification rate, and they measure or estimate oxidative phosphorylation on the basis of the oxygen consumption rate. Assays typically require 3 h after overnight sample preparation. Companion methods are also discussed and provided to aid researchers in developing more sophisticated experimental regimens for extended analyses of cellular bioenergetics.