Accumulation of cholesterol, triglycerides and ceramides in hepatocellular carcinomas of diethylnitrosamine injected mice

Background Dysregulated lipid metabolism is critically involved in the development of hepatocellular carcinoma (HCC). The respective metabolic pathways affected in HCC can be identified using suitable experimental models. Mice injected with diethylnitrosamine (DEN) and fed a normal chow develop HCC. For the analysis of the pathophysiology of HCC in this model a comprehensive lipidomic analysis was performed. Methods Lipids were measured in tumor and non-tumorous tissues by direct flow injection analysis. Proteins with a role in lipid metabolism were analysed by immunoblot. Mann-Whitney U-test or paired Student´s t-test were used for data analysis. Results Intra-tumor lipid deposition is a characteristic of HCCs, and di- and triglycerides accumulated in the tumor tissues of the mice. Peroxisome proliferator-activated receptor gamma coactivator 1 alpha, lipoprotein lipase and hepatic lipase protein were low in the tumors whereas proteins involved in de novo lipogenesis were not changed. Higher rates of de novo lipogenesis cause a shift towards saturated acyl chains, which did not occur in the murine HCC model. Besides, LDL-receptor protein and cholesteryl ester levels were higher in the murine HCC tissues. Ceramides are cytotoxic lipids and are low in human HCCs. Notably, ceramide levels increased in the murine tumors, and the simultaneous decline of sphingomyelins suggests that sphingomyelinases were involved herein. DEN is well described to induce the tumor suppressor protein p53 in the liver, and p53 was additionally upregulated in the tumors. Conclusions Ceramides mediate the anti-cancer effects of different chemotherapeutic drugs and restoration of ceramide levels was effective against HCC. High ceramide levels in the tumors makes the DEN injected mice an unsuitable model to study therapies targeting ceramide metabolism. This model is useful for investigating how tumors evade the cytotoxic effects of ceramides.

Correlation between Actual versus Perceived Body Mass Index using a 3D Avatar on Female Football and Rugby Athletes

Introduction: The pressures of body image can be considered as demoralising, especially within the female sporting environment, where female athletes often express the greatest number of negative thoughts and feelings towards their own body shape, appearance, and dissatisfaction. Research surrounding body image, body composition and body mass index (BMI) has shown that when participants use visual impressions, for perceived body image (PBI) they have failed to produce realistic images and often lacked body stimuli with realistic weight manipulations (Madrigal, 2000). To portray more realistic statistically probable weight manipulations of a personalised stimuli, a 3D Avatar can be used to establish how female athletes perceive their body image. Therefore, the main objective of this investigation is to determine the correlation between actual and perceived BMI using a 3D Avatar within female athletes. Method:n =18 female participants between the ages of 18–23 years of age and competing in football and rugby at club and university level were recruited. Stretched stature (m) and body mass (kg) were taken and values used to calculate actual (kg/m²) and perceived BMI. A computer generated (Unity Player) 3D Avatar, with a visual slide from an underweight to average to obese continuum, was used to assess participants perceived BMI. P value was set at < 0.05 and a Paired Student t-Test was used to test for the difference. A Pearson’s Correlation Coefficient was then used to test the strength of the correlation between the actual and perceived BMI. Results: Actual BMI ranged from 19.5 - 36.9 (x̄ 25.1 ± 4.7), whereas the perceived BMI ranged from 23.2 - 30.8 (x̄ 26.7 ± 2.6). A Paired Student t–test set at P < 0.05 suggested a significant difference between actual and perceived BMI (P = 0.023), and a Pearson’s Correlation Coefficient test confirmed a strong correlation of r = 0.875. Conclusion: Results indicated that perceived BMI was higher than the participants actual BMI and suggested that female athletes competing in football and rugby have a large amount of body dissatisfaction.

A Naturalistic Study of the Maintenance of Gains Made With Treatment of Patients With Profound Treatment-Refractory Obsessive-Compulsive Disorder

Obsessive-compulsive disorder (OCD) generally responds to first-line treatment but patients often relapse. The United Kingdom National OCD Inpatient Service treats patients who have failed to respond to at least two trials of SRI, augmented with a dopamine blocker and two trials of ERP. Despite this, they have profound treatment-refractory OCD and require 24-h nursing care due to severe OCD. We examined patients' Y-BOCS score on admission, discharge and at each follow-up from all patients discharged over 5 years (02/01/2014-31/12/18). Data were analysed using SPSS. Paired student t-tests were used to assess improvement from admission to discharge and each follow-up. Over 5 years, 130 adult patients were treated: 79 male and 51 female with an average age of 42.3 years (20-82; sd14.4). Their ethnic backgrounds were; 115 Caucasian, 11 South Asian, 1 Chinese, and 3 Unspecified. On admission, the average Y-BOCS total score was 36.9 (30-40; sd2.6). At discharge, patients had improved on average by 36% (Y-BOCS reduction to 23.4 = moderate OCD). Similar reduction in Y-BOCS continued throughout the year with an average Y-BOCS of 22.9 at 1 month (n = 69); 23 at 3 months (n = 70); 21.3 at 6 months (n = 78) and 21.9 at 1 year (n = 77). Twenty-seven patients did not attend any follow-up appointment whilst others attended at least one appointment with the majority attending more than 3. Using student t-test, improvements at discharge, 1, 3, 6, and 12 months post-treatment showed a highly significant improvement (p &lt; 0.001). Gains made following inpatient treatment for treatment-refractory OCD were generally maintained until 1 year post-treatment.

Total Knee Arthroplasty in Octogenarians: Should We Still Be so Restrictive?

Demand for total knee arthroplasty (TKA) in octogenarians will increase in subsequent years as society ages. We conducted a retrospective observational study in octogenarians operated on with TKA between 2015 and 2019, comparing preoperative and postoperative Knee Society Score (KSS), Knee Society Function Score (KSFS), extension and flexion balance, and radiologic alignment using a paired Student t-test. A chi-squared test was used to correlate mortality with Charlson comorbidities index score and with ASA scale. Kaplan–Meier analysis was performed to calculate patient survival. In this period 36 patients ≥80 years underwent TKA, with a mean age of 81.6 years. Of these, 24 patients (66.7%) were classified as ASA II and 12 (33.3%) as ASA III. Sixteen patients (44.4%) were Charlson 0, 14 (38.9%) Charlson 1, two (5.6%) Charlson 2, and four (11.1%) Charlson 3. KSS, KSFS, flexion and extension range, and radiologic alignment were statistically significant (p < 0.001) when comparing preoperatory and post-operatory data. No correlation (p > 0.05) was found between mortality and ASA or Charlson score. Seven patients (19.4%) suffered a medical complication and two patients experienced surgical complications. Four patient died (11.1%) during follow-up. The mean patient survival was 67.4 months. Patients ≥80 years achieve clinical improvement after TKA. Comorbidities, not age, are the burden for surgery in older patients.

117 Feasibility and Scar Outcome with a Single-use Adhesive Silicone in Pediatric Burn Survivors

Introduction Silicone sheets are commonly used for prevention and treatment of burn scars and silicone-based products have recently been recommended as the gold standard non-invasive treatment choice. One problem with silicone gel sheets application is that the sheets lose their adhesive qualities making them difficult to affix to the patient and costly to replace when lost. A new ultra-thin, clear and adhesive silicone gel sheet layered on a semi-permeable polyester backing was trialed at our burn center. The silicone is designed to be a one-time application, worn during all activities until it naturally falls off, thus eliminating the need for daily removal and cleaning. The purpose of this study was to evaluate the feasibility and effectiveness of the single-use adhesive silicone sheets with pediatric burn survivors. Methods Medical records of pediatric burn survivors who used the adherent silicone sheets between September 2019 and July 2020 were retrospectively reviewed. Information regarding details of silicone use and scar outcome scores were extracted from the records. Data were analyzed using paired Student t-tests and Wilcoxon Signed Rank to compare scars before and after using the silicone. Results The silicone was applied to 22 body areas of 9 patients. The average age of the patients was 7.4 (±8.5) years and TBSA 53 ±17%. The time from injury or grafting to when the silicone was applied was 200.5 (±13.4) days. The silicone was worn without interruption for an average of 12.8 days. Total Patient Observer Scar Assessment Scale (POSAS) scores for both the patient and observer evaluations significantly improved after use with the single-use adherent silicone (↓13.83 patient p=.005, ↓5.4 observer p=.01). Evaluating individual scar parameters showed 4/7 patient reported and observed areas of the POSAS evaluation improved significantly with adherent silicone use (Figure 1). Only one patient reported a complication of itch at day 5 and the silicone was removed. Conclusions The single-use adhesive silicone investigated in this study appears to adequately adhere to patients over multiple days, thus overcoming common barriers to silicone use of the products falling off, getting lost and needing to be replaced. Use of such a product could improve ease and continuity with scar management. In addition, for this small cohort of patients, scar improvement was noted with silicone use. The results of this study indicate that larger prospective studies evaluating the silicone efficacy may be beneficial.

Subjective Sleep Patterns and Jet Lag Symptoms of Junior Netball Players Prior to and During an International Tournament: A Case Study

Purpose: To assess the impact of long-haul transmeridian travel on subjective sleep patterns and jet lag symptoms in youth athletes around an international tournament. Methods: An observational descriptive design was used. Subjective sleep diaries and perceived responses to jet lag were collected and analyzed for a national junior netball team competing in an international tournament. Sleep diaries and questionnaires were completed daily prior to and during travel, and throughout the tournament. Results were categorized into pretravel, travel, training, and match nights. Means were compared performing a paired Student t test with significance set at P < .05. Data are presented as mean (SD) and median (minimum, maximum). Results: Athletes reported significantly greater time in bed on match days compared with training (P < .001) and travel (P = .002) days, and on pretravel days compared with travel (P < .001) and training (P = .028) days. Sleep ratings were significantly better on pretravel days compared with match (P = .013) days. Perceived jet lag was worse on match (P = .043) days compared with pretravel days. Significant differences were also observed between a number of conditions for meals, mood, bowel activity, and fatigue. Conclusion: Youth athletes experience significantly less opportunity for sleep during long-haul transmeridian travel and face disruptions to daily routines during travel which impact food intake. Young athletes also experience disturbed sleep prior to and during competition. These results highlight the need for practices to alleviate jet lag symptoms and improve the sleep of young athletes traveling for tournaments in an effort to optimize recovery and performance.

Hyperbaric oxygen but not hyperbaric air increases insulin sensitivity in men with type 2 diabetes mellitus

(Wilkinson DC, Chapman IM, Heilbronn LK. Hyperbaric oxygen but not hyperbaric air increases insulin sensitivity in men with type 2 diabetes mellitus. Diving and Hyperbaric Medicine. 2020 December 20;50(4):386–390. doi: 10.28920/dhm50.4.386-390. PMID: 33325020.) Introduction: We have previously shown that hyperbaric oxygen treatment (HBOT) increased insulin sensitivity in men who were obese or overweight, both with and without type 2 diabetes. The aim of this study was to test whether this insulin-sensitising effect is seen in hyperbaric air (HA). Methods: Men with type 2 diabetes who were obese or overweight were randomised to two groups: HBOT (n = 13) or HA (n = 11). A hyperinsulinaemic euglycaemic glucose clamp (80 mU·m-2·min-1) was performed at baseline and during hyperbaric intervention. Both groups were compressed to 203 kPa (two atmospheres absolute) for 90 minutes followed by a linear 30-minute decompression. The HBOT group breathed oxygen via a hood while the HA group breathed chamber air. Insulin sensitivity was assessed from the glucose infusion rate (GIR) during the last 30 minutes in the hyperbaric chamber (SS1) and the first 30 minutes after exit (SS2). Data were analysed for within-group effect by paired student t-test and between-group effect by one-way ANOVA. Results: HBOT increased GIR by a mean 26% at SS1 (P = 0.04) and 23% at SS2 (P = 0.018). There was no significant change in GIR during or after HA. A between-group effect was evident for the change in GIR at SS1 in HBOT vs HA (P = 0.036). Conclusions: The pathway by which insulin sensitivity is increased in men with type 2 diabetes requires the high oxygen partial pressures of HBOT and should be further investigated. Insulin sensitivity was not changed in hyperbaric air.

Air-Drying Llama Sperm Affects DNA Integrity

The objective of this study was to evaluate the effects of air-drying preservation on llama sperm DNA. Semen collections were carried out using electroejaculation under general anesthesia. A total of 16 ejaculates were processed from 4 males (n = 4, r = 4). Each sample was diluted 4:1 in a collagenase solution in TALP media, then incubated and centrifuged at 800 g for 8 min. The pellet was re-suspended to a concentration of 20 million sperm/ml in TALP. Then the samples were placed onto sterile slides forming lines and were left to dry under laminar flow for 15 min. After this, the slides were placed into Falcon centrifuge tubes and kept at 5°C. Sperm characteristics (motility, membrane function, viability and morphology) were evaluated in raw semen and in the air-dried samples kept at 5°C for 30 min. DNA evaluation (integrity and degree of chromatin condensation) was carried out in raw semen and in the air-dried samples after 30 min, 7, 14, 21, 30, and 60 days after preservation. To compare raw semen to the air-dried samples, a Wilcoxon test was used for all sperm characteristics except for DNA, where a paired Student t-test was applied. A split plot design was used to compare chromatin condensation between the different periods of preservation and a Kruskal Wallis test was used to compare DNA integrity. Motility, membrane function, viability and sperm with intact DNA decreased in the air-dried samples (p &lt; 0.05), while morphology and chromatin condensation were not affected (p &gt; 0.05). No significant differences were observed in the percentage of sperm with condensed chromatin between the different periods of preservation (p &gt; 0.05). On the other hand, a significant decrease in the percentage of sperm with intact DNA was observed as from day 7 of preservation (p &lt; 0.05). In conclusion the air-drying process has a negative effect on llama sperm DNA, hence the media used will need to be improved to protect DNA and be able to implement this technique in this species.

Alignment of State Regulations With Breastfeeding and Beverage Best Practices for Childcare Centers and Family Childcare Homes, United States

Objectives Childcare is an important setting for nutrition; nearly half of young children in the United States participate in licensed childcare, where they consume up to two-thirds of their daily dietary intake. We compared state regulations for childcare with best practices to support breastfeeding and healthy beverage provision. Methods We reviewed regulations for childcare centers (centers) and family childcare homes (homes) in effect May–July 2016 and rated all 50 states for inclusion (1 = not included, 2 = partially included, 3 = fully included) of 12 breastfeeding and beverage best practices. We calculated average ratings for 6 practices specific to infants aged 0-11 months, 6 practices specific to children aged 1-6 years, and all 12 practices, by state and across all states. We assessed significant differences between centers and homes for each best practice by using McNemar–Bowker tests for symmetry, and we assessed differences across states by using paired student t tests. Results States included best practices in regulations for centers more often than for homes. Average ratings (standard deviations) in regulations across all states were significantly higher in centers than in homes for infant best practices (2.1 [0.5] vs 1.8 [0.5], P < .001), child best practices (2.1 [0.6] vs 1.8 [0.6], P = .002), and all 12 best practices combined (2.1 [0.5] vs 1.8 [0.6], P < .001). Conclusions Although best practices were more consistently included in regulations for centers than for homes, many state childcare regulations did not include best practices to support breastfeeding and the provision of healthy beverages. Findings can be used to inform efforts to improve regulations and to reduce differences between centers and homes.

The impact of Dapagliflozin, and Empagliflozin on Na+/Li+ countertransport in erythrocyte membranes of healthy volunteers in acute drug tests

Background The formation of the effect of SGLT 2 inhibitors on the cardiovascular system implies inhibition of the Na+/H+exchanger (NHE) that leads to a decrease in the concentration of Na+, which in turn contributes to a decrease in the concentration of Ca2+ in the cardiomyocyte (S. Verma, J. McMurray, 2018). However, this effect may be different against the background of exposure to empagliflozin and dapagliflozin. A study of A. Baartscheer (2017) demonstrated the reduction of [Na+], systolic and diastolic [Ca+] in cardiomyocyte is affected by Empagliflozin. N.N. Hamouda's research shows that Dapagliflozin reduce [Na+], systolic, but not diastolic [Ca2+] in cardiomyocytes. In this regard, studying the Na+/Na+ exchanger (which is the mode of the NHE operation) activity estimated by the Na+/Li+countertransport (SLC) in the erythrocyte membrane (K. Morgan, M. Canessa, 1990; M. Canessa, 1989) is of great interest. Purpose A study of the impact of Dapagliflozin, and Empagliflozin on SLC in erythrocyte membranes of healthy volunteers in acute drug tests. Materials and methods 10 healthy volunteers (7 men and 3 women) were included in the study. The mean age of the study group was 24±0,6 years. The volunteers took singly Dapagliflozin, and Empagliflozin separately in one-month intervals. The activity of SLC was determined before drug consumption, as well as 2, 12, and 24 hours after consumption by the method of M.Canessa (1980). All data were tested for normality using the Kolmogorov–Smirnov test and statistically tested by paired Student t-test. Results Mean speeds of SLC before the drugs were administered was 289.1±33.1 mmol/L cell*h in the Dapagliflozin group and 287.8±37.3 mmol/L cell*h in the Empagliflozin group (p=0.979). Two hours after Dapagliflozin was administered, the speed of SLC increased to 76.3±17.0 mmol/L cell*h (95% CI: 37.9–114.7) and was maximum 365±41.6 mmol/L cell*h (p=0.002) for observation period. Then after 12 and 24 hours decreasing SLC activity was observed. However, the values were higher than the initial level and amounted to 335.5±39.18 mmol/L cell*h (p=0.024) after 12 hours and 331±31.8 mmol/L cell*h (p=0.001) after 24 hours. Two hours after Empagliflozin was administered, the speed of SLC not significantly changed and amounted to 287.8±37.3mmol/L cell*h (p=0.7). After 12 hours, the speed not significantly increased to 23.4±11.9 (95% CI: 4.6–51.4) mmol/L cell*h and amounted to 311.1±39.2 (p=0.08), with the subsequent decrease of speed within 24 hours to 277.6±40.5 mmol/L cell*h (p=0.5). Thus Dapagliflozin and Empagliflozin have different effects on the function of Na+/Na+ exchanger which may differently affect the changing of the [Na+] and [Ca2+] in the cardiomyocyte. Conclusions 1. When Dapagliflozin was administered the SLC speed reached its maximum level after 2h and maintained higher than initial level after 12h and 24h. 2. When Empagliflozin was administered no significant change in SLC speed was observed. Funding Acknowledgement Type of funding source: None