Canonical and Nuclear mTOR Specify Distinct Transcriptional Programs in Androgen-Dependent Prostate Cancer Cells.

mTOR is a serine/threonine kinase that controls prostate cancer cell growth in part by regulating gene programs associated with metabolic and cell proliferation pathways. mTOR-mediated control of gene expression can be achieved via phosphorylation of transcription factors, leading to changes in their cellular localization and activities. mTOR also directly associates with chromatin in complex with transcriptional regulators, including the androgen receptor (AR). Nuclear mTOR (nmTOR) has been previously shown to act as a transcriptional integrator of the androgen signaling pathway in association with the chromatin remodeling machinery, AR, and FOXA1. However, the contribution of cytoplasmic mTOR (cmTOR) and nmTOR and the role played by FOXA1 in this process remains to be explored. Herein, we engineered cells expressing mTOR tagged with nuclear localization and export signals dictating mTOR localization. Transcriptome profiling in AR-positive prostate cancer cells revealed that nmTOR generally downregulates a subset of the androgen response pathway independently of its kinase activity, while cmTOR upregulates a cell cycle-related gene signature in a kinase-dependent manner. Biochemical and genome-wide transcriptomic analyses demonstrate that nmTOR functionally interacts with AR and FOXA1. Ablation of FOXA1 reprograms the nmTOR cistrome and transcriptome of androgen responsive prostate cancer cells. This works highlights a transcriptional regulatory pathway in which direct interactions between nmTOR, AR and FOXA1 dictate a combinatorial role for these factors in the control of specific gene programs in prostate cancer cells. IMPLICATIONS: The finding that canonical and nuclear mTOR signaling pathways control distinct gene programs opens therapeutic opportunities to modulate mTOR activity in prostate cancer cells.

Single-Institution Comparative Study of Magnetic Resonance-Guided Laser Interstitial Thermal Therapy and Open Corpus Callosotomy.

OBJECTIVE: Open corpus callosotomy (CC) poses a higher risk of perioperative morbidity than does magnetic resonance-guided laser interstitial thermal therapy (MRgLITT) for treatment of drop and generalized seizures without documented superiority. We present a single-institution comparison between open and MRgLITT CC. METHODS: A 2-year retrospective review was performed of patients who underwent open and MRgLITT CC (January 2019-January 2021). Demographics, surgical outcome data, hospital costs, and interhemispheric connectivity with diffusion tensor imaging were compared. RESULTS: The average age in years was 9.3 and 11.4 for CC (n = 4) and MRgLITT (n = 9), respectively. Preoperative drop seizure frequency was higher in CC (25 vs. 14.5 seizures/day; P = 0.59). At 10 months follow-up, the reduction in drop seizure frequency was better in open CC, but not statistically significant (93.8% vs. 64.3%; P = 0.21). The extent of CC ablation did not correlate with seizure reduction (Pearson coefficient = 0.09). An inverse correlation between interhemispheric connectivity change (diffusion tensor imaging analysis) and drop seizure frequency reduction was noted (Pearson coefficient = -0.97). Total hospital cost was significantly lower in MRgLITT ($67,754 vs. $107,111; P = 0.004), attributed to lower intensive care unit (1.1 vs. 4 days; P= 0.004) and total hospital stay (1.8 vs. 10.5 days; P = 0.0001). Postoperative hydrocephalus was present in 75% of patients in the CC group compared with zero in the MRgLITT group. CONCLUSIONS: Our middle-volume single-institution experience shows the safety, efficacy, and cost-effective benefit of MRgLITT compared with the traditional CC with therapeutic equipoise. This study is limited by the number of patients and, hence, further patient enrollment or multicenter study is warranted.

Establishment and characterization of patient-derived xenograft of a rare pediatric anaplastic pleomorphic xanthoastrocytoma (PXA) bearing a CDC42SE2-BRAF fusion.

Pleomorphic xanthoastrocytoma (PXA) is a rare subset of primary pediatric glioma with 70% 5-year disease free survival. However, up to 20% of cases present with local recurrence and malignant transformation into more aggressive type anaplastic PXA (AXPA) or glioblastoma. The understanding of disease etiology and mechanisms driving PXA and APXA are limited, and there is no standard of care. Therefore, development of relevant preclinical models to investigate molecular underpinnings of disease and to guide novel therapeutic approaches are of interest. Here, for the first time we established, and characterized a patient-derived xenograft (PDX) from a leptomeningeal spread of a patient with recurrent APXA bearing a novel CDC42SE2-BRAF fusion. An integrated -omics analysis was conducted to assess model fidelity of the genomic, transcriptomic, and proteomic/phosphoproteomic landscapes. A stable xenoline was derived directly from the patient recurrent tumor and maintained in 2D and 3D culture systems. Conserved histology features between the PDX and matched APXA specimen were maintained through serial passages. Whole exome sequencing (WES) demonstrated a high degree of conservation in the genomic landscape between PDX and matched human tumor, including small variants (Pearson's r = 0.794-0.839) and tumor mutational burden (~ 3 mutations/MB). Large chromosomal variations including chromosomal gains and losses were preserved in PDX. Notably, chromosomal gain in chromosomes 4-9, 17 and 18 and loss in the short arm of chromosome 9 associated with homozygous 9p21.3 deletion involving CDKN2A/B locus were identified in both patient tumor and PDX sample. Moreover, chromosomal rearrangement involving 7q34 fusion; CDC42SE-BRAF t (5;7) (q31.1, q34) (5:130,721,239, 7:140,482,820) was identified in the PDX tumor, xenoline and matched human tumor. Transcriptomic profile of the patient's tumor was retained in PDX (Pearson r = 0.88) and in xenoline (Pearson r = 0.63) as well as preservation of enriched signaling pathways (FDR Adjusted P < 0.05) including MAPK, EGFR and PI3K/AKT pathways. The multi-omics data of (WES, transcriptome, and reverse phase protein array (RPPA) was integrated to deduce potential actionable pathways for treatment (FDR < 0.05) including KEGG01521, KEGG05202, and KEGG05200. Both xenoline and PDX were resistant to the MEK inhibitors trametinib or mirdametinib at clinically relevant doses, recapitulating the patient's resistance to such treatment in the clinic. This set of APXA models will serve as a preclinical resource for developing novel therapeutic regimens for rare anaplastic PXAs and pediatric high-grade gliomas bearing BRAF fusions.

Effect of calorie restriction with or without exercise on body composition and fat distribution.

CONTEXT: There is debate over the independent and combined effects of dieting and increased physical activity on improving metabolic risk factors (body composition and fat distribution). OBJECTIVE: The objective of the study was to conduct a randomized, controlled trial (CALERIE) to test the effect of a 25% energy deficit by diet alone or diet plus exercise for 6 months on body composition and fat distribution. DESIGN: This was a randomized, controlled trial. SETTING: The study was conducted at an institutional research center. PARTICIPANTS: Thirty-five of 36 overweight but otherwise healthy participants (16 males, 19 females) completed the study. INTERVENTION: Participants were randomized to either control (healthy weight maintenance diet, n = 11), caloric restriction (CR; 25% reduction in energy intake, n = 12), or caloric restriction plus exercise (CR+EX; 12.5% reduction in energy intake + 12.5% increase in exercise energy expenditure, n = 12) for 6 months. MAIN OUTCOME MEASURES: Changes in body composition by dual-energy x-ray absorptiometry and changes in abdominal fat distribution by multislice computed tomography were measured. RESULTS: The calculated energy deficit across the intervention was not different between CR and CR+EX. Participants lost approximately 10% of body weight (CR: - 8.3 +/- 0.8, CR+EX: - 8.1 +/- 0.8 kg, P = 1.00), approximately 24% of fat mass (CR: - 5.8 +/- 0.6, CR+EX: - 6.4 +/- 0.6 kg, P = 0.99), and 27% of abdominal visceral fat (CR: 0.9 +/- 0.2, CR+EX: 0.8 +/- 0.2 kg, P = 1.00). Both whole-body and abdominal fat distribution were not altered by the intervention. CONCLUSION: Exercise plays an equivalent role to CR in terms of energy balance; however, it can also improve aerobic fitness, which has other important cardiovascular and metabolic implications.

Measurement of children's food intake with digital photography and the effects of second servings upon food intake.

This study tested the reliability and validity of measuring children's food intake with the digital photography method and the effects of second servings upon food intake. Food intake was measured in a school cafeteria for 5 days. Adiposity was assessed with body impedance analysis and body mass index, expressed as percentile rank. Mood and self-esteem were assessed with questionnaires. Estimates of food intake were highly reliable between two registered dietitians who independently estimated food intake. Boys ate more food than girls. A significant association between food intake and adiposity supported convergent validity. Non-significant correlations between food intake and depressed mood and self-esteem supported discriminant validity. When second servings were available, more food was selected and discarded, but mean food intake did not increase. Children who returned for second servings, however, ate more food when second servings were available, and a trend suggested that they also ate more food when second servings were not available. These findings support the reliability and validity of measuring children's food intake using digital photography and demonstrate its utility for studies of food intake and body weight.

Effect of calorie restriction with or without exercise on insulin sensitivity, beta-cell function, fat cell size, and ectopic lipid in overweight subjects.

OBJECTIVE: The purpose of this article was to determine the relationships among total body fat, visceral adipose tissue (VAT), fat cell size (FCS), ectopic fat deposition in liver (intrahepatic lipid [IHL]) and muscle (intramyocellular lipid [IMCL]), and insulin sensitivity index (S(i)) in healthy overweight, glucose-tolerant subjects and the effects of calorie restriction by diet alone or in conjunction with exercise on these variables. RESEARCH DESIGN AND METHODS: Forty-eight overweight volunteers were randomly assigned to four groups: control (100% of energy requirements), 25% calorie restriction (CR), 12.5% calorie restriction +12.5% energy expenditure through structured exercise (CREX), or 15% weight loss by a low-calorie diet followed by weight maintenance for 6 months (LCD). Weight, percent body fat, VAT, IMCL, IHL, FCS, and S(i) were assessed at baseline and month 6. RESULTS: At baseline, FCS was related to VAT and IHL (P < 0.05) but not to IMCL. FCS was also the strongest determinant of S(i) (P < 0.01). Weight loss at month 6 was 1 +/- 1% (control, mean +/- SE), 10 +/- 1% (CR), 10 +/- 1% (CREX), and 14 +/- 1% (LCD). VAT, FCS, percent body fat, and IHL were reduced in the three intervention groups (P < 0.01), but IMCL was unchanged. S(i) was increased at month 6 (P = 0.05) in the CREX (37 +/- 18%) and LCD (70 +/- 34%) groups (P < 0.05) and tended to increase in the CR group (40 +/- 20%, P = 0.08). Together the improvements in S(i) were related to loss in weight, fat mass, and VAT, but not IHL, IMCL, or FCS. CONCLUSIONS: Large adipocytes lead to lipid deposition in visceral and hepatic tissues, promoting insulin resistance. Calorie restriction by diet alone or with exercise reverses this trend.

Psychological adjustment during Army basic training.

This study evaluated changes in depression, anxiety, and stress during Army basic training. During week 1 of training, 139 soldiers from two companies volunteered for participation. In week 8, 93 soldiers were available for retesting. Self-reports of depressive symptoms and perceptions of stressfulness at both assessments fell within the normative range for nonclinical samples, whereas endorsement of anxiety symptoms remained slightly elevated, in the mild range. Women endorsed higher levels of anxiety (F = 8.87, p < 0.01) than did men. No gender or ethnicity differences were noted for changes in psychological distress over time. Regression analyses showed that subjects with the highest levels of initial distress on the anxiety, depression, and stress measures had the most change at the end of 8 weeks of training (r values between 0.61 and 0.39; all p < 0.01). Results suggest that initial levels of psychological distress are slightly elevated for anxiety but most individuals adapt to the stress of basic training, with normal levels of distress by the last week.

Comparison of digital photography to weighed and visual estimation of portion sizes.

OBJECTIVE: The primary goal was to test the validity of digital photography for measuring food portion sizes compared with weighed foods and with direct visual estimation. SAMPLES: A total of 60 test meals consisting of 10 different portion sizes from six different university cafeteria menus were prepared and weighed. DESIGN: Food selections and plate waste, as estimated by digital photography and direct visual estimation, were compared with weighed foods. For each method, three observers independently estimated portion sizes of each food. Observers expressed the portion sizes as a percentage of a standard serving. These percentages were multiplied by the weight of the standard portion to yield estimated weights. Statistical analyses To test validity, the estimates of food weights derived from both methods were compared with weighed foods using correlations and were compared with each other using Bland-Altman regression analysis. RESULTS: For the digital photography and direct visual estimation methods, estimates of the portion sizes for food selections, plate waste, and food intake were highly correlated with weighed foods. Both methods tended to yield small overestimates or underestimates. Bland-Altman regression found the two estimation methods to yield comparable results (bias less than 1.5 g). APPLICATIONS/CONCLUSIONS: These findings support the validity of the digital photography method for measuring portion sizes. Digital photography may be most useful for measuring food intake in settings that allow for the direct observation of food selections and plate waste but require minimum disruption of the eating environment, and allow unhurried estimates of portion sizes.

Changes in food intake and body weight associated with basic combat training.

This research project evaluated changes in food selections, food intake, and body weight during 8 weeks of basic combat training (BCT). During the first week of BCT, 139 soldiers from two companies volunteered for participation in the study. In the eighth week of BCT, 92 soldiers were available for retesting. A digital photography method for measuring food selections and food intake was developed for this study. Fruit intake of soldiers was very low at the beginning and end of BCT. Food intake for grains and milk products was low during the first week of BCT but improved by the end of BCT. Average body weight decreased during the 8 weeks of BCT, but heavier soldiers tended to lose weight and thinner soldiers tended to gain weight. These findings suggest that the overall effect of BCT was a trend toward improvement of healthy eating and healthy body weight.

The surgical reconstruction of the rheumatoid forefoot.

Rheumatoid arthritis is an autoimmune disorder that presents in females more often than males, and may affect people belonging to any age group. This disease shows no regional or ethnic preference. Although genetic and environmental causes have been proposed, the definitive cause of immunologic susceptibility, as well as viral and bacterial infectious processes that may cause rheumatoid arthritis, have not been identified. This article discusses various reconstructive forefoot surgeries to correct rheumatoid arthritis and the perioperative care of the patients who undergo surgery, along with the radiographic and magnetic resonance imaging findings associated with the disease.

Effect of 6-month calorie restriction and exercise on serum and liver lipids and markers of liver function.

OBJECTIVE: Nonalcoholic fatty liver disease (NAFLD) and its association with insulin resistance are increasingly recognized as major health burdens. The main objectives of this study were to assess the relation between liver lipid content and serum lipids, markers of liver function and inflammation in healthy overweight subjects, and to determine whether caloric restriction (CR) (which improves insulin resistance) reduces liver lipids in association with these same measures. METHODS AND PROCEDURES: Forty-six white and black overweight men and women (BMI = 24.7-31.3 kg/m(2)) were randomized to "control (CO)" = 100% energy requirements; "CR" = 25%; "caloric restriction and increased structured exercise (CR+EX)"= 12.5% CR + 12.5% increase in energy expenditure through exercise; or "low-calorie diet (LCD)" = 15% weight loss by liquid diet followed by weight-maintenance, for 6 months. Liver lipid content was assessed by magnetic resonance spectroscopy (MRS) and computed tomography (CT). Lipid concentrations, markers of liver function (alanine aminotransferase (ALT), alkaline phosphatase (ALK)), and whole-body inflammation (tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6), high-sensitivity C-reactive protein (hsCRP)) were measured in fasting blood. RESULTS: At baseline, increased liver lipid content (by MRS) correlated (P < 0.05) with elevated fasting triglyceride (r = 0.52), ALT (r = 0.42), and hsCRP (r = 0.33) concentrations after adjusting for sex, race, and alcohol consumption. With CR, liver lipid content was significantly lowered by CR, CR+EX, and LCD (detected by MRS only). The reduction in liver lipid content, however, was not significantly correlated with the reduction in triglycerides (r = 0.26; P = 0.11) or with the changes in ALT, high-density lipoprotein (HDL)-cholesterol, or markers of whole-body inflammation. DISCUSSION: CR may be beneficial for reducing liver lipid and lowering triglycerides in overweight subjects without known NAFLD.

Comparison of body composition methods in obese African-American women.

OBJECTIVE: To compare the accuracy of percentage body fat (%BF) estimates between bioelectrical impedance analysis (BIA) and DXA in obese African-American women. RESEARCH METHODS AND PROCEDURES: Fifty-five obese African-American women (mean age, 45 years; mean BMI, 38; mean %BF, 48%) were studied. BF was assessed by both BIA (RJL Systems BIA 101Q; RJL Systems, Clinton Township, MI) and DXA (Hologic QDR-2000 Bone Densitometer; Hologic Inc., Bedford, MA). Generalized and ethnicity- and obese-specific equations were used to calculate %BF from the BIA. Bland-Altman analyses were used to compare the agreement between the BIA and the DXA, with the DXA serving as the criterion measure. RESULTS: Two of the generalized equations provided consistent estimates across the weight range in comparison with the DXA estimates, whereas most of the other equations increasingly underestimated %BF as BF increased. One of the generalized and one of the ethnicity-specific equations had mean differences that were not significantly different from the DXA value. DISCUSSION: The findings show that the Lukaski equation provided the most precise and accurate estimates of %BF in comparison with the QDR 2000 and provide preliminary support for the use of this equation for obese African-American women.

Two-year internet-based randomized controlled trial for weight loss in African-American girls.

OBJECTIVE: A randomized controlled trial tested the efficacy of an internet-based lifestyle behavior modification program for African-American girls over a 2-year period of intervention. RESEARCH METHODS AND PROCEDURES: Fifty-seven overweight (mean BMI percentile, 98.3) African-American girls (mean age, 13.2 years) were randomly assigned to an interactive behavioral internet program or an internet health education program, the control condition. Overweight parents were also participants in the study. Forty adolescent-parent dyads (70%) completed the 2-year trial. Outcome data including BMI, body weight, body composition, and weight loss behaviors were collected at baseline and at 6-month intervals. A computer server tracked use of the web sites. RESULTS: An intention-to-treat statistical approach was used, with the last observation carried forward. In comparison with the control condition, adolescents in the behavioral program lost more mean body fat (BF) (-1.12 +/- 0.47% vs. 0.43 +/- 0.47% BF, p < 0.05), and parents in the behavioral program lost significantly more mean body weight (-2.43 +/- 0.66 vs. -0.35 +/- 0.64 kg, p < 0.05) during the first 6 months. This weight loss was regained over the next 18 months. After 2 years, differences in fat for adolescents (-0.08 +/- 0.71% vs. 0.84 +/- 0.72% BF) and weight for parents (-1.1 +/- 0.91 vs. -0.60 +/- 0.89 kg) did not differ between the behavioral and control programs. DISCUSSION: An internet-based weight management program for African-American adolescent girls and their parents resulted in weight loss during the first 6 months but did not yield long-term loss due to reduced use of the web site over time.