OBJECTIVES/GOALS: Clinical tissue specimens are primarily destined for formalin fixed, paraffin embedded processing to create a basis for diagnosis by microscopic examination. Innovations in specimen processing are required to expand its availability for inclusion as the substrate in assays that can contribute to the further development of Precision Medicine. METHODS/STUDY POPULATION: Transurethral resection of bladder tumors were selected for testing based on availability and tissue composition. A wash step was used to generate daughter aliquots composed of dislodged cells and a solution with prior contact to the parent tissue. This wash step served two purposes: 1) reduce the amount of contaminating material from spreading to other cases, a problem known to be associated with this type of specimen; and 2) create aliquots from which additional informative data could be generated. These daughter aliquots were then examined to determine their value as a source for exosome profiling, metabolomic studies, molecular characterization and organoid development. The parent tissue was not compromised, was able to undergo conventional processing and yielded results equivalent to unwashed specimens. RESULTS/ANTICIPATED RESULTS: Exosomes secreted by the tumor cells were identified to be present in the daughter aliquots by a combination of their isolation using CD31 and detection of miR-21 expression. These exosomes were confirmed to be not related to fragmented cells from testing for beta-tubulin. A global/discovery-based approach using mass spectrometry provided insights into early characterization of metabolomic profiles present in these tumor cells. Ample amounts of high quality DNA (226 ng/ul concentrations; 11.3 ug total) were recovered from the dislodged, excess cells in the wash for molecular studies. Finally, from viable cells recovered in one of the daughter wash aliquots, the ability to grow organoids was proven to be possible and reproducible. DISCUSSION/SIGNIFICANCE: Based on these results, the value of the clinical specimen can be markedly expanded for utilization in research and possible clinical use without detracting from the parent tissue. This non-destructive, easy to adopt wash procedure can potentially lead to an influx of data that may ultimately prove useful in improving patient care.

We revisit the problem of estimating the spot volatility of an Itô semimartingale using a kernel estimator. A central limit theorem (CLT) with an optimal convergence rate is established for a general two-sided kernel. A new pre-averaging/kernel estimator for spot volatility is also introduced to handle the microstructure noise of ultra high-frequency observations. A CLT for the estimation error of the new estimator is obtained, and the optimal selection of the bandwidth and kernel function is subsequently studied. It is shown that the pre-averaging/kernel estimator’s asymptotic variance is minimal for two-sided exponential kernels, hence justifying the need of working with kernels of unbounded support. Feasible implementation of the proposed estimators with optimal bandwidth is developed as well. Monte Carlo experiments confirm the superior performance of the new method.

Nutritional Risk Screening index is a standard tool to assess nutritional risk, but epidemiological data are scarce on controlling nutritional status (CONUT) as a prognostic marker in acute haemorrhagic stroke (AHS). We aimed to explore whether the CONUT may predict a 3-month functional outcome in AHS. In total, 349 Chinese patients with incident AHS were consecutively recruited, and their malnutrition risks were determined using a high CONUT score of ≥ 2. The cohort patients were divided into high-CONUT (≥ 2) and low-CONUT (< 2) groups, and primary outcomes were a poor functional prognosis defined as the modified Rankin Scale (mRS) score of ≥ 3 at post-discharge for 3 months. Odds ratios (OR) with 95 % confidence intervals (CI) for the poor functional prognosis at post-discharge were estimated by using a logistic analysis with additional adjustments for unbalanced variables between the high-CONUT and low-CONUT groups. A total of 328 patients (60·38 ± 12·83 years; 66·77 % male) completed the mRS assessment at post-discharge for 3 months, with 172 patients at malnutrition risk at admission and 104 patients with a poor prognosis. The levels of total cholesterol and total lymphocyte counts were significantly lower in high-CONUT patients than low-CONUT patients (P = 0·012 and < 0·001, respectively). At 3-month post discharge, there was a greater risk for the poor outcome in the high-CONUT compared with the low-CONUT patients at admission (OR: 2·32, 95 % CI: 1·28, 4·17). High-CONUT scores independently predict a 3-month poor prognosis in AHS, which helps to identify those who need additional nutritional managements.

Increasing evidence has shown that an active, socially engaged lifestyle in leisure time might protect older adults against the decline of cognitive function. It remains unclear, however, which types of leisure activities are more beneficial to maintain cognitive function, and whether there are gender differences in the association between leisure activities and cognitive function. We used a two-wave of panel data from 1,018 older adults aged 60 and older in rural China to examine the lag effects of different types of leisure activities on cognitive functioning and to identify the gender differences in their impacts on cognition in rural China. Ordinary least-squares regression models showed that high physical activities were associated with better cognitive function. High intensity of cognitive activities and engaging in physical activities have a protective effect on cognitive function among older men rather than older women. Further, we found that cognitive activities had a stronger effect on cognitive function among older men than older women. It is important to consider gender-specific intervention in leisure activities to maintain cognitive function among older adults.

This study examined the association between childhood conditions and arthritis among middle-aged and older adults in China. The data were derived from the 2015 wave and the life-history module of the China Health and Retirement Longitudinal Study. Face-to-face interviews were conducted with respondents age 45 and over across China. Multiple imputation was used to handle the missing data, generating a final analytic sample of 19,800. Doctor-diagnosed arthritis was the main outcome variable. Random-effects logistic regression models were used to test the proposed models. Approximately 8 per cent of the respondents had better family financial status in childhood than their neighbours. Close to 8 per cent had been hospitalised or encountered similar conditions (e.g. confined to bed or home) for at least one month in childhood. Around one-third reported better subjective health in childhood than their peers. The majority of the respondents (80%) reported that they had stable health resources, and that their mothers were illiterate during their childhood. Childhood family financial status, subjective health, mother's education, access to health care and medical catastrophic events were found to be significant factors associated with arthritis in later life, after controlling for adulthood and older-age conditions (family financial status: odds ratio (OR) = 0.885, 95 per cent confidence interval (95% CI) = 0.848–0.924; subjective health: OR = 0.924, 95% CI = 0.889–0.960; mother's education: OR = 0.863, 95% CI = 0.750–0.992; access to health care: OR = 0.729, 95% CI = 0.552–0.964; medical catastrophic events: OR = 1.266, 95% CI = 1.108–1.446). The study results highlight an important role that childhood conditions play in affecting the onset of arthritis in late life in China. Health-care providers may consider childhood conditions as a valuable screening criterion to identify risk populations, which could be used to guide health promotion and prevention programmes, and promote healthy ageing.

Human milk fat is specially tailored to supply the developing infant with adequate and balanced nutrients. The present study aimed to quantify the composition of fatty acids, tocopherols and carotenoids in human milk, with special emphasis on the lactational changes. Colostrum, transitional and mature milk samples were collected longitudinally from the same forty-two healthy, well-nourished Chinese mothers. Fatty acids were quantified by GC with carotenoids (carotenes and xanthophylls) and tocopherols (α-, γ-tocopherol) determined by HPLC. Total fatty acid (TFA) content increased from 15·09 g/l in colostrum to 32·57 g/l in mature milk with the percentages of DHA and arachidonic acid (ARA) decreased. The ratio of n-6:n-3 PUFA and ARA:DHA remained constant during lactation at about 11:1 and 1·3:1, respectively. Both α-tocopherol and γ-tocopherol decreased over lactation with the ratio of α-:γ-tocopherol declined significantly from 7·21:1 to 4·21:1 (P < 0·001). Carotenoids all dropped from colostrum to mature milk as the less polar carotenes dropped by 88·67 %, while xanthophylls only dropped by 35·92 %. Lutein was predominated in both transitional and mature milk carotenoids (51·64–52·49 %), while colostrum carotenoids were mainly composed of lycopene (32·83 %) and β-carotene (30·78 %). The concentrations of tocopherols and xanthophylls but not carotenes were positively associated with TFA content in milk. These results suggested that colostrum and mature milk contained divergent lipid profiles and selective transfer mechanisms related to polarity might be involved. The present outcomes provide new insights for future breast-feeding studies, which also add in scientific evidences for the design of both initial and follow-on infant formulas.

The aim of the present study was to explore the influence of tea consumption on diabetes mellitus in the Chinese population. This multi-centre, cross-sectional study was conducted in eight sites from south, east, north, west and middle regions in China by enrolling 12 017 subjects aged 20–70 years. Socio-demographic and general information was collected by a standardised questionnaire. A standard procedure was used to measure anthropometric characteristics and to obtain blood samples. The diagnosis of diabetes was determined using a standard 75-g oral glucose tolerance test. In the final analysis, 10 825 participants were included and multiple logistic models and interaction effect analysis were applied for assessing the association between tea drinking with diabetes. Compared with non-tea drinkers, the multivariable-adjusted OR for newly diagnosed diabetes were 0·80 (95 % CI 0·67, 0·97), 0·88 (95 % CI 0·71, 1·09) and 0·86 (95 % CI 0·67, 1·11) for daily tea drinkers, occasional tea drinkers and seldom tea drinkers, respectively. Furthermore, drinking tea daily was related to decreased risk of diabetes in females by 32 %, elderly (>45 years) by 24 % and obese (BMI > 30 kg/m2) by 34 %. Moreover, drinking dark tea was associated with reduced risk of diabetes by 45 % (OR 0·55; 95 % CI 0·42, 0·72; P < 0·01). The results imply that drinking tea daily was negatively related to risk of diabetes in female, elderly and obese people. In addition, drinking dark tea was associated with decreased risk of type 2 diabetes mellitus.

This study quantified the fatty acid profile with emphasis on the stereo-specifically numbered (sn) 2 positional distribution in TAG and the composition of main phospholipids at different lactation stages. Colostrum milk (n 70), transitional milk (n 96) and mature milk (n 82) were obtained longitudinally from healthy lactating women in Shanghai. During lactation, total fatty acid content increased, with SFA dominating in fatty acid profile. A high ratio of n-6:n-3 PUFA was observed as 11:1 over lactation due to the abundance of linoleic acid in Chinese human milk. As the main SFA, palmitic acid showed absolute sn-2 selectivity, while oleic acid, linoleic acid and α-linolenic acid, the main unsaturated fatty acids, were primarily esterified at the sn-1 and sn-3 positions. Nervonic acid and C22 PUFA including DHA were more enriched in colostrum with an sn-2 positional preference. A total of three dominant phospholipids (phosphatidylethanolamine (PE), phosphatidylcholine (PC) and sphingomyelin (SM)) were analysed in the collected samples, and each showed a decline in amount over lactation. PC was the dominant compound followed by SM and PE. With prolonged breast-feeding time, percentage of PE in total phospholipids remained constant, but PC decreased, and SM increased. Results from this study indicated a lipid profile different from Western reports and may aid the development of future infant formula more suitable for Chinese babies.

Vision impairment is prevalent and it is strongly associated with depressive symptoms in older adults. This study aimed to investigate the mediating roles of functional limitations and social support on the relationship between vision impairment and depressive symptoms in older adults. This study used data from a probability-based sample of 1,093 adults aged 60 and older in Shanghai, China. Structural equation models were used to examine the structural relationships among sets of variables simultaneously, including vision impairment, activities of daily living ADLs, instrumental ADLs (IADLs), friends support, family support, relatives support and depressive symptoms. The bootstrapping method and the program PRODCLIN were used to test the indirect effects of these variables. This study found that vision impairment was directly associated with a higher level of depressive symptoms, and the association was partially mediated by functional limitations (IADLs) and social support (friends support). The study demonstrates that improving social support from friends and enhancing social participation for visually impaired older adults can reduce depressive symptoms. More importantly, this study contributes to the knowledge of mediating mechanisms between vision impairment and depressive symptoms.

Developing metallic materials with a good combination of strength and ductility has been an unending pursuit of materials scientists. The emergence of high/medium-entropy alloys (HEA/MEA) provided a novel strategy to achieve this. Here, we further strengthened a strong-and-ductile MEA using a traditional solid solution strengthening theory. The selection of solute elements was assisted by mechanical property and microstructure predictive models. Extensive microstructural characterizations and mechanical tests were performed to verify the models and to understand the mechanical behavior and deformation mechanisms of the designated CoCrNi–3W alloy. Our results show good experiment-model agreement. The incorporation of 3 at.% W into the ternary CoCrNi matrix increased its intrinsic strength by ∼20%. External strengthening through microstructural refinement led to a yield strength nearly double that of the parent alloy, CoCrNi. The increase in strength is obtained with still good ductility when tested down to 77 K. Nanoscale twin boundaries are observed in the post-fracture microstructure under 77 K. The combination of strength and ductility after W additions deviate from the traditional strength-ductility-trade-off contour.

We studied how older people describe others with cognitive impairment. Forty-two focus groups represented African Americans, American Indians, Chinese Americans, Latinos, Vietnamese Americans, and Whites other than Latinos (Whites) (N = 396, ages 50+), in nine locations in the United States of America. Axial coding connected categories and identified themes. The constant comparison method compared themes across ethnic groups. African Americans, American Indians and Whites emphasised memory loss. African Americans, American Indians, Latinos and Whites stressed withdrawal, isolation and repetitive speech. African Americans, American Indians, Vietnamese Americans and Whites emphasised ‘slow thinking’. Only Whites described mood swings and personality changes. Many participants attributed dementia to stress. Terms describing others with dementia included ‘Alzheimer's’, ‘dementia’, ‘senile’ and ‘crazy’. Euphemisms were common (‘senior moment’, ‘old timer's disease’). Responses focused on memory, with limited mention of other cognitive functions. Differences among ethnic groups in descriptions of cognitive health and cognitive impairment underscore the need to tailor public health messages about cognitive health to ways that people construe its loss, and to their interest in maintaining it, so that messages and terms used are familiar, understandable and relevant to the groups for which they are designed. Health promotion efforts should develop ethnically sensitive ways to address the widely held misperception that even serious cognitive impairment is a normal characteristic of ageing and also to address stigma associated with cognitive impairment.

Lutein and zeaxanthin are thought to decrease the incidence of age-related macular degeneration (AMD); however, findings have been inconsistent. We conducted a systematic literature review and meta-analysis to evaluate the relationship between dietary intake of lutein and zeaxanthin and AMD risk. Relevant studies were identified by searching five databases up to April 2010. Reference lists of articles were retrieved, and experts were contacted. Literature search, data extraction and study quality assessment were performed independently by two reviewers and results were pooled quantitatively using meta-analysis methods. The potential sources of heterogeneity and publication bias were also estimated. The search yielded six longitudinal cohort studies. The pooled relative risk (RR) for early AMD, comparing the highest with the lowest category of lutein and zeaxanthin intake, was 0·96 (95 % CI 0·78, 1·17). Dietary intake of these carotenoids was significantly related with a reduction in risk of late AMD (RR 0·74; 95 % CI 0·57, 0·97); and a statistically significant inverse association was observed between lutein and zeaxanthin intake and neovascular AMD risk (RR 0·68; 95 % CI 0·51, 0·92). The results were essentially consistent among subgroups stratified by participant characteristics. The findings of the present meta-analysis indicate that dietary lutein and zeaxanthin is not significantly associated with a reduced risk of early AMD, whereas an increase in the intake of these carotenoids may be protective against late AMD. However, additional studies are needed to confirm these relationships.

We studied concerns about cognitive health among ethnically diverse groups of older adults. The study was grounded in theories of health behaviour and the representation of health and illness. We conducted 42 focus groups (N=396, ages 50+) in four languages, with African Americans, American Indians, Chinese Americans, Latinos, Whites other than Latinos (hereafter, Whites) and Vietnamese Americans, in nine United States locations. Participants discussed concerns about keeping their memory or ability to think as they age. Audio recordings were transcribed verbatim. Constant comparison methods identified themes. In findings, all ethnic groups expressed concern and fear about memory loss, losing independence, and becoming ‘a burden’. Knowing someone with Alzheimer's disease increased concern. American Indians, Chinese Americans, Latinos and Vietnamese Americans expected memory loss. American Indians, Chinese Americans and Vietnamese Americans were concerned about stigma associated with Alzheimer's disease. Only African Americans, Chinese and Whites expressed concern about genetic risks. Only African Americans and Whites expressed concern about behaviour changes. Although we asked participants for their thoughts about their ability to think as they age, they focused almost exclusively on memory. This suggests that health education promoting cognitive health should focus on memory, but should also educate the public about the importance of maintaining all aspects of cognitive health.

Alloying effects of iridium on the glass formability (GFA) of the Zr–Ir–Cu–Al system have been investigated, and several new bulk metallic glasses (BMGs) with high GFA have been successfully developed. Additions of Ir in the Zr–Cu–Al system can yield a beneficial distribution in atomic sizes, but the strong chemical interaction of the Zr–Ir atomic pair limits the maximum addable Ir contents and the resultant GFA. Our analyses indicate that the optimum composition for alloying elements is determined by not only topological but also chemical factors. Phase competition upon solidification, rather than effects from individual affecting factors, dictates the GFA of BMG systems.

In vitro studies using highly purified calf thymus RNA polymerase II and a fragment spanning the first intron of H3.3 as template DNA have demonstrated the existence of a strong transcription termination site consisting of thymidine stretches. In this study, nuclear run-on experiments have been performed to assess the extent to which transcription elongation is blocked in vivo using DNA probes corresponding to regions 5′ and 3′ of the in vitro termination sites. These studies suggest that H3.3 expression is stimulated following the inhibition of DNA synthesis through the elimination of the transcription elongation block. Interestingly, both the in vivo and in vitro experiments have revealed that the transcriptional block/termination sites are positioned immediately downstream of a 73 bp region that has been over 90% conserved between the chicken and human H3.3 genes. The extreme conservation of this intronic region suggests a possible role in maintaining cis-acting function. Electrophoretic mobility shift experiments show that HeLa cell nuclear extracts contain protein factors that bind specifically to the region of transcription elongation block. Furthermore, we demonstrate a correlation between the protein binding activity and the transcriptional block in cells that have been either arrested at the initiation of S phase or were replication-interrupted by hydroxyurea. DNA footprinting experiments indicate that the region of protein binding is at the 3′ end of the conserved region and overlaps with one of the three in-vitro-mapped termination sites.