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A key question in current immunology is how the innate immune system generates high levels of specificity. Our previous study in Caenorhabditis elegans revealed that NMUR-1, a neuronal G protein-coupled receptor homologous to mammalian receptors for the neuropeptide neuromedin U (NMU), regulates distinct innate immune responses to different bacterial pathogens. Here, by using quantitative proteomics and functional assays, we discovered that NMUR-1 regulates F 1 F O ATP synthase and ATP production in response to pathogen infection, and that such regulation contributes to NMUR-1-mediated specificity of innate immunity. We further demonstrated that ATP biosynthesis and its contribution to defense is neurally controlled by the NMUR-1 ligand CAPA-1 and its expressing neurons ASG. These findings indicate that NMUR-1 neural signaling regulates the specificity of innate immunity by controlling energy homeostasis as part of defense against pathogens. Our study provides mechanistic insights into the emerging roles of NMU signaling in immunity across animal phyla., Competing Interests: DECLARATION OF INTERESTS The authors declare no competing interests.

Plastic litter is an endemic problem within all regions, particularly cities. Much of this litter reaches the world's oceans with well documented environmental impacts. However, the monitoring of urban litter is often piecemeal at best. Citizen science, the action of utilising the public to support research, has been used to excellent effect for both research and engagement, usually for area clean-ups such as beach cleans. However, to date very few studies have assessed plastic pollution at a city scale. This study presents a novel citizen science approach, using a smartphone application to collect geolocated photographs of plastic litter during 5 city-wide surveys. The study has compiled a significant dataset of photographs (n = 3760), classified by plastic type to assess patterns of plastic pollution Portsmouth, UK. The method is shown to have significant potential for further development to facilitate detailed analysis of plastic litter in urban centres across the world., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Over 25 million individuals living in America are limited English proficient, many of whom live in rural communities. Adequate language accommodations are critical to providing effective healthcare for these populations. An online questionnaire was delivered to 42 rural facilities in Washington State. It included questions about their demand for language services, modalities of interpretation, translated documentation and barriers to providing accommodations. Fifteen of 42 (35.7%) responded. Spanish, Russian, Vietnamese, Japanese, Ukrainian and Mam were encountered daily. Telephonic and virtual remote interpreter services were the most widely available. Not all facilities had vital documents translated to frequently encountered languages. Challenges to providing language access were reported by nearly all participants. The rural facilities surveyed all encountered LEP patient populations and offered oral interpretation. Overall, these facilities were meeting requirements for providing language accommodation services. Even so, many facilities reported experiencing barriers to providing these services., (© 2022. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Understanding the use behaviours of plastic items within households is important to enable informed policy development, particularly with the emerging and developing global plastic treaty. A survey of 400 permanent residents in Portsmouth aimed to identify the general trends in single-use plastic product (SUPP) use and disposal, and their personal motivations and barriers to reducing and recycling plastic. This included identifying common influencers of attitudes such as environmental values, situational characteristics, psychological factors and the individual demographic characteristics of residents. Key factors in consumer behaviour were found to be product availability, affordability and convenience. Often, less conveniently recycled plastics more frequently end up in landfill such as films, shopping bags and personal care items. The age of respondents was found to be the most significantly associated demographic with SUPP consumption, reuse and recycling behaviours. Other demographic variables such as a resident's location within the city, income and vehicle ownership were potential drivers influencing individual attitudes and their incentives towards reducing and recycling their plastic waste. The findings from this study brought to light the importance of effective local plastic governance. This study also identified consumer perceptions and behaviours that could contribute to future holistic plastic policy recommendations., Supplementary Information: The online version contains supplementary material available at 10.1007/s11625-022-01261-5., Competing Interests: Conflict of interestThe authors declare no competing interests., (© The Author(s) 2023, corrected publication 2023.)

This paper describes the synthesis of enamino carbonyl compounds by the copper(i)-catalyzed coupling of acceptor-substituted diazo compounds and tertiary thioamides. We plan to use this method to synthesize indolizidine (-)-237D analogs to find α6-selective antismoking agents. Therefore, we also performed in silico α6-nAchRs binding studies of selected products. Compounds with low root-mean-square deviation values showed more favorable binding free energies. We also report preliminary pharmacokinetic data on indolizidine (-)-237D and found it to have weak activity at CYP3A4. In addition, as enamino carbonyl compounds are also known for antimicrobial properties, we screened previously reported and new enamino carbonyl compounds for antibacterial, antimicrobial, and antifungal properties. Eleven compounds showed significant antimicrobial activities., Competing Interests: There are no conflicts to declare., (This journal is © The Royal Society of Chemistry.)

In recent years, mindfulness-based practices in grade schools have been associated with students' improved cognitive skills and general classroom behavior. In the majority of studies, however, only teacher and parent feedback are elicited, omitting a considerably significant voice - that of the students. Our study aims to fill this gap by exploring student opinions and perceptions regarding the implementation of a classroom-based mindfulness program. Elementary school students ( N  = 51) took part in teacher-facilitated mindfulness activities which were incorporated into their daily classroom routines. Over the course of the 8-week intervention period, students participated in focus groups about their perceptions of the program. Through qualitative content analysis, two major findings emerged from the focus group data: student opinions about the mindfulness program varied substantially and the mindfulness activities were not always liked and enjoyed. Critically, if students do not enjoy classroom-based mindfulness programs, they may be less motivated to engage in mindful activities and in turn may not experience the benefits that mindfulness has to offer. To maximize student engagement with mindfulness while addressing their concerns, the following recommendations are made: A balance between the entertaining and educational aspects of the program, flexible program delivery, and encouraging students to pursue mindful living outside of the classroom. This research is important to educational and clinical practitioners as student insight will benefit the development and modification of classroom-based mindfulness programs to ensure that students are better able to engage with and benefit from these programs., Competing Interests: Conflict of InterestThe authors declare no competing interests., (© The Author(s) 2022, corrected publication 2022.)

Culture shapes children's memories. However, scant attention has been given to the influence of culture on specific memory stages. Thus, we conducted two controlled experiments to examine cultural differences in memory recall at immediate and delayed retrieval phases. In Studies 1 and 2 (n = 217), 7- to 10-year-old Chinese and Euro-Canadian children watched a story involving both social- and individual-focused scenarios. Participants then recalled the story immediately afterwards (Study 1) and 5-7 days later (Studies 1 and 2). Findings reveal that Chinese children accurately reported more details from the social-focused events than did their Euro-Canadian counterparts in the immediate interview, and this result was replicated after a delay in both studies. Moreover, as expected, within-country comparisons showed that Euro-Canadian children had better memory for individual- than for social-focused events in both studies. Chinese children, however, showed better memory for social- than for individual-focused events only in the immediate interview in Study 1; their delayed retrieval was not affected by event focus. These findings reveal that cultural variations in memory are evident in both immediate and delayed retrieval. Implications for future research are discussed., (© 2021 The British Psychological Society.)

One of the many sources of information easily available to children is the internet and the millions of websites providing accurate, and sometimes inaccurate, information. In the current investigation, we examined children's ability to use credibility information about websites when learning about environmental sustainability. In two studies, children studied two different websites and were tested on what they had learned a week later using a multiple-choice test containing both website items and new distracters. Children were given either no information about the websites or were told that one of the websites (the noncredible website) contained errors and they should not use any information from that website to answer the test. In both studies, children aged 7- to 9-years reported information from the noncredible website even when instructed not to, whereas the 10- to 12-year-olds used the credibility warning to 'edit out' information that they had learned from the noncredible website. In Study 2, there was an indication that the older children spontaneously assessed the credibility of the website if credibility markers were made explicit. A plausible explanation is that, although children remembered information from the websites, they needed explicit instruction to bind the website content with the relevant source (the individual websites). The results have implications for children's learning in an open-access, digital age where information comes from many sources, credible and noncredible. Education in credibility evaluation may enable children to be critical consumers of information thereby resisting misinformation provided through public sources.

Destination memory (the ability to remember who one has told information to) has been studied in adult samples, but not with child participants. The goals of the current research were to describe the development of children's destination memory abilities across early to middle childhood and to compare destination memory with source-monitoring abilities within the same age range. In two studies, children aged 5-10 years had conversations with two puppets and were later asked to recall which puppet they had disclosed specific information to. Study 1 (N = 86) revealed age to be a significant predictor of destination memory accuracy. Study 2 (N = 90) demonstrated that source memory and destination memory were related and shared a similar developmental trajectory but that source accuracy was higher on average than destination accuracy. This research contributes to our theoretical understanding of children's memory development, and directions for future research are highlighted in the Discussion. Implications for investigative interviews in which children are asked about prior disclosures are discussed., (Copyright © 2020 Elsevier Inc. All rights reserved.)

The prevalence of "mindfulness" in popular media, academia, and professional circles is difficult to miss. Newspapers, magazines, online articles, clinical programs, podcasts, scholarly and professional meetings, sports organizations, and many other outlets focus on the benefits of mindfulness. Despite the intense focus on mindfulness in Western society, it is astonishing that the evidence base (i.e., documented, scholarly, peer-reviewed evaluations) for these programs is woefully inadequate. Varying definitions of what mindfulness is, what it entails, what specific benefits to psychological functioning are observed (if any), and inadequate scientific testing all contribute to a lean knowledge base. Evaluation of potential benefits of mindful practice with children is even more difficult because children are a more heterogeneous group than adults; the differing developmental levels are likely to have profound effects on the efficacy of mindfulness-based programming with children. We review these issues and provide an explanation of the strength of different kinds of evidence, with suggestions for (a) researchers who study mindfulness with children and (b) clinical professionals and educators interested in developing mindful attitudes and techniques with children., (© 2020 Elsevier Inc. All rights reserved.)

Memory is socially constructed. The types of information that children pay attention to and remember, as well as how children organize and recall their memories can differ as a function of sociocultural background. This chapter presents an overview of cultural variations on children's memory and cognition. We draw attention to the necessity of conducting controlled experiments to examine cultural differences in the specific processes involved in episodic memory (e.g., encoding, retention, discrimination skills). We highlight potential challenges (e.g., language, measurement equivalence) that researchers need to overcome to conduct valid cross-cultural research. In light of cultural transformations in recent decades, we outline promising avenues for future research as well as the applications of this research to important issues for forensics and immigrants and asylum-seekers., (© 2019 Elsevier Inc. All rights reserved.)

Hot electrons established by the absorption of high-energy photons typically thermalize on a picosecond time scale in a semiconductor, dissipating energy via various phonon-mediated relaxation pathways. Here it is shown that a strong hot carrier distribution can be produced using a type-II quantum well structure. In such systems it is shown that the dominant hot carrier thermalization process is limited by the radiative recombination lifetime of electrons with reduced wavefunction overlap with holes. It is proposed that the subsequent reabsorption of acoustic and optical phonons is facilitated by a mismatch in phonon dispersions at the InAs-AlAsSb interface and serves to further stabilize hot electrons in this system. This lengthens the time scale for thermalization to nanoseconds and results in a hot electron distribution with a temperature of 490 K for a quantum well structure under steady-state illumination at room temperature.

The roles of bulk surface states and interfacial defects are probed experimentally using a combination of current-voltage, capacitance-voltage, and impedance measurements. The critical importance of the quality of both the film and interfaces is evident in current-voltage measurements where shunting and interface states result in large dark currents and the subsequent loss of J sc . These properties are shown to be critically related to the nature and role of the PbS QD interface with the (nominally) ohmic gold contact. Specifically, the nonideality of this interface results in the formation of an electric field and therefore a Schottky barrier that opposes the transport of carriers across the conventional ZnO-PbS CQD system. Nonidealities in the structure and absorber layer are also reflected in nonmonotonic behavior and dispersion in C-V measurements with trapping processes on the CQD surfaces, and the ZnO/PbS and PbS/Au interfaces also affecting the carrier dynamics, which is reflected in the response time of these systems under different biases.

The purpose of this study is to elicit guidance from prosecutors across Australia on questioning children about repeated events. Two focus groups were conducted. The first sought broad feedback concerning questioning children about repeated events. The second focused more specifically on eliciting feedback about techniques for aiding children in describing specific instances of repeated events. The techniques used are derived from a combination of empirical research and best practice interview guidelines. Data from both focus groups were compiled because themes were highly similar. Thematic analysis of the focus group discussions revealed three broad themes in prosecutors' perceptions about questioning children about repeated abuse: a) permitting children to provide a full generic account before describing individual episodes of abuse, b) using the information obtained during the generic account to create episode labels, and c) probing incidences of abuse chronologically. These themes are discussed within the context of the child development and mnemonic literature, and implications for interviewing protocols are drawn., Competing Interests: No potential conflict of interest was reported by the authors., (© 2017 The Australian and New Zealand Association of Psychiatry, Psychology and Law.)

Children learn information from a variety of sources and often remember the content but forget the source. Whereas the majority of research has focused on retrieval mechanisms for such difficulties, the present investigation examines whether the way in which sources are encoded influences future source monitoring. In Study 1, 86 children aged 3 to 8 years participated in 2 photography sessions on different days. Children were randomly assigned to either the Difference condition (they were asked to pay attention to differences between the 2 events), the Memory control condition (asked to pay attention with no reference to differences), or the No-Instruction control (no special instructions were given). One week later, during a structured interview about the photography session, the 3- to 4-year-olds in the No-Instruction condition were less accurate and responded more often with 'do not know' than the 7- to 8-year-olds. However, the older children in the Difference condition made more source confusions than the younger children suggesting improved memory for content but not source. In Study 2, the Difference condition was replaced by a Difference-Tag condition where details were pointed out along with their source (i.e., tagging source to content). Ninety-four children aged 3 to 8 years participated. Children in the Difference-Tag condition made fewer source-monitoring errors than children in the Control condition. The results of these 2 studies together suggest that binding processes at encoding can lead to better source discrimination of experienced events at retrieval and may underlie the rapid development of source monitoring in this age range. (PsycINFO Database Record, ((c) 2016 APA, all rights reserved).)

Objectives: One in five children in England are overweight/obese at school entry. Tackling obesity is therefore a priority. Right from the Start with HENRY is a widely-commissioned programme delivered by trained facilitators to small groups of parents over eight weekly sessions. It is designed to provide parents of infants and preschool children with the skills, knowledge and confidence required for a healthier family lifestyle. The aim of this work was to investigate programme impact using data collected routinely for quality control purposes., Study Design: Analysis of routinely collected pre-post data from programmes delivered in the UK from January 2012 to February 2014., Methods: Data were analysed from 144 programmes, including questionnaires relating to parenting, family eating behaviours, dietary intake, and physical activity/screen time., Results: Over 24 months, 1100 parents attended programmes running in 86 locations. 788 (72%) completed >5 sessions of whom 624 (79%) provided baseline and completion questionnaires. Parents reported increases in healthiness of family lifestyle, parenting attributes, and emotional wellbeing following attendance (all P < .001). Both parents and children were reported to have increased their daily fruit/vegetable consumption, and reduced their consumption of high fat/sugar foods (both P < .001). There were also positive changes in eating behaviours, physical activity (P < .001) and children's screen time (P < .001)., Conclusions: Significant changes were reported in all domains similar to those reported in a previous, smaller study in locations selected for experience and quality. The HENRY approach appears to have a beneficial impact even when delivered at scale in non-selected locations. Such changes, if maintained, may serve to protect against later obesity., (Copyright © 2016 The Royal Society for Public Health. Published by Elsevier Ltd. All rights reserved.)

Background: Increased surgical volume is associated with better patient outcomes and shorter lengths of hospitalization. As a consequence, traveling to receive care from a high volume provider may be associated with better outcomes. However, travel may also be associated with a decision by the healthcare provider to increase the length of stay due to a decreased ability to return to the primary hospital should complications arise. Thus, research is needed to understand the relationship between the distance a patient must travel and their outcomes following urologic surgery. Objective: The purpose of this study was to determine whether the distance a patient travels to receive urologic surgery is associated with their length of hospital stay and direct medical hospitalization costs. Methods: This was a retrospective observational cohort study of 12 106 patients over 50 years of age undergoing transurethral resection of the prostate (TURP), radical prostatectomy (RP) or radical cystectomy (RC) in Washington State hospitals between 2009 and 2013. Distance traveled was determined by calculating the linear distance between zip code centroids of patient residence and the hospital performing their procedure. Patients were sorted into four groups classified by distance traveled (≤5 miles, 6-20 miles, 21-50 miles and ≥51 miles) and cost calculated using a charges-to-reimbursement ratio for each hospital. Statistical significance was determined using a Kruskal-Wallis test. Results: Patients traveling greater distances had significantly lower median medical costs compared with patients who lived closer to the hospitals where they underwent TURP and RP (TURP: ≤5 miles, $6243 and ≥51 miles, $5105, p≤0.001; RP: ≤5 miles, $12 407 and ≥51 miles, $11 882, p≤0.001), whereas there was no significant difference for patients undergoing RC (≤5 miles, $27 554 and ≥51 miles, $26 761, p=0.17). Likewise, patients traveling greater distances had significantly lower median lengths of hospitalization for TURP and RP (TURP: p≤0.001, RP: p≤0.001), while there was no difference for RC (p=0.50). Conclusions: Patient travel burden does appear to play a role in cost and length of hospital stay for select urologic procedures with variable levels of morbidity and recovery time. Although these findings are statistically significant, the magnitude of the effect is small.

Objective: To examine hospital-level variation in outcomes following benign urologic surgeries given that hospital-level variation in surgical outcomes can portend quality and appropriateness of care concerns and identify quality improvement opportunities in perioperative care., Materials and Methods: Using the Washington State Comprehensive Hospital Abstract Reporting System, we identified patients who underwent transurethral resection of the prostate (TURP), percutaneous nephrostolithotomy (PCNL), and pyeloplasty from 2003 to 2008. We classified prolonged postoperative length of stay (LOS) as that exceeding the 75th percentile, and we measured the rate of Agency for Healthcare Quality Patient Safety Indicators, readmissions, and death. We calculated hospital-specific observed-to-expected event rates using random effects multilevel multivariable models adjusted for age and comorbidity., Results: We identified 6699 TURP patients at 54 hospitals, 2541 PCNL patients at 45 hospitals, and 584 pyeloplasty patients at 36 hospitals. Complication rates were highest after PCNL (22.9% prolonged LOS vs 17.3% for TURP and 13.9% for pyeloplasty, P < .001; 3.4% 90-day mortality vs 0.6% for TURP and 0% for pyeloplasty). Hospital-level variation was most substantial for LOS after TURP and pyeloplasty (8.1% and 14.3% of variance in prolonged LOS, respectively)., Conclusion: Hospital-level variation is common after benign inpatient urologic surgeries and may relate to difference in perioperative provider practice patterns. The morbidity of PCNL in this study was higher than expected and merits further investigation., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Background: One-quarter of children in England are overweight/obese at school entry. We investigated the impact of a programme designed to provide parents of infants and preschool children with the skills required for a healthier family lifestyle., Method: A cohort of families was followed across the 8-week HENRY (Health Exercise Nutrition for the Really Young) parent course at nine locations in England. Seventy-seven parents enrolled on the course, of which 71 agreed to complete questionnaires addressing eating behaviours, dietary intake and parental self-efficacy. Pre- and post-course data was available from 60 (84.5%) parents (8-week follow-up data from 58 parents) and was analysed using repeated measures analyses., Results: Significant changes were observed, with most sustained at follow-up. Parents reported increased self-efficacy and ability to encourage good behaviour (P < 0.001). Increased consumption of fruits and vegetables was reported in both children and adults, together with reduced consumption of sweets, cakes and fizzy drinks in adults (all P < 0.01). There were also positive changes in eating behaviours (e.g., frequency of family mealtimes and eating while watching television or in response to negative emotion [P < 0.01] ) and reduced screen time in adults (P < 0.001)., Discussion: The results build upon earlier evaluation, indicating that the HENRY intervention has a beneficial impact upon the families of infants and preschool children. Furthermore, the findings suggest that positive changes inspired by the programme can be maintained beyond its completion. Such changes may serve to protect against later obesity., (© 2013 The Authors. Pediatric Obesity © 2013 International Association for the Study of Obesity.)

Previous research on the relationship between executive function and source monitoring in young children has been inconclusive, with studies finding conflicting results about whether working memory and inhibitory control are related to source-monitoring ability. In this study, the role of working memory and inhibitory control in recognition memory and source monitoring with two different retrieval strategies were examined. Children (N = 263) aged 4-8 participated in science activities with two sources. They were later given a recognition and source-monitoring test, and completed measures of working memory and inhibitory control. During the source-monitoring test, half of the participants were asked about sources serially (one after the other) whereas the other half of the children were asked about sources in parallel (considering both sources simultaneously). Results demonstrated that working memory was a predictor of source-monitoring accuracy in both conditions, but inhibitory control was only related to source accuracy in the parallel condition. When age was controlled these relationships were no longer significant, suggesting that a more general cognitive development factor is a stronger predictor of source monitoring than executive function alone. Interestingly, the children aged 4-6 years made more accurate source decisions in the parallel condition than in the serial condition. The older children (aged 7-8) were overall more accurate than the younger children, and their accuracy did not differ as a function of interview condition. Suggestions are provided to guide further research in this area that will clarify the diverse results of previous studies examining whether executive function is a cognitive prerequisite for effective source monitoring.

Developmental differences in the use of social-attention cues to imitation were examined among children aged 3 and 6 years old (n = 58) and adults (n = 29). In each of 20 trials, participants watched a model grasp two objects simultaneously and move them together. On every trial, the model directed her gaze towards only one of the objects. Some object pairs were related and had a clear functional relationship (e.g., flower, vase), while others were functionally unrelated (e.g., cardboard square, ladybug). Owing to attentional effects of eye gaze, it was expected that all participants would more faithfully imitate the grasp on the gazed-at object than the object not gazed-at. Children were expected to imitate less faithfully on trials with functionally related objects than those without, due to goal-hierarchy effects. Results support effects of eye gaze on imitation of grasping. Children's grasping accuracy on functionally related and functionally unrelated trials was similar, but they were more likely to only use one hand on trials where the object pairs were functionally related than unrelated. Implications for theories of imitation are discussed., (© 2013 The British Psychological Society.)

CZE was applied to characterizing a nanoparticle/micellar system, in which CdSe nanoparticles dispersed in a mixture of non-ionic and ionic surfactants are introduced into a capillary to form the sample zone that migrates being sandwiched by the background electrolyte (BGE). Parameters that affect the migration of the surface-modified CdSe nanoparticles and conditions under which they are focused within the micellar zone (or released from it into bulk BGE) were explored, including the sample composition, sample plug length, and applied voltage. The observed migration behavior was analyzed within the framework of a concept of formation of a mixed pseudomicellar system, according to which a nanoparticle coated with an ionic surfactant is treated as a micelle-like entity. It was found out that the nanoparticles are subject to transformation that results in building-up a mixed pseudomicellar system (with regular micelles), with a consequence of exhibiting distinctive migration phenomena. Particularly observed and brought into focus is the event of focusing of the nanoparticles., (Copyright © 2013 Elsevier B.V. All rights reserved.)

We have synthesized CdSe nanocrystals (NCs) in sizes from 2.2 to 5.1 nm passivated with hydrophobic trioctylphosphine oxide (TOPO) in combination trioctylphosphine (TOP) or tributylphosphine (TBP) to obtain particles of the type CdSe/TOPO/TOP or CdSe/TOPO/TBP. These NCs were then dispersed in aqueous solution of ionic or non-ionic surfactants (such as stearate, oleic acid, Tween) using a biphase (water and chloroform or hexane) transfer method. It is found that both the structure of the surfactant and the native surface of the ligand govern the coating of the NCs with surfactants. More specifically, the hydrophobicity-hydrophilicity balance of the surfactant regulates the coating efficacy, thereby transferring the NC from the organic to the aqueous phase. The type of ligand on the NCs and the kind of coating surfactant also affect photoluminescence (PL). The ratio of PL and absorbance unit (defined as PL per 0.1 AU) was implemented as a tool to monitor changes in PL intensity and wavelength as a function of size, coatings and surface defects. Finally, the distribution of CdSe nanocrystals between pseudophases in cloud point extraction was discussed based on experimental results. It was concluded that the size of CdSe nanocrystal present in an appropriate pseudophase is correlated with the way in which the non-ionic surfactant coats CdSe nanocrystals. FigureCoating of CdSe semiconductor nanocrystals with surfactants impacts nanocrystals' spectral features. Absorbance of first exciton absorption band was used to estimate ability of surfactant to disperse CdSe nanocrystals. Photoluminescence (PL) intensity and position of PL band were analysed in terms of nanocrystal's surface phenomena via surfactants applied for coating.

Spermatozoa are highly specialized cells that, when mature, are capable of navigating the female reproductive tract and fertilizing an oocyte. The sperm cell is thought to be largely quiescent in terms of transcriptional and translational activity. As a result, once it has left the male reproductive tract, the sperm cell is essentially operating with a static population of proteins. It therefore is theoretically possible to understand the protein networks contained in a sperm cell and to deduce its cellular function capabilities. To this end, we performed a proteomic analysis of mouse sperm isolated from the cauda epididymis and confidently identified 2850 proteins, which to our knowledge is the most comprehensive sperm proteome for any species reported to date. These proteins comprise many complete cellular pathways, including those for energy production via glycolysis, beta-oxidation and oxidative phosphorylation, protein folding and transport, and cell signaling systems. This proteome should prove a useful tool for assembly and testing of protein networks important for sperm function.

Background: Unexplained variation in outcomes after common surgeries raises concerns about the quality and appropriateness of surgical care. Understanding variation in surgical outcomes may identify processes that could affect the quality of surgical and postoperative care. The authors of this report examined hospital-level variation in outcomes after inpatient urologic oncology procedures., Methods: Patients who underwent radical cystectomy, radical nephrectomy, and radical prostatectomy were identified from the Washington State Comprehensive Hospital Abstract Reporting System for the years 2003 through 2007. The postoperative length of stay (LOS) was measured, and LOS that exceeded the 75th percentile was classified as prolonged. The occurrence of Agency for Healthcare Quality patient safety indicators (PSIs), readmissions, and deaths also were measured. Analyses were adjusted for patient age and comorbidity in random effects, multilevel, multivariable models that assessed hospital-level outcomes., Results: The authors identified 853 patients from 37 hospitals who underwent cystectomy, 3018 patients who underwent nephrectomy from 51 hospitals, and 8228 patients who underwent prostatectomy from 51 hospitals. Complications captured by PSIs were rare. Hospital-level variation was most profound for LOS outcomes after nephrectomy and prostatectomy (variance in prolonged LOS, 8.1% and 26.7%, respectively), thromboembolic events after nephrectomy (8% of variance), and mortality after cystectomy (7.1% of variance)., Conclusions: Hospital-level variation confounds the care of urologic cancer patients in the state of Washington. The authors concluded that transparent reporting of surgical outcomes and local quality-improvement initiatives should be considered to ameliorate the observed variation and improve the quality of cystectomy, nephrectomy, and prostatectomy care., (Copyright © 2011 American Cancer Society.)

We have synthesized CdSe nanocrystals (NCs) possessing a trioctylphosphine surface passivation layer and modified with amphiphilic molecules to form a surface bilayer. The NCs covered with single amphiphiles are not stable in aqueous solution, but a mixed amphiphilic system is shown to provide stability in solution over several months. The solutions of the modified NCs were characterized by UV-Vis absorbance, photoluminescence, and transmission electron microscopy. An electrophoretic study revealed two operational modes. The first relies on the enrichment of NCs using a micellar plug as a tool. The accumulation of NCs at the plug-electrolyte buffer interface results in a sharp peak. By controlling the electrophoretic conditions, nanocrystals were forced to exit a micellar plug into an electrolyte buffer. We conclude that a system consisting of modified nanocrystals and a micellar plug can act as a mixed pseudomicellar system, where modified nanocrystals play the role of pseudomicelles.FigureElectrophoretic focusing of amphiphile coated CdSe nanocrystals using a micellar plug. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s00604-011-0727-8) contains supplementary material, which is available to authorized users.

Children (N = 157) 4 to 8 years old participated 1 time (single) or 4 times (repeated) in an interactive event. Across each condition, half were questioned a week later about the only or a specific occurrence of the event (depth first) and then about what usually happens. Half were prompted in the reverse order (breadth first). Children with repeated experience who first were asked about what usually happens reported more event-related information overall than those asked about an occurrence first. All children used episodic language when describing an occurrence; however, children with repeated-event experience used episodic language less often when describing what usually happens than did those with a single experience. Accuracy rates did not differ between conditions. Implications for theories of repeated-event memory are discussed.

Objective: This study examined the effect of event repetition on the amount and nature of story-grammar produced by children when recalling the event., Method: Children aged 4 years (N=50) and 7 years (N=56) participated in either 1 or 6 occurrences of a highly similar event where details varied across the occurrences. Half the children in each age and event group recalled the last/single occurrence 5-6 days later and the other half recalled the last/single occurrence after 5-6 weeks (the final and single occurrence was the same). Children's free recall responses were classified according to the number and proportion of story-grammar elements (Stein & Glenn, 1979-setting, initiating event, internal response, plan, attempt, direct consequence, and resolution) as well as the prevalence of causal links between the individual story-grammar elements., Results: More story-grammar detail and more links between individual story-grammar elements were reported about the final compared to single occurrence. The amount of story-grammar increased with age and decreased over time. Further, an interaction was revealed such that the effect of retention interval on the production of story-grammar was negligible for older children who experienced the repeated event., Conclusions: Event repetition has a beneficial effect on the production of children's story-grammar content in situations where event details varied from occasion to occasion. This study highlights the importance of eliciting free recall when conducting evidential interviews with child witnesses about repeated events., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

In the present work, CdSe nanocrystals (NCs) synthesized with a trioctylphosphine surface passivation layer were modified using amphiphilic molecules to form a surface bilayer capable of providing stable NCs aqueous solutions. Such modified nanocrystals were used as a test solute in order to analyze new electrophoretic phenomena, by applying a micellar plug as a separation tool for discriminating nanocrystals between micellar and micelle-free zones during electrophoresis. The distribution of NCs between both zones depended on the affinity of nanocrystals towards the micellar zone, and this relies on the kind of surface ligands attached to the NCs, as well as electrophoretic conditions applied. In this case, the NCs that migrated within a micellar zone can be focused using a preconcentration mechanism. By modifying electrophoretic conditions, NCs were forced to migrate outside the micellar zone in the form of a typical CZE peak. In this situation, a two-order difference in separation efficiencies, in terms of theoretical plates, was observed between focused NCs (N ~ 10(7)) and a typical CZE peak for NCs (N ~ 10(5)). By applying the amino-functionalized NCs the preconcentration of NCs, using a micellar plug, was examined, with the conclusion that preconcentration efficiency, in terms of the enhancement factor for peak height (SEF(height)) can be, at least 20. The distribution effect was applied to separate CdSe/ZnS NCs encapsulated in silica, as well as surface-modified with DNA, which allows the estimation of the yield of conjugation of biologically active molecules to a particle surface.

Single-walled carbon nanotubes (SWCNTs) were covalently modified with a redox mediator derived from 2,2'-azino-bis-(3-ethylbenzothiazoline)-6-sulfonic acid (ABTS), and implemented in the construction of electrodes for biocatalytic oxygen reduction. The procedure is based on: covalent bonding of mediator to nanotubes, placing the nanotubes directly on the carbon electrode surface and covering the nanostructured electrode with a Nafion film containing laccase as the biocatalyst. The modified electrode is stable and the problem of mediator (ABTS) leaking from the film is eliminated by binding it covalently to the nanotubes. Three different synthetic approaches were used to obtain ABTS-modified carbon nanotubes. Nanotubes were modified at ends/defect sites or on the nanotube sidewalls and characterized by Raman spectroscopy, TGA and electrochemistry. The accessibility of differently located ABTS units by the laccase active center and mediation of electron transfer were studied by cyclic voltammetry. The surface concentrations of ABTS groups electrically connected with the electrode were compared for each of the electrodes based on the charges of the voltammetric peaks recorded in the deaerated solution. The nanotube modification procedure giving the best parameters of the catalytic process was selected., (Copyright © 2010 Elsevier B.V. All rights reserved.)

Objectives: Idiopathic calcium oxalate (CaOx) stones are believed to develop attached to papillary subepithelial deposits called Randall's plaques. Calcium phosphate (CaP) stones, conversely, are thought to arise within the inner medullary collecting ducts, enlarging and damaging surround tubular structures as they expand. If this is true, we theorize that differences will be seen within the organic portion (matrix) of CaOx stones compared with CaP stones using a mass spectroscopy (MS) approach., Methods: From a cohort of 47 powdered stones, 25 calculi (13 CaOx, 12 CaP) were confirmed to contain a dominant mineral content of >80% by powder x-ray diffraction. Matrix proteins were then extracted, purified, and digested. Peptide tandem MS data were acquired, and spectra were searched against a large human protein database to identify protein matches., Results: No significant differences were seen between pattern profiles of CaOx and CaP stones. However, variations in protein expression patterns were seen within individual CaOx (monohydrate and dihydrate) and CaP (apatite and brushite) mineral subtypes, suggesting a relationship between crystal-surface binding properties and matrix composition. Both groups contain a large number of inflammatory proteins and a catalog of common proteins is included., Conclusions: Calcium kidney stone matrix contains hundreds of proteins and is predominated by proteins associated with inflammatory response. Many of the same proteins were identified in both CaOx and CaP stones, suggesting inflammation as a unifying origin or a common secondary role in calcium stone pathogenesis., (Copyright © 2010 Elsevier Inc. All rights reserved.)

Objective: Young children's descriptions of maltreatment are often sparse thus creating the need for techniques that elicit lengthier accounts. One technique that can be used by interviewers in an attempt to increase children's reports is "paraphrasing," or repeating information children have disclosed. Although we currently have a general understanding of how paraphrasing may influence children's reports, we do not have a clear description of how paraphrasing is actually used in the field., Method: The present study assessed the use of paraphrasing in 125 investigative interviews of allegations of maltreatment of children aged 4-16 years. Interviews were conducted by police officers and social workers. All interviewer prompts were coded into four different categories of paraphrasing. All children's reports were coded for the number of details in response to each paraphrasing statement., Results: "Expansion paraphrasing" was used significantly more often and elicited significantly more details, while "yes/no paraphrasing" resulted in shorter descriptions from children, compared to other paraphrasing styles. Further, interviewers more often distorted children's words when using yes/no paraphrasing, and children rarely corrected interviewers when they paraphrased inaccurately., Conclusions and Practical Implications: Investigative interviewers in this sample frequently used paraphrasing with children of all ages and, though children's responses differed following the various styles of paraphrasing, the effects did not differ by the age of the child. The results suggest that paraphrasing affects the quality of statements by children. Implications for investigative interviewers will be discussed and recommendations offered for easy ways to use paraphrasing to increase the descriptiveness of children's reports of their experiences., (2010 Elsevier Ltd. All rights reserved.)

Dilute boar seminal plasma (SP) has been shown to inhibit in vitro capacitation and cooling-induced capacitation-like changes in boar spermatozoa, as assessed by the ability of the spermatozoa to undergo an ionophore-induced acrosome reaction. We hypothesised that the protein component of SP is responsible for this effect. To test this hypothesis, varying concentrations of total SP protein or SP proteins fractionated by heparin binding were assayed for their ability to inhibit in vitro capacitation, as well as cooling- and cryopreservation-induced capacitation-like changes. In vitro capacitation and cooling-induced capacitation-like changes were prevented by 10% whole SP, as well as by total proteins extracted from SP at concentrations greater than 500 microg mL(-1). No amount of SP protein was able to prevent cryopreservation-induced capacitation-like changes. Total SP proteins were fractionated based on their heparin-binding properties and the heparin-binding fraction was shown to possess capacitation inhibitory activity at concentrations as low as 250 microg mL(-1). The proteins in the heparin-binding fraction were subjected to mass spectrometry and identified. The predominant proteins were three members of the spermadhesin families, namely AQN-3, AQN-1 and AWN, and SP protein pB1. We conclude that one or more of these heparin-binding SP proteins is able to inhibit in vitro capacitation and cooling-induced capacitation-like changes, but not cryopreservation-induced capacitation-like changes, in boar spermatozoa.

Though cryopreservation of mouse sperm yields good survival and motility after thawing, cryopreservation of rat sperm remains a challenge. This study was designed to evaluate the biophysics (membrane permeability) of rat in comparison to mouse to better understand the cooling rate response that contributes to cryopreservation success or failure in these two sperm types. In order to extract subzero membrane hydraulic permeability in the presence of ice, a differential scanning calorimeter (DSC) method was used. By analyzing rat and mouse sperm frozen at 5 degrees C/min and 20 degrees C/min, heat release signatures characteristic of each sperm type were obtained and correlated to cellular dehydration. The dehydration response was then fit to a model of cellular water transport (dehydration) by adjusting cell-specific biophysical (membrane hydraulic permeability) parameters L(pg) and E(Lp). A "combined fit" (to 5 degrees C/min and 20 degrees C/min data) for rat sperm in Biggers-Whitten-Whittingham media yielded L(pg) = 0.007 microm min(-1) atm(-1) and E(Lp) = 17.8 kcal/mol, and in egg yolk cryopreservation media yielded L(pg) = 0.005 microm min(-1) atm(-1) and E(Lp) = 14.3 kcal/mol. These parameters, especially the activation energy, were found to be lower than previously published parameters for mouse sperm. In addition, the biophysical responses in mouse and rat sperm were shown to depend on the constituents of the cryopreservation media, in particular egg yolk and glycerol. Using these parameters, optimal cooling rates for cryopreservation were predicted for each sperm based on a criteria of 5%-15% normalized cell water at -30 degrees C during freezing in cryopreservation media. These predicted rates range from 53 degrees C/min to 70 degrees C/min and from 28 degrees C/min to 36 degrees C/min in rat and mouse, respectively. These predictions were validated by comparison to experimentally determined cryopreservation outcomes, in this case based on motility. Maximum motility was obtained with freezing rates between 50 degrees C/min and 80 degrees C/min for rat and at 20 degrees C/min with a sharp drop at 50 degrees C/min for mouse. In summary, DSC experiments on mouse and rat sperm yielded a difference in membrane permeability parameters in the two sperm types that, when implemented in a biophysical model of water transport, reasonably predict different optimal cooling rate outcomes for each sperm after cryopreservation.

Stress urinary incontinence (SUI) occurs due to anatomic and/or neurologic factors involving connective tissues, muscles and nerves. Although SUI is more common in post-menopausal and multiparous women, studies have also shown a high prevalence of SUI in young, physically fit female athletes. With a goal toward dynamic subject-specific mechanical characterization of the interaction between anatomical structures during physical activities that elicit SUI in females during physical or daily activities, a computer aided design (CAD)-based computer model of the female pelvis has been developed to test the feasibility of the computer modeling approach in understanding the measurable differences between stress-continent and stress-incontinent women. In the present study, a fluid-structure interaction analysis was conducted by using the finite element (FE) analysis technique based on the CAD-based computer model of the female pelvis to investigate the urine leakage in females during jumping. To the best of our knowledge, this is the first application of a fluid-structure interaction FE analysis approach in understanding the mechanisms of SUI in females. Through a series of computer simulations, the effects of varying impact forces determined by jumping height and bladder volume were investigated. The dynamic computer simulation results revealed that jumping heights have a significant influence on the volume of urine leakage caused by the landing impact of jumping. Bladder volume did not have a significant influence on leakage when the jumping heights were smaller than 1 ft, which indicates that normal walking (corresponds to a jumping height smaller than 0.1 ft) is not the primary cause of urine leakage for healthy females. The computer simulation results also showed that the deformation difference between the anterior and posterior portion of the female pelvis causes opening of the urethra and resultant urine leakage. The present study demonstrates the feasibility of using a computer modeling approach to study female SUI during physical and daily activities.

The aim of this study was to determine gross and neuroanatomic features of a novel periurethral neuromuscular electrostimulator. Periurethral leads were placed in eight female cadavers. In two cases, leads were imaged after placement to enhance anatomic understanding. Pelvic viscera were removed en bloc for analysis of lead placement in the six remaining cadavers. Excised tissue was sectioned and immunostained to identify general, afferent, sympathetic, and nitric oxide synthase efferent nerve fibers. The electrodes were found within/lateral (n = 4), within/posterolateral (n = 9), and anterolateral (n = 1) to the external urethral sphincter (distance 0.25 +/- 0.5, 2.9 +/- 3.3, and 1.0 +/- 0.0 mm, respectively). The electrode to the urethra and vagina distance averaged 7.6 +/- 3.4 and 8.8 +/- 4.3 mm, respectively. Variable density staining for all nerve types was found around the electrode. A periurethral electrode interfaces the external urethral sphincter, and the adjacent distribution of nerve fibers supports proposed neuromuscular therapeutic mechanisms.

Cysteine-rich secretory protein 1 (CRISP1) is a secretory glycoprotein produced by the rat epididymal epithelium in two forms, referred to as proteins D and E. CRISP1 has been implicated in sperm-egg fusion and has been shown to suppress capacitation in rat sperm. Several studies have suggested that CRISP1 associates transiently with the sperm surface, whereas others have shown that at least a portion of CRISP1 persists on the surface. In the present study, we demonstrate that protein D associates transiently with the sperm surface in a concentration-dependent manner, exhibiting saturable binding to both caput and cauda sperm in a concentration range that is consistent with its capacitation-inhibiting activity. In contrast, protein E persists on the sperm surface after all exogenous protein D has been dissociated. Comparison of caput and cauda sperm reveal that protein E becomes bound to the sperm in the cauda epididymidis. We show that protein E associates with caput sperm, which do not normally have it on their surfaces, in vitro in a time- and temperature-dependent manner. These studies demonstrate that most CRISP1 interacts with sperm transiently, possibly with a specific receptor on the sperm surface, consistent with its action in suppressing capacitation during epididymal storage of sperm. These studies also confirm a tightly bound population of protein E that could act in the female tract.

The family of mammalian cysteine-rich secretory proteins (CRISP) have been well characterized in the rat, mouse, and human. Here we report the molecular cloning and expression analysis of CRISP1, CRISP2, and CRISP3 in the boar. A partial sequence published in the National Center for Biotechnology Information (NCBI) database was used to derive the full-length sequences for CRISP1 and CRISP2 using rapid amplification of cDNA ends. RT-PCR confirmed the expression of these mRNAs in the boar reproductive tract, and real time RT-PCR showed CRISP1 to be highly expressed throughout the epididymis, with CRISP2 highly expressed in the testis. A search of the porcine genomic sequence in the NCBI database identified a BAC (CH242-199E6) encoding the CRISP1 gene. This BAC is derived from porcine Chromosome 7 and is syntenic with the regions of the mouse, rat, and human genomes encoding the CRISP gene family. This BAC was found to encode a third CRISP protein with a predicted amino acid sequence of high similarity to human CRISP3. Using RT-PCR we show that CRISP3 expression in the boar reproductive tract is confined to the prostate. Recombinant porcine (rp) CRISP2 protein was produced and purified. When incubated with capacitated boar sperm, rpCRISP2 induced an acrosome reaction, consistent with its demonstrated ability to alter the activity of calcium channels.

Background and Purpose: Previous efforts to identify the protein content of stone matrix have been limited by the lack of technology necessary to analyze the highly insoluble protein-crystalline complex. Our study objective is to characterize the matrix of calcium oxalate monohydrate (COM) stones using a comprehensive proteomics approach., Materials and Methods: Seven pure COM stones were powdered, and proteins were extracted using four different buffer solutions. Detergent cleanup spin columns or concentrators were used to remove detergent and to exchange buffers before trypsin digestion. Tryptic peptides were analyzed with reversed-phase, high-performance liquid chromatography (RP-HPLC) and tandem mass spectrometry (MS/MS) using a QSTAR Pulsar i quadrapole time of flight mass spectrometer. Tandem mass spectra were searched against National Center for Biotechnology Information human nonredundant database using ProteinPilot 1.0 software (Applied Biosystems, Inc.) for protein hits; peptide MS/MS spectra were manually inspected., Results: Of the four buffers, only 2% sodium dodecyl sulfate (SDS) samples had normal HPLC and MS/MS elution patterns. We identified 68 distinct proteins with 95% confidence. More than 50 of the proteins have not been previously identified in stone matrix. Of particular note, a significant number of inflammatory proteins were identified, including immunoglobulins, defensin -3, clusterin, complement C3a, kininogen, and fibrinogen., Conclusions: SDS reducing buffer was efficient at solubilizing proteins from stone matrix for further MS-based proteomic analysis. A variety of cellular, structural, and plasma proteins comprise COM stone matrix. Several of the stone proteins are involved in cell injury pathways, which suggests that inflammation plays a role in human COM stone formation.

In this study, a capillary electrophoresis (CE) method was developed as a means to measure levels of penicillin G (PCN G) in Group B Streptococcus (GBS) positive pregnant women during labor and delivery. Volunteers for this developmental study were administered five million units of PCN G at the onset of labor. Urine, blood, and amniotic fluid samples were collected during labor and post delivery. Samples were semi-purified by solid-phase extraction (SPE) using Waters tC18 SepPak 3cc cartridges with a sodium phosphate/methanol step gradient for elution. Capillary electrophoresis or reversed-phase high-performance liquid chromatography (RP-HPLC) with diode-array absorbance detection were used to separate the samples in less than 30 min. Quantification was accomplished by establishing a calibration curve with a linear dynamic range. The tC18 SPE methodology provided substantial sample clean-up with high recovery yields of PCN G ( approximately 90%). It was found that SPE was critical for maintaining the integrity of the separation column when using RP-HPLC, but was not necessary for sample analysis by CE where no stationary phase is present. Quantification results ranged from millimolar concentrations of PCN G in maternal urine to micromolar concentrations in amniotic fluid. Serum and cord blood levels of PCN G were below quantification limits, which is likely due to the prolonged delay in sample collection after antibiotic administration. These results show that CE can serve as a simple and effective means to characterize the pharmacokinetic distribution of PCN G from mother to unborn fetus during labor and delivery. It is anticipated that similar methodologies have the potential to provide a quick, simple, and cost-effective means of monitoring the clinical efficacy of PCN G and other drugs during pregnancy.

Introduction: Historically, exercise performance outcomes may be predicted with anthropometric variables such as body mass. To assess body mass and resistance exercise (REX) performance variable correlations to blood lactate values, subjects performed five different workouts on two devices that do not employ gravitational resistance. One device uses flywheels to impose a low-speed high-resistance exercise stimulus while the other, dubbed a Dual Performance Device (DPD), has a sled mounted on a very low friction track to enable high-speed low-resistance repetitions., Methods: Subjects (n = 18) performed 3 leg press workouts on the flywheel ergometer (FE), which entailed: 1) a 3-set 10-repetition protocol with concentric and eccentric actions; or 2) the same set-repetition paradigm with concentric-only actions; or 3) a 6-set 10-repetition protocol of concentric-only actions. DPD workouts, done by a separate (n = 22) group, involved 2 workouts each composed exclusively of tonic or phasic seated knee and hip extensor repetitions. Multivariate regression was employed, with post-exercise and delta (post/ pre) blood lactate values as criterion measures., Results: While body mass and performance values from FE workouts were weakly correlated to the criterion measures, body mass and average power (AP) variables from DPD tonic workouts explained 99% of the post-exercise and delta[BLa-] variance and yielded two prediction equations: post-REX blood lactate' = 0.06 + 11.21(AP) + -11.53(body mass), delta blood lactate' = 0.03 + 11.83(AP) + -12.00(body mass)., Conclusions: Results were most likely due to differences in central command requirements for each exercise device.

Cysteine-rich secretory protein-1 (CRISP-1) is a glycoprotein secreted by the epididymal epithelium. It is a member of a large family of proteins characterized by two conserved domains and a set of 16 conserved cysteine residues. In mammals, CRISP-1 inhibits sperm-egg fusion and can suppress sperm capacitation. The molecular mechanism of action of the mammalian CRISP proteins remains unknown, but certain non-mammalian CRISP proteins can block ion channels. In the rat, CRISP-1 comprises two forms referred to as Proteins D and E. Recent work in our laboratory demonstrates that the D form of CRISP-1 associates transiently with the sperm surface, whereas the E form binds tightly. When the spermatozoa are washed, the E form of CRISP-1 persists on the sperm surface after all D form has dissociated. Cross-linking studies demonstrate different protein-protein interaction patterns for D and E, although no binding partners for either protein have yet been identified. Mass spectrometric analyses revealed a potential post-translational modification on the E form that is not present on the D form. This is the only discernable difference between Proteins D and E, and presumably is responsible for the difference in behavior of these two forms of rat CRISP-1. These studies demonstrate that the more abundant D form interacts with spermatozoa transiently, possibly with a specific receptor on the sperm surface, consistent with a capacitation-suppressing function during sperm transit and storage in the epididymis, and also confirm a tightly bound population of the E form that could act in the female reproductive tract.

The current study addressed how the timing of interviews affected children's memories of unique and repeated events. Five- to six-year-olds (N=125) participated in activities 1 or 4 times and were misinformed either 3 or 21 days after the only or last event. Although single-experience children were subsequently less accurate in the 21- versus 3-day condition, the timing of the misinformation session did not affect memories of repeated-experience children regarding invariant details. Children were more suggestible in the 21- versus 3-day condition for variable details when the test occurred soon after misinformation presentation. Thus, timing differentially affected memories of single and repeated events and depended on the combination of event-misinformation and misinformation-test delays rather than the overall retention interval.

Porcine seminal plasma (SP) has been shown to contain factors that have a decapacitative or capacitation-inhibiting effect on sperm. The objectives of the present study were to compare the capacitative changes observed in cooled sperm with those seen in sperm after in vitro capacitation and to determine whether SP could prevent these changes. Sperm were subjected to incubation or to slow cooling under noncapacitating or capacitating conditions. The effect of SP on protein tyrosine phosphorylation and the ability of the sperm to undergo an acrosome reaction (AR) were determined. Cooled sperm displayed an increased level of tyrosine phosphorylation and a higher percentage of induced AR sperm compared to incubated sperm. The addition of SP inhibited the number of ARs that occurred during incubation and cooling. These results suggest that cooling of sperm augments the capacitative changes in sperm, and that SP contains a factor(s) that effectively prevents these changes.

Persistent infertility after apparently successful vasectomy reversal is common. One possible etiology is epididymal epithelial dysfunction resulting in improper sperm maturation after vasectomy reversal. The epididymal epithelium secretes a number of proteins that are thought to be required for the maturation of sperm. Ligation of the vas deferens during vasectomy may affect the synthesis of some of these proteins. In the present study, the function of the epididymal epithelium was assessed at early times after vasectomy (1, 4, and 7 days) by measuring the level of mRNA of 4 secreted proteins: Crisp-1, clusterin, osteopontin, and transferrin. In addition, the site of synthesis of these proteins was determined by immunocytochemistry. The results demonstrated that the expression of Crisp-1 and clusterin, representative epididymal secretory proteins, was largely unaffected by vasectomy. However, osteopontin mRNA increased in the vas deferens in response to vasectomy. Immunocytochemical localization of osteopontin suggested that both infiltrating immune cells and deferential luminal epithelium were responsible for this up-regulation. Transferrin expression was viewed as a marker for immune cells at the site of injury. However, both the caput epididymis and deferential epithelia were found to express transferrin, in addition to immune cells. In conclusion, there appear to be only minor changes in expression of genes encoding epididymal secretory proteins acutely after vasectomy, but, not surprisingly, there was evidence of an inflammatory response after vasectomy.

Crisp-1 is a member of the cysteine-rich secretory protein family. This family of proteins is characterized by the presence of 16 conserved cysteine residues, the characteristic from which the family name is derived. Members of the Crisp protein family are found in the secretions of the reproductive tract and salivary glands, including venom toxins from several species of snakes and lizards. The Crisp proteins are modular, each containing an amino terminal pathogenesis-related (PR)-like domain and a carboxyl terminal cysteine-rich domain (CRD) connected by a hinge region. Sequence and structural similarities to proteins with known functions suggest that the Crisp family of proteins may act by regulating cellular ion channels. Rat Crisp-1 is synthesized as two distinct isoforms (referred to as Proteins D and E) by the epididymal epithelium and both are secreted into the luminal fluid where they interact with spermatozoa. Our laboratory has correlated Crisp-1 binding to sperm with inhibiting the signaling cascades that initiate capacitation while others have shown that blocking Crisp-1 binding sites on oocytes interferes with sperm-egg fusion. We hypothesize that the D and E populations of rat Crisp-1 have different interactions with sperm that modulate these distinct biological activities. Through tandem mass spectrometry (MS/MS) and monosaccharide composition analyses, we have identified at least one difference between the D and E forms as an additional single O-linked N-acetyl galactosamine on an amino terminal threonine residue in Protein E. This post-translational modification appears to account for the unique 'E' epitope bound by monoclonal antibody 4E9 developed in our laboratory, and may also lead to differential processing and localization of Protein E on sperm, when compared to Protein D. These findings are the first step in distinguishing the molecular basis of the biological activities of the D and E forms of rat Crisp-1. The epididymal-specific expression of Crisp-1, combined with its role in regulation of sperm capacitation and oocyte interaction, make it an attractive target for post-testicular contraceptive development.

Participants (6- and 7-year-olds, N=130) participated in classroom activities four times. Children were interviewed about the final occurrence (target event) either 1 week or 4 weeks later, during which half of the event items were described inaccurately. Half of these suggestions were consistent with the theme of the detail across the occurrences (e.g., always sat on a kind of floor mat) or were inconsistent (e.g., sat on a chair). When memory for the target event was tested 1 day later, children falsely recognized fewer inconsistent suggestions than consistent suggestions, especially compared with a control group of children who experienced the event just one time. Furthermore, the longer delay reduced accuracy only for consistent suggestions. Source-monitoring ability was strongly and positively related to resistance to suggestions, and encouraging children to identify the source of false suggestions allowed them to retract a significant proportion of their reports of inconsistent suggestions but not of consistent suggestions. The results suggest that the gist consistency of suggestions determines whether event repetition increases or decreases suggestibility.