Development of a thiol-reactive fluorescent probe for the identification of human spermatozoa

Current methods of identifying semen include preliminary methods such as an alternate light source (ALS) and color tests which test for the presence of acid phosphatase (AP). Confirmatory methods for identifying semen include the microscopic identification of spermatozoa which typically use staining methods such as the Christmas Tree Stain (KPIC) or hematoxylin and eosin stain (H&E). Other methods include immunoassay cards that test for the components of semen such as prostate specific antigen (PSA) and semenogelin (Sg). One common fluorescent staining method used to identify the presence of spermatozoa is SPERM HY-LITERTM which uses an anti-human sperm-specific mouse monoclonal antibody coupled to an AlexaFluor 488 dye. This causes the entire head of the sperm cell to fluoresce when a FITC filter is used. It also utilizes a 4,6-diamidino-2- phenylindole (DAPI) fluorescent dye that stains all cell nuclei non-specifically. This allows the analyst to easily identify whether sperm is present in the sample. A drawback of the antibody labeling procedure is that there are many washing and transfer steps that can lead to sample loss; thus a need for a new, optimized staining method exists. During spermiogenesis, protamines replace histones to further condense the DNA of the sperm nucleus. Humans express two proamines, protamine 1 (P1) and protamine 2 (P2). The protamines contain an arginine-rich core as well as cysteine residues. The high levels of arginine create a net positive charge that allows for stronger binding to DNA. The cysteine residues allow for the formation of inter and intra-protamine disulfide bonds which allow for the chromatin compaction. If the disulfide bonds found in protamines can be reduced to produce free thiols, a thiol-reactive probe can bind and label the protamines in the sperm nucleus. Reduction of the disulfide bonds can be performed by use of reagents such as dithiothreitol (DTT) or tris-(2-carboxyethyl)phosphine (TCEP). From the literature on thiol-reactive probes, it is suggested that TCEP be used as a reducing agent due to its structure. DTT contains thiol groups and when removed from the sample, thiol groups may be oxidized back into disulfide bond. In addition, TCEP is more stable than DTT at a higher pH and higher temperatures. Once disulfide bonds have been reduced, a thiol-reactive probe may be introduced. There are many different types of probes that may be used. Maleimides are commonly used for thiol modification and quantitation. When the compound encounters a thiol group, the thiol group is added across the double bond, yielding a thioether. The reagent used is N-7-dimethylamino-4-methylcoumarin-3-yl)maleimide (DACM). It absorbs light at 376 nm and emits light at 476 nm, producing a blue fluorescence. The dye is nonfluorescent until it reacts with a thiol group. In this research, both sperm and epithelial cells were added to the slides in order to develop a novel staining procedure. The initial protocol used TCEP to break the disulfide bonds followed by DACM to bind free thiol groups. Sodium dodecyl sulfate (SDS) was then added to the protocol to lyse cells. DTT was tested for use as a reducing agent as well. The purpose of establishing this protocol was to design a procedure for rapid labeling of sperm that does not require antibody labeling. In the antibody labeling procedure, there are many washing and transfer steps. The proposed method may limit the number of procedural steps resulting in less loss of biological material. It can also help to limit the time-consuming methods of the current staining techniques, such as KPIC and H&E

New Clinical Faculty Training Program: Transforming Practicing Dentists into Part-Time Dental Faculty Members

At Indiana University School of Dentistry, a New Clinical Faculty Training (NCFT) program was created with the primary goals of informing new part-time faculty members of clinical policies and assessment guidelines and thus developing qualified and satisfied faculty members. The aim of this study was to determine if participation in the training program improved the participants’ satisfaction and competence in comparison to their colleagues who did not participate in the program. Two cohorts were compared: a control group of part-time faculty members who did not receive formal training when they were hired (n=21; response rate 58.3%); and the intervention group, who had participated in the NCFT program (n=12; response rate 80%). A survey of faculty members in the control group gathered information on their experiences when initially hired, and a pretest was administered to measure their knowledge of clinical policies. After the control group was given an overview of the program, their feedback was collected through post surveys, and a posttest identical to the pretest was given that found statistically significant increases on questions one (p=0.003) and four (p=0.025). In February 2014, 15 new faculty members participated in the pilot implementation of the NCFT program. Of those 15, 12 (the intervention group) completed follow-up surveys identical to the pre survey used with the control group. Statistically significant differences were found for the factors clinical teaching (p=0.005) and assessment training (p=0.008) with better responses for the NCFT group. These results suggest that participation in the program was associated with improved clinical teaching knowledge and job satisfaction

NICU Discharge Readiness

After days, weeks or months in the neonatal intensive care unit (NICU), families can be overwhelmed by the discharge preparation and the transition to home. Literature supports that inadequate preparation for discharge from the NICU increases family anxiety, risk for hospital readmission and emergency room visits, as well as infant care issues at home such as feeding difficulty. This quality improvement project was identified to improve and enhance discharge readiness in the NICU. The project started with the development of a discharge readiness committee. After reviewing the literature, the committee met to identify areas in need of improvement which included improving communication of discharge plan, family/caregiver preparedness, completion of education before day of discharge and nursing documentation of completed education. Initial actions items incorporated the initiation of weekly interdisciplinary health team rounds, development of a standardized discharge teaching tool, inclusion of requested circumcision awareness in OB morning huddle, creation of a badge backer to include a QR code for easy access to required discharge videos, and identification/education of necessary educational documentation required within first 24-48 hours of admission in NICU. The committee plans to measure length of stay, parent satisfaction and documentation of education to determine the impact of the identified action items. The goal of this quality improvement project is for families to be feel supported during their NICU journey and prepared for discharge well before the discharge day, resulting in increased patient satisfaction, decreased length of stay and decrease nursing workload on the day of discharge.https://scholar.rochesterregional.org/nursingresearchday/1004/thumbnail.jp

Is a Picture Worth a Thousand Words? Effectiveness of iPad Technology in Preclinical Dental Laboratory Courses

Dental educators should consider alternative modalities of instruction when experiencing difficulties conveying feedback to students. The aim of this study was to determine if integrating iPad technology as a visual learning tool would enhance the exchange of assessment information and improve academic performance in Indiana University School of Dentistry's preclinical curriculum. In 2016, the first‐year Tooth Morphology (TM) and second‐year Fixed Prosthodontics (Fixed) courses implemented a project using iPad images that allowed instructors to annotate acceptable and deficient areas of students' tooth wax‐ups and preparations. In the two courses, all students (TM n=106 and Fixed n=105) and instructors (TM n=21 and Fixed n=17) were given pre‐intervention surveys to report their perceived effectiveness of verbal feedback and were given post‐intervention surveys to rate their experiences with iPad image feedback. Response rates for students in the two courses on the pre surveys were TM 87.7% and Fixed 85.7% and on the post surveys were TM 26.4% and Fixed 76.2%. Response rates for instructors on the pre surveys were TM 52.4% and Fixed 82.4% and on the post surveys were TM 76.2% and Fixed 76.5%. The results showed that a majority of both groups preferred the combination of verbal and iPad image feedback: 53% of responding students in TM and 51% in Fixed, and 75% of instructors in TM and 77% in Fixed. In the TM course, responding instructors had a statistically significantly higher agreement than students that feedback with iPad images was superior to verbal feedback alone (p=0.008). Furthermore, a multi‐year analysis of TM practical examination grades found statistically significant lower change scores for the first and second exams in 2014 and 2015 compared to the 2016 scores when the iPad intervention occurred. These results suggest that verbal feedback combined with iPad images resulted in an enhanced exchange of information and increased student grades, particularly in the first‐year dental curriculum

Bias effects on the electro-optic response of Ge-on-Si waveguide photodetectors

Silicon photonics, allowing for the integration of optoelectronic components into CMOS-compatible platforms, has shown great promise in the development of next-generation telecommunication systems and fast, low-power data interconnects. Within this framework, the increasing transistor integration and power dissipation density has led, during the last decades, to a steady decrease of the CMOS bias voltages. This trend may also affect the operation of silicon photonics components: the present work is focused on the effects of lower bias in Ge-on-Si waveguide photodetectors (WPDs), using the electro-optic (EO) frequency response as a figure of merit

More Rapid Increase in BMI from Age 5–15 is Associated with Elevated Weight Status at Age 24 among Non-Hispanic White Females

Background: A rapidly increasing BMI trajectory throughout childhood is associated with negative health outcomes in adulthood such as obesity, cardiovascular disease, and diabetes. The purpose of the current study was to assess whether BMI trajectories from age 5–15 predicted changes in weight and BMI from adolescence to adulthood, and dieting-related behaviors in young adulthood. Methods: Non-Hispanic White female participants from Early Dieting in Girls (n=182), a longitudinal cohort study, were followed from age 5 to 15 and completed a follow-up survey at age 24. Participants were classified by age 5–15 BMI trajectory groups: UPC, accelerated weight gain from age 5–9; DDPC, accelerated weight gain from 5 to 9 followed by a decrease; 60PT, weight tracked along 60th percentile; 50PT, weight tracked along 50th percentile. Data at age 24 included self-reported weight, height, dietary restraint, disinhibition, and dieting. Results: Majority of participants (80.8%) completed the follow-up survey; of these participants, 60% in UPC group had obesity at age 24, compared to\u3c10% in the other 3 groups. Participants in the UPC group had greater increases in BMI since age 15, compared to the 50PT group, and trend-level greater weight increases than those in the DDPC and 60PT groups. Dietary restraint, but not disinhibition, differed across the groups. Conclusions: Children with accelerated weight gain continued to have the greatest weight gain from adolescence to adulthood and the highest prevalence of obesity in adulthood

3D multiphysics transient modeling of vertical Ge-on-Si pin waveguide photodetectors

We report transient simulations of Ge-on-Si vertical pin waveguide photodetectors (WPDs), where the optical generation term used by the time-domain model is the FDTD solution of the electromagnetic problem treated as a spatially-distributed pulsed signal. This approach, validated against experimental measurements of the frequency response, paves the way to future studies of the dynamic response of WPDs, enabling the description of complex modulation schemes including saturation effects and current tails due to slow carriers

Modeling the frequency response of vertical and lateral Ge-on-Si waveguide photodetectors: Is 3D simulation unavoidable?

Using a 3D multiphysics model as a reference, we investigate the achievements and limitations of a simpler 2D drift-diffusion model to reproduce and optimize the electrooptical frequency response of vertical and lateral Ge-on-Si waveguide photodetectors

Evaluation of a peer coach-led intervention to improve pain symptoms (ECLIPSE): Rationale, study design, methods, and sample characteristics

Chronic pain is prevalent, costly, and a leading cause of disability. Pain self-management (i.e., employing self-management strategies including behavioral modifications) is an effective, evidence-based treatment. However, implementation and delivery of a pain self-management model is challenging because of time and resources. Peer supported pain self-management offers a promising approach to implementing pain self-management programs using fewer clinical resources. Evaluation of a Peer Coach-Led Intervention for the Improvement of Pain Symptoms (ECLIPSE) is a randomized controlled trial testing effectiveness of peer coach-delivered pain self-management intervention versus controls receiving a class on pain and pain self-management. ECLIPSE is a Hybrid Type 1 study testing effectiveness while examining implementation factors. ECLIPSE enrolled 215 veterans randomly assigned to the peer coaching (N = 120) or control (N = 95) arm. The peer coaching intervention lasts 6 months, with patient-peer coach pairs instructed to talk twice per month. Coaches attend initial training, are provided a detailed training manual, and attend monthly booster sessions. Outcomes are assessed at baseline, 6 months, and 9 months. The primary outcome is overall pain (intensity and interference), measured by the Brief Pain Inventory (BPI). Secondary outcomes are self-efficacy, social support, pain catastrophizing, patient activation, health-related quality of life, and health care utilization. To maximize implementation potential of pain self-management, innovative delivery methods are needed that do not require additional resources from healthcare teams. A novel and promising approach is a peer-coaching model, in which patients who are successfully managing their pain offer information, ongoing support, and advice to other patients with pain