Inspiring a convergent engineering approach to measure and model the tissue microenvironment.

Understanding the molecular and physical complexity of the tissue microenvironment (TiME) in the context of its spatiotemporal organization has remained an enduring challenge. Recent advances in engineering and data science are now promising the ability to study the structure, functions, and dynamics of the TiME in unprecedented detail; however, many advances still occur in silos that rarely integrate information to study the TiME in its full detail. This review provides an integrative overview of the engineering principles underlying chemical, optical, electrical, mechanical, and computational science to probe, sense, model, and fabricate the TiME. In individual sections, we first summarize the underlying principles, capabilities, and scope of emerging technologies, the breakthrough discoveries enabled by each technology and recent, promising innovations. We provide perspectives on the potential of these advances in answering critical questions about the TiME and its role in various disease and developmental processes. Finally, we present an integrative view that appreciates the major scientific and educational aspects in the study of the TiME.

On-demand Milifluidic Synthesis of Quantum Dots in Digital Droplet Reactors.

Colloidal quantum dots (QDs) offer dramatic potential due to their size-dependent optical properties. Lack of facile synthesis methods for precise and reproducible size and composition, however, present an extant barrier to their widespread use. Here we report the use of droplet microfluidics for the simple and highly reproducible synthesis of cadmium sulfide (CdS) and cadmium selenide (CdSe) QDs without the use of harsh solvents and in ambient conditions. Our approach uses a liquid-liquid barrier between two immiscible liquids to generate a digital droplet reactor. This reaction droplet is easily controlled and manipulated and offers enhanced mixing when coupled to a helical mixer, resulting in a significant reduction in size distribution compared to benchtop procedures. Furthermore, QD characteristics have modeled and predicted based on the parameters of the microfluidic device. We believe this method overcomes the current manufacturing challenges with synthesizing nanostructures, which is required for the next generation of nanosensors.

An fMRI investigation of the neural correlates underlying the autonomous sensory meridian response (ASMR).

Introduction : The "autonomous sensory meridian response" (ASMR) is a neologism used to describe an internal sensation of deep relaxation and pleasant head tingling which is often stimulated by gentle sounds, light touch, and personal attention. Methods : An fMRI-based methodology was employed to examine the brain activation of subjects prescreened for ASMR-receptivity (n=10) as they watched ASMR videos and identified specific moments of relaxation and tingling. Results : Subjects who experienced ASMR showed significant activation in regions associated with both reward (NAcc) and emotional arousal (dACC and Insula/IFG). Brain activation during ASMR showed similarities to patterns previously observed in musical frisson as well as affiliative behaviors. Conclusion : This is the first study to measure the activation of various brain regions during ASMR and these results may help to reveal the mechanistic underpinnings of this sensation.

Pharmacy students' use and perceptions of Apple mobile devices incorporated into a basic health science laboratory.

OBJECTIVE: To describe pharmacy students' use of mobile devices in a basic health science laboratory and to report the students' perceptions on how solving cases with their mobile devices influenced their attitudes, abilities, and view on the use of mobile devices as tools for pharmacists. METHODS: First-year pharmacy students utilized mobile devices to solve clinical case studies in a basic health sciences laboratory. A pre-survey and two post-surveys were administered to assess the students' comfort, awareness, use, and perceptions on the use of their mobile devices and apps. RESULTS: The pre-survey and first post-survey each had a response rate of 99%, and the second post-survey had a response rate of 100%. In comparing the pre-survey and first post-survey data, there was a statistically significant increase in the number of students that agreed or strongly agreed that they were more comfortable utilizing their mobile device (p = 0.025), they were more aware of apps for pharmacists (p < 0.005), and they have used more apps that can be useful for pharmacists (p < 0.005). The second post-survey demonstrated that over 78% of students agreed or strongly agreed that completing the case studies influenced them to be more comfortable with their mobile devices, to be more aware of apps that can be useful for pharmacists, and to be more agreeable with mobile device utilization by pharmacists in improving patient care. In addition, the second post-survey also demonstrated that 84% of students responded that using their mobile devices to solve the cases influenced them to either use their mobile device in a clinical setting for a clinical and/or pharmacy-related purpose for the first time or to use it more frequently for this purpose. CONCLUSIONS: The use of mobile devices to solve clinical cases in a first-year basic health science laboratory course was perceived as beneficial by students and influenced them to utilize their mobile device even more in a pharmacy practice setting.

A Retinotopic Basis for the Division of High-Level Scene Processing between Lateral and Ventral Human Occipitotemporal Cortex.

In humans, there is a repeated category-selective organization across the lateral and ventral surfaces of the occipitotemporal cortex. This apparent redundancy is often explained as a feedforward hierarchy, with processing within lateral areas preceding the processing within ventral areas. Here, we tested the alternative hypothesis that this structure better reflects distinct high-level representations of the upper (ventral surface) and lower (lateral surface) contralateral quadrants of the visual field, consistent with anatomical projections from early visual areas to these surfaces in monkey. Using complex natural scenes, we provide converging evidence from three independent functional imaging and behavioral studies. First, population receptive field mapping revealed strong biases for the contralateral upper and lower quadrant within the ventral and lateral scene-selective regions, respectively. Second, these same biases were observed in the position information available both in the magnitude and multivoxel response across these areas. Third, behavioral judgments of a scene property strongly represented within the ventral scene-selective area (open/closed), but not another equally salient property (manmade/natural), were more accurate in the upper than the lower field. Such differential representation of visual space poses a substantial challenge to the idea of a strictly hierarchical organization between lateral and ventral scene-selective regions. Moreover, such retinotopic biases seem to extend beyond these regions throughout both surfaces. Thus, the large-scale organization of high-level extrastriate cortex likely reflects the need for both specialized representations of particular categories and constraints from the structure of early vision. SIGNIFICANCE STATEMENT: One of the most striking findings in fMRI has been the presence of matched category-selective regions on the lateral and ventral surfaces of human occipitotemporal cortex. Here, we focus on scene-selective regions and provide converging evidence for a retinotopic explanation of this organization. Specifically, we demonstrate that scene-selective regions exhibit strong biases for different portions of the visual field, with the lateral region representing the contralateral lower visual field and the ventral region the contralateral upper visual field. These biases are consistent with the retinotopy found in the early visual areas that lie directly antecedent to category-selective areas on both surfaces. Furthermore, these biases extend beyond scene-selective cortex and provide a retinotopic basis for the large-scale organization of occipitotemporal cortex.

Pharmacy students' preference for using mobile devices in a clinical setting for practice-related tasks.

OBJECTIVE: To examine pharmacy students' ownership of, use of, and preference for using a mobile device in a practice setting. METHODS: Eighty-one pharmacy students were recruited and completed a pretest that collected information about their demographics and mobile devices and also had them rank the iPhone, iPad mini, and iPad for preferred use in a pharmacy practice setting. Students used the 3 devices to perform pharmacy practice-related tasks and then completed a posttest to again rank the devices for preferred use in a pharmacy practice setting. RESULTS: The iPhone was the most commonly owned mobile device (59.3% of students), and the iPad mini was the least commonly owned (18.5%). About 70% of the students used their mobile devices at least once a week in a pharmacy practice setting. The iPhone was the most commonly used device in a practice setting (46.9% of students), and the iPod Touch was the least commonly used device (1.2%). The iPad mini was the most preferred device for use in a pharmacy practice setting prior to performing pharmacy practice-related tasks (49.4% of students), and was preferred by significantly more students after performing the tasks (70.4%). CONCLUSION: Pharmacy students commonly use their mobile devices in pharmacy practice settings and most selected the iPad mini as the preferred device for use in a practice setting even though it was the device owned by the fewest students.

Warfarin Dosing in a Patient with CYP2C9(∗)3(∗)3 and VKORC1-1639 AA Genotypes.

Genetic factors most correlated with warfarin dose requirements are variations in the genes encoding the enzymes cytochrome P450 2C9 (CYP2C9) and vitamin K epoxide reductase (VKOR). Patients receiving warfarin who possess one or more genetic variations in CYP2C9 and VKORC1 are at increased risk of adverse drug events and require significant dose reductions to achieve a therapeutic international normalized ratio (INR). A 74-year-old white female with atrial fibrillation was initiated on a warfarin dose of 2 mg PO daily, which resulted in multiple elevated INR measurements and three clinically significant hemorrhagic events and four vitamin K antidote treatments over a period of less than two weeks. Genetic analysis later revealed that she had the homozygous variant genotypes of CYP2C9∗3∗3 and VKORC1-1639 AA. Warfarin dosing was subsequently restarted and stabilized at 0.5 mg PO daily with therapeutic INRs. This is the first case report of a white female with these genotypes stabilized on warfarin, and it highlights the value of pharmacogenetic testing prior to the initiation of warfarin therapy to maximize efficacy and minimize the risk of adverse drug events.

First report of warfarin dose requirements in patients possessing the CYP2C9*12 allele.

BACKGROUND: Warfarin is the most frequently prescribed anticoagulant in North America and Europe. It is administered as a racemate, but S-warfarin is principally responsible for its anticoagulant activity. Cytochrome P450 (CYP) 2C9 is the enzyme primarily responsible for the metabolism of S-warfarin. Numerous variant alleles of CYP2C9 have been identified. The CYP2C9*12 (rs9332239) allele harbors a P489S substitution in CYP2C9 which has been shown to result in a 40% decline in catalytic activity in vitro. CASES: Four Caucasian patients with a low mean weekly warfarin dose (MWWD) were genotyped for CYP2C9, VKORC1 and APOE variant alleles. None of the four patients carried the common CYP2C9 variant alleles (*2, *3, *5, *6, *7, *8, *9, *11, *13) despite a relatively low MWWD (23.4±7.94 mg) compared to 208 patients carrying the CYP29C9*1 genotype (32.2±12.65 mg). Given that CYP2C9*12 confers decreased in vitro activity to the enzyme, we investigated whether these patients carried this allele. All four patients were CYP2C9*12 CT heterozygotes. Individual comparisons with patients possessing the same VKORC1 and APOE genotypes also demonstrated lower dose requirements in the patients that possessed CYP2C9*12 allele. CONCLUSIONS: There are no reports of the clinical impact of rs9332239 on CYP2C9 substrates. This is the first report of patients with the rare CYP2C9*12 genotype and lower warfarin dose requirements.

Student pharmacists' use and perceived impact of educational technologies.

OBJECTIVE: To assess the frequency of use by and perceived impact of various educational technologies on student pharmacists. METHODS: Data were obtained using a validated, Web-based survey instrument designed to evaluate the frequency of use and impact on learning of various technologies used in educating first-, second-, and third-year student pharmacists. Basic demographic data also were collected and analyzed. RESULTS: The majority (89.4%) of the 179 respondents were comfortable with the technology used in the academic program. The most frequently used technologies for educational purposes were in class electronic presentations, course materials posted on the school Web site, and e-mail. The technologies cited as having the most beneficial impact on learning were course materials posted on the Web site and in-class electronic presentations, and those cited as most detrimental were video-teleconferencing and online testing. Compared to the course textbook, students reported more frequent use of technologies such as electronic course materials, presentations, digital lecture recordings, e-mail, and hand-held devices. CONCLUSIONS: Because students' opinions of educational technologies varied, colleges and schools should incorporate educational technologies that students frequently use and that positively impact learning.

Comparison of cell cycle regulatory gene mRNA in three different types of human trophoblasts and effect of transforming growth factor.

AIM: Identifying the factors responsible for reducing the proliferation, syncytialization, and invasiveness of trophoblast tissues, as seen with preeclampsia, intrauterine growth restriction, and spontaneous miscarriage, is a current challenge in reproductive biology. These factors, transforming growth factor (TGF)-beta as an example, can work by altering trophoblast differentiation or proliferation. We therefore investigated and compared specific markers of trophoblast proliferation and differentiation in three commonly used trophoblast tissue cell models, and also investigated the influence of TGF-beta on these markers. METHODS: In this study, we isolated human trophoblasts from first trimester and term placentas, and additionally used human choriocarcinoma cells (JEG-3). Baseline values of human chorionic gonadotropin (hCG) secretion and relative mRNA levels of cell cycle regulators (cyclin E, p21, p27, and p57) were investigated for each cell type. We also investigated the influence of TGF-beta on these parameters. RESULTS: Quantitative and longitudinal production of hCG differed between the three cell types. Significantly different amounts of cyclin E, p21, p27, and p57 mRNA were demonstrated within each cell type, as well as between all the cell types, throughout the culture time period. Each trophoblast type demonstrated a reduction of hCG secretion in response to TGF-beta. TGF-beta did not show a consistent effect on the cell cycle mRNA of any of the cell types. CONCLUSION: We were able to characterize and compare the differential production of hCG, as well as the differential expression of cell cycle-associated mRNA of early trophoblasts, term trophoblasts, and choriocarcinoma cells. The production of hCG was altered by TGF-beta, although mRNA levels were not markedly altered by TGF-beta.

Enzymatic removal of asparagine-linked carbohydrate chains from heterodimer human chorionic gonadotrophin and effect on bioactivity.

The native form of human chorionic gonadotropin (hCG) is a heterodimer protein with two asparagine (Asn)-linked carbohydrate chains on each subunit. Removal of the Asn-linked carbohydrate chains from hCG has resulted in hCG variants with consistent antagonistic properties on isolated murine cells. Specific and direct enzymatic removal of these carbohydrate chains from native hCG with resultant antagonistic properties has not been reported. An antagonist to the hCG/luteinising hormone (LH) receptor could be used as an anticancer therapy, emergency contraceptive or for therapeutic resolution of ectopic pregnancies. Therefore, our aim was to use enzymes to specifically remove Asn-linked carbohydrate chains from hCG in the heterodimer form and analyse the resultant bioactivity. Native hCG was treated with endoglycosidases, carbohydrate removal was analysed with electrophoresis and the hCG variants were tested for altered bioactivity with human and murine cells. Endoglycosidases were able to cleave most of the Asn-linked carbohydrate chains from the native hCG. The deglycosylated hCG demonstrated a 75% reduction in bioactivity on a murine Leydig cell line and a 65% reduction in bioactivity on human granulosa cells. These results exemplify a simple and efficient method for creating deglycosylated hCG and provide the most direct evidence for the importance of Asn-linked carbohydrate chains in maintaining hCG bioactivity.

Monocytes are progressively activated in the circulation of pregnant women.

Pregnancy is characterized by the presence of generalized leukocyte activation. We used flow cytometry to investigate changes in phenotype and intracellular cytokines of circulating granulocytes, monocytes, and T lymphocytes of pregnant women during gestation. We report that peripheral circulation of pregnancy is characterized by an increased percentage of granulocytes and a decrease in lymphocytes. The proportion of monocytes remains stable throughout gestation; however, a progressive up-regulation of surface markers CD11a, CD54, and CD64 was detected. Monocytes also showed higher production of interleukin (IL)-12 and IL-1beta compared with the nonpregnant state, and granulocytes had greater potential to synthesize IL-8. All these changes were particularly marked in late gestation. T lymphocytes did not have any characteristics of the activated state and showed a decreased IL-6 production. These findings demonstrate that activation of maternal monocytes and granulocytes increases during pregnancy and support the idea that pregnancy results in an elevation of the innate immune system and suppression of the adaptive immune system.