Dual Supramolecular Nanoparticle Vectors Enable CRISPR/Cas9-Mediated Knockin of Retinoschisin 1 Gene-A Potential Nonviral Therapeutic Solution for X-Linked Juvenile Retinoschisis.

The homology-independent targeted integration (HITI) strategy enables effective CRISPR/Cas9-mediated knockin of therapeutic genes in nondividing cells in vivo, promising general therapeutic solutions for treating genetic diseases like X-linked juvenile retinoschisis. Herein, supramolecular nanoparticle (SMNP) vectors are used for codelivery of two DNA plasmids-CRISPR-Cas9 genome-editing system and a therapeutic gene, Retinoschisin 1 (RS1)-enabling clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein 9 (CRISPR/Cas9) knockin of the RS1 gene with HITI. Through small-scale combinatorial screenings, two SMNP vectors, with Cas9 and single guide RNA (sgRNA)-plasmid in one and Donor-RS1 and green fluorescent protein (GFP)-plasmid in the other, with optimal delivery performances are identified. These SMNP vectors are then employed for CRISPR/Cas9 knockin of RS1/GFP genes into the mouse Rosa26 safe-harbor site in vitro and in vivo. The in vivo study involves intravitreally injecting the two SMNP vectors into the mouse eyes, followed by repeated ocular imaging by fundus camera and optical coherence tomography, and pathological and molecular analyses of the harvested retina tissues. Mice ocular organs retain their anatomical integrity, a single-copy 3.0-kb RS1/GFP gene is precisely integrated into the Rosa26 site in the retinas, and the integrated RS1/GFP gene is expressed in the retinas, demonstrating CRISPR/Cas9 knockin of RS1/GFP gene.

Public perception of pharmacist-prescribed self-administered non-emergency hormonal contraception: An analysis of online social discourse.

BACKGROUND: Increasing access to hormonal contraception reduces unintended pregnancies. One strategy gaining momentum is allowing patients to access hormonal contraception directly from pharmacists. Commentary on online news articles provides a naturalistic and real-time data source to assess public perceptions on timely and often controversial issues. OBJECTIVE: Characterize public perceptions of pharmacist-prescribed self-administered non-emergency hormonal contraception using comments posted in response to online news articles. METHODS: Retrospective, cross-sectional, mixed methods analysis of public comments posted in response to articles published by major media outlets between January 1 to December 31, 2015 on pharmacist-prescribed hormonal contraception. Comments were then extracted and reviewed through a two-step process to identify emerging themes using a thematic analysis. RESULTS: A total of 1060 public online comments were collected from eight articles. Of these, 757 comments (71.4%) were not related to pharmacist-prescribed hormonal contraception. Thematic analysis of the 303 relevant comments (28.6%) identified three overarching messages. First, a wide variety of reasons for how pharmacist-prescribed hormonal contraception improves healthcare and/or supports patient preferences were cited, but there was no consensus on the primary vantage. Second, individuals have varying opinions about the role of the pharmacist which creates both opportunities and challenges for pharmacist-prescribed hormonal contraception. Third, practical and logistical considerations will need to be addressed by healthcare systems, pharmacies, and payers prior to and alongside implementation. CONCLUSION: Commenters were generally positive and cited several benefits, such as increasing access to healthcare, reducing unintended pregnancies, and supporting individual autonomy. However, it was acknowledged that these benefits would need to be balanced with potential safety concerns and logistical issues associated with delivering clinical services in a community pharmacy setting. Study results can help understand public concerns and may be useful in addressing barriers hindering public acceptance of expanded pharmacist roles.

Guideline concordant venous ulcer care predicts healing in a tertiary care Veterans Affairs Medical Center.

This study describes the impact of 80% adherence to guideline concordant care for compression therapy, moist wound-healing environment, and debridement on venous ulcer outcomes. The retrospective cohort design included patients from a tertiary care Veterans Affairs Medical Center from October 2003 to September 2007. During this 5-year interval, 155 patients with 400 venous ulcers met study inclusion. A majority of ulcers (n=362) healed, with an average time to healing of 18.1 weeks (range 2-209 weeks, median 10.4 weeks). From the multivariate Poisson regression, the likelihood of ulcer healing increased when compression therapy was provided during at least 80% of visits (relative risk [RR], 1.93; 95% confidence interval [CI], 1.27-2.92) or when a moist wound-healing environment was provided during at least 80% of visits (RR, 1.63; 95% CI, 1.09-2.42). Debridement alone was not significantly associated with ulcer healing (RR, 1.0; 95% CI, 0.61-1.64). Patients who received all three treatments during at least 80% of their visits were more likely to heal than those who received < 80% treatment (RR, 2.52; 95% CI, 1.53-4.16). Guideline concordant venous ulcer care was significantly associated with venous ulcer healing, when provided at 80% or more of patient visits.

Cutaneous manifestations of Corynebacterium jeikeium sepsis.

Corynebacterium jeikeium is a rare but increasingly important cause of septicemia in neutropenic patients that can present with characteristic cutaneous findings. A 61-year-old man with myelodysplastic syndrome developed C. jeikeium sepsis and an erythematous papular eruption while neutropenic from induction chemotherapy. A review of the English language literature shows only six reported cases of hemorrhagic or erythematous papular eruption in C. jeikeium sepsis. The case is presented with an updated literature review of the history, risk factors, and treatment.

Structure determination of chiral sulfoxide in diastereomeric bicalutamide derivatives.

We report on the synthesis and investigation of two diastereomers (5a and 5b) of a new bicalutamide analog with an asymmetric carbon atom and a chiral sulfoxide group. These bicalutamide analogs are novel androgen receptor antagonists with biological activities that depend significantly on the configuration of their stereogenic centers. We determined the absolute configuration at the SO center by combining X-ray and NMR measurements with quantum chemical calculations. Since 5a and 5b failed to yield satisfactory crystals for X-ray crystal structure determination, analogs 6a and 6b differing in only one remote functional group relative to the chiral sulfoxide were synthesized, which yielded satisfactory crystals. X-ray structure determination of 6a and 6b provided the absolute configuration at the chiral sulfoxide. Since the structural difference between 5 and 6 is remote from the chiral sulfoxide, the structural assignment was extended from the diastereomers of 6 to those of 5 provisionally. These assignments were verified with the help of measured and DFT-calculated proton and carbon NMR chemical shifts.

Skin problems in individuals with lower-limb loss: literature review and proposed classification system.

Problems with skin integrity can disrupt daily prosthesis use and lead to decreased mobility and function in individuals with lower-limb loss. This study reviewed the literature to examine how skin problems are defined and diagnosed and to identify the prevalence and types of skin problems in individuals with lower-limb loss. We searched the literature for terms related to amputation and skin problems. We identified 777 articles. Of the articles, 90 met criteria for review of research methodology. Four clinical studies met our selection criteria. The prevalence rate of skin problems was 15% to 41%. The most commonly reported skin problems were wounds, abscesses, and blisters. Given the lack of standardized definitions of skin problems on residual limbs, we conclude this article with a system for classification.

Tetracycline protects myocardium against ischemic injury.

Stress pretreatments protect myocardium from ischemic injury. We hypothesized that tetracycline, an antibiotic, may induce a stress response via the inhibition of mitochondrial translation as it induces the cold stress response by translational inhibition in E. coli. If so, tetracycline may protect myocardium from ischemic injury as stress pretreatments do. Thus, we investigated the effects of tetracycline on myocardial ischemia and its association with stress response. In a dog model of acute ischemia, 4mg/kg tetracycline injected 30 min prior to the occlusion improved the functional recovery from stunning of myocardium caused by ischemia. The same dosage of tetracycline dramatically reduced the size of infarct area in murine hearts analyzed by tetrazolium staining. In HeLa cells, tetracycline induced molecules that were increased by cold stress, which suggests that tetracycline may induce a cold stress-like response in mammalian cells. These molecules were also induced by ischemic stress in murine hearts, suggesting that the stress response caused by translational inhibition in mitochondria may be associated with the cardioprotection by tetracycline. Our results suggest that a subclinical dosage of tetracycline may protect heart from ischemic injury. Therefore, tetracycline may be of great use in suppressing the development of infarction caused by myocardial ischemia. This study is also important for providing new insights into the non-antimicrobial effects of tetracycline and its derivatives.

Transcriptional activation of human CYP17 in H295R adrenocortical cells depends on complex formation among p54(nrb)/NonO, protein-associated splicing factor, and SF-1, a complex that also participates in repression of transcription.

The first 57 bp upstream of the transcription initiation site of the human CYP17 (hCYP17) gene are essential for both basal and cAMP-dependent transcription. EMSA carried out by incubating H295R adrenocortical cell nuclear extracts with radiolabeled -57/-38 probe from the hCYP17 promoter showed the formation of three DNA-protein complexes. The fastest complex contained steroidogenic factor (SF-1) and p54(nrb)/NonO, the intermediate complex contained p54(nrb)/NonO and polypyrimidine tract-binding protein-associated splicing factor (PSF), and the slowest complex contained an SF-1/PSF/p54(nrb)/NonO complex. (Bu)(2)cAMP treatment resulted in a cAMP-inducible increase in the binding intensity of only the upper complex and also activated hCYP17 gene transcription. SF-1 coimmunoprecipitated with p54(nrb)/NonO, indicating direct interaction between these proteins. Functional assays revealed that PSF represses basal transcription. Further, the repression of hCYP17 promoter-reporter construct luciferase activity resulted from PSF interacting with the corepressor mSin3A. Trichostatin A attenuated the inhibition of basal transcription, suggesting that a histone deacetylase interacts with the SF-1/PSF/p54(nrb)/NonO/mSin3A complex. Our studies lend support to the idea that the balance between transcriptional activation and repression is essential in the control of adrenocortical steroid hormone biosynthesis.