In situ production and secretion of proteins endow therapeutic benefit against psoriasiform dermatitis and melanoma.

Genetic medicines have the potential to treat various diseases; however, certain ailments including inflammatory diseases and cancer would benefit from control over extracellular localization of therapeutic proteins. A critical gap therefore remains the need to develop and incorporate methodologies that allow for posttranslational control over expression dynamics, localization, and stability of nucleic acid-generated protein therapeutics. To address this, we explored how the body's endogenous machinery controls protein localization through signal peptides (SPs), including how these motifs could be incorporated modularly into therapeutics. SPs serve as a virtual zip code for mRNA transcripts that direct the cell where to send completed proteins within the cell and the body. Utilizing this signaling biology, we incorporated secretory SP sequences upstream of mRNA transcripts coding for reporter, natural, and therapeutic proteins to induce secretion of the proteins into systemic circulation. SP sequences generated secretion of various engineered proteins into the bloodstream following intravenous, intramuscular, and subcutaneous SP mRNA delivery by lipid, polymer, and ionizable phospholipid delivery carriers. SP-engineered etanercept/TNF-α inhibitor proteins demonstrated therapeutic efficacy in an imiquimod-induced psoriasis model by reducing hyperkeratosis and inflammation. An SP-engineered anti-PD-L1 construct mediated mRNA encoded proteins with longer serum half-lives that reduced tumor burden and extended survival in MC38 and B16F10 cancer models. The modular nature of SP platform should enable intracellular and extracellular localization control of various functional proteins for diverse therapeutic applications.

Characterization of ALTO-encoding circular RNAs expressed by Merkel cell polyomavirus and trichodysplasia spinulosa polyomavirus.

Circular RNAs (circRNAs) are a conserved class of RNAs with diverse functions, including serving as messenger RNAs that are translated into peptides. Here we describe circular RNAs generated by human polyomaviruses (HPyVs), some of which encode variants of the previously described alternative large T antigen open reading frame (ALTO) protein. Circular ALTO RNAs (circALTOs) can be detected in virus positive Merkel cell carcinoma (VP-MCC) cell lines and tumor samples. CircALTOs are stable, predominantly located in the cytoplasm, and N6-methyladenosine (m6A) modified. The translation of MCPyV circALTOs into ALTO protein is negatively regulated by MCPyV-generated miRNAs in cultured cells. MCPyV ALTO expression increases transcription from some recombinant promoters in vitro and upregulates the expression of multiple genes previously implicated in MCPyV pathogenesis. MCPyV circALTOs are enriched in exosomes derived from VP-MCC lines and circALTO-transfected 293T cells, and purified exosomes can mediate ALTO expression and transcriptional activation in MCPyV-negative cells. The related trichodysplasia spinulosa polyomavirus (TSPyV) also expresses a circALTO that can be detected in infected tissues and produces ALTO protein in cultured cells. Thus, human polyomavirus circRNAs are expressed in human tumors and infected tissues and express proteins that have the potential to modulate the infectious and tumorigenic properties of these viruses.

A Protein Kinase C Phosphorylation Motif in GLUT1 Affects Glucose Transport and is Mutated in GLUT1 Deficiency Syndrome.

Protein kinase C has been implicated in the phosphorylation of the erythrocyte/brain glucose transporter, GLUT1, without a clear understanding of the site(s) of phosphorylation and the possible effects on glucose transport. Through in vitro kinase assays, mass spectrometry, and phosphospecific antibodies, we identify serine 226 in GLUT1 as a PKC phosphorylation site. Phosphorylation of S226 is required for the rapid increase in glucose uptake and enhanced cell surface localization of GLUT1 induced by the phorbol ester 12-O-tetradecanoyl-phorbol-13-acetate (TPA). Endogenous GLUT1 is phosphorylated on S226 in primary endothelial cells in response to TPA or VEGF. Several naturally occurring, pathogenic mutations that cause GLUT1 deficiency syndrome disrupt this PKC phosphomotif, impair the phosphorylation of S226 in vitro, and block TPA-mediated increases in glucose uptake. We demonstrate that the phosphorylation of GLUT1 on S226 regulates glucose transport and propose that this modification is important in the physiological regulation of glucose transport.

Caregiving experiences of Korean family caregivers of cancer patients: An integrative literature review.

OBJECTIVE: The purpose of this study was to describe the experiences of Korean family caregivers of patients with cancer by reviewing recent literature. METHODS: Five electronic databases were searched-MEDLINE, the Cumulative Index to Nursing and Allied Health Literature (CINAHL), and PubMed for English literature, and Korean Information Service System (KISS), and Research Information Sharing Service (RISS) for Korean literature for articles published from January 2010 to March 2020 using the key words cancer, caregiver, and Korean. Twenty-six articles met the inclusion criteria and remained in the final review. RESULTS: No intervention study was found and most of studies were quantitative without theoretical/conceptual framework. All the studies were conducted with Koreans living in Korea. No previous study has been conducted with Koreans living in the U.S. or other countries. Most studies focused on caregivers' quality of life, burden, unmet needs, and resilience/adaptation/post-traumatic growth. CONCLUSIONS: Comprehensive intervention studies focused on improving quality of life, burden, and adaptation to their complex roles as caregivers in the context of Korean culture would be helpful. Further research is needed in examining the caregiver-patient dyad interactions longitudinally to understand the dynamic complicated processes of caregiving.

Perceptions of illness severity in adults with drug-resistant temporal lobe epilepsy.

OBJECTIVE: The purpose of this study was to explore how subjective perceptions of illness severity were described by a sample of participants with drug-resistant epilepsy (DRE) who were considering surgery. METHODS: A qualitative methodology, constructivist grounded theory, guided all aspects of the study. Data were collected via 51 semi-structured interviews with 35 adults in our multiethnic sample. At interview, the 20 women (57%) and 15 men (43%) ranged in age from 18 to 68 years (mean = 35.6 years) and had lived with epilepsy for an average of 15.4 y (range = 2-44 years). RESULTS: A grounded theory with four interrelated categories was developed to reflect the process by which participants arrived at an explanation of illness severity. Illness severity for participants evolved as participants reflected upon the burdensome impact of uncontrolled seizures on self and others. Epilepsy, when compared with other chronic conditions, was described as less serious, and participants imagined that other peoples' seizures were comparatively worse than their own. Illness severity was not uppermost in participants' minds but emerged as a concept that was both relative and linked to social burden. Perceptions of overall disease severity expanded upon determinants of seizure severity to offer a more complete explanation of what patients themselves did about longstanding, uncontrolled epilepsy. CONCLUSIONS: Perceptions of illness severity played a vital role in treatment decision-making with the potential to impact the illness trajectory. How to measure components of illness severity represents a new challenge for outcomes research in DRE.

Adaptation and validation of the health belief model scale for colorectal cancer screening.

Korea has the second highest incidence of colorectal cancer in the world. Instruments that are culturally and contextually sensitive, as well as valid and reliable, for determining health beliefs regarding colorectal cancer screening are essential for obtaining accurate information. The purpose of this study was to adapt and validate the health belief model scale for Koreans regarding colorectal cancer and fecal occult blood test utilization. Individual and cognitive interviews (also known as cognitive debriefing) with 33 Koreans, expert reviews with seven nursing practitioners and professors, and a pilot test with 18 Koreans were conducted to make the existing health belief model scale culturally and contextually sensitive. Subsequently, a cross-sectional survey with 728 Koreans aged >50 years was conducted. Exploratory and confirmatory factor analyses of the construct validity and internal consistency reliability supported the adapted health belief model scale. The adapted and validated health belief model scale in this study could contribute to the assessment of health beliefs regarding the fecal occult blood test among Koreans with a greater degree of accuracy with respect to Korean culture and context.

Trichodysplasia spinulosa in a child: Identification of trichodysplasia spinulosa-associated polyomavirus in skin, serum, and urine.

A 6-year-old girl with a history of chronic immunosuppression following small bowel and colon transplantation for tufting enteropathy presented with a diffuse, facial-predominant eruption composed of pink-to-skin-colored papules with central white dystrophic spicules. Histology from a punch biopsy and polymerase chain reaction (PCR) from plucked spicules confirmed a diagnosis of trichodysplasia spinulosa (TS). Additional molecular studies identified several strains of the trichodysplasia spinulosa-associated polyomavirus infecting multiple tissues of the patient, confirming the systemic nature of trichodysplasia spinulosa infections.

Cancer screening in Koreans: a focus group approach.

BACKGROUND: Cancer is the greatest disease burden in Korea. Cancer screening can reduce the burden of cancer but cancer screening rates among Koreans remain low. The purposes of this study were to a) understand Koreans' beliefs and knowledge about cancer screening, and b) explore preferred strategies for increasing cancer screening utilization. METHODS: We conducted a descriptive, qualitative study using eight face-to-face focus groups with a total of 64 Koreans aged 40 and over. Participants answered semi-structured, open-ended questions assessing their experiences with, and beliefs, knowledge, and opinions about, cancer screening. All interview data were recorded and analyzed in the context of the health belief model (HBM). RESULTS: The most important themes that emerged from the focus group data were (a) perceived susceptibility (most of the participants believed they were not susceptible to cancer; those who perceived themselves susceptible to cancer were reluctant to express it); (b) perceived benefits (early detection and feelings of relief after cancer screening were benefits; participants had screening because they wanted to take advantage of the Korean government's Medical Payment Support program for cancer patients who have participated in the National Cancer Screening program); (c) perceived barriers (no symptoms; self-care when having symptoms; widespread distrust of tests, doctors, and hospitals; unkind health care providers; the financial burdens of advanced cancer screening tests; and the discomfort during cancer screening); and (d) knowledge of the causes of cancer (incorrect knowledge including beliefs that stress, personality, and body overuse cause cancer). Almost all of the participants were very knowledgeable about the seriousness of cancer and were confident that they were able to have cancer screening. Participants preferred strategies of cancer screening using group interventions with family or friends; various information delivery methods; information emphasizing the importance of cancer prevention; convenient, free, or inexpensive services; and kind health care providers. CONCLUSIONS: This HBM-based research suggests that beliefs in low susceptibility to cancer, many barriers to cancer screening, and incorrect knowledge should be the foci for increasing cancer screening rates in Koreans. Interventions could change individual cultural beliefs and increase knowledge as well as the quality of health care for Koreans.

Glutathione Depletion, Pentose Phosphate Pathway Activation, and Hemolysis in Erythrocytes Protecting Cancer Cells from Vitamin C-induced Oxidative Stress.

The discovery that oxidized vitamin C, dehydroascorbate (DHA), can induce oxidative stress and cell death in cancer cells has rekindled interest in the use of high dose vitamin C (VC) as a cancer therapy. However, high dose VC has shown limited efficacy in clinical trials, possibly due to the decreased bioavailability of oral VC. Because human erythrocytes express high levels of Glut1, take up DHA, and reduce it to VC, we tested how erythrocytes might impact high dose VC therapies. Cancer cells are protected from VC-mediated cell death when co-cultured with physiologically relevant numbers of erythrocytes. Pharmacological doses of VC induce oxidative stress, GSH depletion, and increased glucose flux through the oxidative pentose phosphate pathway (PPP) in erythrocytes. Incubation of erythrocytes with VC induced hemolysis, which was exacerbated in erythrocytes from glucose-6-phosphate dehydrogenase (G6PD) patients and rescued by antioxidants. Thus, erythrocytes protect cancer cells from VC-induced oxidative stress and undergo hemolysis in vitro, despite activation of the PPP. These results have implications on the use of high dose VC in ongoing clinical trials and highlight the importance of the PPP in the response to oxidative stress.

Human polyomavirus 6 and 7 are associated with pruritic and dyskeratotic dermatoses.

BACKGROUND: Human polyomavirus (HPyV)6 and HPyV7 are shed chronically from human skin. HPyV7, but not HPyV6, has been linked to a pruritic skin eruption of immunosuppression. OBJECTIVE: We determined whether biopsy specimens showing a characteristic pattern of dyskeratosis and parakeratosis might be associated with polyomavirus infection. METHODS: We screened biopsy specimens showing "peacock plumage" histology by polymerase chain reaction for HPyVs. Cases positive for HPyV6 or HPyV7 were then analyzed by immunohistochemistry, electron microscopy, immunofluorescence, quantitative polymerase chain reaction, and complete sequencing, including unbiased, next-generation sequencing. RESULTS: We identified 3 additional cases of HPyV6 or HPyV7 skin infections. Expression of T antigen and viral capsid was abundant in lesional skin. Dual immunofluorescence staining experiments confirmed that HPyV7 primarily infects keratinocytes. High viral loads in lesional skin compared with normal-appearing skin and the identification of intact virions by both electron microscopy and next-generation sequencing support a role for active viral infections in these skin diseases. LIMITATION: This was a small case series of archived materials. CONCLUSION: We have found that HPyV6 and HPyV7 are associated with rare, pruritic skin eruptions with a distinctive histologic pattern and describe this entity as "HPyV6- and HPyV7-associated pruritic and dyskeratotic dermatoses."

Web-Based Study for Improving Mammography Among Korean American Women.

Web-based culture-specific interventions for Korean American (KA) women to improve mammography utilization are not available. An established intervention developed to improve mammography utilizations for Korean American (KA) women was tested via the Web for its efficacy, feasibility, and acceptability. A randomized controlled trial, with a pretest-posttest control group design, was conducted with 136 KA women and their spouses. Intention to have a mammogram within the next 12 months increased significantly in the intervention group compare to controls. Among women whose mammograms had not been updated, 22 % of women in the intervention and 13 % of women in the control group obtained a mammogram at 2-month post-baseline, even though the difference was not statistically significant. The Web-based study educating couples was feasible and could improve KA women's breast cancer screening intention and behaviors. Combining off-line contact for recruitment/data collection with online intervention material could decrease the attrition rate in the future study because the attrition rate in this study was higher than the original study.

Cancer Information Seeking Behaviors of Korean American Women: A Mixed-Methods Study Using Surveys and Focus Group Interviews.

Despite the high risk of cancer to the population, Korean Americans are known to have lower knowledge about cancer related information and a lower level of adherence to cancer prevention guidelines. This indicates the necessity of cancer interventions targeting the Korean American population. To reach this population effectively, it is imperative to understand Korean Americans' cancer information seeking behaviors. This study (a) identified cancer information sources that are trusted and used by Korean American women and (b) examined how general media exposure and trust in cancer information sources are related to the use of these sources. It also (c) explored perceived usefulness and limitations of cancer information sources. A mixed methods study using seven focus group interviews with 34 Korean American women and surveys with 152 Korean American women was conducted in the Washington, DC, metropolitan area from 2011 to 2012. The results indicate that Korean American women viewed health care professionals as the most trusted cancer information source but used the Internet and Korean ethnic media more often for cancer information seeking because of language, cultural, and economic barriers. Korean American women were most likely to obtain cancer information from media they used frequently for general purposes. Correlations between usage frequency and trust in doctor/health providers and the Internet as cancer information sources were negligible. When seeking cancer information, important factors for Korean American women were accessibility, affordability, and language proficiency, cultural sensitivity, meeting immediate needs, understandability, convenience, and reliability of cancer information sources. Findings from this study support developing interventions using Korean language media, including print, television and the Internet for health promotion and cancer prevention targeting Korean American women.

Red blood cells as glucose carriers to the human brain: Modulation of cerebral activity by erythrocyte exchange transfusion in Glut1 deficiency (G1D).

Red blood cells circulating through the brain are briefly but closely apposed to the capillary endothelium. We hypothesized that this contact provides a nearly direct pathway for metabolic substrate transfer to neural cells that complements the better characterized plasma to endothelium transfer. While brain function is considered independent of normal fluctuations in blood glucose concentration, this is not borne out by persons with glucose transporter I (GLUT1) deficiency (G1D). In them, encephalopathy is often ameliorated by meal or carbohydrate administration, and this enabled us to test our hypothesis: Since red blood cells contain glucose, and since the red cells of G1D individuals are also deficient in GLUT1, replacing them with normal donor cells via exchange transfusion could augment erythrocyte to neural cell glucose transport via mass action in the setting of unaltered erythrocyte count or plasma glucose abundance. This motivated us to perform red blood cell exchange in 3 G1D persons. There were rapid, favorable and unprecedented changes in cognitive, electroencephalographic and quality-of-life measures. The hypothesized transfer mechanism was further substantiated by in vitro measurement of direct erythrocyte to endothelial cell glucose flux. The results also indicate that the adult intellect is capable of significant enhancement without deliberate practice. ClinicalTrials.gov registration: NCT04137692 https://clinicaltrials.gov/ct2/show/NCT04137692.

GLUT3 promotes macrophage signaling and function via RAS-mediated endocytosis in atopic dermatitis and wound healing.

The facilitative GLUT1 and GLUT3 hexose transporters are expressed abundantly in macrophages, but whether they have distinct functions remains unclear. We confirmed that GLUT1 expression increased after M1 polarization stimuli and found that GLUT3 expression increased after M2 stimulation in macrophages. Conditional deletion of Glut3 (LysM-Cre Glut3fl/fl) impaired M2 polarization of bone marrow-derived macrophages. Alternatively activated macrophages from the skin of patients with atopic dermatitis showed increased GLUT3 expression, and a calcipotriol-induced model of atopic dermatitis was rescued in LysM-Cre Glut3fl/fl mice. M2-like macrophages expressed GLUT3 in human wound tissues as assessed by transcriptomics and costaining, and GLUT3 expression was significantly decreased in nonhealing, compared with healing, diabetic foot ulcers. In an excisional wound healing model, LysM-Cre Glut3fl/fl mice showed significantly impaired M2 macrophage polarization and delayed wound healing. GLUT3 promoted IL-4/STAT6 signaling, independently of its glucose transport activity. Unlike plasma membrane-localized GLUT1, GLUT3 was localized primarily to endosomes and was required for the efficient endocytosis of IL-4Rα subunits. GLUT3 interacted directly with GTP-bound RAS in vitro and in vivo through its intracytoplasmic loop domain, and this interaction was required for efficient STAT6 activation and M2 polarization. PAK activation and macropinocytosis were also impaired without GLUT3, suggesting broader roles for GLUT3 in the regulation of endocytosis. Thus, GLUT3 is required for efficient alternative macrophage polarization and function, through a glucose transport-independent, RAS-mediated role in the regulation of endocytosis and IL-4/STAT6 activation.

Breast cancer screening beliefs among older Korean American women.

Korean American women's breast cancer screening rates are low, and the rates among older Korean American women are even lower.This article describes health beliefs related to older Korean American women's screening behaviors, comparing them to beliefs of younger Korean American women. The 73 women age 65 and older had significantly dif-ferent health beliefs than the 114 women between ages 40 and 64. Further, older women's perceptions of the seriousness of the disease and benefits of and barriers to taking action to prevent the disease predated their screening behaviors. Interventions to change the health beliefs of older Korean American women are suggested.

Evaluating Differential Item Functioning of the CES-D Scale According to Caregiver Status and Cultural Context in Korean Women.

BACKGROUND: Proper evaluation of the generalizability of an instrument is critical for its use across different social contexts such as caregiver status. OBJECTIVE: To examine the differential item functioning of the Center for Epidemiologic Studies Depression (CES-D) Scale, patterns of response to each item of the CES-D Scale attributable to caregiver status was assessed. STUDY DESIGN: Using a cross-study comparison method, a sample of 58 matched pairs of Korean American caregivers and noncaregivers was used for matched moderated regression analysis on the CES-D Scale. RESULTS: The authors identified three items that vary according to caregiver status in the present study: Item 2 (My appetite was poor), Item 4 (I felt that I was as good as other people), and Item 14 (I felt lonely). CONCLUSIONS: Beyond assessing the level of depression through total CES-D scores, it is important to examine variations in the items of the CES-D Scale across different social contexts. J Am Psychiatr Nurses Assoc, 2009; 15(4), 240-248. DOI: 10.1177/1078390309343713.

Transforming activity of an oncoprotein-encoding circular RNA from human papillomavirus.

Single-stranded circular RNAs (circRNAs), generated through 'backsplicing', occur more extensively than initially anticipated. The possible functions of the vast majority of circRNAs remain unknown. Virus-derived circRNAs have recently been described in gamma-herpesviruses. We report that oncogenic human papillomaviruses (HPVs) generate circRNAs, some of which encompass the E7 oncogene (circE7). HPV16 circE7 is detectable by both inverse RT-PCR and northern blotting of HPV16-transformed cells. CircE7 is N6-methyladenosine (m6A) modified, preferentially localized to the cytoplasm, associated with polysomes, and translated to produce E7 oncoprotein. Specific disruption of circE7 in CaSki cervical carcinoma cells reduces E7 protein levels and inhibits cancer cell growth both in vitro and in tumor xenografts. CircE7 is present in TCGA RNA-Seq data from HPV-positive cancers and in cell lines with only episomal HPVs. These results provide evidence that virus-derived, protein-encoding circular RNAs are biologically functional and linked to the transforming properties of some HPV.

Assessment of circularized E7 RNA, GLUT1, and PD-L1 in anal squamous cell carcinoma.

Anal squamous cell carcinoma (ASCC) is a rare, potentially fatal malignancy primarily caused by high-risk human papillomaviruses (HPV). The prognostic implication of programmed death-ligand 1 (PD-L1) expression remains controversial, and glucose transporter 1 (GLUT1) expression has never been examined in ASCC. Covalently closed circular RNAs have recently been shown to be widespread in cancers and are proposed to be biomarkers. We discovered HPV16 expresses a circular E7 RNA (circE7) which has not been assessed as a potential biomarker. A retrospective, translational case series at UT Southwestern was conducted to analyze PD-L1, GLUT1, HPV-ISH, and HPV circE7 in relation to the clinical features and overall survival of patients with ASCC. Twenty-two (22) subjects were included in the study. Improved overall survival was predicted by basaloid histology ( p = 0.013), PD-L1 expression ( p = 0.08), and HPV-ISH positivity ( p & 0.001), but not GLUT1 expression. High levels of circE7 by quantitative RT-PCR predicted improved overall survival in ASCC ( p = 0.023) and analysis of The Cancer Genome Atlas sequencing from HPV-positive head and neck cancer and cervical cancer suggested high circE7 marked improved survival in 875 subjects ( p = 0.074). While our study suggests that circE7 levels correlate with improved survival in ASCC, larger, prospective studies are necessary to confirm the potential role of circE7 as a biomarker.