Let-7 derived from endometrial extracellular vesicles is an important inducer of embryonic diapause in mice.

Embryonic diapause is a maternally controlled phenomenon. The molecule controlling the onset of the phenomenon is unknown. We demonstrated that overexpression of microRNA let-7a or incubation with let-7g-enriched extracellular vesicles from endometrial epithelial cells prolonged the in vitro survival of mouse blastocysts, which developed into live pups after having been transferred to foster mothers. Similar to in vivo dormant blastocysts, let-7-induced dormant blastocysts exhibited low level of proliferation, apoptosis, and nutrient metabolism. Let-7 suppressed c-myc/mTORC1 and mTORC2 signaling to induce embryonic diapause. It also inhibited ODC1 expression reducing biosynthesis of polyamines, which are known to reactivate dormant embryos. Furthermore, the overexpression of let-7 blocked trophoblast differentiation and implantation potential of human embryo surrogates, and prolonged survival of human blastocysts in vitro, supporting the idea that embryonic diapause was an evolutionary conserved phenomenon. In conclusion, let-7 is the main factor inducing embryonic diapause.

Trans-stromal epithelial cyst after multiple lamellar keratoplasties - Case report and review of literature.

DESIGN: Single Observational Case Report. SETTING: A 67-year-old male of Persian descent had a complex systemic and ocular history prior to a right penetrating keratoplasty (PK) reported here. The clinical diagnoses leading to the PK included Cogan's syndrome, chronic uveitis, secondary glaucoma, and corneal stromal scarring, presumed secondary to a corneal ulcer diagnosed on the second visit to our clinic. The specimen described here had been in place for 11 months and 17 days after the third failed Descemet stripping endothelial keratoplasty (DSEK). Visual acuities had ranged from 20/100 to 20/400 in both eyes. Visual acuity in the right eye just before surgery was 20/400. Intraocular pressures were 22 mmHg in both eyes with functioning Ahmed glaucoma shunts. The stromal cyst was not suspected preoperatively and no clinical imaging was performed. STUDY METHODS: Histopathology including serial sections and immunohistochemistry. RESULTS: Histologic study demonstrated a trans-stromal corneal epithelial cyst without goblet cells that extended through a 500 µm gap in the donor tissue surface and edge. CONCLUSIONS: This large stromal cyst was an unusual complication of serial posterior lamellar keratoplasties and we postulate that multiple prior posterior lamellar grafts may have been a risk factor for this complication. Anterior segment imaging with either anterior segment optical coherence tomography or high-resolution ultrasound would likely have detected this stromal cyst.

The gelling effect of platelet-rich fibrin matrix when exposed to human tenocytes from the rotator cuff in small-diameter culture wells and the design of a co-culture device to overcome this phenomenon.

OBJECTIVES: Platelet-rich fibrin matrix (PRFM) has been proved to enhance tenocyte proliferation but has mixed results when used during rotator cuff repair. The optimal PRFM preparation protocol should be determined before clinical application. To screen the best PRFM to each individual's tenocytes effectively, small-diameter culture wells should be used to increase variables. The gelling effect of PRFM will occur when small-diameter culture wells are used. A co-culture device should be designed to avoid this effect. METHODS: Tenocytes harvested during rotator cuff repair and blood from a healthy volunteer were used. Tenocytes were seeded in 96-, 24-, 12-, and six-well plates and co-culture devices. Appropriate volumes of PRFM, according to the surface area of each culture well, were treated with tenocytes for seven days. The co-culture device was designed to avoid the gelling effect that occurred in the small-diameter culture well. Cell proliferation was analyzed by water soluble tetrazolium-1 (WST-1) bioassay. RESULTS: The relative quantification (condition/control) of WST-1 assay on day seven revealed a significant decrease in tenocyte proliferation in small-diameter culture wells (96 and 24 wells) due to the gelling effect. PRFM in large-diameter culture wells (12 and six wells) and co-culture systems induced a significant increase in tenocyte proliferation compared with the control group. The gelling effect of PRFM was avoided by the co-culture device. CONCLUSION: When PRFM and tenocytes are cultured in small-diameter culture wells, the gelling effect will occur and make screening of personalized best-fit PRFM difficult. This effect can be avoided with the co-culture device.Cite this article: C-H. Chiu, P. Chen, W-L. Yeh, A. C-Y. Chen, Y-S. Chan, K-Y. Hsu, K-F. Lei. The gelling effect of platelet-rich fibrin matrix when exposed to human tenocytes from the rotator cuff in small-diameter culture wells and the design of a co-culture device to overcome this phenomenon. Bone Joint Res 2019;8:216-223. DOI: 10.1302/2046-3758.85.BJR-2018-0258.R1.

Pre-stroke physical activity is associated with fewer post-stroke complications, lower mortality and a better long-term outcome.

BACKGROUND AND PURPOSE: Physical activity is associated with a reduced incidence of first-time stroke. However, few studies have examined the effect of pre-stroke physical activity on post-stroke complications and clinical outcomes. METHODS: A total of 39 835 cases of stroke registered in the nationwide stroke registry system of Taiwan between 2006 and 2009 were analyzed according to five levels of severity as determined by National Institutes of Health Stroke Scale score upon hospital admission. Pre-stroke physical activity was defined in the Taiwan Stroke Registry as dedicated leisure-time physical activity for at least 30 min/day for 3 days/week for more than 6 months. A Cox model was used to compare complications and outcomes between active and inactive groups. RESULTS: The active and inactive groups were similar in age distribution and stroke type distribution, but the active group had better National Institutes of Health Stroke Scale scores upon admission. The active group also had significantly fewer post-stroke complications. Active patients had lower hospital mortality and better functional outcomes upon discharge as per the modified Rankin Scale. Improved functional status in the active group was significant at 1, 3 and 6 months post-stroke. CONCLUSION: Dedicated leisure-time physical activity for at least 30 min/day, at least three times per week for more than 6 months was associated with decreased stroke severity, fewer post-stroke complications, lower mortality and better outcomes.

Hyperglycemia impedes definitive endoderm differentiation of human embryonic stem cells by modulating histone methylation patterns.

Exposure to maternal diabetes during fetal growth is a risk factor for the development of type II diabetes (T2D) in later life. Discovery of the mechanisms involved in this association should provide valuable background for therapeutic treatments. Early embryogenesis involves epigenetic changes including histone modifications. The bivalent histone methylation marks H3K4me3 and H3K27me3 are important for regulating key developmental genes during early fetal pancreas specification. We hypothesized that maternal hyperglycemia disrupted early pancreas development through changes in histone bivalency. A human embryonic stem cell line (VAL3) was used as the cell model for studying the effects of hyperglycemia upon differentiation into definitive endoderm (DE), an early stage of the pancreatic lineage. Hyperglycemic conditions significantly down-regulated the expression levels of DE markers SOX17, FOXA2, CXCR4 and EOMES during differentiation. This was associated with retention of the repressive histone methylation mark H3K27me3 on their promoters under hyperglycemic conditions. The disruption of histone methylation patterns was observed as early as the mesendoderm stage, with Wnt/ß-catenin signaling being suppressed during hyperglycemia. Treatment with Wnt/ß-catenin signaling activator CHIR-99021 restored the expression levels and chromatin methylation status of DE markers, even in a hyperglycemic environment. The disruption of DE development was also found in mouse embryos at day 7.5 post coitum from diabetic mothers. Furthermore, disruption of DE differentiation in VAL3 cells led to subsequent impairment in pancreatic progenitor formation. Thus, early exposure to hyperglycemic conditions hinders DE development with a possible relationship to the later impairment of pancreas specification.

Estimating active layer thickness and volumetric water content from ground penetrating radar measurements in Barrow, Alaska.

Ground penetrating radar (GPR) has emerged as an effective tool for estimating active layer thickness (ALT) and volumetric water content (VWC) within the active layer. In August 2013, we conducted a series of GPR and probing surveys using a 500 MHz antenna and metallic probe around Barrow, Alaska. We collected about 15 km of GPR data and 1.5 km of probing data. Here, we describe the GPR data processing workflow from raw GPR data to the estimated ALT and VWC. We include the corresponding uncertainties for each measured and estimated parameter. The estimated average GPR-derived ALT was 41 cm, with a standard deviation of 9 cm. The average probed ALT was 40 cm, with a standard deviation of 12 cm. The average GPR-derived VWC was 0.65, with a standard deviation of 0.14.

Population-based comparison of prognostic factors in invasive micropapillary and invasive ductal carcinoma of the breast.

BACKGROUND: Invasive micropapillary carcinoma (IMPC) is a variant of breast carcinoma with a higher propensity for lymph node metastases compared with invasive ductal carcinoma (IDC). METHODS: Retrospective analysis of 636 IMPC and 297 735 IDC cases in the Surveillance, Epidemiology and End RESULTS database comparing disease-specific survival (DSS) and overall survival (OS) between IMPC and IDC. RESULTS: A higher percentage of IMPC cases (52.0%) had nodal metastases compared with IDC cases (34.6%). The 5-year DSS and OS for IMPC was 91.8% and 82.9%, respectively compared with 88.6% and 80.5% for IDC, respectively. For both IMPC and IDC, oestrogen-receptor positivity was associated with better survival, while having four or more positive lymph nodes or larger tumour size correlated with worse survival. Radiotherapy provided a survival benefit for both histological types. CONCLUSIONS: Despite IMPC's higher propensity for lymph node metastasis, IMPC has DSS and OS that compare favourably with IDC.

Chinese American adolescents: perceived parenting styles and adolescents' psychosocial health.

BACKGROUND: Asian Americans are one of the fastest-growing minority groups in the USA, and Chinese constitute the largest group. Evidence suggests that Asian American adolescents experience higher levels of depressive symptoms than their same-gender white counterparts. Quantitative findings suggest associations between parenting factors and Chinese American adolescents' mental health. A qualitative understanding regarding Chinese American adolescents' perceived parenting styles and its relationship with adolescents' psychosocial health is warranted. AIM: To gain an in-depth understanding of Chinese American adolescents' perceived parenting styles and how parenting styles might influence adolescents' psychosocial health. METHODS: In this qualitative study, we recruited 15 Chinese American adolescents aged 12-17 years in a southwest metropolitan area. We conducted two focus group interviews. Participants also filled out a brief questionnaire that included their socio-demographic information, immigration history and level of acculturation. RESULTS: Participants reported perceiving that parents had high expectations about academic performance and moral values. They also perceived stricter family rules regarding choices of friends compared with their non-Asian peers. Parents tended to be more protective of girls than of boys. Both Chinese American boys and girls reported poor or ineffective communication with their parents, which contributed to increased conflict between parents and adolescents and emotional distress of the adolescents. CONCLUSIONS: The findings provide evidence for nurses to develop linguistically and culturally tailored resources (e.g. parent support groups, programs aimed to improving parent-child communication) or connect these families with existing resources to enhance parenting skills and consequently reduce emotional distress of their adolescent children.

Microstructural remodeling of articular cartilage following defect repair by osteochondral autograft transfer.

OBJECTIVE: To assess collagen network alterations occurring with flow and other abnormalities of articular cartilage at medial femoral condyle (MFC) sites repaired with osteochondral autograft (OATS) after 6 and 12 months, using quantitative polarized light microscopy (qPLM) and other histopathological methods. DESIGN: The collagen network structure of articular cartilage of OATS-repaired defects and non-operated contralateral control sites were compared by qPLM analysis of parallelism index (PI), orientation angle (α) relative to the local tissue axes, and retardance (Γ) as a function of depth. qPLM parameter maps were also compared to ICRS and Modified O'Driscoll grades, and cell and matrix sub-scores, for sections stained with H&E and Safranin-O, and for Collagen-I and II. RESULTS: Relative to non-operated normal cartilage, OATS-repaired regions exhibited structural deterioration, with low PI and more horizontal α, and unique structural alteration in adjacent host cartilage: more aligned superficial zone, and reoriented deep zone lateral to the graft, and matrix disorganization in cartilage overhanging the graft. Shifts in α and PI from normal site-specific values were correlated with histochemical abnormalities and co-localized with changes in cell organization/orientation, cloning, or loss, indicative of cartilage flow, remodeling, and deterioration, respectively. CONCLUSIONS: qPLM reveals a number of unique localized alterations of the collagen network in both adjacent host and implanted cartilage in OATS-repaired defects, associated with abnormal chondrocyte organization. These alterations are consistent with mechanobiological processes and the direction and magnitude of cartilage strain.

Int6 regulates both proteasomal degradation and translation initiation and is critical for proper formation of acini by human mammary epithelium.

INT6/EIF3E has been implicated in breast tumorigenesis, but its functional activities remain poorly defined. We found that, repressing INT6 expression induced transformed properties in normal human mammary epithelium (MCF10A); in contrast, Int6 silencing induced apoptosis in HeLa cells. As in fission yeast, Int6 in human cells was required for assembly of active proteasomes. A reverse-phase protein array screen identified SRC3/AIB1 as one oncoprotein the level and stability of which increased when Int6 was silenced in MCF10A cells. Our data further show that Int6 binds SRC3 and its ubiquitin ligase Fbw7, thus perhaps mediating the interaction between SRC3-Fbw7 and proteasomes. Consistent with this, Int6 silencing did not increase SRC3 levels in HeLa cells, which have low Fbw7 levels. It is surprising that, however, polyubiquitylated proteins do not accumulate or may even decrease in Int6-silenced cells that contain defective proteasomes. Considering that decreased ubiquitin might explain this observation and that Int6 might control ubiquitin levels in its role as a subunit of eIF3 (eukaryote translation initiation factor 3), we found that silencing Int6 reduced monoubiquitin protein levels, which correlated with a shift of ubiquitin mRNAs from larger polysomes to non-translating ribosomes. In contrast, levels of many housekeeping proteins did not change. This apparent reduction in the translation of ubiquitin genes correlated with a modest reduction in protein synthesis rate and formation of large polysomes. To further determine whether Int6 can selectively control translation, we analyzed translation of different 5'-untranslated region reporters and found that indeed, loss of Int6 had differential effects on these reporters. Together the data suggest that Int6 depletion blocks ubiquitin-dependent proteolysis by decreasing both ubiquitin levels and the assembly of functional proteasome machinery, leading to accumulation of oncoproteins, such as SRC3 that can transform mammary epithelium. Our data also raise the possibility that Int6 can further fine-tune protein levels by selectively controlling translation of specific mRNAs.

Shared and persistent asymptomatic cutaneous human papillomavirus infections in healthy skin.

Cutaneous human papillomavirus (HPV) types are commonly found in normal skin, and some of them have been suspected to play a role in the development of non-melanoma skin cancer. This present study is divided into three sections, the aims of this study were to examine if certain HPV-types persist over time and if HPV-types are shared within families. From the first part of the study, swab samples from foreheads were collected for three longitudinal studies from one family with a newborn baby. Five specific HPV-types were isolated from the family with a newborn, with HPV-5 and FA67 being found at various time points and prevalence rates in all four members of the family. Part 2 consisted of a followed up study from two families with a 6 years interval. Six of the family members were found to have at least one of the HPV-types identified in the family 6 years earlier. Many of the HPV-types identified were shared within the families studied. Part 3 of this study involved weekly samples from four healthy females for 4 months. Among the four healthy individuals, 11%, 65%, and 56% of the weekly samples were HPV-DNA positive with one individual HPV-negative. All specimens were tested for HPV-DNA by PCR using the broad range HPV-type primer pair FAP59/64. The positive samples were HPV-type determined by cloning and sequencing. Specific cutaneous HPV-types persist over long periods of time in healthy skin in most individuals investigated and certain HPVs are shared between family members.

Autofluorescence reveals menstrual phase in the endometrium.

OBJECTIVE: Tissue contains fluorophores that autofluoresce without additional dye or photosensitizer with the appropriate light excitation. This technique has been widely applied for discrimination between normal and precancerous tissue. The aim of this study was to explore the capability and reliability of autofluorescence phase determination in samples of human endometrium. METHODS: A total of 70 measurement sites from 31 endometrial tissue samples from hysterectomy were enrolled. Xenon light (330 nm) was directed at the endometrial tissue and the resultant autofluorescence intensity recorded. Spectra were then grouped according to the proliferative and secretory phase, with multivariant analysis, partial least square (PLS) and analysis of variance (ANOVA) used for evaluation of the statistical significance of phase determination. RESULTS: Both proliferative and secretory autofluorescence spectra showed a similar characteristic triple-peak curve shape pattern, however, each of the intensities at the three peaks between the two phases varied markedly (p < 0.01). PLS analysis confirmed that collagen, NADH and FAD autofluorescence were the principle determinants of endometrial spectrum; the sensitivity and specificity of phase determination by autofluorescent was 100% and 97%, respectively. CONCLUSION: Autofluorescence measurement provides real-time information on endometrial phase status and, based on our results, it appears reasonable to suggest that it may be promising as a clinical tool for prompt phase interpretation.

Hurricane Katrina: prior trauma, poverty and health among Vietnamese-American survivors.

BACKGROUND: The flooding of New Orleans after Hurricane Katrina revealed the disproportionate vulnerability of ethnic minority communities for emergency preparedness, disaster relief and health. Nurses need to analyse Katrina's health consequences for the most vulnerable segments of our society. AIM: To examine factors contributing to differential health outcomes among the New Orleans Vietnamese community in response to Katrina. METHODS: A sample of 113 adult Vietnamese Katrina survivors from New Orleans was recruited. A mixed-method approach, including survey and focus groups, was used to collect data. Survey questions were modified from standardized instruments to evaluate survivors' health status and factors contributing to health outcomes. Multivariate and content analysis were used to investigate effects of prior trauma, financial strain, social support and acculturation level in predicting survivors' health outcomes. RESULTS: Findings suggested financial strain was the strongest risk factor for Vietnamese survivors' post-traumatic stress disorder (PTSD) symptoms, and physical and mental health post-disaster; while social support was a strong protective factor for health. Survivors who perceived higher impact from previous traumatic experiences had poorer physical health, but not PTSD symptoms or poor mental health after controlling for financial strain and social support, suggesting complex relationships among these measures in predicting PTSD symptoms and health. Less-acculturated individuals also reported higher levels of PTSD symptoms and poorer physical health. CONCLUSIONS: Catastrophic events like Katrina can result in disproportionate risk of negative health outcomes among vulnerable populations. Nurses should take into account prior trauma, financial strain, social support network and acculturation level, to adequately address survivors' needs.

Effects of hyperbaric oxygen and platelet derived growth factor on medial collateral ligament fibroblasts.

PURPOSE: This study investigated hyperbaric oxygen (HBO2) and platelet-derived growth factor-BB (PDGF-BB) to determine their combined effects on fibroblasts from rabbit medial collateral ligament (MCL). METHOD: Cells were divided into four groups: (I) Control, (II) HBO2 treatment, (III) PDGF-BB treatment and (IV) HBO2 combined with PDGF-BB treatment. All hyperoxic cells were exposed to 100% O2 at 2.5 atmospheres absolute (ATA) in a hyperbaric chamber for 120 minutes per 48 hours. Measurement of cell growth was based on increase in cell number. Cell cycle modulations were analyzed by fluorescence-activated cell sorter (FACS). Quantity of Type I and Type III collagen was determined by western blotting and image analyzer. RESULTS: Treatment doses of HBO2 alone or PDGF-bb alone dependently increased cell growth. A combination of HBO2 treatment plus PDGF-bb treatment had an additive effect on cell growth in comparison with HBO2 treatment alone or PDGF-bb treatment alone. FACS analysis revealed that HBO2 alone, PDGF-bb alone and PDGF-bb plus HBO2 treatment increase the percentage of cells accumulated in S-phase. Western blotting analysis revealed that Type III collagen content was decreased significantly after HBO2 treatment alone or HBO2 plus PDGF-bb treatment but not in PDGF-bb treatment alone. In contrast, although Type I collagen content was increased after HBO2 treatment, the increase in Type I collagen (increase /original) was not statistically significant. CONCLUSION: HBO2 or HBO2 plus PDGF-bb treatment decreases the Type III collagen/Type I collagen content, which could result in mechanically stronger collagen fibrils. We propose HBO2 therapy as a potentially effective treatment for MCL healing.

Serial analysis of gene expression in the southern cattle tick following acaricide treatment of larvae from organophosphate resistant and susceptible strains.

Organophosphate resistant and susceptible tick larvae from laboratory strains of the southern cattle tick, Rhipicephalus (Boophilus) microplus were exposed to low doses of the organophosphate (OP) acaricide, coumaphos. Serial analysis of gene expression (SAGE) was used to analyse differential gene expression in response to OP treatment and to compare the responses of OP-treated and -untreated resistant and susceptible tick larvae. An R. microplus Gene Index was used as an EST database to identify genes which corresponded to SAGE tags whose abundance changed in response to acaricide exposure. Relative quantitative RT-PCR was used to confirm the differential expression results from the SAGE experiments. Of particular interest is a SAGE tag which corresponds to a cytochrome P450-like EST in the Gene Index which was more abundant in untreated OP resistant larvae compared to untreated OP susceptible larvae. This SAGE tag was also more abundant in OP resistant larvae treated with OP compared to OP susceptible larvae treated with OP.

Lasers and other thermal stimulators for activation of skin nociceptors in humans.

Pain can be induced by thermal, chemical, and mechanical stimulation in animals and man. Of the thermal stimulation modalities, heat is the most commonly used, as a variety of reliable stimulation techniques are available. Heat is a natural stimulus modality to evoke pain, and it has been used to study animal nociception and human pain perception for (a) examining the mechanisms of tissue injury and sensitisation and (b) quantifying the therapeutic effects of pharmacological, physical, and psychological interventions. This paper summarises the current understanding of the physiology and psychophysical response to painful heat stimulation in humans. By understanding the underlying mechanisms, new methods of heat stimulation may be developed for basic and clinical applications. Traditionally, contact heat, indirect thermal heat by focused light bulb, and laser pulses have been the methods used to induce heat pain in humans for experimental and clinical studies. The following lasers have been used in pain research: argon (488-515 nm), copper vapour (510-577 nm), semiconductor (e.g. 970 nm), neodymium-YAG (1064 nm), thulium-YAG (2000 nm), and CO(2) (10,600 nm).

The effects of one-week fluticasone propionate inhalation therapy for Tc-99m DTPA radioaerosol distribution in asthma of children: a preliminary report.

This study evaluated the effects of fluticasone propionate inhalation therapy for the distribution pattern of Tc-99m DTPA radioaerosols in 10 children with asthma. The homogeneous degree of depositing Tc-99m DTPA radioaerosol was evaluated using a modified standard score system over the bilateral lungs. The baseline scores were calculated from Tc-99m DTPA radioaerosol inhalation lung scintigraphy before inhalation therapy (100 microg fluticasone propionate two times daily for one week), and the scores were recalculated after inhalation therapy to evaluate the effects of one-week of fluticasone propionate inhalation therapy for Tc-99m DTPA radioaerosol distribution patterns. After one week of fluticasone propionate inhalation therapy, the scores were decreased in all of the 10 children, which may mean that the bronchial constriction degree due to asthma is decreased. In addition, there was a significantly statistical difference in the scores before and after one-week fluticasone propionate inhalation therapy (p < 0.05). In conclusion, one-week fluticasone propionate inhalation therapy could significantly improve the bronchial constriction due to asthma in children based on the evidence of Tc-99m DTPA radioaerosol inhalation lung scintigraphic findings.