Stromal PTEN inhibits the expansion of mammary epithelial stem cells through Jagged-1.

This corrects the article DOI: 10.1038/onc.2016.383.

Stromal PTEN inhibits the expansion of mammary epithelial stem cells through Jagged-1.

Fibroblasts within the mammary tumor microenvironment are active participants in carcinogenesis mediating both tumor initiation and progression. Our group has previously demonstrated that genetic loss of phosphatase and tensin homolog (PTEN) in mammary fibroblasts induces an oncogenic secretome that remodels the extracellular milieu accelerating ErbB2-driven mammary tumor progression. While these prior studies highlighted a tumor suppressive role for stromal PTEN, how the adjacent normal epithelium transforms in response to PTEN loss was not previously addressed. To identify these early events, we have evaluated both phenotypic and genetic changes within the pre-neoplastic mammary epithelium of mice with and without stromal PTEN expression. We report that fibroblast-specific PTEN deletion greatly restricts mammary ductal elongation and induces aberrant alveolar side-branching. These mice concomitantly exhibit an expansion of the mammary epithelial stem cell (MaSC) enriched basal/myoepithelial population and an increase in in vitro stem cell activity. Further analysis revealed that NOTCH signaling, specifically through NOTCH3, is diminished in these cells. Mechanistically, JAGGED-1, a transmembrane ligand for the NOTCH receptor, is downregulated in the PTEN-null fibroblasts leading to a loss in the paracrine activation of NOTCH signaling from the surrounding stroma. Reintroduction of JAGGED-1 expression within the PTEN-null fibroblasts was sufficient to abrogate the observed increase in colony forming activity implying a direct role for stromal JAGGED-1 in regulation of MaSC properties. Importantly, breast cancer patients whose tumors express both low stromal JAG1 and low stromal PTEN exhibit a shorter time to recurrence than those whose tumors express low levels of either alone suggesting similar stromal signaling in advanced disease. Combined, these results unveil a novel stromal PTEN-to-JAGGED-1 axis in maintaining the MaSC niche, and subsequently inhibiting breast cancer initiation and disease progression.

Reprogramming of the tumour microenvironment by stromal PTEN-regulated miR-320.

PTEN (Phosphatase and tensin homolog deleted on chromosome 10) expression in stromal fibroblasts suppresses epithelial mammary tumours, but the underlying molecular mechanisms remain unknown. Using proteomic and expression profiling, we show that Pten loss from mammary stromal fibroblasts activates an oncogenic secretome that orchestrates the transcriptional reprogramming of other cell types in the microenvironment. Downregulation of miR-320 and upregulation of one of its direct targets, ETS2 (v-ets erythroblastosis virus E26 oncogene homolog 2) are critical events in Pten-deleted stromal fibroblasts responsible for inducing this oncogenic secretome, which in turn promotes tumour angiogenesis and tumour-cell invasion. Expression of the Pten-miR-320-Ets2-regulated secretome distinguished human normal breast stroma from tumour stroma and robustly correlated with recurrence in breast cancer patients. This work reveals miR-320 as a critical component of the Pten tumour-suppressor axis that acts in stromal fibroblasts to reprogramme the tumour microenvironment and curtail tumour progression.

How well do skinfold equations predict percent body fat in elite soccer players?

The use of generic equations for estimating percent body fat from skinfold thicknesses can be criticised when applied to specific sports. The present aims were to compare existing methods of using skinfold data and to derive an equation for predicting body fat values in professional soccer players. Forty-five professional soccer players (24.2 +/- 5.0 years; 82.0 +/- 8.5 kg; 1.82 +/- 0.07 m) participated. Skinfold thicknesses were assessed at eight sites for the application of existing prediction equations. Skinfold data were also utilised to determine a novel soccer-specific equation. All players had a reference estimate of percent fat by dual-energy x-ray absorptiometry (DXA). The existing skinfold equations differed from the DXA-referenced values by varying degrees, the equation of Withers et al. (1987) demonstrating the lowest bias and highest relationship and agreement with DXA. Regression analysis resulted in an equation incorporating anterior thigh, abdominal, triceps and medial calf sites, accounting for 78.4% variance in DXA criterion values.

Stat3 activation in human endometrial and cervical cancers.

The activation of signal transducer and activator of transcription 3 (Stat3) has been implicated in the oncogenesis of cancer and is regarded as a novel target for cancer therapy. Stat3 is classified as a proto-oncogene, because an activated form of Stat3 can mediate oncogenic transformation in cultured cells and tumour formation in nude mice. The constitutive activation of Stat3 has been frequently detected in various types of human cancers. However, the constitutive activation of Stat3 in endometrial and cervical cancers has not been studied. We examined tyrosine phosphorylation of Stat3 (activated form of Stat3) in multiple endometrial and cervical cancer tissues using tissue microarray slides as well as cancer cell lines to explore the possible activation of Stat3. Our results indicated that elevated phosphorylation of Stat3 was detected in cervical and endometrial cancer cell lines. Our results also showed that elevated levels of phosphorylation of Stat3 protein were detected in the endometrial and cervical cancer specimens. This is the first study to demonstrate that Stat3 is activated in human endometrial and cervical cancer tissues. Immunohistochemical staining showed that activated Stat3 is associated with increased expression of downstream antiapoptotic genes, Bcl-xL, survivin, and Mcl-1 in these tissues. Expression of a dominant-negative Stat3 mutant using adenovirus-mediated gene transfer inhibited cell growth and induced apoptosis in HeLa and SiHa cervical cancer cell lines expressing elevated levels of Stat3 phosphorylation. Further, a JAK/Stat3 small molecular inhibitor, JSI-124, induced apoptosis more selectively in HeLa and SiHa cancer cell lines than Ishikawa cell line without elevated levels of Stat3 phosphorylation. These results indicate that Stat3 is activated in human endometrial and cervical cancers and the inhibition of constitutive Stat3 signaling may be an effective target for cancer intervention in these two cancers.

A comparative analysis of Mineral Trioxide Aggregate and Portland cement.

The purpose of this study was to compare the composition of Portland cement and Mineral Trioxide Aggregate (MTA). Samples of MTA and Portland cement were analysed for fifteen different elements by inductively coupled plasma emission spectrometry (ICP-ES). Comparative analysis revealed there was significant similarity except there was no detectable quantity of Bismuth in Portland cement. Quantitative results are given in both parts per million (p.p.m.) and wt%. It was concluded that there is no significant difference between the 14 different elements in both Portland cement and MTA.

Telemedicine for the Medicare population: pediatric, obstetric, and clinician-indirect home interventions.

BACKGROUND: This report is a supplement to an earlier evidence report, Telemedicine for the Medicare Population, which was intended to help policymakers weigh the evidence relevant to coverage of telemedicine services under Medicare. That report focused on telemedicine programs and clinical settings that had been used with or were likely to be applied to Medicare beneficiaries. While we prepared that report, it became apparent that there are also telemedicine studies among non-Medicare beneficiaries--e.g., children and pregnant women--that could inform policymakers and provide more comprehensive evidence of the state of the science regarding telemedicine applications. In addition, the first evidence report only partially included a class of telemedicine applications (called self-monitoring/testing telemedicine) in which the beneficiary used a home computer or modern-driven telephone system to either report information or access information and support from Internet resources and indirectly interact with a clinician. Self-monitoring/testing applications in the first report required direct interaction with a clinician. The goal of this report is to systematically review the evidence in the clinical areas of pediatric and obstetric telemedicine as well as home-based telemedicine where there is indirect involvement of the health care professional. (In this report, we will refer to the latter as clinician-indirect home telemedicine.) Specifically, the report summarizes scientific evidence on the diagnostic accuracy, access, clinical outcomes, satisfaction, and cost-effectiveness of services provided by telemedicine technologies for these patient groups. It also identifies gaps in the evidence and makes recommendations for evaluating telemedicine services for these populations in the future. The evidence is clustered according to three categories of telemedicine service defined in our original report: store-and-forward, self-monitoring/testing, and clinician-interactive services. The three clinical practice areas reviewed in this report are defined as follows. The term pediatric applies to any telemedicine study in which the sample consisted wholly or partially of persons aged 18 or younger, including studies with neonatal samples. The term obstetric applies to any telemedicine study in which the sample consisted entirely of women seeking pregnancy-related care. The term clinician-indirect home telemedicine applies to home-based telemedicine (called self-monitoring/testing in our original report) where a telemedicine application used in the home has only indirect involvement by the health care professional. Interactive home telemedicine was applied in this report to all patient populations. KEY QUESTIONS: The key questions that served as a guide for reviewing the literature in the evaluation of pediatric, obstetric, and clinician-indirect home telemedicine applications were derived by consensus among the evidence-review team based on the analytic framework established for the original evidence report. For the current report, the questions were applied to studies in all three practice areas as a whole group within each of the three categories of telemedicine services: store-and-forward; self-monitoring/testing; and clinician-interactive. The specific key questions were: 1. Does telemedicine result in comparable diagnosis and appropriateness of recommendations for management? 2. Does the availability of telemedicine provide comparable access to care? 3. Does telemedicine result in comparable health outcomes? 4. Does telemedicine result in comparable patient or clinician satisfaction with care? 5. Does telemedicine result in comparable costs of care and/or cost-effectiveness? METHODS: We searched for peer-reviewed literature using several bibliographic databases. In addition, we conducted hand searches of leading telemedicine journals and identified key papers from the reference lists of journal articles. For our original evidence report on telemedicine for the Medicare population, we designed a search to find any publications about telemedicine and used it to search the MEDLINE, CINAHL, and HealthSTAR databases for all years the databases were available. Through this process, we captured studies of pediatric, obstetric, and clinician-indirect home telemedicine; however, they were excluded from the original report since they were outside its scope. For this supplemental report, we reviewed our original search results and identified studies relevant to this report. We identified additional studies from the reference lists of included papers and from hand searching two peer-reviewed telemedicine publications, the Journal of Telemedicine and Telecare and Telemedicine Journal. We critically appraised the included studies for each study area and key question and discussed the strengths and limitations of the most important studies at weekly meetings of the research team. We also developed recommendations for research to address telemedicine knowledge gaps. To match these gaps with the capabilities of specific research methods, we classified the telemedicine services according to the type of evidence that would be needed to determine whether the specific goals of covering such services had been met. We emphasized the relationship between the type and level of evidence found in the systematic review of effectiveness and the types of studies that might be funded to address the gaps in knowledge in this growing field of research. FINDINGS: We identified a total of 28 eligible studies. In the new clinical areas, we found few studies in store-and-forward telemedicine. There is some evidence of comparable diagnosis and management decisions made using store-and-forward telemedicine from the areas of pediatric dental screening, pediatric ophthalmology, and neonatalogy. In self-monitoring/testing telemedicine for the areas of pediatrics, obstetrics, and clinician-indirect home telemedicine, there is evidence that access to care can be improved when patients and families have the opportunity to receive telehealth care at home rather than in-person care in a clinic or hospital. Access is particularly enhanced when the telehealth system enables timely communication between patients or families and care providers that allows self-management and necessary adjustments that may prevent hospitalization. There is some evidence that this form of telemedicine improves health outcomes, but the study sample sizes are usually small, and even when they are not, the treatment effects are small. There is also some evidence for the efficacy of clinician-interactive telemedicine, but the studies do not clearly define which technologies provide benefit or cost-efficiency. Some promising areas for diagnosis include emergency medicine, psychiatry, and cardiology. Most of the studies measuring access to care provide evidence that it is improved. Although none of these studies were randomized controlled trials, they provide some evidence of access improvement over prior conditions. Clinician-interactive telemedicine was the only area for which any cost studies were found. The three cost studies did not adequately demonstrate that telemedicine reduces costs of care (except comparing only selected costs). No study addressed cost-effectiveness. CONCLUSIONS: This supplemental report covering the areas of pediatrics, obstetrics, and indirect-clinician home telemedicine echoes the findings of our initial report for the Medicare domain, which is that while the use of telemedicine is small but growing, the evidence for its efficacy is incomplete. Many of the studies are small and/or methodologically limited, so it cannot be determined whether telemedicine is efficacious. Future studies should focus on the use of telemedicine in conditions where burden of illness and/or barriers to access for care are significant. Use of recent innovations in the design of randomized controlled trials for emerging technologies would lead to higher quality studies. Journals publishing telemedicine evaluation studies must set high standards for methodologic quality so that evidence reports need not rely on studies with marginal methodologies.

A comparative evaluation of the diagnostic efficacy of film and digital sensors for detection of simulated periapical lesions.

OBJECTIVE: The purpose of this study was to evaluate the diagnostic efficacy of Ektaspeed Plus film (EPF), a charge-coupled device (CCD), and photostimulable phosphor (PSP)-based digital images for detection of simulated periapical lesions. STUDY DESIGN: Lesions were simulated in the periapical areas of 24 human mandibular sections invested in acrylic using burs of sizes #1, 2, 4, and 6 and imaged using EPF, CCD, and PSP sensors. Percent correct response scores, sensitivity, and specificity values were computed for all variables. Repeated-measures analysis of variance and post hoc testing were performed to determine the effects of imaging modality, observer, and lesion sizes with respect to lesion detection. RESULTS: EPF displayed the highest sensitivity and specificity, followed by PSP and CCD images (P <.001). Percent correct score was the highest for 3 of 4 observers when EPF was used. Analysis of variance revealed significance (P <.001) with respect to all variables. Observers with experience in digital image-viewing performed better than those without such experience (P <.001). Intraobserver agreement was fair (kappa = 0.5). CONCLUSIONS: EPF outperformed CCD and PSP images when observers could manipulate image characteristics.

Magnetic resonance lipid signals in rat brain after experimental stroke correlate with neutral lipid accumulation.

Proton magnetic resonance spectroscopy (MRS) signals from lipids in brain have been observed to increase after ischemic brain injury. However, neither the chemical identity nor the cellular location of these lipids has been established. The aim of the present study was to identify the origin of MRS lipid signals in rat brain after temporary (90 min) middle cerebral artery occlusion (MCAO). Fatty acyl proton signals were detected by short-echo one and two dimensional (1)H MRS in superfused brain slices from the infarcted hemisphere 1-5 days after MCAO. The intensities of these signals were strongly correlated with the amount of triacylglyceride and cholesterol ester in lipid extracts from the samples (r(2)=0.96, P<0.05) and were not correlated with the amount of free fatty acids in the tissue. Histological staining of tissue revealed the presence of neutral lipid droplets in infarcted regions. Dual labeling by immunohistochemistry demonstrated that these droplets were localized to microglia/macrophage (OX-42-labeled cells). These results strongly suggest that (1)H MRS lipid signals from brain after stroke arise from microglia/macrophage phagocytosis of cellular membranes.

Effect of regimen complexity on patient satisfaction and compliance with warfarin therapy.

This study sought to determine patient satisfaction with anticoagulation care in a pharmacist-managed clinic and assess the effect of warfarin regimen complexity on patient satisfaction, compliance, and international normalized ratios (INRs). Retrospective chart reviews of 476 anticoagulation clinic patients were conducted. Patients were divided into groups by complexity of warfarin regimen (whole tablet, split tablet, alternating dose) for comparisons of compliance and INRs. An oral Likert scale satisfaction survey was administered to a convenience sample of 100 patients. Degree of satisfaction was compared based on warfarin regimen. Compliant patients were more likely to have therapeutic INRs (p < 0.01) and to be taking whole tablet dosage regimens (p < 0.01). Patients were generally satisfied with the pharmacist-managed clinic. There was a significantly (p < 0.025) higher score for patients on split tablet regimens, indicating a greater likelihood for agreement that alternating doses made taking their medication more difficult. A relationship exists between patient compliance and prescribed dosage regimen, as well as with therapeutic INR values.

Detection of artificially induced vertical radicular fractures using tuned aperture computed tomography.

This study compared the accuracy of three imaging modalities for the detection of artificially induced vertical root fractures (VRF) on teeth in cadaver mandibles. Fifty-four single-rooted, endodontically treated mandibular teeth being prepared to carry posts were evaluated using direct digital radiography (DDI) with a Schick sensor, unprocessed Tuned Aperture Computed Tomography (TACT-U) images and iteratively restored TACT (TACT-IR) images. Twenty-eight of these teeth had been subjected to fracture induction using an apically driven force. Nine basis images were used for each TACT image generation. Eight observers used a five-point confidence rating scale to record the confidence with which they considered a fracture to be present or not. Sensitivity and specificity values were computed and receiver operating characteristic (ROC) curves were generated. The areas under the curves (Az) used as an indication of the diagnostic accuracy of the imaging system were as follows: DDI: 0.37; TACT-U: 0.77 and TACT-IR: 0.81. DDI was significantly inferior to the TACT modalities. Differences in detection efficacy based on observers and observation sessions were noted on ANOVA and post-hoc Tukey's tests. This study indicates that TACT is the imaging modality of choice for VRF in endodontically treated teeth.

Clinical and economic effects of pharmacy services in geriatric ambulatory clinic.

Pharmacy services were introduced in an established multidisciplinary geriatric ambulatory clinic. The pharmacist collaborated with primary care providers to optimize patients' drug regimens. Over 8 months there were 250 patient visits to the clinic. Traditional medical care was provided at 144 (57.6%) of these visits and traditional medical care plus pharmacist evaluation was provided at 106 (42.4%). The pharmacist identified 220 potential and actual drug-related problems. Acceptance of pharmacist-recommended changes in drug therapy was 98.6%. A mean reduction of 3.4 agents/patient was achieved in the intervention group (p<0.0001). Clinical outcomes of changes in drug therapy were neutral or positive in 99.5% of cases. Pharmacy services resulted in net savings of $7,788 annually.

Clinical significance of rare and benign side effects: propofol and green urine.

A 55-year-old man requiring airway protection for esophagogastroduodenoscopy was sedated with propofol. On the third day of propofol infusion his urine was dark green. Although he was afebrile and his white blood cell count was within normal limits, the green urine was suspected to be of infectious etiology. Laboratory tests were ordered and broad-spectrum antibiotics were considered. Antibiotics were avoided when propofol was recognized as a rare and benign potential cause of the green urine. Earlier recognition of this side effect may have averted unnecessary laboratory monitoring. Prompt recognition of such side effects is important in limiting medical expenditures, inordinate drug exposure, and distress among patients and clinicians.

Differential regulation of the Kar3p kinesin-related protein by two associated proteins, Cik1p and Vik1p.

The mechanisms by which kinesin-related proteins interact with other proteins to carry out specific cellular processes is poorly understood. The kinesin-related protein, Kar3p, has been implicated in many microtubule functions in yeast. Some of these functions require interaction with the Cik1 protein (Page, B.D., L.L. Satterwhite, M.D. Rose, and M. Snyder. 1994. J. Cell Biol. 124:507-519). We have identified a Saccharomyces cerevisiae gene, named VIK1, encoding a protein with sequence and structural similarity to Cik1p. The Vik1 protein is detected in vegetatively growing cells but not in mating pheromone-treated cells. Vik1p physically associates with Kar3p in a complex separate from that of the Kar3p-Cik1p complex. Vik1p localizes to the spindle-pole body region in a Kar3p-dependent manner. Reciprocally, concentration of Kar3p at the spindle poles during vegetative growth requires the presence of Vik1p, but not Cik1p. Phenotypic analysis suggests that Cik1p and Vik1p are involved in different Kar3p functions. Disruption of VIK1 causes increased resistance to the microtubule depolymerizing drug benomyl and partially suppresses growth defects of cik1Delta mutants. The vik1Delta and kar3Delta mutations, but not cik1Delta, partially suppresses the temperature-sensitive growth defect of strains lacking the function of two other yeast kinesin-related proteins, Cin8p and Kip1p. Our results indicate that Kar3p forms functionally distinct complexes with Cik1p and Vik1p to participate in different microtubule-mediated events within the same cell.

Efficacy of cobalt chelates in the rabbit eye model for epithelial herpetic keratitis.

PURPOSE: A new class of antiviral agent, cobalt chelates (the CTC series), was evaluated for treating epithelial herpetic keratitis, consequent stromal disease being the major infectious cause of blindness in industrial nations. METHODS: Effects of CTC complexes were monitored in cell cultures and in a rabbit eye model, either infected with herpes simplex virus type 1 (HSV-1) or uninfected. Several antiviral concentrations of CTC complexes nontoxic to Vero cells were administered to rabbit eyes with HSV-1-induced keratitis. Corneal surface virus titers were measured, and corneal lesions of epithelial keratitis were monitored by slit-lamp microscopy and scored. Recovery rates and incidence were compared in eyes treated with CTC complexes, placebo, or clinically formulated trifluorothymidine (Viroptic), using nonparametric statistics. RESULTS: All CTC complexes inhibited HSV-1 replication in vitro, CTC-96 being best. CTC-96, CTC-23, and CTC-67 eliminated (<1 plaque-forming unit[pfu]) corneal surface HSV-1 (otherwise >10(5) pfu) in order of descending potency, but CTC-82 was ineffective. CTC-96 (either 5 microg/ml six times daily or 10 microg/ml five times daily) accelerated herpetic dendritic keratitis recovery better than or the same as trifluorothymidine (10 mg/ml nine times daily). CTC complexes were nontoxic to Vero cells continuously exposed to < or =25 microg/ml; 50 microg/ml of CTC 96 nine times daily did not irritate uninfected rabbit eyes. CONCLUSION: Topical CTC-96 applications were at least as effective as Viroptic in diminishing disease signs and corneal surface virus at concentrations less than one-thousandth that of Viroptic.

Tyrosine hydroxylase-containing neurons in the spinal cord of the chicken. I. Development and analysis of catecholamine synthesis capabilities.

1. The development of tyrosine hydroxylase-immunoreactive (TH-IR) neurons was examined in the spinal cord of the chick embryo and hatchling. 2. Two groups of TH-IR cells are described, both of which appear to reach their full complement in number relatively late in embryonic development. One group is comprised of numerous cells located ventral to the central canal which make direct contact with the lumen of the canal. The other group consists of large multipolar neurons that reside in the dorsal horn, more commonly along the outer margin of the gray matter within lamina I and II, and less frequently deeper in the dorsal horn within medial portions of laminae V, VI or VII. 3. TH-IR cells ventral to the central canal in the chick are comparable in location to dopamine (DA)-containing spinal cord cells in lower vertebrate species. In contrast, the dorsally-suited TH-IR cells in the chick are known only to occur in similar positions in higher vertebrates. Therefore, the chick is novel in that the presence of both groups of TH-IR cells appearing together in significant numbers within the spinal cord has not been shown in any other species studied to date. 4. The TH-containing cells in the chick cord do not appear to contain the catecholamine biosynthesis enzymes, DBH or PNMT. Moreover, using anti-DA immunocytochemistry, neither group of TH-IR cells demonstrated detectable levels of DA in control animals nor in animals pretreated with inhibitors of MAO (MAO-I). 5. However, a difference was noted though between the two TH-IR cell groups in terms of their responses to exogenously supplied L-DOPA, the immediate precursor to DA. With the administration of L-DOPA and a MAO-I to chick hatchlings, cells in the region ventral to the central canal stained intensely for DA. In contrast, the same treatment failed to produce DA-immunoreactive cells in the dorsal horn. 6. One reasonable hypothesis for these results is that the TH-IR cells ventral to the central canal contain an active form of AADC, the enzyme that converts L-DOPA to DA. With this interpretation, if these cells can produce DA from L-DOPA, yet do not appear to synthesize DA endogenously, it would appear that the TH enzyme contained in these cells occurs in an inactive form. Whether the TH enzyme in the dorsally located immunoreactive cells is also inactive is uncertain since it remains unclear whether they contain AADC.

Transantral endodontic surgery.

A case is presented in which periradicular maxillary molar surgery was performed with the transantral approach. Included is a discussion of the related anatomy, physiology, and pathology of the maxillary sinus.

Blood mercury levels with amalgam retroseals: a longitudinal study.

A clinical study was performed to determine if placement of an amalgam retroseal resulted in elevated blood mercury levels. Ten subjects had blood drawn 7 days before and immediately before placement of an amalgam retroseal. Postoperative blood draws occurred at 7 and 30 days. Blood samples were analyzed for mercury content by Cold-Vapor Atomic Absorption Spectrophotometry. No statistically significant increase in blood mercury levels was detected at 7 and 30 days after placement of an amalgam retroseal as compared with preoperative levels (p = 0.97). Findings support the hypothesis that placement of an amalgam retroseal does not result in significant elevations of blood mercury levels.