Southern Illinois University School of Medicine

There are books-few and far between-which carefully, delightfully, and genuinely turn your head inside out. This is one of them. It ranges over some central issues in Western philosophy and begins the long overdue job of giving us a radically new account of meaning, rationality, and objectivity.-Yaakov Garb, San Francisco Chronicle

Importance: Elevated systolic blood (SBP) pressure is a leading global health risk. Quantifying the levels of SBP is important to guide prevention policies and interventions. Objective: To estimate the association between SBP of at least 110 to 115 mm Hg and SBP of 140 mm Hg or higher and the burden of different causes of death and disability by age and sex for 195 countries and territories, 1990-2015. Design: A comparative risk assessment of health loss related to SBP. Estimated distribution of SBP was based on 844 studies from 154 countries (published 1980-2015) of 8.69 million participants. Spatiotemporal Gaussian process regression was used to generate estimates of mean SBP and adjusted variance for each age, sex, country, and year. Diseases with sufficient evidence for a causal relationship with high SBP (eg, ischemic heart disease, ischemic stroke, and hemorrhagic stroke) were included in the primary analysis. Main Outcomes and Measures: Mean SBP level, cause-specific deaths, and health burden related to SBP (≥110-115 mm Hg and also ≥140 mm Hg) by age, sex, country, and year. Results: Between 1990-2015, the rate of SBP of at least 110 to 115 mm Hg increased from 73 119 (95% uncertainty interval [UI], 67 949-78 241) to 81 373 (95% UI, 76 814-85 770) per 100 000, and SBP of 140 mm Hg or higher increased from 17 307 (95% UI, 17 117-17 492) to 20 526 (95% UI, 20 283-20 746) per 100 000. The estimated annual death rate per 100 000 associated with SBP of at least 110 to 115 mm Hg increased from 135.6 (95% UI, 122.4-148.1) to 145.2 (95% UI 130.3-159.9) and the rate for SBP of 140 mm Hg or higher increased from 97.9 (95% UI, 87.5-108.1) to 106.3 (95% UI, 94.6-118.1). For loss of DALYs associated with systolic blood pressure of 140 mm Hg or higher, the loss increased from 95.9 million (95% uncertainty interval [UI], 87.0-104.9 million) to 143.0 million (95% UI, 130.2-157.0 million) [corrected], and for SBP of 140 mm Hg or higher, the loss increased from 5.2 million (95% UI, 4.6-5.7 million) to 7.8 million (95% UI, 7.0-8.7 million). The largest numbers of SBP-related deaths were caused by ischemic heart disease (4.9 million [95% UI, 4.0-5.7 million]; 54.5%), hemorrhagic stroke (2.0 million [95% UI, 1.6-2.3 million]; 58.3%), and ischemic stroke (1.5 million [95% UI, 1.2-1.8 million]; 50.0%). In 2015, China, India, Russia, Indonesia, and the United States accounted for more than half of the global DALYs related to SBP of at least 110 to 115 mm Hg. Conclusions and Relevance: In international surveys, although there is uncertainty in some estimates, the rate of elevated SBP (≥110-115 and ≥140 mm Hg) increased substantially between 1990 and 2015, and DALYs and deaths associated with elevated SBP also increased. Projections based on this sample suggest that in 2015, an estimated 3.5 billion adults had SBP of at least 110 to 115 mm Hg and 874 million adults had SBP of 140 mm Hg or higher.

We have systematically optimized the concentrations of 20 components of a previously published serum-free medium (Brewer and Cotman, Brain Res 494: 65-74, 1989) for survival of rat embryonic hippocampal neurons after 4 days in culture. This serum-free medium supplement, B27, produced neuron survival above 60%, independent of plating density above 160 plated cells/mm2. For isolated cells (< 100 cells/mm2), survival at 4 days was still above 45%, but could be rescued to the 60% level at 40 cells/mm2 by simply applying a coverslip on top of the cells. This suggests a need for additional trophic factors. High survival was achieved with osmolarity lower than found in Dulbecco's Modified Eagle's Medium (DMEM), and by reducing cysteine and glutamine concentrations and by the elimination of toxic ferrous sulphate found in DME/F12. Neurobasal is a new medium that incorporates these modifications to DMEM. In B27/Neurobasal, glial growth is reduced to less than 0.5% of the nearly pure neuronal population, as judged by immunocytochemistry for glial fibrillary acidic protein and neuron-specific enolase. Excellent long-term viability is achieved after 4 weeks in culture with greater than 90% viability for cells plated at 640/mm2 and greater than 50% viability for cells plated at 160/mm2. Since the medium also supports the growth of neurons from embryonic rat striatum, substantia nigra, septum, and cortex, and neonatal dentate gyrus and cerebellum (Brewer, in preparation), support for other neuron types is likely. B27/Neurobasal should be useful for in vitro studies of neuronal toxicology, pharmacology, electrophysiology, gene expression, development, and effects of growth factors and hormones.

The increasingly popular term 'problem-based learning' does not refer to a specific educational method. It can have many different meanings depending on the design of the educational method employed and the skills of the teacher. The many variables possible can produce wide variations in quality and in the educational objectives that can be achieved. A taxonomy is proposed to facilitate an awareness of these differences and to help teachers choose a problem-based learning method most appropriate for their students.

Abstract This chapter reviews the motivation for the change to problem‐based learning, its definition, and the educational objectives it can serve. It discusses changing an established curriculum to problem‐based learning and asks whether problem‐based learning is worth the trouble.

BACKGROUND: Consensus criteria for classifying tremor disorders were published by the International Parkinson and Movement Disorder Society in 1998. Subsequent advances with regard to essential tremor, tremor associated with dystonia, and other monosymptomatic and indeterminate tremors make a significant revision necessary. OBJECTIVES: Convene an international panel of experienced investigators to review the definition and classification of tremor. METHODS: Computerized MEDLINE searches in January 2013 and 2015 were conducted using a combination of text words and MeSH terms: "tremor", "tremor disorders", "essential tremor", "dystonic tremor", and "classification" limited to human studies. Agreement was obtained using consensus development methodology during four in-person meetings, two teleconferences, and numerous manuscript reviews. RESULTS: Tremor is defined as an involuntary, rhythmic, oscillatory movement of a body part and is classified along two axes: Axis 1-clinical characteristics, including historical features (age at onset, family history, and temporal evolution), tremor characteristics (body distribution, activation condition), associated signs (systemic, neurological), and laboratory tests (electrophysiology, imaging); and Axis 2-etiology (acquired, genetic, or idiopathic). Tremor syndromes, consisting of either isolated tremor or tremor combined with other clinical features, are defined within Axis 1. This classification scheme retains the currently accepted tremor syndromes, including essential tremor, and provides a framework for defining new syndromes. CONCLUSIONS: This approach should be particularly useful in elucidating isolated tremor syndromes and syndromes consisting of tremor and other signs of uncertain significance. Consistently defined Axis 1 syndromes are needed to facilitate the elucidation of specific etiologies in Axis 2. © 2017 International Parkinson and Movement Disorder Society.

Reduced signaling of insulin-like peptides increases the life-span of nematodes, flies, and rodents. In the nematode and the fly, secondary hormones downstream of insulin-like signaling appear to regulate aging. In mammals, the order in which the hormones act is unresolved because insulin, insulin-like growth factor-1, growth hormone, and thyroid hormones are interdependent. In all species examined to date, endocrine manipulations can slow aging without concurrent costs in reproduction, but with inevitable increases in stress resistance. Despite the similarities among mammals and invertebrates in insulin-like peptides and their signal cascade, more research is needed to determine whether these signals control aging in the same way in all the species by the same mechanism.

Older adults and special populations (living with disability and/or chronic illness that may limit mobility and/or physical endurance) can benefit from practicing a more physically active lifestyle, typically by increasing ambulatory activity. Step counting devices (accelerometers and pedometers) offer an opportunity to monitor daily ambulatory activity; however, an appropriate translation of public health guidelines in terms of steps/day is unknown. Therefore this review was conducted to translate public health recommendations in terms of steps/day. Normative data indicates that 1) healthy older adults average 2,000-9,000 steps/day, and 2) special populations average 1,200-8,800 steps/day. Pedometer-based interventions in older adults and special populations elicit a weighted increase of approximately 775 steps/day (or an effect size of 0.26) and 2,215 steps/day (or an effect size of 0.67), respectively. There is no evidence to inform a moderate intensity cadence (i.e., steps/minute) in older adults at this time. However, using the adult cadence of 100 steps/minute to demark the lower end of an absolutely-defined moderate intensity (i.e., 3 METs), and multiplying this by 30 minutes produces a reasonable heuristic (i.e., guiding) value of 3,000 steps. However, this cadence may be unattainable in some frail/diseased populations. Regardless, to truly translate public health guidelines, these steps should be taken over and above activities performed in the course of daily living, be of at least moderate intensity accumulated in minimally 10 minute bouts, and add up to at least 150 minutes over the week. Considering a daily background of 5,000 steps/day (which may actually be too high for some older adults and/or special populations), a computed translation approximates 8,000 steps on days that include a target of achieving 30 minutes of moderate-to-vigorous physical activity (MVPA), and approximately 7,100 steps/day if averaged over a week. Measured directly and including these background activities, the evidence suggests that 30 minutes of daily MVPA accumulated in addition to habitual daily activities in healthy older adults is equivalent to taking approximately 7,000-10,000 steps/day. Those living with disability and/or chronic illness (that limits mobility and or/physical endurance) display lower levels of background daily activity, and this will affect whole-day estimates of recommended physical activity.

MicroRNAs are small noncoding RNA molecules that control expression of target genes. Our previous studies show that mir-21 is overexpressed in tumor tissues compared with the matched normal tissues. Moreover, suppression of mir-21 by antisense oligonucleotides inhibits tumor cell growth both in vitro and in vivo. However, it remains largely unclear as to how mir-21 affects tumor growth, because our understanding of mir-21 targets is limited. In this study, we performed two-dimensional differentiation in-gel electrophoresis of tumors treated with anti-mir-21 and identified the tumor suppressor tropomyosin 1 (TPM1) as a potential mir-21 target. In agreement with this, there is a putative mir-21 binding site at the 3′-untranslated region (3′-UTR) of TPM1 variants V1 and V5. Thus, we cloned the 3′-UTR of TPM1 into a luciferase reporter and found that although mir-21 down-regulated the luciferase activity, anti-mir-21 up-regulated it. Moreover, deletion of the mir-21 binding site abolished the effect of mir-21 on the luciferase activity, suggesting that this mir-21 binding site is critical. Western blot with the cloned TPM1-V1 plus the 3′-UTR indicated that TPM1 protein level was also regulated by mir-21, whereas real-time quantitative reverse transcription-PCR revealed no difference at the mRNA level, suggesting translational regulation. Finally, overexpression of TPM1 in breast cancer MCF-7 cells suppressed anchorage-independent growth. Thus, down-regulation of TPM1 by mir-21 may explain, at least in part, why suppression of mir-21 can inhibit tumor growth, further supporting the notion that mir-21 functions as an oncogene. MicroRNAs are small noncoding RNA molecules that control expression of target genes. Our previous studies show that mir-21 is overexpressed in tumor tissues compared with the matched normal tissues. Moreover, suppression of mir-21 by antisense oligonucleotides inhibits tumor cell growth both in vitro and in vivo. However, it remains largely unclear as to how mir-21 affects tumor growth, because our understanding of mir-21 targets is limited. In this study, we performed two-dimensional differentiation in-gel electrophoresis of tumors treated with anti-mir-21 and identified the tumor suppressor tropomyosin 1 (TPM1) as a potential mir-21 target. In agreement with this, there is a putative mir-21 binding site at the 3′-untranslated region (3′-UTR) of TPM1 variants V1 and V5. Thus, we cloned the 3′-UTR of TPM1 into a luciferase reporter and found that although mir-21 down-regulated the luciferase activity, anti-mir-21 up-regulated it. Moreover, deletion of the mir-21 binding site abolished the effect of mir-21 on the luciferase activity, suggesting that this mir-21 binding site is critical. Western blot with the cloned TPM1-V1 plus the 3′-UTR indicated that TPM1 protein level was also regulated by mir-21, whereas real-time quantitative reverse transcription-PCR revealed no difference at the mRNA level, suggesting translational regulation. Finally, overexpression of TPM1 in breast cancer MCF-7 cells suppressed anchorage-independent growth. Thus, down-regulation of TPM1 by mir-21 may explain, at least in part, why suppression of mir-21 can inhibit tumor growth, further supporting the notion that mir-21 functions as an oncogene. MicroRNAs (miRNAs) 2The abbreviations used are: miRNA, microRNA; siRNA, short interfering RNA; RT, reverse transcription; qRT, quantitative reverse transcription; GFP, green fluorescent protein; EGFP, enhanced green fluorescent protein; TPM, tropomyosin; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazoliumbromide; 5-aza-dC, 5-aza-2′-deoxycytidine; UTR, untranslated region; 2-DIGE, two-dimensional differentiation in-gel. are a class of naturally occurring small noncoding RNAs that regulate gene expression by targeting mRNAs for translational repression or cleavage (1Pillai R.S. RNA. 2005; 11: 1753-1761Crossref PubMed Scopus (604) Google Scholar, 2Zamore P.D. Haley B. Science. 2005; 309: 1519-1524Crossref PubMed Scopus (1132) Google Scholar). Like protein-coding mRNAs, miRNAs are transcribed as long primary transcripts in the nucleus. However, unlike protein-coding mRNAs, miRNAs are subsequently cleaved to produce stem-loop-structured precursor molecules of ∼70 nucleotides in length (pre-miRNAs) by the nuclear RNase III enzyme Drosha (3Kim V.N. Nat. Rev. Mol. Cell. Biol. 2005; 6: 376-385Crossref PubMed Scopus (2013) Google Scholar). The pre-miRNAs are then exported to the cytoplasm, where the RNase III enzyme Dicer further processes them into mature miRNAs (∼22 nucleotides). Thus, miRNAs are related to, but distinct from, short inferring RNAs (siRNAs) (4Bartel D.P. Cell. 2004; 116: 281-297Abstract Full Text Full Text PDF PubMed Scopus (29863) Google Scholar, 5Fitzgerald K. Curr. Opin. Drug Discovery Dev. 2005; 8: 557-566PubMed Google Scholar). A key difference between siRNAs and miRNAs is that siRNAs require almost identical sequences to targets to exert their silencing function, whereas miRNAs bind through partial sequence homology to the 3′-untranslated region (3′-UTR) of target genes. Because of this unique feature, a single miRNA has multiple targets. Thus, miRNAs could regulate a large fraction of protein-coding genes, and as high as 30% of all genes could be miRNA targets (6Lewis B.P. Burge C.B. Bartel D.P. Cell. 2005; 120: 15-20Abstract Full Text Full Text PDF PubMed Scopus (9936) Google Scholar). As a new layer of gene regulation mechanism, miRNAs have diverse functions, including the regulation of cellular differentiation, proliferation, and apoptosis (7Croce C.M. Calin G.A. Cell. 2005; 122: 6-7Abstract Full Text Full Text PDF PubMed Scopus (1221) Google Scholar, 8Chen C.Z. Li L. Lodish H.F. Bartel D.P. Science. 2004; 303: 83-86Crossref PubMed Scopus (2804) Google Scholar). Hence, deregulation of miRNA expression may lead to a variety of disorders. Aberrant expression of miRNAs in cancer has been well documented (7Croce C.M. Calin G.A. Cell. 2005; 122: 6-7Abstract Full Text Full Text PDF PubMed Scopus (1221) Google Scholar). Apparently, miRNAs may function as tumor suppressors or oncogenes by targeting oncogenes or tumor suppressor genes (9Chen C.Z. N. Engl. J. Med. 2005; 353: 1768-1771Crossref PubMed Scopus (693) Google Scholar). In this regard, tumor-suppressive miRNAs are usually underexpressed in tumors. For instance, let-7 is down-regulated in lung cancer (10Takamizawa J. Konishi H. Yanagisawa K. Tomida S. Osada H. Endoh H. Harano T. Yatabe Y. Nagino M. Nimura Y. Mitsudomi T. Takahashi T. Cancer Res. 2004; 64: 3753-3756Crossref PubMed Scopus (2164) Google Scholar, 11Johnson S.M. Grosshans H. Shingara J. Byrom M. Jarvis R. Cheng A. Labourier E. Reinert K.L. Brown D. Slack F.J. Cell. 2005; 120: 635-647Abstract Full Text Full Text PDF PubMed Scopus (3119) Google Scholar). Furthermore, more than 60% of investigated patients suffering from B-cell chronic lymphocytic leukemia (B-CLL) have been reported to show a deletion at chromosome 13q14 where the mir-15 and mir-16 genes are located; these genes are under-represented in many B-CLL patients (12Calin G.A. Dumitru C.D. Shimizu M. Bichi R. Zupo S. Noch E. Aldler H. Rattan S. Keating M. Rai K. Rassenti L. Kipps T. Negrini M. Bullrich F. Croce C.M. Proc. Natl. Acad. Sci. U. S. A. 2002; 99: 15524-15529Crossref PubMed Scopus (4274) Google Scholar). Deregulation of miRNAs has also been reported in many other types of cancers. However, although miRNAs have been the subject of extensive research in recent years, the molecular basis of miRNA-mediated gene regulation and the effect of these genes on tumor growth remain largely unknown because of our limited understanding of miRNA target genes. Identification of miRNA target genes has been a great challenge. Computational algorithms have been the major driving force in predicting miRNA targets (13Stark A. Brennecke J. Russell R.B. Cohen S.M. PLoS Biol. 2003; 1: e60Crossref PubMed Scopus (628) Google Scholar, 14Lewis B.P. Shih I.H. Jones-Rhoades M.W. Bartel D.P. Burge C.B. Cell. 2003; 115: 787-798Abstract Full Text Full Text PDF PubMed Scopus (4250) Google Scholar, 15Kiriakidou M. Nelson P.T. Kouranov A. Fitziev P. Bouyioukos C. Mourelatos Z. Hatzigeorgiou A. Genes Dev. 2004; 18: 1165-1178Crossref PubMed Scopus (644) Google Scholar). The approaches are mainly based on base pairing of miRNA and target gene 3′-UTR, emphasizing the location of miRNA complementary elements in 3′-UTR of target mRNAs, the concentration in the seed (6-8 bp) of continuous Watson-Crick base pairing in the 5′ proximal half of the miRNA, and the phylogenetic conservation of the complementary sequences in 3′-UTRs of orthologous genes. However, evidence suggests that perfect seed pairing may not necessarily be a reliable predictor for miRNA-target interactions (16Didiano D. Hobert O. Nat. Struct. Mol. Biol. 2006; 13: 849-851Crossref PubMed Scopus (354) Google Scholar), which may explain why many predicted target sites are nonfunctional. A recent study also suggests that there may be at least three types of miRNA-mRNA interactions in mammals (17Smalheiser N.R. Torvik V.I. Methods Mol. Biol. 2006; 342: 115-127PubMed Google Scholar). Hence, with few exceptions, large portion of the physiologic targets for miRNAs remain to be identified or verified experimentally. In this study, we analyzed tumors derived from breast cancer MCF-7 cells treated with antisense mir-21 oligonucleotide (anti-mir-21) or the negative control by two-dimensional differentiation in-gel (2-DIGE) and identified the tumor suppressor tropomyosin 1 (TPM1) as a putative mir-21 target. Subsequent experiments confirmed that mir-21 down-regulated expression of TPM1, whereas anti-mir-21 up-regulated its expression through the mir-21 binding site at the 3′-UTR region. Furthermore, ectopic expression of TPM1 suppressed anchorage-independent growth. Cell Culture—MCF-7 cells (obtained from American Type Cell Collection, Manassas, VA) were grown in RPMI 1640 (Cambrex, Walkersville, MD) supplemented with 10% fetal bovine serum (Sigma-Aldrich), 2 mm glutamine, 100 units of penicillin/ml, and 100 μg of streptomycin/ml (Cambrex). MCF10A cells (ATCC) were grown in serum-free mammary epithelial growth medium (from Cambrex) supplemented with 100 ng/ml cholera toxin (EMD Biosciences, San Diego, CA). 293T cells (ATCC) were grown in Dulbecco’s modified Eagle’s medium (Cambrex) supplemented with 10% fetal bovine serum. All cells were incubated at 37 °C in a humidified chamber supplemented with and the negative control were from was from CA). of MCF-7 cells was performed with the the cells were in at 30% on the μg of or control was used for in medium of 293T cells was performed the as Cell Res. PubMed Scopus Google Scholar). The negative control oligonucleotide or anti-mir-21 oligonucleotide from at or μg of was used for was by or of mir-21 by miRNA used the C. Z. M. Res. 2005; PubMed Scopus Google Scholar, K. C. Res. 2006; PubMed Scopus Google to the expression level of mature For RNA was used in and with the The was the °C for °C for °C for and then on the the were at and of the was used for with The was at °C for by of °C for and °C for in the real-time The real-time were analyzed and as miRNA expression of which was then to C. Z. M. Res. 2005; PubMed Scopus Google Scholar). The and for mir-21 C. Z. M. Res. 2005; PubMed Scopus Google were from or RNA was used for of TPM1 of TPM1 was performed the the °C for by of °C for °C for 1 and °C for were and were from a mir-21, we modified by the by then a from MCF10A where the site is and where the site is The was cloned into and was subsequently cloned into this modified at the and TPM1 plus 3′-UTR was from MCF-7 cells and then cloned into and into the The 3′-UTR region of TPM1 was also from MCF-7 cells and the and was cloned into control at the a the protein we also used and as indicated was cloned into at the and sites with the region. the 3′-UTR of TPM1 into a which was from the we modified the by a in the of the multiple sites by and then the into site of this modified All were verified by into the cells were in and with luciferase the as the cells were into and for luciferase a luciferase to the was used for Cell with control or the cells were into at The was used to cell growth as S. H. Z. F. in 2006; Google Scholar). anchorage-independent growth of the cells were grown in to a S. J. M. PubMed Google Scholar). 1 with TPM1, cells were and with medium to in a concentration of of this cell were in with in medium and at 37 °C with cell in cell was by in CA). Western protein was from tumor or 293T cells with an in cell mm 100 mm 1 mm concentration was the protein The was with or and then with were with or were the Biosciences, were from and were in the of All studies were in with of and a by the MCF-7 cells were with at and into mammary of tumor growth, a of was the usually 1 anti-mir-21 or negative control oligonucleotide was to tumor sites by of the oligonucleotide of of the was was other of MCF-7 tumors were and in a °C of that were and at °C were for and a by CA). protein was and with or in the was at and in the was in were and to are as and is as by of by have that of MCF-7 cells with anti-mir-21 tumor growth in a S. H. Z. F. in 2006; Google Scholar). Thus, we of anti-mir-21 has the effect on tumor growth. treated with anti-mir-21 in than treated with the negative tumors treated with anti-mir-21 revealed a level of compared with the control is with the previous S. H. Z. F. in 2006; Google Scholar), suggesting that suppression of tumor growth because of cell proliferation, or both as S. H. Z. F. in 2006; Google Scholar, Cancer Res. 2005; PubMed Scopus Google Scholar). these not the notion that mir-21 is an miRNA but also that anti-mir-21 has a TPM1 in as by mir-21 is overexpressed in many types of suggesting its in cancer the of is unclear largely because of limited mir-21 targets. algorithms have predicted many putative mir-21 these targets have not been experimentally. Because miRNAs are to regulate gene expression mainly through translational repression in we to the expression of from the tumor with was from tumors derived from MCF-7 cells treated with the negative control with or anti-mir-21 with two-dimensional in which are in this with fluorescent in a single and for of expression the by and we found that were up-regulated or down-regulated as by or green in is in agreement with the that also or down-regulation of many J. N. R. T. M. M. 2005; PubMed Scopus Google Scholar), because of these may be to the effect of miRNA regulation. of of tumor from revealed an almost identical to that of suggesting the of this are in up-regulated by anti-mir-21 because are potential targets for protein that were up-regulated more than in the tumor treated with anti-mir-21 compared with the negative these are in identified of them with a three have been in TPM1 N. M. Mol. Cell. Biol. PubMed Scopus Google Scholar), protein F. F. S. M. E. F. A. S. Cancer Res. Google and H. S.M. J. 2004; PubMed Scopus Google Scholar). we these three genes by their into a luciferase Western blot of the tumor also indicated that the TPM1 was in the tumors by almost and identified TPM1 as a target for mir-21 as TPM1 a mir-21 for by or are a of that bind to the of there are at least TPM1, and by genes 13: PubMed Scopus Google Scholar). TPM1 has variants through TPM1 variants 1 and a putative mir-21 binding as predicted by the miRNA base target 1 from in a sequence for and also by an nucleotides of the 3′-UTR The potential base pairing between mir-21 and TPM1 3′-UTR is in Thus, we to this region of both variants from MCF-7 to TPM1 Hence, we cloned this 1 3′-UTR into control As in the luciferase in 293T cells for was than that of control suggesting that TPM1 3′-UTR a that this region is mir-21 we 293T cells with the with the or the The ectopic expression of mir-21 was confirmed by real-time which revealed a mir-21 expression in the cells than in control In anti-mir-21 mir-21 by almost as by the then the 293T cells with of As in of luciferase by mir-21 was suggesting that this regulation is to In mir-21 no effect on which is derived from and the mir-21 binding site In we the effect of anti-mir-21 on the luciferase of As mir-21 suppressed the luciferase activity, whereas anti-mir-21 the luciferase further suggesting that expression of TPM1 is regulated by the of the mir-21 binding site in its we the mir-21 binding site in 1 As in mir-21 anti-mir-21 effect on the luciferase activity, the of this mir-21 binding of TPM1 expression by mir-21 at the translational expression of in 293T cells as by Western The was with and then with as The effect of mir-21 or anti-mir-21 on the protein of and or and in which the mir-21 binding site was is and are of at least three and are the of three experiments mir-21 or anti-mir-21 has no effect on the mRNA of as by real-time negative control that down-regulation of luciferase by mir-21 was not to the reporter we we reporter in In this we cloned into the site of the modified with the luciferase the level of was by mir-21 but was by as by Western blot or mir-21 TPM1 at the repression is a major of miRNAs to regulate gene expression 2006; PubMed Scopus Google Scholar). mir-21 also TPM1 through translational we cloned the TPM1 plus the 3′-UTR into of TPM1 was confirmed by although ectopic expression of mir-21 TPM1 anti-mir-21 enhanced TPM1 protein and further the of the mir-21 binding we experiments with in which the site was of this site abolished the effect of mir-21 or anti-mir-21 on TPM1 expression at the protein level and However, the effect of mir-21 or anti-mir-21 on TPM1 at the protein level, no effect on the TPM1 mRNA level was by real-time for these that the mir-21 binding site in the region is for regulation at the translational In we a with the TPM1 plus the 3′-UTR confirmed its expression of its protein by Western blot a of we the green protein all the cell for protein was to the Moreover, mir-21 also the protein as by which was confirmed by Western blot In this we the cells with and a between with and with Thus, the mir-21 binding site is from the by TPM1 it is further that TPM1 is a mir-21 of expression of the protein by expression of or in 293T The and were into 293T cells were on and grown for were with and a that protein is in the as compared with which is the and effect of mir-21 on expression of the protein as by or Western blot with indicated a between with control and with of TPM1 Cell in and previous studies have indicated that suppression of TPM1 is a of many and TPM1 functions as a tumor suppressor S. T. 2003; PubMed Scopus Google Scholar), we overexpression of TPM1-V1 affects cell growth. Thus, was to MCF-7 cells and their growth by found that overexpression of TPM1-V1 suppressed cell growth in a For instance, although there was no difference between control and TPM1-V1 the 2 by and we that cells with TPM1-V1 more than the with TPM1-V1 affects anchorage-independent growth, we MCF-7 cells with control or in the As in the of from MCF-7 cells with was than that of although in vitro cell growth was a effect was on of Furthermore, the of the from the cells with was than of control and are with the that expression of TPM1 S. T. 2003; PubMed Scopus Google Scholar), further evidence that TPM1 is a tumor of TPM1 as mir-21 target gene may explain at least in why suppression of mir-21 can inhibit tumor growth, as we have S. H. Z. F. in 2006; Google Scholar). is well that miRNAs regulate a variety of cellular through regulation of expression of multiple target genes (4Bartel D.P. Cell. 2004; 116: 281-297Abstract Full Text Full Text PDF PubMed Scopus (29863) Google Scholar). In this regard, mir-21 has been to function as an because it is overexpressed in many types of tumors compared with the normal tissues S. H. Z. F. in 2006; Google Scholar, Cancer Res. 2005; PubMed Scopus Google Scholar, M. A. R. S. E. M. M. M. S. A. P. S. Calin G.A. P. Negrini M. Croce C.M. Cancer Res. 2005; PubMed Scopus Google Scholar, C. E. M. P. S. Calin G.A. S. A. Croce C.M. J. 2006; PubMed Scopus Google Scholar). Furthermore, suppression of mir-21 inhibits cell growth, through of apoptosis S. H. Z. F. in 2006; Google Scholar, Cancer Res. 2005; PubMed Scopus Google Scholar). However, it largely remains to be as to how a miRNA affects these in because is the physiologic targets of Our study that TPM1 is target. As a tumor TPM1 has been to a in suppression of the N. M. Mol. Cell. Biol. PubMed Scopus Google Scholar, J. Proc. Natl. Acad. Sci. U. S. A. PubMed Scopus Google Scholar, K. S. B. D. Cell Res. 2002; PubMed Scopus Google Scholar). Thus, of TPM1 as a mir-21 target gene a of why suppression of mir-21 can inhibit tumor growth S. H. Z. F. in 2006; Google Scholar). In miRNAs are to bind through partial sequence to a target gene at 3′-UTR, translational notion is by well miRNA target genes that a in and In the let-7 to the 3′-UTR of and its translational repression by of homology S.M. Grosshans H. Shingara J. Byrom M. Jarvis R. Cheng A. Labourier E. Reinert K.L. Brown D. Slack F.J. Cell. 2005; 120: 635-647Abstract Full Text Full Text PDF PubMed Scopus (3119) Google Scholar). mir-16 targets at the 3′-UTR by a A. Calin G.A. M. M. Shimizu M. Zupo S. M. Rassenti L. H. S. Kipps Negrini M. Croce C.M. Proc. Natl. Acad. Sci. U. S. A. 2005; PubMed Scopus Google Scholar). Apparently, both are tumor-suppressive to a limited of target genes has been although there is on putative targets predicted by For instance, the miRNA base target targets for mir-21, which is not with the of 100 target genes single miRNA J. A. Russell R.B. Cohen S.M. PLoS Biol. 2005; PubMed Scopus Google Scholar). Furthermore, we of the putative mir-21 targets as and by Western but of them to be regulated by mir-21 S. H. Z. F. in 2006; Google Scholar). it is that a small fraction of predicted targets may be and it be a to we an because a major of miRNAs is to be at the 2006; PubMed Scopus Google Scholar). of evidence that TPM1 is a mir-21 target. TPM1 expression is in tumors treated with the of mir-21 to regulate TPM1 protein expression is as it to the 3′-UTR region of TPM1 mRNA with to the mir-21 seed region is to mir-21 overexpression or Finally, deletion of the mir-21 site its mir-21 regulation. miRNAs may regulate protein expression by RNA from RNA J. R. Genes Dev. 2006; PubMed Scopus Google Scholar), our that mir-21 inhibits TPM1 protein as TPM1 mRNA are not by mir-21 or are in all cell types with that as and J. Res. Cell PubMed Scopus Google Scholar). In tropomyosin genes for diverse that are in a and regulated by an 13: PubMed Scopus Google Scholar). of TPM1 and has been reported in suggesting a for these in N. M. Mol. Cell. Biol. PubMed Scopus Google Scholar, B. Cancer Res. Google Scholar). In of into cells K. A. Cancer PubMed Scopus Google Scholar, Proc. Natl. Acad. Sci. U. S. A. PubMed Scopus Google Scholar). Moreover, regulate both and anchorage-independent growth, the of in cell J. Proc. Natl. Acad. Sci. U. S. A. PubMed Scopus Google Scholar). to the class tumor suppressor genes Nat. PubMed Scopus Google Scholar), because expression of is subject to regulation L. Li K. 2005; PubMed Scopus Google Scholar, S. Cancer 2002; PubMed Scopus Google these genes are in their sequences but are underexpressed or to down-regulation or silencing in or of expression to cellular is For instance, of cancer cells with mRNA of TPM1 L. Li K. 2005; PubMed Scopus Google Scholar). can growth of TPM1 and of the growth tumor suppressor function L. Li K. 2005; PubMed Scopus Google Scholar). MCF-7 cells TPM1 K. S. B. D. Cell Res. 2002; PubMed Scopus Google Scholar), and of with not TPM1 expression S. Cancer 2002; PubMed Scopus Google Scholar). However, of the A and in expression of TPM1 S. Cancer 2002; PubMed Scopus Google Scholar), suggesting that may also be in TPM1 Thus, this study potential of regulation of TPM1 cell and tumor cell growth. the 3′-UTR region of may also a in tumor For instance, expression of RNA from the 3′-UTR anchorage-independent growth and tumor in a cell F. Cell. Full Text PDF PubMed Scopus Google Scholar), although the 3′-UTR of may not be to tumor suppression or may not be for tumor suppression in other types of cells Mol. Biol. Cell. 8: PubMed Scopus Google Scholar, 13: Google Scholar). it be of to miRNAs as mir-21 with this region. this we that overexpression of this 3′-UTR region a of miRNAs in the to tumor In TPM1 expression can be regulated by study our the regulation of TPM1, a tumor suppressor Thus, in to as TPM1 is also regulated at the translational level by that a single miRNA has multiple we that mir-21 also has many targets. is our that more mir-21 targets be identified in the with the that we be to the molecular basis of

PURPOSE: This article provides a critical overview of problem-based learning (PBL), its effectiveness for knowledge acquisition and clinical performance, and the underlying educational theory. The focus of the paper is on (1) the credibility of claims (both empirical and theoretical) about the ties between PBL and educational outcomes and (2) the magnitude of the effects. METHOD: The author reviewed the medical education literature, starting with three reviews published in 1993 and moving on to research published from 1992 through 1998 in the primary sources for research in medical education. For each study the author wrote a summary, which included study design, outcome measures, effect sizes, and any other information relevant to the research conclusion. RESULTS AND CONCLUSION: The review of the literature revealed no convincing evidence that PBL improves knowledge base and clinical performance, at least not of the magnitude that would be expected given the resources required for a PBL curriculum. The results were considered in light of the educational theory that underlies PBL and its basic research. The author concludes that the ties between educational theory and research (both basic and applied) are loose at best.

The author defines the term standardized patient (SP), the umbrella term for both a simulated patient (a well person trained to simulate a patient's illness in a standardized way) and an actual patient (who is trained to present his or her own illness in a standardized way). He first discusses the many values of simulated patients over actual patients as teaching and assessment tools in the classroom and refutes a few myths about the use of SPs. Then he recounts the origin and development of SPs over a three-decade period, beginning with his work as a neurologist at the Los Angeles County Hospital, where he trained a model from the art department to simulate a neurological patient and assist in the assessment of clinical clerks. He then describes additional roles of SPs that have developed, including: (1) their use in the Clinical Practice Examination created at Southern Illinois University School of Medicine and (2) the major use that has come into being over the last 10-15 years; facilitating the comprehensive assessment of clinical competence using multiple stations in examinations such as the objective structured clinical examination. He concludes with information about recent and current work on SPs, who are becoming more and more accepted in the assessment process, and urges skeptics not to make judgments about the value of SPs until they have experienced the technique firsthand and reviewed the literature concerning the extensive and often high-quality research about this assessment tool.

The tumor suppressor p53 negatively regulates a number of genes, including the proto-oncogene c-Myc, in addition to activating many other genes. One mechanism of the p53-mediated c-Myc repression may involve transcriptional regulation. However, it is not clear whether microRNAs (miRNAs) play a role in the p53-mediated posttranscriptional regulation of c-Myc. In this study, we show that a putative tumor suppressor, miR-145, is expressed through the phosphoinositide-3 kinase (PI-3K)/Akt and p53 pathways. Importantly, p53 transcriptionally induces the expression of miR-145 by interacting with a potential p53 response element (p53RE) in the miR-145 promoter. We further show that c-Myc is a direct target for miR-145. Although miR-145 silences the expression of c-Myc, anti-miR-145 enhances its expression. This specific silencing of c-Myc by miR-145 accounts at least in part for the miR-145-mediated inhibition of tumor cell growth both in vitro and in vivo. Finally, the blockade of miR-145 by anti-miR-145 is able to reverse the p53-mediated c-Myc repression. Together, these results define the role of miR-145 in the posttranscriptional regulation of c-Myc by p53 and suggest that, as a new member of the p53 regulatory network, miR-145 provides a direct link between p53 and c-Myc in this gene regulatory network.

Aging is characterized by a deterioration in the maintenance of homeostatic processes over time, leading to functional decline and increased risk for disease and death. The aging process is characterized metabolically by insulin resistance, changes in body composition, and physiological declines in growth hormone (GH), insulin-like growth factor-1 (IGF-1), and sex steroids. Some interventions designed to address features of aging, such as caloric restriction or visceral fat depletion, have succeeded in improving insulin action and life span in rodents. Meanwhile, pharmacologic interventions and hormonal perturbations have increased the life span of several mammalian species without necessarily addressing features of age-related metabolic decline. These interventions include inhibition of the mammalian target of rapamycin and lifetime deficiency in GH/IGF-1 signaling. However, strategies to treat aging in humans, such as hormone replacement, have mostly failed to achieve their desired response. We will briefly discuss recent advances in our understanding of the complex role of metabolic pathways in the aging process and highlight important paradoxes that have emerged from these discoveries. Although life span has been the major outcome of interest in the laboratory, a special focus is made in this study on healthspan, as improved quality of life is the goal when translated to humans.

BACKGROUND: The use of central venous catheters impregnated with either minocycline and rifampin or chlorhexidine and silver sulfadiazine reduces the rates of catheter colonization and catheter-related bloodstream infection as compared with the use of unimpregnated catheters. We compared the rates of catheter colonization and catheter-related bloodstream infection associated with these two kinds of antiinfective catheters. METHODS: We conducted a prospective, randomized clinical trial in 12 university-affiliated hospitals. High-risk adult patients in whom central venous catheters were expected to remain in place for three or more days were randomly assigned to undergo insertion of polyurethane, triple-lumen catheters impregnated with either minocycline and rifampin (on both the luminal and external surfaces) or chlorhexidine and silver sulfadiazine (on only the external surface). After their removal, the tips and subcutaneous segments of the catheters were cultured by both the roll-plate and the sonication methods. Peripheral-blood cultures were obtained if clinically indicated. RESULTS: Of 865 catheters inserted, 738 (85 percent) produced culture results that could be evaluated. The clinical characteristics of the patients and the risk factors for infection were similar in the two groups. Catheters impregnated with minocycline and rifampin were 1/3 as likely to be colonized as catheters impregnated with chlorhexidine and silver sulfadiazine (28 of 356 catheters [7.9 percent] vs. 87 of 382 [22.8 percent], P<0.001), and catheter-related bloodstream infection was 1/12 as likely in catheters impregnated with minocycline and rifampin (1 of 356 [0.3 percent], vs. 13 of 382 [3.4 percent] for those impregnated with chlorhexidine and silver sulfadiazine; P<0.002). CONCLUSIONS: The use of central venous catheters impregnated with minocycline and rifampin is associated with a lower rate of infection than the use of catheters impregnated with chlorhexidine and silver sulfadiazine.

The skin is a known target organ for the proopiomelanocortin (POMC)-derived neuropeptides alpha-melanocyte stimulating hormone (alpha-MSH), beta-endorphin, and ACTH and also a source of these peptides. Skin expression levels of the POMC gene and POMC/corticotropin releasing hormone (CRH) peptides are not static but are determined by such factors as the physiological changes associated with hair cycle (highest in anagen phase), ultraviolet radiation (UVR) exposure, immune cytokine release, or the presence of cutaneous pathology. Among the cytokines, the proinflammatory interleukin-1 produces important upregulation of cutaneous levels of POMC mRNA, POMC peptides, and MSH receptors; UVR also stimulates expression of all the components of the CRH/POMC system including expression of the corresponding receptors. Molecular characterization of the cutaneous POMC gene shows mRNA forms similar to those found in the pituitary, which are expressed together with shorter variants. The receptors for POMC peptides expressed in the skin are functional and include MC1, MC5 and mu-opiate, although most predominant are those of the MC1 class recognizing MSH and ACTH. Receptors for CRH are also present in the skin. Because expression of, for example, the MC1 receptor is stimulated in a similar dose-dependent manner by UVR, cytokines, MSH peptides or melanin precursors, actions of the ligand peptides represent a stochastic (predictable) nonspecific response to environmental/endogenous stresses. The powerful effects of POMC peptides and probably CRH on the skin pigmentary, immune, and adnexal systems are consistent with stress-neutralizing activity addressed at maintaining skin integrity to restrict disruptions of internal homeostasis. Hence, cutaneous expression of the CRH/POMC system is highly organized, encoding mediators and receptors similar to the hypothalamic-pituitary-adrenal (HPA) axis. This CRH/POMC skin system appears to generate a function analogous to the HPA axis, that in the skin is expressed as a highly localized response which neutralizes noxious stimuli and attendant immune reactions.

OBJECTIVE: Tinnitus is the perception of sound without an external source. More than 50 million people in the United States have reported experiencing tinnitus, resulting in an estimated prevalence of 10% to 15% in adults. Despite the high prevalence of tinnitus and its potential significant effect on quality of life, there are no evidence-based, multidisciplinary clinical practice guidelines to assist clinicians with management. The focus of this guideline is on tinnitus that is both bothersome and persistent (lasting 6 months or longer), which often negatively affects the patient's quality of life. The target audience for the guideline is any clinician, including nonphysicians, involved in managing patients with tinnitus. The target patient population is limited to adults (18 years and older) with primary tinnitus that is persistent and bothersome. PURPOSE: The purpose of this guideline is to provide evidence-based recommendations for clinicians managing patients with tinnitus. This guideline provides clinicians with a logical framework to improve patient care and mitigate the personal and social effects of persistent, bothersome tinnitus. It will discuss the evaluation of patients with tinnitus, including selection and timing of diagnostic testing and specialty referral to identify potential underlying treatable pathology. It will then focus on the evaluation and treatment of patients with persistent primary tinnitus, with recommendations to guide the evaluation and measurement of the effect of tinnitus and to determine the most appropriate interventions to improve symptoms and quality of life for tinnitus sufferers. ACTION STATEMENTS: The development group made a strong recommendation that clinicians distinguish patients with bothersome tinnitus from patients with nonbothersome tinnitus. The development group made a strong recommendation against obtaining imaging studies of the head and neck in patients with tinnitus, specifically to evaluate tinnitus that does not localize to 1 ear, is nonpulsatile, and is not associated with focal neurologic abnormalities or an asymmetric hearing loss. The panel made the following recommendations: Clinicians should (a) perform a targeted history and physical examination at the initial evaluation of a patient with presumed primary tinnitus to identify conditions that if promptly identified and managed may relieve tinnitus; (b) obtain a prompt, comprehensive audiologic examination in patients with tinnitus that is unilateral, persistent (≥ 6 months), or associated with hearing difficulties; (c) distinguish patients with bothersome tinnitus of recent onset from those with persistent symptoms (≥ 6 months) to prioritize intervention and facilitate discussions about natural history and follow-up care; (d) educate patients with persistent, bothersome tinnitus about management strategies; (e) recommend a hearing aid evaluation for patients who have persistent, bothersome tinnitus associated with documented hearing loss; and (f) recommend cognitive behavioral therapy to patients with persistent, bothersome tinnitus. The panel recommended against (a) antidepressants, anticonvulsants, anxiolytics, or intratympanic medications for the routine treatment of patients with persistent, bothersome tinnitus; (b) Ginkgo biloba, melatonin, zinc, or other dietary supplements for treating patients with persistent, bothersome tinnitus; and (c) transcranial magnetic stimulation for the routine treatment of patients with persistent, bothersome tinnitus. The development group provided the following options: Clinicians may (a) obtain an initial comprehensive audiologic examination in patients who present with tinnitus (regardless of laterality, duration, or perceived hearing status); and (b) recommend sound therapy to patients with persistent, bothersome tinnitus. The development group provided no recommendation regarding the effect of acupuncture in patients with persistent, bothersome tinnitus.

OBJECTIVE: To describe trends of primary efficacy and safety outcomes of islet transplantation in type 1 diabetes recipients with severe hypoglycemia from the Collaborative Islet Transplant Registry (CITR) from 1999 to 2010. RESEARCH DESIGN AND METHODS: A total of 677 islet transplant-alone or islet-after-kidney recipients with type 1 diabetes in the CITR were analyzed for five primary efficacy outcomes and overall safety to identify any differences by early (1999-2002), mid (2003-2006), or recent (2007-2010) transplant era based on annual follow-up to 5 years. RESULTS: Insulin independence at 3 years after transplant improved from 27% in the early era (1999-2002, n = 214) to 37% in the mid (2003-2006, n = 255) and to 44% in the most recent era (2007-2010, n = 208; P = 0.006 for years-by-era; P = 0.01 for era alone). C-peptide ≥0.3 ng/mL, indicative of islet graft function, was retained longer in the most recent era (P < 0.001). Reduction of HbA(1c) and resolution of severe hypoglycemia exhibited enduring long-term effects. Fasting blood glucose stabilization also showed improvements in the most recent era. There were also modest reductions in the occurrence of adverse events. The islet reinfusion rate was lower: 48% by 1 year in 2007-2010 vs. 60-65% in 1999-2006 (P < 0.01). Recipients that ever achieved insulin-independence experienced longer duration of islet graft function (P < 0.001). CONCLUSIONS: The CITR shows improvement in primary efficacy and safety outcomes of islet transplantation in recipients who received transplants in 2007-2010 compared with those in 1999-2006, with fewer islet infusions and adverse events per recipient.

The classical observations of the skin as a target for melanotropins have been complemented by the discovery of their actual production at the local level. In fact, all of the elements controlling the activity of the hypothalamus-pituitary-adrenal axis are expressed in the skin including CRH, urocortin, and POMC, with its products ACTH, alpha-MSH, and beta-endorphin. Demonstration of the corresponding receptors in the same cells suggests para- or autocrine mechanisms of action. These findings, together with the demonstration of cutaneous production of numerous other hormones including vitamin D3, PTH-related protein (PTHrP), catecholamines, and acetylcholine that share regulation by environmental stressors such as UV light, underlie a role for these agents in the skin response to stress. The endocrine mediators with their receptors are organized into dermal and epidermal units that allow precise control of their activity in a field-restricted manner. The skin neuroendocrine system communicates with itself and with the systemic level through humoral and neural pathways to induce vascular, immune, or pigmentary changes, to directly buffer noxious agents or neutralize the elicited local reactions. Therefore, we suggest that the skin neuroendocrine system acts by preserving and maintaining the skin structural and functional integrity and, by inference, systemic homeostasis.

Mathematical ability is related to both activation of the prefrontal cortex in neuroimaging studies of adults and to executive functions in school-age children. The purpose of this study was to determine whether executive functions were related to emergent mathematical proficiency in preschool children. Preschool children (N = 96) were administered an executive function battery that was reduced empirically to working memory (WM), inhibitory control (IC), and shifting abilities by calculating composite scores derived from principal component analysis. Both WM and IC predicted early arithmetic competency, with the observed relations robust after controlling statistically for child age, maternal education, and child vocabulary. Only IC accounted for unique variance in mathematical skills, after the contribution of other executive functions were controlled statistically as well. Specific executive functions are related to emergent mathematical proficiency in this age range. Longitudinal studies using structural equation modeling are necessary to better characterize these ontogenetic relations.

Cancer associated fibroblasts (CAFs) is one of the most crucial components of the tumor microenvironment which promotes the growth and invasion of cancer cells by various mechanisms. CAFs demonstrate a high degree of heterogeneity due to their various origins; however, many distinct morphological features and physiological functions of CAFs have been identified. It is becoming clear that the crosstalk between the cancer cells and the CAFs plays a key role in the progression of cancer, and understanding this mutual relationship would eventually enable us to treat cancer patients by targeting CAFs. In this review, we will discuss the latest findings on the role of CAFs in tumorigenesis and metastasis as well as potential therapeutic implication of CAFs.