The 3' untranslated region of human Cyclin-Dependent Kinase 5 Regulatory subunit 1 contains regulatory elements affecting transcript stability

<p>Abstract</p> <p>Background</p> <p><it>CDK5R1 </it>plays a central role in neuronal migration and differentiation during central nervous system development. <it>CDK5R1 </it>has been implicated in neurodegenerative disorders and proposed as a candidate gene for mental retardation. The remarkable size of <it>CDK5R1 </it>3'-untranslated region (3'-UTR) suggests a role in post-transcriptional regulation of <it>CDK5R1 </it>expression.</p> <p>Results</p> <p>The bioinformatic study shows a high conservation degree in mammals and predicts several AU-Rich Elements (AREs). The insertion of <it>CDK5R1 </it>3'-UTR into luciferase 3'-UTR causes a decreased luciferase activity in four transfected cell lines. We identified 3'-UTR subregions which tend to reduce the reporter gene expression, sometimes in a cell line-dependent manner. In most cases the quantitative analysis of luciferase mRNA suggests that CDK5R1 3'-UTR affects mRNA stability. A region, leading to a very strong mRNA destabilization, showed a significantly low half-life, indicating an accelerated mRNA degradation. The 3' end of the transcript, containing a class I ARE, specifically displays a stabilizing effect in neuroblastoma cell lines. We also observed the interaction of the stabilizing neuronal RNA-binding proteins ELAV with the CDK5R1 transcript in SH-SY5Y cells and identified three 3'-UTR sub-regions showing affinity for ELAV proteins.</p> <p>Conclusion</p> <p>Our findings evince the presence of both destabilizing and stabilizing regulatory elements in <it>CDK5R1 </it>3'-UTR and support the hypothesis that <it>CDK5R1 </it>gene expression is post-transcriptionally controlled in neurons by ELAV-mediated mechanisms. This is the first evidence of the involvement of 3'-UTR in the modulation of <it>CDK5R1 </it>expression. The fine tuning of <it>CDK5R1 </it>expression by 3'-UTR may have a role in central nervous system development and functioning, with potential implications in neurodegenerative and cognitive disorders.</p

PENGETAHUAN IBU HAMIL TENTANG TANDA-TANDA BAHAYA KEHAMILAN PADA TRIMESTER III

Pada umumnya kehamilan berkembang dengan normal dan menghasilkan kelahiran bayi sehat cukup bulan melalui jalan lahir namun kadang-kadang tidak sesuai dengan yang diharapkan. Sulit diketahui sebelumnya bahwa kehamilan akan menjadi masalah (Prawirohardjo, 2006).Kurangnya pengetahuan ibu dan keluarga tentang kehamilan resiko tinggi dapat meningkatkan kejadian resiko tinggi pada kehamilan. Keterlambatan dalam mengenali secara dini tanda bahaya kehamilan dapat meningkatkan kejadian resiko tinggi kehamilan karena dalam keadaan kehamilan normal pun dapat secara tiba-tiba menjadi resiko tinggi (Depkes, 2009).Tanda-tanda bahaya pada kehamilan yang terjadi pada seorang ibu hamil merupakan suatu pertanda telah terjadinya suatu masalah yang serius pada ibu atau janin yang dikandungnya. Tanda-tanda bahaya ini dapat terjadi pada awal kehamilan (hamil muda) atau pada pertengahan atau pada akhir kehamilan (hamil tua) (Depkes RI, 2009). Adapun macam-macam tanda bahaya kehamilan pada trimester tiga antara lain: Perdarahan pervaginam, keluar air ketuban sebelum waktunya, demam tinggi, bengkak dikaki, muka dan tangan, sakit kepala yang hebat, gerakan janin tidak ada atau kurang (minimal 3 kali dalam 1 jam) (Azrul, 2005).Pengetahuan ibu dan keluarga tentang tanda-tanda bahaya kehamilan dapat memberikan landasan yang kuat agar terwujudnya perilaku sehat dalam menekankan upaya kesehatan promotif selama kehamilan. Di mana pengetahuan merupakan salah satu faktor predisposisi yang dapat mempengaruhi perilaku manusia, termasuk di dalamnya adalah perilaku ibu hamil dalam memanfaatkan secara optimal fasilitas pelayanan kesehatan (Notoatmodjo, 2007).Banda Ace

What kind of surgery for cervical carcinoma?

The incidence and mortality of cervical cancer have changed over the past 50 years in developed countries, but this kind of tumor still remains a significant clinical problem because it is the second most common cause of morbidity and mortality from cancer among women. After histological confirmation of invasive cervical cancer, the extent of disease was determined using clinical criteria to assign a stage. This assessment is important because, while for the other gynecologic cancers clinical information obtained by surgery and histopathological examination is implemented and concurs to define the staging of the disease, the cervical cancer tumor stage is given after the primary diagnosis. In this review we discuss how the surgical approach to cervical cancer has been evolved, in order to modulate the radicality of the intervention itself and thus to preserve the pelvic innervation. This step has been achieved by deepening knowledge of functional pelvic anatomy and modulating the radicality of hysterectomy according to well defined surgical landmarks

Il carcinoma ovarico avanzato: ruolo della chemioipertermia intraperitoneale

La diagnosi del carcinoma ovarico nella maggior parte dei casi avviene quando la neoplasia è ormai in fase avanzata e dunque, indipendentemente dall'estensione agli organi viciniori e/o dalla metastatizzazione, in una fase in cui il peritoneo risulta essere coinvolto in maniera estesa. La strategia terapeutica migliore in tali casi è quella della massima citoriduzione chirurgica, con una migliore sopravvivenza quando il residuo neoplastico è inferiore ai 3 cm. Rimane da discutere il ruolo della micro-disseminazione peritoneale, quindi della persistenza della malattia sul peritoneo. La somministrazione di farmaci antiblastici direttamente nella cavità peritoneale ha permesso di ottenere concentrazioni 20-100 volte superiori rispetto ai protocolli sistemici “ad alte dosi”, esponendo direttamente le cellule neoplastiche residue all'azione dei chemioterapici. La chemioterapia intraperitoneale in ipertermia è una tecnica innovativa che combina la somministrazione di chemioterapici ad elevate temperature alla chirurgia citoriduttiva. La chemioipertemia intraperitoneale eseguita dopo chirurgia citoriduttiva del cancro ovarico incrementa sia la sopravvivenza a 2 anni, rallentando la progressione tumorale, sia l’intervallo libero da malattia

Controlled nitrogen atmosphere for the preservation of functional molecules during silos storage: A case study using old Italian wheat cultivars

The use of a controlled atmosphere preserves the nutraceutical features of old wheat varieties. After 12 months of silos storage, the content of bound antioxidants drops. The content of free antioxidants and polyphenols, both free and bound, is not affected by long-term storage. The content of vitamin E follows the trend of bound antioxidants. The use of a controlled atmosphere slows down the loss of vitamin E

Relationship between plasma concentrations of the l-enantiomer of methadone and response to methadone maintenance treatment

This study evaluated the relationship between the plasma concentration of l-methadone and response to methadone in real-world patients, in order to identify a minimum plasma concentration above which methadone treatment is effective. Ninety-four patients with opioid dependence under maintenance methadone treatment were consecutively recruited. Response was defined as negative urine analyses in the three weeks prior to the blood sampling. The percentage of participants with a plasma l-methadone concentration between 100 and 250 ng/ml was 54.2% among those with a methadone dosage

The 3' untranslated region of human contains regulatory elements affecting transcript stability-4

<p><b>Copyright information:</b></p><p>Taken from "The 3' untranslated region of human contains regulatory elements affecting transcript stability"</p><p>http://www.biomedcentral.com/1471-2199/8/111</p><p>BMC Molecular Biology 2007;8():111-111.</p><p>Published online 3 Dec 2007</p><p>PMCID:PMC2222623.</p><p></p> element (nt 2637–2687) was deleted (hatched bar) from the pGL4.71P-C6 in construct pGL4.71P-C6del by PCR. B) Luciferase activity of the two chimeric constructs in SK-N-BE, SH-SY5Y, HEK-293 and MCF-7 cell lines. Cells, transiently co-transfected with the pGL4.71P- constructs (Renilla luciferase) and the pGL3 (Firefly luciferase) vector were harvested 24 hours post-transfection. Luciferase activity of the chimeric constructs normalized as described, is represented as a percentage of the activity observed in cells transfected with pGL4.71P (defined as 100%). Means ± s.d. luciferase values were obtained from at least four independent experiments (* p < 0.01 compared with the corresponding pGL4.71P value). C) mRNA levels of the chimeric transcripts. Total RNA was extracted from the cells harvested 24 hours post-transfection, and the reporter gene mRNA levels were analyzed by RealTime PCR. mRNA levels of the chimeric reporter gene normalized as described, are represented as a percentage of the mRNA levels observed in cells transfected with pGL4.71P (defined as 100%). Means ± s.d. luciferase mRNA values were obtained from at least three independent experiments (* p < 0.01 compared with the corresponding pGL4.71P value)

The 3' untranslated region of human contains regulatory elements affecting transcript stability-3

<p><b>Copyright information:</b></p><p>Taken from "The 3' untranslated region of human contains regulatory elements affecting transcript stability"</p><p>http://www.biomedcentral.com/1471-2199/8/111</p><p>BMC Molecular Biology 2007;8():111-111.</p><p>Published online 3 Dec 2007</p><p>PMCID:PMC2222623.</p><p></p>nds). SK-N-BE cells were transiently transfected with reporter gene constructs and the pGL3 (firefly luciferase) vector. 24 hours after transfection (time = 0 h), cells were treated with DRB. Total RNA was extracted after various time-points. RNA was reverse-transcribed and RealTime RT-PCR was performed. Renilla luciferase transcript levels were normalized to GAPDH and firefly luciferase mRNAs. Data are expressed as percentage of RNA remaining after DRB addition and are representative of three independent experiments. B) Schematic representation of luciferase constructs carrying C2 fragment and sub-fragments. C) Luciferase activity of the chimeric constructs in SK-N-BE, SH-SY5Y, HEK-293 and MCF-7 cell lines. Cells, transiently co-transfected with the pGL4.71P- constructs (Renilla luciferase) and the pGL3 (Firefly luciferase) vector were harvested 24 hours post-transfection. Luciferase activity of the chimeric constructs normalized as described, is represented as a percentage of the activity observed in cells transfected with pGL4.71P (defined as 100%). Means ± s.d. luciferase values were obtained from at least four independent experiments (* p < 0.01 compared with the corresponding pGL4.71P value). D) mRNA levels of the chimeric transcripts. Total RNA was extracted from the cells harvested 24 hours post-transfection, and the reporter gene mRNA levels were analyzed by RealTime PCR. mRNA levels of the chimeric reporter gene normalized as described, are represented as a percentage of the mRNA levels observed in cells transfected with pGL4.71P (defined as 100%). Means ± s.d. luciferase mRNA values were obtained from at least three independent experiments (* p < 0.01 compared with the corresponding pGL4.71P value)