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Fei Li

Publications and source records attributed to Fei Li.

At least 55 records · Page 3Linked to original sources

Classification of real and pseudo microRNA precursors using local structure-sequence features and support vector machine.

BACKGROUND: MicroRNAs (miRNAs) are a group of short (approximately 22 nt) non-coding RNAs that play important regulatory roles. MiRNA precursors (pre-miRNAs) are characterized by their hairpin structures. However, a large amount of similar hairpins can be folded in many genomes. Almost all current methods for computational prediction of miRNAs use comparative genomic approaches to identify putative pre-miRNAs from candidate hairpins. Ab initio method for distinguishing pre-miRNAs from sequence segments with pre-miRNA-like hairpin structures is lacking. Being able to classify real vs. pseudo pre-miRNAs is important both for understanding of the nature of miRNAs and for developing ab initio prediction methods that can discovery new miRNAs without known homology. RESULTS: A set of novel features of local contiguous structure-sequence information is proposed for distinguishing the hairpins of real pre-miRNAs and pseudo pre-miRNAs. Support vector machine (SVM) is applied on these features to classify real vs. pseudo pre-miRNAs, achieving about 90% accuracy on human data. Remarkably, the SVM classifier built on human data can correctly identify up to 90% of the pre-miRNAs from other species, including plants and virus, without utilizing any comparative genomics information. CONCLUSION: The local structure-sequence features reflect discriminative and conserved characteristics of miRNAs, and the successful ab initio classification of real and pseudo pre-miRNAs opens a new approach for discovering new miRNAs.

Animals↗

Alterations of intestinal immune function and regulatory effects of L-arginine in experimental severe acute pancreatitis rats.

AIM: To discuss the changes of intestinal mucosal immune function in rats with experimental severe acute pancreatitis (SAP) and the regulatory effect of L-arginine. METHODS: Male adult Wistar rats were randomly divided into pancreatitis group, sham-operation group, and L-arginine treatment group. Animals were killed at 24, 48, and 72 h after SAP models were developed and specimens were harvested. Endotoxin concentration in portal vein was determined by limulus endotoxin analysis kit. CD3+, CD4+, CD8+ T lymphocytes in intestinal mucosal lamina propria were examined by immunohistochemistry. Secretory immunoglobulin A (SIgA) in cecum feces was examined by radioimmunoassay. RESULTS: Compared to the control group, plasma endotoxin concentration in the portal vein increased, percentage of CD3+ and CD4+ T lymphocyte subsets in the end of intestinal mucosal lamina propria reduced significantly, CD4+/CD8+ ratio decreased, and SIgA concentrations in cecum feces reduced at 24, 48, and 72 h after SAP developed. Compared to SAP group, the L-arginine treatment group had a lower level of plasma endotoxin concentration in the portal vein, a higher CD3+ and CD4+ T lymphocyte percentage in the end of intestinal mucosal lamina propria, an increased ratio of CD4+/CD8+ and a higher SIgA concentration in cecum feces. CONCLUSION: Intestinal immune suppression occurs in the early stage of SAP rats, which may be the main reason for bacterial and endotoxin translocation. L-arginine can improve the intestinal immunity and reduce bacterial and endotoxin translocation in SAP rats.

Acute Disease↗

Two novel proteins, dos1 and dos2, interact with rik1 to regulate heterochromatic RNA interference and histone modification.

BACKGROUND: Chromosomal behavior during mitosis and meiosis depends in part on heterochromatic modifications such as histone H3 lysine-9 methylation (H3K9me). In fission yeast, the Heterochromatin Protein 1 homolog Swi6 recognizes H3K9me, silences transcription, and retains cohesin at pericentromeric repeats. Heterochromatin formation also depends on processing of transcripts derived from centromeric repeats by the RNAi machinery. The DDB1 homolog, Rik1, and histone methyltransferase, Clr4, act in a complex to promote H3K9me. However, the mechanism underlying this interaction is poorly understood. RESULTS: Using a cytological screen, we have identified two novel genes, dos1(+) and dos2(+), which are required for localization of Swi6. Deletion of either of these genes results in mitotic and meiotic chromosome missegregation, defects in mitotic centromeric cohesion and meiotic telomere clustering, and loss of heterochromatic silencing. Dos1 is predominantly located in the nucleus in a Dos2-dependent manner and directly interacts with Rik1. Each of these genes is required for the association of H3K9me with centromeric repeats, as well as for the production of small interfering RNAs. CONCLUSIONS: Dos1 and Dos2 are required for the formation of heterochromatin in fission yeast. We hypothesize that the physical interaction between Dos1 and Rik1 represents a role in regulating activity of the Rik1/Clr4 complex. Dos2 contributes to this role by regulating Dos1 localization. Our findings suggest a mechanism for recruitment of Clr4 in the RNAi-dependent heterochromatin pathway, in which Dos1 and Dos2 are essential.

Carrier Proteins↗

The effect of U1 snRNA binding free energy on the selection of 5' splice sites.

The importance of U1 snRNA binding free energy in the regulation of alternative splicing has been studied in some genes with site-directed mutagenesis. Here we report a large-scale analysis of its impact on 5' splice site (5'ss) selection in human genome. The results show that free energy exerts different effects on alternative 5'ss choice in different situations and -8.1 kcal/mol is a threshold. When both free energies of two competing 5'ss are larger than -8.1 kcal/mol, the 5'ss with lower free energy is more frequently used. However, in other pairs of 5'ss, lower-free-energy 5'ss does not seem to be favored and even the other 5'ss is used more frequently, which suggests that very low binding free energy would impair splicing. Some observations hold true only for those alternative 5' splicing with short alternative exons (<50nt), which implies a complex mechanism of 5'ss selection involving both U1 snRNA binding free energy and regulatory factors.

Alternative Splicing↗

MicroRNA identification based on sequence and structure alignment.

MOTIVATION: MicroRNAs (miRNA) are approximately 22 nt long non-coding RNAs that are derived from larger hairpin RNA precursors and play important regulatory roles in both animals and plants. The short length of the miRNA sequences and relatively low conservation of pre-miRNA sequences restrict the conventional sequence-alignment-based methods to finding only relatively close homologs. On the other hand, it has been reported that miRNA genes are more conserved in the secondary structure rather than in primary sequences. Therefore, secondary structural features should be more fully exploited in the homologue search for new miRNA genes. RESULTS: In this paper, we present a novel genome-wide computational approach to detect miRNAs in animals based on both sequence and structure alignment. Experiments show this approach has higher sensitivity and comparable specificity than other reported homologue searching methods. We applied this method on Anopheles gambiae and detected 59 new miRNA genes. AVAILABILITY: This program is available at http://bioinfo.au.tsinghua.edu.cn/miralign. SUPPLEMENTARY INFORMATION: Supplementary information is available at http://bioinfo.au.tsinghua.edu.cn/miralign/supplementary.htm.

Algorithms↗

An epileptic seizure prediction algorithm based on second-order complexity measure.

The quality of life of many epilepsy patients may be improved significantly if the occurrence of epileptic seizures can be successfully forecasted and clinical intervention, such as electrical stimulation or drug delivery, can then be used to suppress their emergence, or warn the patient of the forthcoming events. In this paper, a prediction algorithm based on the second-order complexity measure was proposed to predict the impending seizures. Through the analysis of long-term intracranial EEG recordings from two frontal lobe epilepsy patients, the results indicated that the sensitivity of prediction was 77.8% (14/18) and 66.7% (4/6) and the number of false warnings was 3 and 2 for the two patients, respectively. Because only the information of past seizures was utilized to predict the current seizure and the computation load was low, the prediction algorithm could possibly be applied to clinical practice.

Adolescent↗

Supramolecular structures and assembly and luminescent properties of quinacridone derivatives.

The synthesis and single-crystal X-ray structures of two quinacridone derivatives, N,N'-di(n-butyl)quinacridone (1) and N,N'-di(n-butyl)-1,3,8,10-tetramethylquinacridone (2), are reported, and the 1H NMR, absorption, photoluminescent (PL), and electroluminescent (EL) characteristics are presented. Both these crystal structures are characterized by intermolecular pi...pi and hydrogen bonding interactions. The intermolecular pi...pi interactions lead to the formation of molecular columns in the solids of 1 and 2, and the interplanar contact distances between two adjacent molecules are 3.48 and 3.55 angstroms, respectively. Crystals of 1 display shorter intermolecular pi...pi contacts and higher density than 2. These results suggest that tighter intermolecular interactions exist in 1. The 1H NMR, absorption, and PL spectra of 1 and 2 in solutions exhibit concentration-dependent properties. The PL quantum yields of 1 in solutions decrease more quickly with the increase of concentration compared to that of 2 in solutions. For solid thin films of Alq3:1 (Alq3 = tris(8-hydroxyquinolinato)aluminum), emission intensities dramatically decrease and obvious red shifts are observed when the dopant concentration is above 4.2%, while for films of Alq3:2, a similar phenomenon occurs when the concentration is above 6.7%. EL devices with Alq3:1 as emitting layer only show high efficiencies (20.3-14.5 cd/A) within the narrow dopant concentration range of 0.5-1.0%. In contrast, high efficiencies (21.5-12.0 cd/A) are achieved for a wider dopant concentration range of 0.5-5.0% when Alq3:2 films are employed as emitting layer. The different PL and EL concentration-dependent properties of the solid thin films Alq3:1 and Alq3:2 are attributed to their different molecular packing characteristics in the solid state.

Journal Article↗

Structure, assembly, and topology of the G185R mutant of the fourth transmembrane domain of divalent metal transporter.

The mammalian iron transporter, divalent metal transporter (DMT1), is a 12-transmembrane domain integral protein, responsible for dietary iron uptake in the duodenum and iron acquisition from transferrin in peripheral tissues. Two disease-causing mutants in animals have been found and attributed to the same missense mutation (G185R), which occurs within the putative transmembrane domain 4 (TM4) of DMT1. We have characterized a synthetic 24-mer peptide, corresponding to the sequence of the TM4 of DMT1 with G185R mutation using circular dichroism (CD) and NMR spectroscopy and show that the G185R peptide assumes mainly alpha-helical conformations in various membrane-mimetic environments. Solution structures derived from NMR and molecular dynamics/simulated annealing calculations demonstrate that the peptide exhibits a highly defined alpha-helix in its middle portion, flanked by a highly flexible N-terminus and a relatively ordered C-terminus. Both the folding and location of the C-terminus in SDS micelles are regulated by pH values. Paramagnetic broadening on peptide NMR signals by spin-labeled 5- and 16-doxylstearic acids and Mn(2+) ion suggests that both the N-terminus and the helical region of the peptide are embedded in SDS micelles. Surprisingly, self-association of the peptides for both the wild type and the G185R mutant studied by CD, electrospray ionization mass spectrometry, and NMR diffusion-ordered spectroscopy demonstrated that mutation of the Gly185 to a bulky and positively charged arginine causes a different self-assembly of the peptide, e.g., from a trimer to a hexamer, which implies that the quaternary structure of integral DMT1 may be crucial for its function in vivo.

Amino Acid Sequence↗

NMR structures and orientation of the fourth transmembrane domain of the rat divalent metal transporter (DMT1) with G185D mutation in SDS micelles.

DMT1, also known as Nramp2, is an iron transporter, and belongs to the family of Nramp proteins. Disease-causing mutations both in Nramp1 and Nramp2 occurring at the conserved two adjacent glycine residues located within the fourth transmembrane domain (TM4) suggest that TM4 may serve an important biological function. In the present study, we have determined the high-resolution structures of a synthetic peptide, corresponding to the sequence of the fourth transmembrane domain of rat DMT1 with G185D mutation, in membrane-mimetic environments (e.g., SDS micelles) using NMR spectroscopy and distance-geometry/simulated annealing calculations. The spatial structures showed alpha-helices without a kink in the middle portion of the peptide, with a highly flexible and poorly defined N-terminus. Both the N-terminus and the helical core of the peptide were embedded into the SDS micelles. Interestingly, the folding and membrane location of the C-terminus was pH dependent, being well-folded and inserted into SDS micelles only at a low pH value (4.0). The peptide exhibited amphipathic characteristics, with hydrophilic residues (Asp7, Thr11, Asp14, and Thr15) lying in one side of the helix, which provide a basis for the formation of water-filled channel architectures through self-associations. The significant broadening of the resonances of the hydrophilic residues Asp7, Thr11, and Asp14, which are buried inside SDS micelles, upon addition of Mn2+ further verified the possibility of the formation of a channel through which metal ions pass. The substitution of Gly7 by an aspartate residue neither significantly altered the structure and membrane location of the peptide nor abolished its properties of channel forming and metal permeation compared with the wild-type peptide.

Amino Acid Sequence↗

Effect of cyclophilin A on gene expression in human pancreatic cancer cells.

BACKGROUND: We previously found that cyclophilin A (CypA) is overexpressed in human pancreatic cancer cells and stimulates cell proliferation through CD147. In this study, we further investigated the effect of CypA on gene expression of several key molecules that are involved in pancreatic cancer cell proliferation. METHODS: Human pancreatic cancer cell lines (Panc-1, MIA PaCa-2, and BxPC-3) and human pancreatic ductal epithelial (HPDE) cells were used. The messenger RNA (mRNA) levels of CypA, CypB, CD147, neuropilins (NRPs), vascular endothelial growth factor (VEGF), and VEGF receptors upon the treatment of exogenous recombinant human CypA were determined by real-time reverse-transcription polymerase chain reaction. RESULTS: Exogenous human recombinant CypA reduced the mRNA levels of NRP-1 and VEGF, but not endogenous CypA, CypB, and CD147, in Panc-1, MIA PaCa-2, and BxPC-3 cells. In contrast, HPDE cells showed a decrease of endogenous CypA and CD147 mRNA, but not detectable changes of CypB, NRPs, and VEGF mRNA levels upon exogenous CypA treatment. CONCLUSIONS: These data show that exogenous CypA downregulates NRP-1 and VEGF expression in pancreatic cancer cells. This effect is different in normal HPDE cells. Thus, soluble CypA may affect cell growth of pancreatic cancer.

Antigens, CD↗

Transfection of SSTR-1 and SSTR-2 inhibits Panc-1 cell proliferation and renders Panc-1 cells responsive to somatostatin analogue.

BACKGROUND: Somatostatin inhibits cell proliferation through interaction with its cellular receptor, somatostatin receptors (SSTRs). We have previously demonstrated that overexpression of individual SSTR-1 or SSTR-2 genes in receptor-negative pancreatic cancer cells inhibited cell proliferation. We hypothesize that reintroduction of SSTR genes back into pancreatic cancer cells might be an effective gene therapy strategy for pancreatic cancer. STUDY DESIGN: We transfected human pancreatic cancer cell line (Panc-1) with human SSTR-1 and SSTR-2 genes and examined the expression by real-time reverse transcriptase-polymerase chain reaction and immunofluorescence. Panc-1 cell proliferation was determined by [(3)H]-thymidine incorporation assay. Activation of phosphorylated c-Jun N-terminal protein kinase (JNK) and cytokine secretion after SSTR-1 and SSTR-2 transfection were detected by Bio-Plex 4-plex phosphoprotein assay and cytokine assay. RESULTS: Panc-1 cells did not express SSTR-1 or SSTR-2, although Panc-1 cells transfected with SSTR-1 and SSTR-2 genes showed a significant amount of SSTR expression. Cell growth rate in Panc-1 cells transfected with SSTR-1 and SSTR-2 was inhibited about 41%, and the cell proliferation of Panc-1 expressing SSTR-1 and SSTR-2 was further reduced about 12% on treatment with somatostatin analogue as compared with the control group. SSTR-1 and SSTR-2 cotransfected Panc-1 cells activated phosphorylation of JNK and increased secretion of interferon-gamma and interleukin-5. CONCLUSIONS: These findings suggest, for the first time, a synergistic inhibitory effect of multiple SSTRs in response to a somatostatin analogue in Panc-1 cells. These studies may improve our understanding of the mechanism by which SSTR inhibits cell growth and lead to novel gene therapies for pancreatic cancer.

Cell Proliferation↗

Taurine replacement attenuates hyperalgesia and abnormal calcium signaling in sensory neurons of STZ-D rats.

The etiology of painful diabetic neuropathy is poorly understood, but may result from neuronal hyperexcitability secondary to alterations of Ca2+ signaling in sensory neurons. The naturally occurring amino acid taurine functions as an osmolyte, antioxidant, Ca2+ modulator, inhibitory neurotransmitter, and analgesic such that its depletion in diabetes may predispose one to neuronal hyperexcitability and pain. This study reports the effects of taurine replacement on hyperalgesia and sensory neuron Ca2+ homeostasis in streptozotocin-diabetic (STZ-D) rats. Nondiabetic and STZ-D rats were treated with a 2% taurine-supplemented diet for 6-12 wk. Thermal hyperalgesia and mechanical allodynia were determined by measuring hindpaw withdrawal latency to radiant heat and the withdrawal threshold to the von Frey anesthesiometer. Intracellular Ca2+ signaling was explored in neurons from L4-L6 dorsal root ganglia (DRG), using fura 2 fluorescence. Taurine replacement of diabetic rats attenuated deficits of nerve conduction and prevented reductions of mechanical and thermal withdrawal threshold and latency, respectively. In small DRG sensory neurons from diabetic rats, recovery of intracellular Ca2+ concentration ([Ca2+]i) in response to KCl was slowed and 73% corrected by taurine. The amplitudes of caffeine and ATP-induced [Ca2+]i transients were decreased by 47 and 27% (P < 0.05), respectively, in diabetic rat DRG sensory neurons and corrected by 74 and 93% (P < 0.05), respectively, by taurine replacement. These data indicate that taurine is important in the regulation of neuronal Ca2+ signaling and that taurine deficiency may predispose one to nerve hyperexcitability and pain, complicating diabetes.

Adenosine Triphosphate↗

Low-dose poly(ADP-ribose) polymerase inhibitor-containing combination therapies reverse early peripheral diabetic neuropathy.

Poly(ADP-ribose) polymerase (PARP) inhibition has recently been identified as a novel approach to treatment of experimental peripheral diabetic neuropathy (PDN). However, long-term inhibition of PARP, an enzyme involved in DNA repair, can potentially result in premature aging, loss of genome stability, and other side effects. This study evaluated potential synergistic interactions between low doses of the potent and specific PARP inhibitor 1,5-isoquinolinediol (ISO) and one of two vasodilators, the ACE inhibitor lisinopril (LIS) and the beta2-adrenoceptor agonist salbutamol (SAL) in the model of early PDN. Control and streptozotocin (STZ)-induced diabetic rats were treated with either ISO plus LIS or ISO plus SAL for 2 weeks after an initial 2 weeks without treatment. ISO (intraperitoneally) and LIS and SAL (both in the drinking water) were used in subtherapeutic doses, resulting in a minor correction of diabetes-associated sciatic motor and hind-limb digital sensory nerve conduction deficits when administered as monotherapies. Both combination treatments corrected endoneurial blood flow and vascular conductance deficits in STZ-induced diabetic rats. ISO plus SAL corrected all other changes of PDN, i.e., motor nerve conduction velocity (MNCV) and sensory nerve conduction velocity (SNCV) deficits as well as thermal and mechanical hyperalgesia. With ISO plus LIS, no significant correction of MNCV was observed, and the effect on thermal hyperalgesia was quite modest. SNCV and mechanical hyperalgesia were corrected. In vitro studies in human endothelial and Schwann cells showed early accumulation of poly(ADP-ribosyl)ated proteins (Western blot analysis) in response to high glucose, thus suggesting the importance of PARP activation in human PDN. In conclusion, low-dose PARP inhibitor-containing combination therapies may constitute a new approach for treatment of PDN.

Albuterol↗

[Long-term effects and mortality of biventricular pacing therapy in patients with congestive heart failure].

OBJECTIVE: To study the long-term effects and mortality of biventricular pacing therapy in patients with congestive heart failure. METHODS: Twenty-five patients, 18 men and 7 women, aged 34-75 [mean aged of (61.42 +/- 10.36)] years, with a cardiac function of New York Heart Association (NYHA) class III (n = 10) or IV (n = 15) received biventricular pacing therapy from Mar. 2001 to Feb. 2005. The etiologies of heart failure were idiopathic dilated cardiomyopathy (16 cases), hypertensive heart disease (3 cases) and ischemic heart disease (6 cases). Left ventricular end-diastolic dimension (LVEDD) was > 60 mm, Left ventricular ejection fraction (LVEF) was < 0.40 and QRS duration was > 130 ms in all the patients. Heart function parameters were repeatedly measured before and 3 months, 6 months, 1 year, 2 years and 3 years after pacemaker implantation. Mortality was also determined. The average follow up period was (20.88 +/- 11.51) months. RESULTS: (1) Mortality: 5 patients died during follow-up (3 non-cardiac and 1 cardiac sudden death and 1 acute myocardial infarction). (2) The mean 6-min walking distance was increased significantly (P < 0.05) at 3 months to 3 years of follow-up. (3) NYHA class: The cardiac function of all patients improved significantly, with a reduction of mean NYHA class of more than one grade at 3 months to 3 years follow-up. (4) LVEDD: LVEDD reduced significantly (P < 0.05) at 3 months to 3 years follow-up. (5) LVEF: LVEF increased significantly (P < 0.05) at 3 months to 2 years follow-up. LVEF also improved at third year's follow-up, but the difference was not significant statistically. CONCLUSIONS: Cardiac resynchronization, a pacemaker-based therapy for heart failure, may enhance quality of life and heart function and reverse LV remodeling. The long-term effects of treatment were stable, leading to the reduction of mortality from advanced heart failure.

Adult↗

[Immunoregulatory effects of the Lonicera aquatic extract in the ovalbulmin-sensitized BALB/c mice].

OBJECTIVE: To evaluate the immunoregulatory effects of the Lonicera water extract in the ovalbulmin (OVA)-sensitized BALB/c mice and to explore feasibility of treating food allergy with the traditional Chinese medicine (TCM). METHODS: Forty female BALB/c mice aged 6 weeks fed with ovalbulmin-free feed, were randomly divided into 5 groups with 8 mice in each. Four groups were sensitized with OVA intraperitoneally two times and challenged intragastrically four times. Groups H, M and L were treated respectively with high (100 mg/100 ml), medium (50 mg/100ml) and low (25 mg/100 ml) concentration of the Lonicera water extract at a dose of 0.3 ml/10 g body weight just 4 hours after the first challenge and then twice daily for 10 consecutive days. The mice in group Ch were used as positive control and were sensitized intraperitoneally and treated with normal saline solution intragastrically daily. The mice in NS group were used as negative control without sensitization and challenge. Just 1 hour after the last challenge, the mice in each group were sacrificed and specimens of jejunum were taken. Histological examinations on the jejunum specimens were performed after either HE or toluidine blue staining, the levels of histamine in gut of the mice were assayed with a fluorescent method; the IFN-gamma and IL-4 production in peripheral lymph node mononuclear cell (PLNMC) and the OVA-specific IgE levels in serum were measured by using ELISA; the mRNA expression of IL-12p40 in PLNMC of the mice was evaluated by RT-PCR; the footpad swelling reactions were assessed for the OVA-induced delayed hypersensitivity. RESULTS: (1) The inflammatory reactions were significantly inhibited in the mice of group H and M; the accumulated and degranulated mast cells in lamina propria were significantly reduced in the mice by gavage with 100% or 50% of the Lonicera extract, concomitant with the increased percentage of the intact mast cells. (2) The release of histamine in gut in the mice of group H and M was significantly reduced. (3) Either the IL-4 production and the ratio of IL-4/IFN-gamma in PLNMC or the IFN-gamma generation was significantly reduced in group H and M. (4) IL-12p40 mRNA expression in PLNMC was significantly reduced in group H and M. (5) The levels of OVA-specific IgE in serum were reduced in the mice of group H and M. (6) The footpad swelling reactions induced in the allergic mice were significantly inhibited after giving the Lonicera extract of the three different concentrations. CONCLUSION: The Lonicera extract showed significant immunoregulatory effects in OVA-induced allergic mice model in this study. Lonicera extract may be of potential research value in treatment of both IgE and none IgE mediated food allergy.

Animals↗

Spindle pole body duplication in fission yeast occurs at the G1/S boundary but maturation is blocked until exit from S by an event downstream of cdc10+.

The regulation and timing of spindle pole body (SPB) duplication and maturation in fission yeast was examined by transmission electron microscopy. When cells are arrested at G1 by nitrogen starvation, the SPB is unduplicated. On release from G1, the SPBs were duplicated after 1-2 h. In cells arrested at S by hydroxyurea, SPBs are duplicated but not mature. In G1 arrest/release experiments with cdc2.33 cells at the restrictive temperature, SPBs remained single, whereas in cells at the permissive temperature, SPBs were duplicated. In cdc10 mutant cells, the SPBs seem not only to be duplicated but also to undergo partial maturation, including invagination of the nuclear envelope underneath the SPB. There may be an S-phase-specific inhibitor of SPB maturation whose expression is under control of cdc10(+). This model was examined by induction of overreplication of the genome by overexpression of rum1p or cdc18p. In cdc18p-overexpressing cells, the SPBs are duplicated but not mature, suggesting that cdc18p is one component of this feedback mechanism. In contrast, cells overexpressing rum1p have large, deformed SPBs accompanied by other features of maturation and duplication. We propose a feedback mechanism for maturation of the SPB that is coupled with exit from S to trigger morphological changes.

CDC2 Protein Kinase↗

Evidence and characteristics of putative human alpha recombination hotspots.

Understanding recombination rate variation is very important for studying genome diversity and evolution, and for investigation of phenotypic association and genetic diseases. Recombination hotspots have been observed in many species and are well studied in yeast. Recent study demonstrated that recombination hotspots are also a ubiquitous feature of the human genome. But the nature of human hotspots remains largely unknown. We have developed and validated a novel computational method for testing the existence of hotspots as well as for localizing them with either unphased or phased genotyping data. To study the characteristics of hotspots within or close to genes, we scanned for unusually high levels of recombination using the European population samples in the SeattleSNPs database, and found evidence for the existence of human alpha hotspots similar to those of yeast. This type of hotspots, found at promoter regions, accounts for about half of the total detected and appears to depend on some specific transcription factor binding sites (such as CGCCCCCGC). These characteristics can explain the observed weak correlation between hotspots and GC-content, and their variation may contribute to the diversity of hotspot distribution among different individuals and species. These long-sought putative human alpha recombination hotspots should deserve further experimental investigations.

Binding Sites↗

Decreased expression of brain cAMP response element-binding protein gene following pentylenetetrazol seizure.

The present study investigated whether the expression of the cAMP response element-binding protein (CREB) in the rat brain is altered following an acute self-limited seizure induced by pentylenetetrazol (PTZ). Male rats were injected intraperitoneally with a single convulsive dose (45 mg/kg) of PTZ, and the matched controls were given saline. For immunohistochemistry, animals were perfused with 4% parafomaldehyde at 24 h following PTZ seizures, and CREB immunoreactivity was examined in rat brain. For real-time RT-PCR, animals were sacrificed at 2 and 24 h and 1 week following PTZ seizures. Tissues from different rat brain regions were micropunched and subjected to real-time RT-PCR using Taqman probe. The CREB immunoreactive profiles were significantly decreased in CA3 and dentate gyrus of hippocampal formation, sensory cerebral cortex and thalamus at 24 h after PTZ seizures. Consistent with changes in CREB immunoreactivity, levels of CREB mRNA were significantly decreased in the hippocampus, cerebral cortex, amygdala and thalamus at 24 h after PTZ seizures. No significant change was found for CREB mRNA expression in these regions at 2 h or 1 week following PTZ seizures. These results show that a brief seizure caused a decline in CREB expression up to 24 h later.

Animals↗