Similar results were observed in the contralateral nuclei The ex

Similar results were observed in the contralateral nuclei. The expression levels of GABA(A) beta 2/3 were unchanged.

These findings suggest that, following longer periods of monaural conductive hearing loss, the synthesis and subsequent composition of specific glutamate and glycine receptors in projection neurons and their synapses are altered; these changes may contribute to abnormal auditory processing. (C) 2011 IBRO. Published by Elsevier Ltd. All rights reserved.”
“There is currently no blood-based test that can rapidly and objectively distinguish Niraparib between chest pain which is initiated by increased myocardial oxygen demand (stable angina pectoris (SAP)) and chest pain initiated due to decreased coronary blood flow (unstable angina pectoris (UAP)). Since leukocytes play an active role in the progression of coronary artery disease (CAD), we hypothesize these can

provide novel markers of SAP and UAP. Here we use a microarray of 82 cluster of differentiation (CD) antibodies (plus controls) to selectively immobilize peripheral blood mononuclear cells. We find that the pattern of leukocyte immobilization from patients with CAD significantly differs from healthy donors. Within the see more CAD group, 15 SAP patients exhibited significant (p<0.05) changes in 8 of 82 CD antibody spots compared to 19 age-matched healthy blood donors. An additional ten CD antigens differed between healthy donors and patients with UAP (p<0.05). Furthermore, seven CD antibody spots are significantly different between SAP and UAP patients. These preliminary data suggest it is now appropriate to undertake a larger clinical trial to test the hypothesis that these antibody microarrays can monitor the progression from SAP to UAP.”
“Neuropathic pain management is challenging for physicians and a vexing problem for basic researchers. Recent studies reveal that activated spinal astrocytes may play a vital role in nerve Non-specific serine/threonine protein kinase injury-induced neuropathic pain, although the mechanisms are

not fully understood. We have found increased glial fibrillary acidic protein (GFAP) expression, a hallmark of reactive gliosis, and elevated brain-derived neurotrophic factor (BDNF) expression in the dorsal horn in a rat model of allodynia induced by spinal nerve ligation (SNL). The high GFAP expression and mechanical allodynia that SNL induces were prevented by the intrathecal injection of the BDNF-sequestering fusion protein TrkB/Fc. Additionally, mechanical allodynia and GFAP overexpression was induced by the spinal administration of exogenous BDNF to naive rats, and exogenous BDNF given together with fluorocitrate, an astrocytic metabolism inhibitor, inhibited allodynia and GFAP upregulation. Exogenous BDNF also activated the astrocytes directly when tested in vitro. Furthermore, intrathecal administration of BDNF-stimulated astrocytes also induced mechanical allodynia in naive rats.

Imaging and genetic technologies make it possible to safely and n

Imaging and genetic technologies make it possible to safely and non-invasively test these hypotheses directly in humans and can help guide clinical trial efforts designed to correct myelination abnormalities. Such efforts may provide insights into novel avenues for treatment and prevention of some of the most prevalent and devastating human diseases. Published by Elsevier Ltd.”
“The medial prefrontal cortex (mPFC), hippocampus, and amygdala are implicated in the regulation of affect and physiological processes, including hypothalamic-pituitary-adrenal (HPA) axis function. Anhedonia is likely associated with dysregulation

of these processes. Dense-array resting electroencephalographic

and cortisol were obtained from healthy and anhedonic groups. Low-resolution electromagnetic tomography was EX 527 clinical trial used to compute intracerebral current density. For the control group, voxelwise analyses found a relationship between current density in beta and gamma bands and steeper cortisol slope (indicative of more adaptive HPA axis functioning) in regions of the hippocampus, parahippocampal gyrus, and mPFC. For the anhedonic selleck chemicals llc group, the mPFC finding was absent. Anhedonia may be characterized by disruptions of mPFC-mediated neuroendocrine regulation, which could constitute a vulnerability to the development of stress- related disorders.”
“Transmission of pathogenic avian influenza viruses (AIV) from wild birds to domestic poultry and humans is continuing in multiple countries around the world. In preparation for a potential AIV pandemic, multiple vaccine candidates are under development. In the case of H5N1 AIV, a clear shift in transmission from clade 1 to clade 2 viruses occurred in

recent years. The virus-like particle (VLP) represents an economical approach to pandemic vaccine development. In the current study, we evaluated the humoral immune response in humans vaccinated with H5N1 A/Indonesia/05/2005 (clade 2.1) VLP vaccine manufactured in Sf9 insect cells. almost The VLPs were comprised of the influenza virus hemagglutinin (HA), neuraminidase (NA), and matrix 1 (M1) proteins. In an FDA-approved phase I/II human clinical study, two doses of H5N1 VLPs at 15, 45, or 90 mu g HA/dose resulted in seroconversion and production of functional antibodies. Moreover, cross-reactivity against other clade 2 subtypes was demonstrated using virus neutralization assays. H5N1 whole-genome fragment phage display libraries (GFPDL) were used to elucidate the antibody epitope repertoire in postvaccination human sera. Diverse epitopes in HA1/HA2 and NA were recognized by postvaccination sera from the two high-dose groups, including large segments spanning the HA1 receptor binding domain.

Membrane immobilization is specifically highlighted as a route to

Membrane immobilization is specifically highlighted as a route to

maximize process performance.”
“Optokinetic stimulation (OKS) modulates many facets of the neglect syndrome. This sensory stimulation technique is known to activate multiple brain regions (temporo-parietal cortex, basal ganglia, brain stem, cerebellum) some of which are involved in auditory and visual space coding. Here, learn more we evaluated whether OKS modulates auditory neglect transiently and induces a sustained effect (Study 1), and whether repetitive OKS permanently recovers auditory neglect (Study 2). In Study 1,20 patients with visuospatial neglect and auditory neglect in an auditory midline task following rightsided stroke were randomly allocated to an experimental

and a control group matched for neglect severity and socio-demographic factors. Both groups showed a stable, pathological shift of their auditory subjective median plane (ASMP) in front space to the right side. During leftward OKS the experimental group showed a complete normalization of the shift of the ASMP, which endured until 30 min poststimulation, and returned almost to baseline values 24h after OKS. In contrast, the control group who viewed the identical but static dot pattern, showed neither change in their ASMP during this condition, nor any significant change at 30 min or 24 h poststimulation. In Study 2, we show in two samples of neglect patients (N=3 Selleck SHP099 each) that repetitive leftward OKS with smooth pursuit eye movements as a therapy induces lasting improvements in auditory (the ASMP) and visual neglect while visual

scanning therapy yielded no measurable effects on auditory and significantly smaller effects on visual neglect. In conclusion, the experiments show that a single session of OKS induces rapid though transient recovery from auditory neglect including a sustained effect mafosfamide after termination of stimulation, while repetitive OKS therapy yields enduring and multimodal recovery from auditory and visual neglect. OKS therapy with pursuit eye movements therefore represents a multimodally effective and easily applicable technique for the treatment of auditory and visual neglect. (C) 2011 Elsevier Ltd. All rights reserved.”
“While the human leukocyte antigen (HLA) genotype has been associated with the rate of HIV disease progression in untreated patients, little is known regarding these relationships in patients using highly active antiretroviral therapy (HAART). The limited data reported to date identified few HLA-HIV disease associations in patients using HAART and even occasional associations that were opposite of those found in untreated patients. We conducted high-resolution HLA class I and II genotyping in a random sample (n = 860) of HIV-seropositive women enrolled in a long-term cohort initiated in 1994. HLA-HIV disease associations before and after initiation of HAART were examined using multivariate analyses.

This [(3)H]-NPD1/PD1 also displayed specific and selective high a

This [(3)H]-NPD1/PD1 also displayed specific and selective high affinity binding with isolated human neutrophils (K(d) similar to 25nM). Neither resolvin E1 nor lipoxin A(4) competed for [(3)H]-NPD1/PD1 specific binding with human neutrophils. Together, these results provide evidence for stereoselective specific binding of NPD1/PD1

with retinal pigment epithelial cells as well as human neutrophils. Moreover, they suggest specific receptors for this novel mediator in both the immune and visual systems. (C) 2009 Elsevier Ltd. All rights reserved.”
“Animal models that mimic human diabetic nephropathy are useful to identify key factors in pathogenesis of this disease, as well as the development of new therapies. Several mouse TPX-0005 cell line models of diabetes have features of human diabetic nephropathy, yet none of these completely fulfill the Animal Models of Diabetes Complications Consortium criteria and completely reproduce pathological and functional features of the human disease. The Akita mouse carries a mutation in the insulin-2 gene and, to date, only survives as heterozygotes that develop spontaneous type 1 diabetes. Here we show that Akita mice with mutation of both insulin-2 alleles (Akita knockout (KO)) survive if crossed onto

the Balb/c background. These mice develop hyperglycemia, more severe albuminuria, and mesangial sclerosis compared with heterozygous mice on the same genetic background. Interestingly, crossing these AkitaKO mice with integrin alpha

1KO mice, a check details model of exacerbated glomerulosclerosis after injury and also on the Balb/c background, resulted in a 16-fold increase in albuminuria, significant mesangial matrix expansion, nodular and diffuse glomerulosclerosis, and a 2-fold increase in glomerular basement membrane thickening when Sirolimus order compared with nondiabetic mice. Moreover, a significant decline in glomerular filtration was evident in the alpha 1KOAkitaKO mice at 6 months of age. Thus, the integrin a1KOAkitaKO Balb/c mouse represents a promising model presenting with most features of human diabetic nephropathy. Kidney International (2012) 81, 1086-1097; doi: 10.1038/ki.2011.474; published online 1 February 2012″
“The pontine micturition center, or Barrington’s nucleus, is an essential component in the micturition reflex. The purpose of the present study is to examine the connections between Barrington’s nucleus and sacral preganglionic neurons with an electrophysiological method using an intracellular recording technique. When the bladder pressure was near 0 mmH(2)O, electrical stimulation of Barrington’s nucleus either evoked no postsynaptic potential or evoked very small excitatory postsynaptic potentials (EPSPs) with ambiguous onset, in sacral preganglionic neurons. However, when the bladder showed micturition contraction, electrical stimulation of the nucleus evoked clear EPSPs in sacral preganglionic neurons. Latencies of EPSPs ranged from 21.9 to 47.5 ms.

Thus, use of a focal demyelination model suggests that progestero

Thus, use of a focal demyelination model suggests that progesterone exerts promyelinating and anti-inflammatory effects at the spinal cord level. (C) 2011 IBRO. Published by Elsevier Ltd. All rights reserved.”
“Objective: Motor-evoked potential monitoring is used to prevent paraplegia during thoracic aortic surgery.

Multidetector computed tomography has been used preoperatively to detect the Adamkiewicz artery, but the hemodynamic significance of the Adamkiewicz artery is controversial. This study aims to evaluate whether the multidetector computed tomography-defined Adamkiewicz artery is hemodynamically essential and needs to be reconstructed with cold blood spinoplegia under motor-evoked potential monitoring.

Methods: GNS-1480 supplier Protein Tyrosine Kinase inhibitor From 2005 to 2008, both preoperative multidetector computed tomographic analysis and intraoperative neurogenic motor-evoked potential monitoring with cold blood infusion into the clamped segment of the

aorta were done in 15 patients. A motor-evoked potential decrease to less than 50% of the initial value at 3 minutes after cold blood infusion determined the hemodynamic significance of the multidetector computed tomography-defined Adamkiewicz artery. Adamkiewicz arteries determined to be essential were reconstructed, and those determined to be nonessential were sacrificed.

Results: The Adamkiewicz artery was involved in the clamped segment of the aorta in 11 cases. After cold blood infusion, 8 patients experienced no significant motor-evoked potential decrease, and Adamkiewicz artery ligation was undertaken, whereas a moderate motor-evoked potential decrease was noted in 1 patient, prompting reconstruction. None of these 9 patients had permanent neurologic deficits. In 2 patients, the Adamkiewicz artery was reconstructed based on motor-evoked potential findings, with paraparesis occurring in 1 patient. In 4 patients without Adamkiewicz artery involvement in Resveratrol the clamped segment, there was no neurologic deficit.

Conclusions: Cold blood infusion accelerates motor-evoked potential

changes and might enable decision making regarding the need for reconstruction of multidetector computed tomography-defined Adamkiewicz arteries. Cold blood-loaded motor-evoked potential is beneficial to minimize Adamkiewicz artery reconstruction time and limit spinal cord ischemia. (J Thorac Cardiovasc Surg 2011;141:755-61)”
“Rotenone, a widely used pesticide and an environmental risk factor for Parkinson’s disease (PD), induces nigrostriatal injury, Lewy body-like inclusions, and Parkinsonian symptoms in rat models for PD. Our previous data indicated that glyceraldehyde-3-phosphate dehydrogenase (GAPDH) overexpression and glycolytic inhibition were co-current in rotenone-induced PC12 (rat adrenal pheochromocytoma cells) cell death.

“The study’s objective was to empirically assess cognitive

“The study’s objective was to empirically assess cognitive and emotional empathy in patients with narcissistic personality disorder (NPD). To date, “”lack of empathy”" is a core feature of NPD solely based on clinical observation. The study’s method was that forty-seven patients

with NPD, 53 healthy controls, and 27 clinical controls with borderline personality disorder (BPD) were included in the study. Emotional and cognitive empathy were assessed with traditional questionnaire measures, the newly developed Multifaceted Empathy Test (MET), and the Movie for the Assessment of Social Cognition (MASC). The study’s results were that individuals with NPD displayed significant impairments in emotional empathy on the MET. Furthermore, relative to BPD patients and healthy controls, NPD patients did not show deficits in cognitive empathy on the BLZ945 ic50 MET or MASC. Crucially, this empathic profile of NPD is not captured by the Structured Clinical Interview for DSM-IV for Axis II Disorders (SCID-II). The study’s conclusions were that while NPD involves deficits in emotional empathy, cognitive empathy seems grossly unaffected. (C) 2010 Elsevier Ireland Ltd. All rights reserved.”
“Purpose: We described population level trends in radical prostatectomy for patients

Selleck PF477736 with prostate cancer by hospitals with robotic surgery, and assessed whether socioeconomic disparities exist in access to such hospitals.

Materials and Methods: After merging the NIS (Nationwide Inpatient Sample) and the AHA (American Hospital Association) survey from 2006 to 2008, we identified 29,837 patients with prostate cancer who underwent radical prostatectomy. The primary

outcome was treatment with radical prostatectomy at hospitals that have adopted robotic surgery. Multivariate logistic regression was used to identify patient and hospital characteristics Edoxaban associated with radical prostatectomy performed at hospitals with robotic surgery.

Results: Overall 20,424 (68.5%) patients were surgically treated with radical prostatectomy at hospitals with robotic surgery, while 9,413 (31.5%) underwent radical prostatectomy at hospitals without robotic surgery. There was a marked increase in radical prostatectomy at hospital adopters from 55.8% in 2006 and 70.7% in 2007 to 76.1% in 2008 (p < 0.001 for trend). After adjusting for patient and hospital features, lower odds of undergoing radical prostatectomy at hospitals with robotic surgery were seen in black patients (OR 0.81, p < 0.001) and Hispanic patients (OR 0.77, p < 0.001) vs white patients. Compared to having private health insurance, being primarily insured with Medicaid (OR 0.70, p < 0.001) was also associated with lower odds of being treated at hospitals with robotic surgery.

Conditioned media from estradiol treated neurons applied to the g

Conditioned media from estradiol treated neurons applied to the glioma cell line resulted in a significant 7-fold CHIR98014 price increase in L-PGDS promoter

activity supporting the possibility that neuronal-glial interactions are involved in estradiol regulation of L-PGDS. (C) 2010 Elsevier Ireland Ltd. All rights reserved.”
“Monoclonal B-cell lymphocytosis (MBL) is a preclinical hematologic syndrome characterized by small accumulations of CD5+ B lymphocytes. Most MBL share phenotypic characteristics with chronic lymphocytic leukemia (CLL). Although some MBL progress to CLL, most MBL have apparently limited potential for progression to CLL, particularly those MBL with normal absolute B-cell counts (‘low-count’ MBL). Most CLL are monoclonal and it is not known whether MBL are monoclonal

or oligoclonal; this is important AZD2171 because it is unclear whether MBL represent indolent CLL or represent a distinct premalignant precursor before the development of CLL. We used flow cytometry analysis and sorting to determine immunophenotypic characteristics, clonality and molecular features of MBL from familial CLL kindreds. Single-cell analysis indicated four of six low-count MBL consisted of two or more unrelated clones; the other two MBL were monoclonal. 87% of low-count MBL clones had mutated immunoglobulin genes, and no immunoglobulin heavy-chain rearrangements of VH family 1 were observed. Some MBL were diversified, clonally related populations with evidence of antigen drive. We conclude that although low-count MBL share many phenotypic characteristics with CLL, many MBL are oligoclonal. This supports a model for step-wise development of MBL into CLL. Leukemia (2010) 24, 133-140; doi:10.1038/leu.2009.192; published online 15 October 2009″
“Oxidative damage to mitochondrial DNA (mtDNA) has been implicated

as an important mechanism underlying mitochondrial deficiency in epileptic seizures. In focusing on the role of the DNA repair pathway, we determined the response of the mitochondrial base excision repair (mtBER) pathway in pilocarpine-induced status epilepticus (SE) in hippocampi of male Wistar rats. The expression of 8-oxoguanine DNA glycosylase (OGG1) and polymerase gamma (pol gamma) was decreased at both the cellular mRNA and mitochondrial protein levels at DOCK10 3, 9 and 25 h after the onset of SE. The mRNA and protein levels of APE1 were maintained, but the mitochondrial protein level decreased at 3 and 9 h and recovered at 25 h. Therefore, the mtBER pathway failed to respond to SE induced by pilocarpine. The failure of mitochondrial import might be an important factor responsible for the lowered mtBER enzymes in mitochondria. We hypothesize that the down-regulation of mtBER enzymes may aggravate mtDNA damage and mitochondrial deficiency after the onset of SE. (C) 2010 Elsevier Ireland Ltd. All rights reserved.”
“ZAP-70 is a key signaling molecule in T cells.

Cerebral blood flow was lower in WMH areas relative to normal app

Cerebral blood flow was lower in WMH areas relative to normal appearing white matter, which in turn, was lower than grey matter. Regions with consistently lower CBF across individuals were more likely to appear as WMH. Results are consistent with an emerging

literature linking diminished regional perfusion with the risk of developing WMH. (C) 2008 Elsevier Ireland Ltd. All rights reserved.”
“Purpose: We histologically examined the urethral anatomy to Angiogenesis inhibitor assess whether the surgical procedure for radical cystectomy should be modified in females.

Materials and Methods: Anatomical and histological studies were performed on 20 adult female cadavers. Semiserial sections were processed for histological examination and immunohistochemistry. To assess the clinical value of the antegrade approach we examined blood loss and function in 12 consecutive patients who underwent radical cystectomy by this approach.

Results: Vaginal wall smooth muscle contributed to urethral wall formation, in addition to a thin layer of proper urethral smooth muscle, particularly when the bladder detrusor was poorly developed or degenerated. The middle urethra was attached tightly to the vaginal

smooth muscles with PRT062607 abundant veins running at the interface. The urethral sphincter and its inferoposterior continuation (urethrovaginal sphincter) were embedded in the elastic fiber rich perineal membrane. The membrane was U shaped, wrapping

around the anterior aspect of the middle urethra and extending posterior along the distal vagina to end at the lateral extension of the perineal body near the external anal sphincter. Mean estimated blood loss was 965 ml. Of patients who received a neobladder hyper-continence was observed in 14.3% and 57.1% achieved continence.

Conclusions: There is topographical variation in the anatomy of tissues surrounding the female urethra. Care should be taken when dissecting the tissues dorsal or lateral to the urethra. The antegrade approach is useful since the urethra can be dissected under direct vision and traction can be applied to these structures.”
“Non-alcoholic fatty liver disease (NAFLD) has emerged as a major health problem worldwide. Whereas overnutrition and obesity are crucially involved in the development of a simple fatty liver, it remains unclear Vitamin B12 why approximately 10% of all affected individuals develop the ‘inflammatory’ phenotype so-called non-alcoholic steatohepatitis (NASH). A link between the intestinal microbiota and the development of obesity and its metabolic consequences including NAFLD is becoming clearer. First clinical, but especially experimental, studies are suggesting that microbiotal factors are driving forces of hepatic steatosis and inflammation that involve Toll-like receptors and proinflammatory cytokines such as tumor necrosis factor-alpha (TNF alpha).

Oxidative stress has been proposed to be an important factor

Oxidative stress has been proposed to be an important factor

in the pathogenesis of AD. Cyanidin 3-O-glucoside (Cy3G) is a neuroprotective antioxidant. However, the effects of Cy3G on cognition are unclear. In this paper, we show that Cy3G is protective against the A beta-induced impairment of learning and memory, but has no effect on normal learning and memory. Moreover, we found that Gy3G attenuated the A beta-induced tau hyperphosphorylation and GSK-3 beta hyperactivation observed in AD. Taken together, these results demonstrated AZD3965 solubility dmso that Cy3G can rescue the cognitive impairments that are induced by A beta via the modulation of GSK-3 beta/tau, suggesting a potential therapeutic role of Cy3G in AD. (C) 2013 Elsevier Ireland Ltd. All rights reserved.”

To investigate the effect of age on heart rate recovery (HRR) from cognitive challenge. Background: Aging is an independent risk factor for the development of cardiovascular disease. HRR from exercise is an established predictor of cardiac morbidity and mortality, and evidence suggests that HRR from cognitive challenge is predictive of cardiac morbidity as well. Aging is associated with delayed HRR from exercise stress, but little is known about the effect of aging on HRR from psychological stress. We tested the hypothesis that age would be related to delayed HRR from psychological stress. Methods: HRR post exposure to cognitive challenge ( mental arithmetic and Stroop) was investigated 4-Hydroxytamoxifen in a sample of 436 participants aged 35 to 84 years in MIDUS II, a national study of health and well-being. HRR was measured as 1) the amount of change from the stress level; 2) time to recover; and 3) the area under the curve. The analyses were controlled for medical comorbidities and medications that for influence HR, such as body mass index, smoking, sex, menopausal status, and amount of physical activity/exercise. Results: There was no effect for age on HRR as evaluated by all three

recovery assessment methods. Conclusions: Contrary to expectation and in contrast to findings concerning HRR from exercise, HRR from cognitive challenge was preserved with age. These findings require further inquiry into differential mechanism(s) underlying HRR from psychological versus exercise stress, including any role for improved emotion regulation with greater age.”
“Seasonal epidemics caused by antigenic variations in influenza A virus remain a public health concern and an economic burden. The isolation and characterization of broadly neutralizing anti-hemagglutinin monoclonal antibodies (MAb) have highlighted the presence of highly conserved epitopes in divergent influenza A viruses.

8 mM final concentration The culture was grown for an additional

8 mM final concentration. The culture was grown for an additional 4 h, and then the biomass was collected by 10 min centrifugation at 4,000× g. All of the isolation steps were carried out at 4°C. The collected biomass was treated with DNase, RNase and lysozyme buy BIBF 1120 on ice for 1 h, as described by the manufacturer (QIAgen), and complete EDTA-free protease inhibitor cocktail (Roche) was added. The cells were ruptured with 12 consecutive ultrasonication bursts (alternating 30 s pulse, 30 s pause) at the 55 setting (Sonics Vibra Cell). The cell lysates were cleared by three

20 min centrifugations at 20,000× g. All of the other protein isolation steps were carried out. When needed, Imu3 was further purified with size-exclusion FPLC chromatography (Superdex 75 HR 10/30, Amersham Biosciences) equilibrated with 50 mM Tris-HCl, pH 7.5, containing 0.15 M NaCl. Buffer exchanges were carried out using Amicon MWCO 3 kDa microconcentrators (Millipore). The his-tag was removed with the Thrombin Cleavage Capture Kit (Novagen) as described by the manufacturer. Actual mass of Imu3 protein was determined via mass spectrometry ESI + and Q-Tof (Waters-Micromass, United Kingdom). The degree of Usp-producing cell protection provided by each of the three individual immunity BLZ945 proteins (Imu1-3) was examined in E. coli BL21(DE3) pLysE cells that were

transformed with the plasmid pET8c carrying the combination of Usp and either Imu1, Imu2 or Imu3. The transformants were isolated on LB Ap plates with IPTG (0.8 mM final concentration) after being grown overnight at 37°C. Imu3 and Usp binding Formation of a Imu3 dimer was checked using the cross-linking glutaraldehyde assay as previously described [20], native PAGE Interleukin-3 receptor and size exclusion chromatography (HPLC). Imu3 samples (2 mg/mL) with or learn more without the addition of 2.7 kbp double-stranded linear DNA (pUC19/EcoRI) were initially incubated at 37°C for 30 min, to allow for potential multimerization. Samples were then subjected to either native PAGE resolution or to the glutaraldehyde cross-linking procedure and SDS-PAGE resolution, with the LexA protein as a dimerisation-positive control. Aditionally, Imu3 was checked for

dimerisation with size exclusion chromatography (HPLC, Phenomenex Biosep SEC-S2000 column, flow rate: 1 mL/min, 50 mM NaH2PO4, 300 mM NaCl, pH8), self-cleaved LexA protein was used as a standard (11 kDa, 13 kDa and 26 kDa). Formation of the Imu3–USP complex was also investigated using the glutaraldehyde assay, after Imu3 and Usp had been mixed in equimolar ratios. DNA/RNA binding Various concentrations of either EcoRI linearised pUC19 DNA or total RNA (isolated from E. coli) and the Imu3 protein were used to establish the nucleic-acid-binding ability of Imu3. The Imu3 was incubated with either the DNA or RNA in TE buffer (10 mM Tris, 1 mM EDTA, pH 8) at 37°C for 30 min, prior to the electromobility shift assays (EMSAs) with 0.8% agarose gels.