Top PDF Expression of Caytaxin protein in Cayman Ataxia mouse models correlates with phenotype severity.

Expression of Caytaxin protein in Cayman Ataxia mouse models correlates with phenotype severity.

Expression of Caytaxin protein in Cayman Ataxia mouse models correlates with phenotype severity.

A construct to create a GST-human Caytaxin fusion protein was generated by excising a 1.3 kb fragment from human ATCAY cDNA clone 4153341 (Life Technologies) with SalI and EcoRI restriction enzymes, and cloning it into the pGEX vector (GE Healthcare Life Sciences). The GST-human Caytaxin fusion protein was created by expressing this vector in bacteria and purifying it by preparative SDS-PAGE. The purified human Caytaxin fusion protein was used as an immunogen in mice and spleen cells from an immunized mouse were fused with myeloma cells for generating anti-Caytaxin secreting hybridoma cell lines according to the University of Michigan Hybridoma Core Facility protocols (http://www.med.umich.edu/mdrtc/cores/ hybridomaCore/services.htm). Hybridoma supernatants were screened by Western blot analysis on total wild type mouse brain protein extracts and selected for specific reactivity compared with positive and negative control antibodies (Fig.S1). Three hybrid- oma lines 1E2, 4E3, and 8F4 were selected, sub-cloned and used for ascites production by The University of Michigan Hybridoma Core Facility (http://www.med.umich.edu/mdrtc/cores/ hybridomaCore/index.html). Except where indicated, all anti- Caytaxin monoclonal antibodies were used at a dilution of 1:500. The anti-actin antibody was used for Western blotting according to the manufacturer’s protocol (monoclonal mouse anti-beta-actin from Sigma, Cat. No. A1978). Mouse-anti-beta tubulin E7 antibody was donated by Dr. Diane Fingar at the University of Michigan and used at 1:1000 or 1:2000. Peroxidase-conjugated AffiniPure Goat anti-mouse IgG antibodies were used per manufacturer’s protocol (Jackson ImmunoResearch Laboratories, Inc. Cat. No. 115-035-062).
Mostrar mais

12 Ler mais

Comparative Magnetic Resonance Imaging and Histopathological Correlates in Two SOD1 Transgenic Mouse Models of Amyotrophic Lateral Sclerosis.

Comparative Magnetic Resonance Imaging and Histopathological Correlates in Two SOD1 Transgenic Mouse Models of Amyotrophic Lateral Sclerosis.

An interesting, if surprising, result of this study is the fact that the 129Sv SOD1G93A mice, with a faster disease progression, exhibited a remarkable delay in the alterations of the cranial nuclei detected by both MRI and histopathological analysis, as compared to C57 SOD1G93A mice. In fact, this suggests that the degeneration of brainstem nuclei is not predictive of a more severe phenotype in the SOD1G93A mouse models. Recently, the comparative analysis of the spinal cord and motor neurons of these two SOD1G93A mouse strains highlighted differences associated with the disease severity which were unrelated to the degree of motor neuron loss [13]. In fact, the loss of motor neurons was similar two weeks after the symptom onset in both the slow and fast progressing ALS mouse model. However, an earlier accumulation of ubiquiti- nated protein aggregates was found in the motor neurons of the 129Sv SOD1G93A compared to C57 SOD1G93A mice clearly indicating a dysfunction of remaining motor neurons [13]. A differential vulnerability of cranial motoneurons among different MND animal models has already been reported [23], suggesting that there is a selective motoneuron loss in this region which depends upon the particular mouse model and disease stage. However, since 129Sv SOD1G93A mice die much earlier than the C57 SOD1G93A mice we cannot exclude that the preservation of their brainstem motor neurons is only due to the premature death of these mice rather than their different vulnerability.
Mostrar mais

19 Ler mais

Motor dysfunctions and neuropathology in mouse models of spinocerebellar ataxia type 2: A comprehensive review

Motor dysfunctions and neuropathology in mouse models of spinocerebellar ataxia type 2: A comprehensive review

2011). Interestingly, electrophysiological impairments in Q58 PCs have also been reported, nearly concomitant with the onset of behavioral anomalies, suggesting that the burst firing of these neurons is directly linked with the ataxic symptoms (Kasumu et al., 2012b). These behavioral and neuropathological traits are common to several SCAs, as it was evidenced by similar results in mouse models of SCA1 (Burright et al., 1995), MJD (Chou et al., 2008; Torashima et al., 2008), or SCA7 (Garden et al., 2002). The diversity of the Q58 phenotype and its extensive characterization make this mouse model quite practical to study the effect of new therapeutic strategies in different aspects of the SCA2 phenotype, as it was demonstrated already by some interesting works: Liu et al. (2009) treated the Q58 mice with a pharmacological inhibitor of the Ryanodine receptor, successfully improving the performance of the mutant animals in the beamwalk and the rotarod, and establishing this receptor as a potential therapeutic target for SCA2; Chang et al. (2011), rescued some of the Q58 motor symptoms by injecting these mice intravenously with human mesenchymal stem cells; Kasumu et al. (2012a) used a selective modulator of the SK2/3 channels, administered orally, to alleviate the motor dysfunctions of this model, achieving a significant improvement in the beamwalk and accelerated rotarod; on a different work, Kasumu et al. (2012b) relied on a phosphatase, delivered by adeno-associated viruses, to inhibit the 1,4,5-triphosphate receptor-mediated calcium release. This strategy was successful in preventing the PC dysfunction and rescuing some of the motor impairments of transgenic mice. Nevertheless, there are some issues worth of notice in this model. First, it uses a heterologous promoter that, unlike the human patients that have expression of mutant ATXN2 across the whole body, directs the expression exclusively to the PCs. This feature, although useful for the study of cerebellar dysfunctions like protein aggregation in PCs, hamper the analysis of other brain regions and non-cerebellar symptoms such as Parkinsonism and ALS-related degeneration of motor neurons (Shan et al., 2001; Furtado et al., 2004; Elden et al., 2010). Second, the method of pronuclear injection by which this model was generated results in the integration of the transgene in a random location of the genome, which might be disrupting the function of endogenous genes. Finally, the three Q58 founder mice integrated two, three or four copies of the transgene, leading to non-physiological levels of ataxin-2 expression.
Mostrar mais

14 Ler mais

Population-specific regulation of Chmp2b by Lbx1 during onset of synaptogenesis in lateral association interneurons.

Population-specific regulation of Chmp2b by Lbx1 during onset of synaptogenesis in lateral association interneurons.

Figure 7. Funicular Localization of Chmp2b Protein. (A) Chmp2b staining between the cellular region and basal lamina was observed in a dorsolateral zone (between arrowheads) and a ventrolateral zone (between arrows)in heterozygotes. (B) The dorsolateral zone is lost in mutants while the ventrolateral zone is retained. (C–E) ß-tubulin Stains longitudinal axons in the lateral white matter as blue puncta. (C) The dorsolateral zone of Chmp2b expression is located between the tubulin stained DLF (*) and lateral funiculus (LF; {) in heterozygotes. It appears smaller in this section than in panel A. (D) Tubulin staining at the sulcus limitans (horizontal line) and in the VLF is severely reduced in mutants. The LF appears to be absent and Chmp2b staining in the VLF has an altered appearance. (E) Chmp2b stained projections enter the heterozygote DLF at E12.5. Afferent projections in this funiculus project longitudinally and have not entered the grey matter at this stage. They are labeled with tubulin (blue). Single channel images of this panel are shown in Fig. S3 A–C. GFP/Chmp2b double labels of the insets are enlarged in Fig. S3D–F. They show coincident labeling of GFP and Chmp2b in the white matter (F–K) The red channel of the color images in F, H, J were converted to greyscale images in G, I, and K, respectively. This allows Chmp2b staining in the circumferential trajectory (arrows) to be seen at thoracic levels at E11.5 (G) and E12.5 (I). No such staining can be seen at cervical levels at E12.5 (K). Note that Chmp2b staining is only seen in GFP stained areas at cervical levels (J), whereas it is observed outside of the GFP stained areas at thoracic levels (F, H).
Mostrar mais

17 Ler mais

Transposon-Based Reporter Marking Provides Functional Evidence for Intercellular Bridges in the Male Germline of Rabbits.

Transposon-Based Reporter Marking Provides Functional Evidence for Intercellular Bridges in the Male Germline of Rabbits.

Formation of intercellular bridges in the rabbit male germline were published [22,32], how- ever evidence for the functionality of intercellular bridges in this species has not been reported to date. The testis-expressed gene 14 (TEX-14) is a marker for sperm cell intercellular bridges. TEX-14-/- mutant mice showed drastically lower numbers of spermatocytes, postmeiotic sper- matids and spermatozoa [33]. Here we showed the presence of TEX-14 protein in the testes intercellular bridges of transgenic rabbits, and provide the first evidence of their function. We showed that hemi- and homozygote SB-transgenic spermatozoa carrying the ectopic Venus protein fulfilled the standard semen quality requirements. Indeed, these semen samples were used for artificial insemination through four generations, and neither reduced fertility nor smaller litter sizes were experienced.
Mostrar mais

15 Ler mais

The KISS1 Receptor as an In Vivo Microenvironment Imaging Biomarker of Multiple Myeloma Bone Disease.

The KISS1 Receptor as an In Vivo Microenvironment Imaging Biomarker of Multiple Myeloma Bone Disease.

Currently, diagnosis of MM relies on the detection of excessive monoclonal immunoglobu- lins in the blood and urine and the degree of bone marrow infiltration, though this technique is often insufficient to monitor disease progression [18] and fails to localize aberrant malignant plasma cell clones. Whole body radiography was previously the standard practice for site-spe- cific assessment of MM bone disease. However, because this technique requires at least 30% bone loss prior to detection [19], patients frequently already suffer from severe skeletal involve- ment at the time of diagnosis. In recent years, more sensitive magnetic resonance imaging- or computed tomography-based techniques have been utilized to detect up to 80% more osteolytic lesions. These techniques, however, are expensive, complicated to perform, and yield mixed results depending on the location of the lesion [20]. In order to overcome these limitations, other sensitive, simple, cost-effective assays are needed to easily and conclusively identify MM bone lesions. Disease localization using advanced nuclear medicine imaging approaches may be suited if a specific and sensitive targeting molecule could be identified. Diagnostic methods that allow monitoring of early events in myeloma-affected bone lesions may provide informa- tion for individualized therapies and may offer a survival advantage, as treatments are currently only recommended for patients with active disease.
Mostrar mais

17 Ler mais

Dement. neuropsychol.  vol.9 número4

Dement. neuropsychol. vol.9 número4

TBI occurs when an outside force hits the head hard enough to cause the brain to move violently within the skull disrupting normal brain function. According to whether the injury causes unconsciousness and how long unconsciousness lasts, brain injury may be classiied as mild (mTBI) when no loss of consciousness occurs or unconsciousness lasts less than 30 minutes, moderate when unconsciousness lasts more than 30 minutes and severe if unconsciousness lasts more than 24 hours. TBI can also be classiied using the Glasgow coma scale (GCS) as mild (GCS=13-15), moderate (GCS=9-12) and severe (GCS<8). However, deining minor versus major head injuries is complex. Some authors suggest major injury as worsening level of consciousness, loss of consciousness for more than 5 min, focal neurological indings, seizure, failure to improve in mental status over time, penetrating skull injuries, signs of a basal or depressed skull fracture, and confusion or aggression on examination. 3-8
Mostrar mais

13 Ler mais

Eradication of Aedes aegypti on Cayman Brac and Little Cayman, West Indies, with Abate (Temephos) in 1970-1971

Eradication of Aedes aegypti on Cayman Brac and Little Cayman, West Indies, with Abate (Temephos) in 1970-1971

Relative abundance of container types and Aedes ae@ti container indexes at 289 premises on Cayman Brat and Little Cayman before application of insecticide.. Contain[r]

12 Ler mais

MutLα heterodimers modify the molecular phenotype of Friedreich ataxia.

MutLα heterodimers modify the molecular phenotype of Friedreich ataxia.

Pms2 showed an increased mutability frequency, biased towards GAA repeat expansion in an allele-dose dependent manner. In the current study, the effect of Mlh1 on intergenerational transmission of the YG22 FXN GAA mouse model was similarly assessed by comparing GAA repeat sizes of offspring with those of parents containing different Mlh1 genotypes: FXN GAA /Mlh1 +/+ (par- ents = 2, offspring = 10) or FXN GAA /Mlh1 +/2 (parents = 4, off- spring = 21; Fig. 1). However, study of Mlh1 2/2 male and female parents was not feasible, since they are sterile due to lack of the Mlh1 protein [31]. The transmitted GAA repeat sizes were determined (see Materials and Methods) and categorized into ‘GAA repeat expansions’, ‘no change’ or ‘GAA repeat contrac- tions’ subsets and the percentage of frequencies, as well as mean size changes of each subset, were determined to give an overall ‘transmission profile’. Analysis of data demonstrated a mutability level of 86.7% in the FXN GAA /Mlh1 +/+ transmission profile with a bias towards GAA repeat expansions (Fig. 2). With loss of one Mlh1 allele, the mutability level of the FXN GAA /Mlh1 +/2 intergenerational transmission profile increased to 93.7% with a bias towards GAA repeat contractions (Fig. 2, Table 1). This suggests that down-regulation of Mlh1 protein leads to further GAA repeat instability, with a switch from expansions to contractions. Furthermore, the mean variations of GAA repeat size (i.e. the summation of total GAA repeat changes divided by the entire number of GAA repeat transmissions) for Mlh1 +/2 shows a tendency towards contraction compared with Mlh1 +/+ (Table 2). These findings suggest that Mlh1 protein can protect against intergenerational GAA repeat contractions and may also promote GAA repeat expansions.
Mostrar mais

10 Ler mais

Effects of propofol on lipopolysaccharide-induced expression and release of HMGB1 in macrophages

Effects of propofol on lipopolysaccharide-induced expression and release of HMGB1 in macrophages

Sepsis is a systemic inflammatory response syndrome (SIRS) that develops when the host organism immune response to severe infection by various pathogenic micro- organisms in blood or tissue triggers a life-threatening whole- body inflammatory response. In clinical settings, sepsis has a higher morbidity rate than myocardial infarction and is the major cause of death for patients in intensive care units without heart disease. Sepsis in patients can be detected by identifying the pathogenic microorganism or observing highly suspicious infection foci. The occurrence and development of the disease is followed by pathological processes and eventual regulation. In recent years, although anti-infective therapy and organ function support technology have been improved greatly, the mortality rate of sepsis remains bet- ween 30 and 70%. In particular, the mortality rate of patients with septic shock might be as high as 50% (1). Because of the expense of treatments and large consumption of medical resources, sepsis seriously affects the quality of human life and poses a great threat to human health.
Mostrar mais

6 Ler mais

De Novo Assembly of the Donkey White Blood Cell Transcriptome and a Comparative Analysis of Phenotype-Associated Genes between Donkeys and Horses.

De Novo Assembly of the Donkey White Blood Cell Transcriptome and a Comparative Analysis of Phenotype-Associated Genes between Donkeys and Horses.

Prior to the mechanization of agriculture and labor-intensive tasks, humans used donkeys (Equus africanus asinus) for farm work and packing. However, as mechanization increased, donkeys have been increasingly raised for meat, milk, and fur in China. To maintain the development of the donkey industry, breeding programs should focus on traits related to these new uses. Compared to conventional marker-assisted breeding plans, genome- and transcriptome-based selection methods are more efficient and effective. To analyze the coding genes of the donkey genome, we assembled the transcriptome of donkey white blood cells de novo. Using transcriptomic deep-sequencing data, we identified 264,714 distinct donkey unigenes and predicted 38,949 protein fragments. We annotated the don- key unigenes by BLAST searches against the non-redundant (NR) protein database. We also compared the donkey protein sequences with those of the horse (E. caballus) and wild horse (E. przewalskii), and linked the donkey protein fragments with mammalian pheno- types. As the outer ear size of donkeys and horses are obviously different, we compared the outer ear size-associated proteins in donkeys and horses. We identified three ear size- associated proteins, HIC1, PRKRA, and KMT2A, with sequence differences among the donkey, horse, and wild horse loci. Since the donkey genome sequence has not been released, the de novo assembled donkey transcriptome is helpful for preliminary investiga- tions of donkey cultivars and for genetic improvement.
Mostrar mais

11 Ler mais

Chagas' disease: an update on immune mechanisms and therapeutic strategies

Chagas' disease: an update on immune mechanisms and therapeutic strategies

During the chronic phase of Chagas’ disease, the majority of indi- viduals show potent cellular and humuoral immune responses [77, 78]. The relevance of a strong immune response for parasite control has been shown by the fact that chemically immunosup- pressed individuals [79] and AIDS patients can develop sympto- matic forms of the disease [80]. The mechanisms underlying the transition from asymptomatic to clinically symptomatic are still unclear. Several factors may be involved, such as differences in parasite strain, parasite load, infection time, host genetic back- ground and immune response. In animal models, different para- site strains, mouse backgrounds and re-infections can play a role in the development of heart pathology and/or protection [81–83]. Regarding human infections, a study with patients acutely infected with T. cruzi has shown that CD4 ⫹ and CD8 ⫹ T cells are present in endomyocardial biopsies where myocarditis was also detected in 100% of the cases, reinforcing the role of the immune response in acute pathology [84]. With respect to the chronic phase, a predominance of activated CD8 ⫹ T cells was found in myocardial biopsy fragments from patients with CCC [85, 86]. It is also of interest that peripheral blood mononuclear cells (PBMCs) from chronically infected chagasic patients were able to produce IFN-␥ upon stimulation with recombinant T. cruzi derived proteins [87, 88]. Furthermore, PBMCs from CCC patients pro- duced more IFN-␥ and less IL-10 [87, 89]. IL-10 expression in monocytes from patients with the indeterminate form was higher than in cardiac patients [89, 90]. By contrast, analysis of IFN-␥- producing CD8 ⫹ T cells present in infected patients (with unde- tectable, mild or more severe forms of clinical disease) showed that there was a negative correlation between the capacity of their cells to respond to T. cruzi amastigote antigens and disease sever- ity [91]. A careful analysis of these cells showed that in respond- ing individuals (individuals with milder heart disease), there were early differentiated (CD27 ⫹ CD28 ⫹ ) and effector memory (CD45RA ⫺ CCR7 ⫺ ) CD8 ⫹ T cells. On the other hand, individuals with more severe forms of the disease presented fully differenti- ated (CD27 ⫺ CD28 ⫺ ) CD8 ⫹ T cells [77]. The authors suggest that
Mostrar mais

12 Ler mais

Extraction of matrine from Sophora flavescens Ait. and evaluation of its inhibitory effects on human nasopharyngeal carcinoma CNE-2 cells

Extraction of matrine from Sophora flavescens Ait. and evaluation of its inhibitory effects on human nasopharyngeal carcinoma CNE-2 cells

The present study investigated the extraction and purification technology of matrine from Sophora flavescens Ait. and the inhibitory effects of matrine on human nasopharyngeal carcinoma (NPC) CNE-2 cells. The matrine was extracted from Sophora flavescens Ait. by decocting using ethanol-aqueous solution as the solvent, followed by purification using ion exchange resin adsorption. The extraction and purification conditions were investigated. The effects of matrine on growth, cell apoptosis and cycle of CNE-2 cells and expressions of Bac-2 and caspase-3 protein in cells were detected. Results showed that, the optimal extraction and purification technologies of matrine from Sophora flavescens Ait. were obtained, under which the purity of matrine was 81.56%. Matrine could obtain inhibit the growth of CNE-2 cells, promote the cell apoptosis, and arrest most cells in G0/G1 phase. On addition, after treated matrine, the relative expression level of Bcl-2 protein in CNE-2 cells was decreased, and the expression level of Caspase-3 protein was increased. On conclusion, matrine has inhibitory effects on human NPC CNE-2 cells. The mechanism may be related to its down-regulation of Bcl-2 protein and up-regulation of Caspase-3 protein expression. Keywords: matrine; extraction; nasopharyngeal carcinoma; CNE-2.
Mostrar mais

5 Ler mais

Adult onset sporadic ataxias: a diagnostic challenge

Adult onset sporadic ataxias: a diagnostic challenge

Multiple system atrophy (MSA) is an adult onset sporadic neurodegenerative disease whose clinical spectrum is com- prised by parkinsonian features (MSA-P) or cerebellar signs (MSA-C). In both, pyramidal signs are frequent and auto- nomic dysfunction is mandatory. Autonomic dysfunction is variable, and mostly related to urinary disturbances and pos- tural hypotension. The estimated annual prevalence of the disease is 0.6:100.000 per year, reaching 3:100.000 in popula- tions over 50 years. Median survival is 8-10 years. The under- lying neuropathological findings in MSA are the presence of glial cytoplasmatic inclusions in oligodendroglial cells at aut- opsy. Due to the abundant presence of alpha-synuclein in this disease, similar to Parkinson’s disease and dementia with Lewy bodies, MSA is currently classified as synucleino- pathie. Patients with MSA present a widespread involvement of the central nervous system, with degeneration including basal ganglia, brainstem, cerebellum, pyramidal tracts and spinal cord 37 .
Mostrar mais

9 Ler mais

Over Restriction of Dietary Protein Allowance: The Importance of Ongoing Reassessment of Natural Protein Tolerance in Phenylketonuria

Over Restriction of Dietary Protein Allowance: The Importance of Ongoing Reassessment of Natural Protein Tolerance in Phenylketonuria

The challenge with extra dietary protein was performed in preparation for a BH4 loading test in order to examine in a controlled way the maximum Phe tolerance of patients before a trial with BH4 [17]. Even in patients who were proven not to respond to BH4, many were still able to increase NP intake without deterioration of blood Phe control. A median NP increase of 12 g/day natural protein is equivalent to three slices of regular bread or 60 g of cheese daily. Therefore, this provided an important improvement in both the amount and quality types of NP offered, which possibly would ease some of the dietary burden, improve patient ability to socialise more easily and contribute to better dietary adherence.
Mostrar mais

10 Ler mais

On the pressure response in the brain due to short duration blunt impacts.

On the pressure response in the brain due to short duration blunt impacts.

When the head is subject to non-penetrating (blunt) impact, contusion-type injuries are commonly identified beneath the impact site (the coup) and, in some instances, at the opposite pole (the contre-coup). This pattern of injury has long eluded satisfactory explanation and blunt head injury mechanisms in general remain poorly understood. There are only a small number of studies in the open literature investigating the head’s response to short duration impacts, which can occur in collisions with light projectiles. As such, the head impact literature to date has focussed almost exclusively on impact scenarios which lead to a quasi-static pressure response in the brain. In order to investigate the response of the head to a wide range of impact durations, parametric numerical studies were performed on a highly bio-fidelic finite element model of the human head created from in vivo magnetic resonance imaging (MRI) scan data with non-linear tissue material properties. We demonstrate that short duration head impacts can lead to potentially deleterious transients of positive and negative intra-cranial pressure over an order of magnitude larger than those observed in the quasi-static regime despite reduced impact force and energy. The onset of this phenomenon is shown to be effectively predicted by the ratio of impact duration to the period of oscillation of the first ovalling mode of the system. These findings point to dramatically different pressure distributions in the brain and hence different patterns of injury depending on projectile mass, and provide a potential explanation for dual coup/contre-coup injuries observed clinically.
Mostrar mais

13 Ler mais

Possible roles of COX-1 in learning and memory impairment induced by traumatic brain injury in mice

Possible roles of COX-1 in learning and memory impairment induced by traumatic brain injury in mice

People who suffer from traumatic brain injury (TBI) often experience cognitive deficits in spatial reference and working memory. The possible roles of cyclooxygenase-1 (COX-1) in learning and memory impairment in mice with TBI are far from well known. Adult mice subjected to TBI were treated with the COX-1 selective inhibitor SC560. Performance in the open field and on the beam walk was then used to assess motor and behavioral function 1, 3, 7, 14, and 21 days following injury. Acquisition of spatial learning and memory retention was assessed using the Morris water maze on day 15 post-TBI. The expressions of COX-1, prostaglandin E2 (PGE2), interleukin (IL)-6, brain-derived neurotrophic factor (BDNF), platelet-derived growth factor BB (PDGF-BB), synapsin-I, and synaptophysin were detected in TBI mice. Administration of SC560 improved performance of beam walk tasks as well as spatial learning and memory after TBI. SC560 also reduced expressions of inflammatory markers IL-6 and PGE2, and reversed the expressions of COX-1, BDNF, PDGF-BB, synapsin-I, and synaptophysin in TBI mice. The present findings demonstrated that COX-1 might play an important role in cognitive deficits after TBI and that selective COX-1 inhibition should be further investigated as a potential therapeutic approach for TBI.
Mostrar mais

7 Ler mais

Variation in MSRA modifies risk of neonatal intestinal obstruction in cystic fibrosis.

Variation in MSRA modifies risk of neonatal intestinal obstruction in cystic fibrosis.

The initial starting point in our modifier gene search was a 9 Mb region on chromosome 8 within a region linked to risk for MI in 30 concordant sibling pairs [12]. Association analysis of 133 families using transmission disequilibrium testing identified a single SNP (rs614197) that achieved region-wide significance. Lack of significant association between MI and this SNP in the CGS sample motivated us to search for evidence of untyped alleles using haplotype analysis. The association of both ‘protective’ and ‘risk’ haplotypes with MI led us to surmise that alleles of different effect exist in or near MSRA. We then tested whether the ‘protective’ MSRA haplotype had a similar influence on MI in an independent CF sample. While the TSS sample was potentially subject to ascertainment bias due to its recruitment criteria (i.e. having a sibling with CF), the CGS sample was ideal for replication as recruitment was based only on a diagnosis of CF, and the sample comprises 70% of the patients with CF in Canada. The significant correlation between the ‘protective’ haplotype and reduced incidence of MI in the CGS sample and conformity with an additive model provided reassurance that variants in MSRA modified MI risk. Finally, as several studies, including the present study, have indicated that the CFTR genotype can affect the rate of MI [14–16], we evaluated whether CFTR allelic variation contributed to the association between the MSRA haplotype and MI. The T-G-A haplotype associated with MI in an additive fashion in individuals with identical CFTR genotypes (p.Phe508del homozygotes), thereby demonstrating that MSRA modifies MI independently of variation in CFTR, the disease-causing gene.
Mostrar mais

10 Ler mais

Leishmania major infection in humanized mice induces systemic infection and provokes a nonprotective human immune response.

Leishmania major infection in humanized mice induces systemic infection and provokes a nonprotective human immune response.

However, we detected a 10-fold increase in the expression of murine iNOS mRNA in the footpad tissue of humanized mice three weeks after infection with L. major whereas no increase of iNOS mRNA was found in the spleen. This clearly indicates that mouse macrophages somehow get activated during infection, although the upregulation of iNOS mRNA in humanized mice is ,factor 100 lower than that seen in infected BALB/c mice. In accordance, no iNOS mRNA increase was visible in the spleen of BALB/c mice upon infection [unpublished data, U. Schleicher and C. Bogdan, Erlangen). With regard to parasite control this lack of iNOS increase in the spleen is of minor importance as rather NADPH-oxidase-dependent than iNOS-dependent mech- anism are required for the resolution of the parasites in this organ [65]. In contrast to mouse iNOS mRNA no induction of human iNOS mRNA was detectable in infected humanized mice. Since we demonstrated that human macrophages get infected in humanized mice this result rather supports the idea that iNOS expression by macrophages or other cells in response to infections does not occur in the human immune system [15]. That this statement might not hold true in human cutaneous leishmaniasis is supported by results demonstrating iNOS mRNA and protein
Mostrar mais

12 Ler mais

The expression of p53 protein in patients with multiple myeloma

The expression of p53 protein in patients with multiple myeloma

Gen p53 je jedan od kqučnih gena u složenoj mre­ ži signalnih puteva odgovornih za regulaciju ćelij­ skog ciklusa i apoptoze. Wegov genski proizvod, pro­ tein p53, kontroliše ćelijski ciklus (u G1iG2 fa­ zi) i apoptozu ćelija odgovarajući na nenormalne proliferacione signale i stres, ukqučujući ošte­ ćewe DNK [1]. Po svojoj strukturi je jedarni fosfo­ protein čiji se gen nalazi na kratkom kraku hromo­ zoma 17 [2]. U normalnim ćelijama koje nisu izlože­ ne stresu protein p53 se vezuje za MDM­2, JNKili Pirh­2, koji promovišu wegovu degradaciju preko pu­ ta ubikvitin­proteozoma [3]. Posle izlagawa ćelije stresu ili oštećewa DNK različitim mehanizmima, dolazi do aktivacije gena p53, koji učestvuje u otkla­ wawu nastalog oštećewa ili dela DNK kada je ošte­ ćewe veliko [3]. Čiwenice da je p53 najčešće deleti­ ran ili mutiran gen u tumorima kod čoveka i da po­ rodice sa germinativnim mutacijama 53 imaju pove­ ćanu sklonost za malignitete u mlađem životnom do­ bu ukazuju na značajnu ulogu gena p53 u sprečavawu ma­ lignog preobražaja [3, 4]. Inaktivacija proteina p53 može biti posledica mutacije wegovog gena ili gu­ bitka alela mehanizmom insercije i delecije. Muta­ cije gena p53 utiču na interakciju proteina p53sa DNK, odnosno narušavaju wegov strukturni integri­ tet, vodeći delimičnom ili potpunom gubitku wego­
Mostrar mais

5 Ler mais

Show all 10000 documents...