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1.
Poult Sci ; 102(5): 102621, 2023 May.
Article in English | MEDLINE | ID: mdl-37003172

ABSTRACT

The current shift from conventional cage (CC) hen housing facilities towards cage-free (CF) hen housing in the egg industry has left many questions regarding how level of fecal exposure and exposure to other hens may influence the hen intestinal microbiota. In a previous publication we reported differences in the bacterial ileal communities and ileal morphology between hens in CC and CF production environments at a single commercial site. Here, we present the first 18S rRNA gene amplicon sequencing-based characterization of the eukaryotic ileal microbiota of adult layer hens, and their associations with intestinal health parameters and the bacterial microbiota. DNA was extracted from the ileal digesta of hens (n = 32 CC, n = 48 CF) using the Qiagen Powerlyzer Powersoil kit, followed by amplification of the V9 region of the 18S rRNA gene. Paired end sequencing was performed with the Illumina MiSeq platform, and the resulting reads were processed according to the Mothur MiSeq protocol in Mothur v1.43.0. De novo operational taxonomic unit (OTU) clustering was performed in mothur with a 99% similarity threshold, and OTUs were taxonomically classified with the SILVA SSU v138 reference database. OTUs classified as vertebrate, plant, or arthropod were removed, resulting in 3,136,400 high quality reads and 1,370 OTUs. Associations between OTUs and intestinal parameters were calculated using PROC GLIMMIX. PERMANOVA over Bray-Curtis distances revealed differences between CC and CF eukaryotic ileal microbiota at the whole community level, but no OTUs were differentially abundant after correcting for false discovery (P > 0.05; q > 0.1). Kazachstania and Saccharomyces, closely related genera of yeast, represented 77.1% and 9.7% of sequences respectively. Two Kazachstania OTUs and 1 Saccharomycetaceae OTU were positively correlated with intestinal permeability (r2 ≥ 0.35). Eimeria accounted for 7.6% of sequences across all samples. Intriguingly, 15 OTUs classified as Eimeria were inversely associated with intestinal permeability (r2 ≤ -0.35), suggesting Eimeria may play a more complex role in the microbiota of healthy birds than has been observed in disease challenges.


Subject(s)
Eimeria , Microbiota , Animals , Female , Chickens , Eukaryota , Housing , Ileum , Bacteria/genetics
2.
Transl Anim Sci ; 6(2): txac047, 2022 Apr.
Article in English | MEDLINE | ID: mdl-35663613

ABSTRACT

Forty sows (PIC Camborough 1050) from a single farm were randomly selected at 112 days of gestation to evaluate if gut bacteria transverse the blood system of the sow to deposit gut microbiota into the colostrum for piglet gut inoculation via the entero-mammary pathway. Fourteen first-parity gilts and 20 third-parity sows were used for the study. At the time of farrowing, colostrum, fecal samples, and blood samples were collected to evaluate the presence of bacteria in each sample. Colostrum and blood samples were processed via centrifugation to separate the immune cell fraction. Total DNA was extracted from fecal, colostrum, and white blood cell fractions. 16S ribosomal RNA gene amplicon sequencing was conducted at the Iowa State University DNA Facility (Ames, IA) to further characterize the bacterial and archaeal taxa present within each sample. Data were analyzed using Mothur and using R v4.0.3 (R Core Team, 2020). The experimental unit was the sow. Tables were generated to demonstrate the relative abundances of bacteria and archaea present in each type of sample and also identify organisms differentially abundant between sample types. Firmicutes were the most abundant phylum in colostrum and fecal samples and Tenericutes had the greatest abundance in blood comparative to other phyla. Further evaluation of the classification of bacteria present demonstrated that a few genera of bacteria are present in all three samples. Clostridum_sensu_stricto 1 was present in high relative abundance in colostrum and moderate abundance in the feces while also being present within the blood. Other genera present in all three sample types include Ruminococcus and Mycoplasma. In conclusion, the data suggest that there are bacteria present in all three locations of the sow at the time of farrowing and that first parity sows have different microbial populations than third parity sows.

3.
J Robot Surg ; 15(4): 571-577, 2021 Aug.
Article in English | MEDLINE | ID: mdl-32885379

ABSTRACT

Partial nephrectomy (PN) for small renal masses is common, but outcomes are not reported in a standard manner. Traditionally, parameters such as 90-day mortality, blood loss, transfusion rates, length of stay, nephrometry scoring and complications are published but their collective impact on warm ischemia time (WIT) and post-surgery GFR is rarely determined. Thus, our aim was to assess if "Trifecta" and "Pentafecta" outcomes could be used as useful surgical outcome markers. A prospective database of 252 Robotic-Assisted PN (RAPN) cases (2008-2019) was analysed. "Pentafecta" was defined as achievement of "Trifecta" (negative surgical margin, no postoperative complications and WIT of < 25 min) plus over 90% estimated GFR preservation and no CKD stage upgrading at 1 year. Binary logistic regression analysis was conducted to predict factors which may prevent achieving a Trifecta/Pentafecta. Median tumour size was 3 cm and mean WIT was 15 min. Positive surgical margins (PSM) occurred in 2 cases. Overall, the intra-operative complication rate was 7%. One recurrence conferred 5-year cancer-free survival of 97%. Trifecta outcome was achieved in 169 (67%) and Pentafecta in 141 (56%) of cases. At logistic regression analysis, intraoperative blood loss was the only factor to affect Trifecta achievement (p = 0.018). Advanced patient age negatively impacted Pentafecta achievement (p = 0.010). The Trifecta and Pentafecta outcomes are easily applicable to PN data, and offer an internationally comparable PN outcome, quality measure. We recommend applying this standardization to national data collection to improve the quality of reporting and ease of interpretation of surgeon/centres' outcomes.


Subject(s)
Kidney Neoplasms , Robotic Surgical Procedures , Humans , Kidney Neoplasms/surgery , Neoplasm Recurrence, Local , Nephrectomy , Reference Standards , Retrospective Studies , Robotic Surgical Procedures/methods , Treatment Outcome
4.
BMC Biol ; 15(1): 63, 2017 07 31.
Article in English | MEDLINE | ID: mdl-28756777

ABSTRACT

BACKGROUND: Helicoverpa armigera and Helicoverpa zea are major caterpillar pests of Old and New World agriculture, respectively. Both, particularly H. armigera, are extremely polyphagous, and H. armigera has developed resistance to many insecticides. Here we use comparative genomics, transcriptomics and resequencing to elucidate the genetic basis for their properties as pests. RESULTS: We find that, prior to their divergence about 1.5 Mya, the H. armigera/H. zea lineage had accumulated up to more than 100 more members of specific detoxification and digestion gene families and more than 100 extra gustatory receptor genes, compared to other lepidopterans with narrower host ranges. The two genomes remain very similar in gene content and order, but H. armigera is more polymorphic overall, and H. zea has lost several detoxification genes, as well as about 50 gustatory receptor genes. It also lacks certain genes and alleles conferring insecticide resistance found in H. armigera. Non-synonymous sites in the expanded gene families above are rapidly diverging, both between paralogues and between orthologues in the two species. Whole genome transcriptomic analyses of H. armigera larvae show widely divergent responses to different host plants, including responses among many of the duplicated detoxification and digestion genes. CONCLUSIONS: The extreme polyphagy of the two heliothines is associated with extensive amplification and neofunctionalisation of genes involved in host finding and use, coupled with versatile transcriptional responses on different hosts. H. armigera's invasion of the Americas in recent years means that hybridisation could generate populations that are both locally adapted and insecticide resistant.


Subject(s)
Genome, Insect , Herbivory , Moths/genetics , Animals , Gene Expression Profiling , Genomics , Introduced Species , Larva/genetics , Larva/growth & development , Moths/classification , Moths/growth & development , Sequence Analysis, DNA
6.
Mol Ecol ; 25(21): 5296-5311, 2016 11.
Article in English | MEDLINE | ID: mdl-27661785

ABSTRACT

Helicoverpa armigera is a major agricultural pest that is distributed across Europe, Asia, Africa and Australasia. This species is hypothesized to have spread to the Americas 1.5 million years ago, founding a population that is at present, a distinct species, Helicoverpa zea. In 2013, H. armigera was confirmed to have re-entered South America via Brazil and subsequently spread. The source of the recent incursion is unknown and population structure in H. armigera is poorly resolved, but a basic understanding would highlight potential biosecurity failures and determine the recent evolutionary history of region-specific lineages. Here, we integrate several end points derived from high-throughput sequencing to assess gene flow in H. armigera and H. zea from populations across six continents. We first assemble mitochondrial genomes to demonstrate the phylogenetic relationship of H. armigera with other Heliothine species and the lack of distinction between populations. We subsequently use de novo genotyping-by-sequencing and whole-genome sequences aligned to bacterial artificial chromosomes, to assess levels of admixture. Primarily, we find that Brazilian H. armigera are derived from diverse source populations, with strong signals of gene flow from European populations, as well as prevalent signals of Asian and African ancestry. We also demonstrate a potential field-caught hybrid between H. armigera and H. zea, and are able to provide genomic support for the presence of the H. armigera conferta subspecies in Australasia. While structure among the bulk of populations remains unresolved, we present distinctions that are pertinent to future investigations as well as to the biosecurity threat posed by H. armigera.


Subject(s)
Gene Flow , Genetics, Population , Moths/genetics , Animals , Brazil , Genome, Insect , Genome, Mitochondrial , Genotype , Hybridization, Genetic , Phylogeny
7.
mBio ; 7(1): e02051-15, 2016 Feb 16.
Article in English | MEDLINE | ID: mdl-26884434

ABSTRACT

UNLABELLED: Salmonella enterica is an intracellular pathogen that causes diseases ranging from gastroenteritis to typhoid fever. Salmonella bacteria trigger an autophagic response in host cells upon infection but have evolved mechanisms for suppressing this response, thereby enhancing intracellular survival. We recently reported that S. enterica serovar Typhimurium actively recruits the host tyrosine kinase focal adhesion kinase (FAK) to the surface of the Salmonella-containing vacuole (SCV) (K. A. Owen et al., PLoS Pathog 10:e1004159, 2014). FAK then suppresses autophagy through activation of the Akt/mTORC1 signaling pathway. In FAK(-/-) macrophages, bacteria are captured in autophagosomes and intracellular survival is attenuated. Here we show that the cell-autonomous bacterial suppression of autophagy also suppresses the broader innate immune response by inhibiting production of beta interferon (IFN-ß). Induction of bacterial autophagy (xenophagy), but not autophagy alone, triggers IFN-ß production through a pathway involving the adapter TRIF and endosomal Toll-like receptor 3 (TLR3) and TLR4. Selective FAK knockout in macrophages resulted in rapid bacterial clearance from mucosal tissues after oral infection. Clearance correlated with increased IFN-ß production by intestinal macrophages and with IFN-ß-dependent induction of IFN-γ by intestinal NK cells. Blockade of either IFN-ß or IFN-γ increased host susceptibility to infection, whereas experimental induction of IFN-ß was protective. Thus, bacterial suppression of autophagy not only enhances cell-autonomous survival but also suppresses more-systemic innate immune responses by limiting type I and type II interferons. IMPORTANCE: Salmonella enterica serovar Typhimurium represents one of the most commonly identified bacterial causes of foodborne illness worldwide. S. Typhimurium has developed numerous strategies to evade detection by the host immune system. Autophagy is a cellular process that involves the recognition and degradation of defective proteins and organelles. More recently, autophagy has been described as an important means by which host cells recognize and eliminate invading intracellular pathogens and plays a key role in the production of cytokines. Previously, we determined that Salmonella bacteria are able to suppress their own autophagic capture and elimination by macrophages. Building on that study, we show here that the inhibition of autophagy by Salmonella also prevents the induction of a protective cytokine response mediated by beta interferon (IFN-ß) and IFN-γ. Together, these findings identify a novel virulence strategy whereby Salmonella bacteria prevent cell autonomous elimination via autophagy and suppress the activation of innate immune responses.


Subject(s)
Autophagy , Interferon-beta/biosynthesis , Interferon-beta/immunology , Macrophages/immunology , Macrophages/microbiology , Salmonella Infections, Animal/immunology , Salmonella typhimurium/pathogenicity , Adaptor Proteins, Vesicular Transport/metabolism , Animals , Focal Adhesion Protein-Tyrosine Kinases/genetics , Focal Adhesion Protein-Tyrosine Kinases/metabolism , Immunity, Innate , Interferon-gamma/biosynthesis , Interferon-gamma/immunology , Intestines/cytology , Killer Cells, Natural/immunology , Mice , Mice, Knockout , Salmonella Infections, Animal/microbiology , Salmonella typhimurium/immunology , Toll-Like Receptor 3/metabolism , Toll-Like Receptor 4/metabolism
8.
Cardiovasc Intervent Radiol ; 36(3): 682-9, 2013 Jun.
Article in English | MEDLINE | ID: mdl-23354958

ABSTRACT

PURPOSE: To study the feasibility of percutaneous radiofrequency ablation (RFA) of large angiomyolipomas (AMLs) using saline-cooled electrodes. MATERIALS AND METHODS: Institutional Review Board approval for the study was received. Four patients (all female, age range 33-67 years) with large AMLs (maximal axis 6.1-32.4 cm) not suitable for embolotherapy or surgery consented to a trial of RFA. Procedures were performed under computerized tomographic guidance using 14G saline-infused electrodes. Two ablations (diameter 4-7 cm) were undertaken in each patient. Variables studied were technical success, treatment safety, alteration of tumor consistency, tumor size, effect on renal function, and medium-term freedom from haemorrhage. RESULTS: All four patients underwent successful RFA without any intraprocedural complications. There has been no haemorrhage, or new renal specific symptom, during a minimum 48-month period, and normal renal function has been normal. On follow-up radiological imaging, the tumors have become fattier with involution of the soft-tissue elements (soft tissue-to-total tumor ratio decreased mean [range] of 0.26 [0.14-0.48] to 0.17 [0.04-0.34] U; p = 0.04 [paired Student t test]). Further evidence of treatment effect was the development of a capsule around the ablation zone, but there was no change in overall tumor volume (mean [range] 1,120 [118-2,845] to 1150 [90-3,013] ml; p = 1 [paired Student t test]). CONCLUSION: RFA of large AMLs is technically feasible using saline-infused electrodes. The soft-tissue elements decreased in volume; the tumors become fattier; and there has been no renal haemorrhage during a 48-month period.


Subject(s)
Angiomyolipoma/surgery , Catheter Ablation/methods , Kidney Neoplasms/surgery , Adult , Aged , Aged, 80 and over , Angiomyolipoma/pathology , Catheter Ablation/instrumentation , Feasibility Studies , Female , Follow-Up Studies , Humans , Kidney Neoplasms/pathology , Middle Aged , Radiography, Interventional , Tomography, X-Ray Computed , Treatment Outcome
9.
Environ Pollut ; 172: 200-7, 2013 Jan.
Article in English | MEDLINE | ID: mdl-23063995

ABSTRACT

Exposures of Lumbricus rubellus to a series of arsenic concentrations in soil were used to assess life-stage (juvenile, adult) and genotype specific sensitivities, to calculate population growth rate (λ) and to assess patterns of As accumulation. Significant mortality was seen in juveniles at 125 mg/kg As, while growth and maturation was affected from 36 mg/kg and above. In adults, cocoon production at the highest concentration (125 mg/kg) was significantly reduced. Phylogenetic analysis was performed by comparison of mitochondrial sequences to establish genotypic variation among juveniles. Three clades with more than 7.5% divergent were described, with 70% of earthworms belonging to a single clade. Date of and mass at maturation was significantly different between clades, but clades were not differentially As sensitive. Parameter λ was reduced at 36 mg/kg As and was negative at 125 mg/kg As, suggesting impacts and population stability and potential extinction at environmentally relevant concentrations.


Subject(s)
Arsenic/toxicity , Life Cycle Stages/drug effects , Soil Pollutants/toxicity , Animals , Genotype , Oligochaeta , Phylogeny , Soil/chemistry
10.
Spinal Cord ; 50(3): 213-9, 2012 Mar.
Article in English | MEDLINE | ID: mdl-22124348

ABSTRACT

STUDY DESIGN: Qualitative study. OBJECTIVE: To determine categories of coping the first year after injury used by 24 young adults who sustained a spinal cord injury (SCI) during adolescence (11-15 years). SETTING: Sweden. METHODS: Content analysis using the existing theories of coping as a framework, including the instrument BriefCOPE-a deductive category application. The analysis looked critically at comments in the interviews that reflected attempts to cope with the injury during the first post-injury year. RESULTS: All 14 of the categories of coping described by the BriefCOPE were included in the interviews at least once, except 'self-blame', which was not used by any interviewee. In addition to the predefined categories of the BriefCOPE, three new coping categories emerged from the interviews: fighting spirit, downward comparison and helping others. CONCLUSIONS: Adolescents who sustain SCIs use a variety of strategies to help them to cope with the consequences of the injury. Many of these coping strategies are similar to those used by others facing stresses, but it is instructive to hear, in their own words, how young adults recall the coping strategies they used as adolescents when they were injured and also how they conceptualized the process of coping. This information can be useful in helping future patients.


Subject(s)
Adaptation, Psychological , Spinal Cord Injuries/psychology , Adolescent , Adult , Female , Humans , Male , Stress, Psychological/etiology , Surveys and Questionnaires , Sweden , Young Adult
11.
Spinal Cord ; 49(2): 200-5, 2011 Feb.
Article in English | MEDLINE | ID: mdl-20603632

ABSTRACT

STUDY DESIGN: Cross-sectional survey. OBJECTIVES: To describe anxiety and depression among caregivers of youth with spinal cord injury (SCI), examine predictors of caregiver psychological functioning and evaluate relationships between caregiver and child psychological outcomes. The protective factor of youth social relationships was also included to examine its impact on relationships between caregiver and child psychological functioning. SETTING: Families received services at one of three pediatric specialty hospitals within a single hospital system in the United States. METHODS: The study included English-speaking youth with SCI, aged between 7 and 17 years, who had been injured at least one year before, and their self-identified primary caregivers. Participants completed surveys assessing their anxiety, depression and youth's perceived social relationships. RESULTS: The study included 203 youth with an average age of 12.70 years (s.d.=3.15), and among them 70% had paraplegia. Seventy-eight percent of caregivers were mothers, 14% fathers and 8% other family members. In all, 16 and 21% of caregivers scored in the range of moderate/severe anxiety and depression, respectively. Being female and having a child with mental health problems predicted caregiver anxiety and depression. In addition, having a child who was older at the time of injury predicted caregiver depression. Poor social relationships, having a caregiver with mental health problems and having a caregiver with less education predicted both child anxiety and depression. CONCLUSION: Caregiver sex, child age at injury and child mental health were related to caregiver outcomes; caregiver education, marital status and child age were not. Caregiver mental health and education and child social relationships predicted child outcomes. Neither injury level nor injury severity was related to caregiver or child outcomes.


Subject(s)
Anxiety/epidemiology , Caregivers/psychology , Depression/epidemiology , Spinal Cord Injuries/nursing , Spinal Cord Injuries/psychology , Adolescent , Age Factors , Anxiety/diagnosis , Child , Cross-Sectional Studies , Female , Humans , Male , Predictive Value of Tests , Prevalence , Psychology , United States/epidemiology
12.
Q J Nucl Med Mol Imaging ; 52(2): 185-92, 2008 Jun.
Article in English | MEDLINE | ID: mdl-18043536

ABSTRACT

Copper-64 (t(1/2)=12.7 h; beta+: 17.4%; E(beta+max)=656 keV; beta-: 39%; E(beta-max)=573 keV) has emerged as an important non-standard positron-emitting radionuclide for positron emission tomography imaging of diseased tissues. A significant challenge of working with copper radionuclides is that they must be delivered to the living system as a stable complex that is attached to a biological targeting molecule for effective imaging and therapy. Significant research has been devoted to the development of ligands that can stably chelate (64)Cu, in particular, the cross-bridged (CB) macrocyclic chelators. This review describes the coordination chemistry and biological behavior of (64)Cu-labeled CB complexes.


Subject(s)
Chelating Agents/chemistry , Copper/chemistry , Cross-Linking Reagents/chemistry , Macrocyclic Compounds/chemistry , Radioisotopes/chemistry , Isotope Labeling/methods , Positron-Emission Tomography/methods , Radiopharmaceuticals/chemistry
13.
Curr Pharm Des ; 13(1): 3-16, 2007.
Article in English | MEDLINE | ID: mdl-17266585

ABSTRACT

Molecular imaging is an important scientific discipline that plays a major role in clinical medicine and pharmaceutical development. While several imaging modalities including X-ray computed tomography (CT) and magnetic resonance imaging (MRI) generate high-resolution anatomical images, positron emission tomography (PET) and single photon emission computed tomography (SPECT) offer insight into the physiological processes that occur within a living organism. Of these two nuclear medicine imaging techniques, PET has advantages with respect to sensitivity and resolution, and this has led to the production and development of many positron emitting radionuclides that include non-traditional radionuclides of the transition metals. Copper-64 (t(1/2) = 12.7 h, beta(+): 17.4%, E(beta+max) = 656 keV; beta(-): 39%, E(beta-max) = 573 keV) has emerged as an important positron emitting radionuclide that has the potential for use in diagnostic imaging and radiotherapy. However, (64)Cu must be delivered to the living system as a stable complex that is attached to a biological targeting molecule for effective imaging and therapy. Therefore, significant research has been devoted to the development of ligands that can stably chelate (64)Cu. This review discusses the necessary characteristics of an effective (64)Cu chelator, while highlighting the development and evaluation of (64)Cu-complexes attached to biologically-targeted ligands.


Subject(s)
Chelating Agents/chemistry , Copper Radioisotopes/chemistry , Positron-Emission Tomography , Radiopharmaceuticals/chemical synthesis , Animals , Cyclams , Cyclohexylamines/chemistry , Dipeptides/chemical synthesis , Drug Design , Drug Stability , Heterocyclic Compounds/chemistry , Humans , Ligands , Molecular Structure , Radiopharmaceuticals/chemistry
14.
Prostate ; 67(1): 74-82, 2007 Jan 01.
Article in English | MEDLINE | ID: mdl-17044087

ABSTRACT

BACKGROUND: Benign prostatic hyperplasia is an age- and androgen-dependent condition of urethral compression caused by prostatic contractility and glandular enlargement. In this study we investigate whether testosterone, dihydrotestosterone and estradiol modulate the ability of human cultured prostatic stromal cells (HCPSCs) to respond to the adrenoceptor agonists, noradrenaline (30 microM) and phenylephrine (100 microM), the protein kinase C activating phorbol ester, phorbol diacetate (PDA, 10 microM), and the L-type Ca(2+) channel activator, (-)-Bay K8644 (Bay K, 10 microM) with elevations of intracellular Ca(2+) ([Ca(2+)](i)). METHODS: Cells were loaded with the Ca(2+) sensitive fluorophore, FURA-2AM (10 microM) and changes in intracellular Ca(2+) determined before and 8-12 min after ligand addition. RESULTS: Compared to steroid-free (SF) controls, the incubation of HCPSC with testosterone (30 and 300 pM) significantly increased responses to both noradrenaline and phenylephrine. Responses to Bay K were significantly reduced between 30 nM to 300 pM but responses to PDA were not greatly affected. Compared to SF the addition of estradiol (E(2), 100 pM) did not affect responses to phenylephrine. The concomitant addition of dihydrotestosterone (DHT) and E(2) (to give ratios from 1:1 to 1,000:1) elevated the responses to noradrenaline and phenylephrine at the extreme ranges. Responses to PDA and Bay K generally increased as DHT:E(2) approached unity. CONCLUSIONS: These results indicate that sex steroids modulate the activities of HCPSCs through the regulation of both receptors and signal transduction processes.


Subject(s)
Calcium/metabolism , Gonadal Steroid Hormones/pharmacology , Prostate/cytology , Prostate/metabolism , Receptors, Adrenergic, alpha-1/metabolism , Adrenergic alpha-1 Receptor Agonists , Aged , Cells, Cultured , Gonadal Steroid Hormones/metabolism , Humans , Male , Phenylephrine/pharmacology , Prostate/drug effects , Stromal Cells/cytology , Stromal Cells/drug effects , Stromal Cells/metabolism
15.
Oncogene ; 25(1): 61-9, 2006 Jan 05.
Article in English | MEDLINE | ID: mdl-16170363

ABSTRACT

Basal telomerase activity is dependent on expression of the hTERT and hTR genes and upregulation of telomerase gene expression is associated with tumour development. It is therefore possible that signal transduction pathways involved in tumour development and features of the tumour environment itself may influence telomerase gene regulation. The majority of solid tumours contain regions of hypoxia and it has recently been demonstrated that hypoxia can increase telomerase activity by mechanisms that are still poorly defined. Here, we show that hypoxia induces the transcriptional activity of both hTR and hTERT gene promoters. While endogenous hTR expression is regulated at the transcriptional level, hTERT is subject to regulation by alternative splicing under hypoxic conditions, which involves a switch in the splice pattern in favour of the active variant. Furthermore, analysis of the chromatin landscape of the telomerase promoters reveals dynamic recruitment of a transcriptional complex involving the hypoxia-inducible factor-1 transcription factor, p300, RNA polymerase II and TFIIB, to both promoters during hypoxia, which traffics along and remains associated with the hTERT gene as transcription proceeds. These studies show that hTERT and hTR are subject to similar controls under hypoxia and highlight the rapid and dynamic regulation of the telomerase genes in vivo.


Subject(s)
DNA-Binding Proteins/biosynthesis , DNA-Binding Proteins/genetics , Gene Expression Regulation, Neoplastic , Gene Expression Regulation , RNA Processing, Post-Transcriptional , RNA/biosynthesis , RNA/genetics , Telomerase/biosynthesis , Telomerase/genetics , Transcription, Genetic , Alternative Splicing , Blotting, Western , Cell Line, Tumor , Chromatin/metabolism , Chromatin Immunoprecipitation , DNA, Complementary/metabolism , Exons , Genetic Variation , Humans , Hypoxia , Hypoxia-Inducible Factor 1, alpha Subunit/metabolism , Luciferases/metabolism , Polymerase Chain Reaction , Promoter Regions, Genetic , RNA Polymerase II/metabolism , Signal Transduction , Telomere/metabolism , Time Factors , Transcription Factor TFIIB/metabolism , Transfection , Vascular Endothelial Growth Factor A , p300-CBP Transcription Factors/metabolism
17.
Vet Comp Oncol ; 2(2): 82-90, 2004 Jun.
Article in English | MEDLINE | ID: mdl-19379188

ABSTRACT

Copper-64 emits beta(+) and beta(-) particles suitable for positron emission tomography and radioimmunotherapy (RIT) of cancer. Copper-64-labelled antibodies have caused complete responses in laboratory animal RIT studies at far lower radiation doses than traditionally prescribed. The intracellular localization of copper radioisotopes may lead to cytotoxic effects by mechanisms beyond ionizing radiation damage. The purpose of this research was to develop a model using both internalizing and non-internalizing antibodies for direct comparison in future RIT studies using the same animal model of cancer. The monoclonal antibodies, cBR96 and cT84.66, were conjugated with N-hydroxysulfosuccinimidyl DOTA. All conjugates retained high immunoreactivity and labelled efficiently with (64)Cu with high specific activity and radiochemical purity. Twenty-four hour biodistributions determined in LS174T tumour-bearing nude mice demonstrated low organ and high tumour uptakes for both monoclonal antibodies. This model constitutes a promising system for elucidating whether internalization of (64)Cu is responsible for an enhanced tumour cytotoxicity in vivo.

18.
Spinal Cord ; 41(12): 684-91, 2003 Dec.
Article in English | MEDLINE | ID: mdl-14639448

ABSTRACT

STUDY DESIGN: Interview using a structured questionnaire and standardized measures. OBJECTIVES: To determine domain-specific satisfaction levels in adults with pediatric-onset spinal cord injuries (SCI), to determine factors associated with these levels of satisfaction, and to determine the relationship of domain-specific satisfaction to overall life satisfaction in this population. SETTING: US and Canada. METHODS: The participants were adults who sustained SCI at age 18 years or younger and were 24 years of age or older at the time of interview and did not have significant head injury. In addition to providing information about themselves, including education level, employment, marital status, and community participation, they were asked to rate their level of satisfaction in seven domains: transportation in the community, educational achievement, employment opportunities, income, social/recreational opportunities, dating opportunities, and sexual experience. They also completed the satisfaction with life scale (SWLS), the Craig handicap assessment and reporting technique, the functional independence measure, and the short-form-12 perceived health scale. RESULTS: A total of 216 individuals were interviewed. The mean age at injury was 14 years and the mean age at interview was 29 years. From most satisfied to least satisfied, the domains were ranked in the following order: satisfaction with transportation in the community, educational achievement, social and recreational opportunities, sexual experiences, dating opportunities, job opportunities, and income. Age at interview, gender, and perceived health were identified in regression analyses as predictors of some of the domain-specific satisfactions, but the primary predictive factors were in the area of participation. Neither severity of neurologic impairment nor level of functional independence were predictors for any of the domains. Satisfaction in each of the domains was significantly associated with SWLS and satisfaction with dating, job opportunities, education, and income were identified as predictive factors in a regression analysis. CONCLUSIONS: Dating opportunities, job opportunities, and income are the three domains in which adults with pediatric-onset SCI are least satisfied and those domains have a significant impact on overall satisfaction.


Subject(s)
Activities of Daily Living , Patient Satisfaction , Physical Therapy Modalities/methods , Quality of Life , Spinal Cord Injuries/epidemiology , Spinal Cord Injuries/rehabilitation , Adolescent , Adult , Age of Onset , Canada/epidemiology , Child , Female , Health Care Surveys , Humans , Injury Severity Score , Linear Models , Male , Middle Aged , Multivariate Analysis , Outcome Assessment, Health Care , Paraplegia/diagnosis , Paraplegia/rehabilitation , Probability , Quadriplegia/diagnosis , Quadriplegia/rehabilitation , Sickness Impact Profile , Spinal Cord Injuries/diagnosis , Surveys and Questionnaires , United States/epidemiology
19.
Q J Nucl Med ; 47(3): 201-8, 2003 Sep.
Article in English | MEDLINE | ID: mdl-12897711

ABSTRACT

metastatic Matrix metalloproteinases (MMPs) are zinc-dependent secreted or transmembrane enzymes constituting a family of over 21 proteolytic members that are capable of selectively digesting a wide spectrum of both extracellular matrix (ECM) and nonmatrix proteins. MMPs play a critical role in tumor growth, angiogenesis, and metastatic processes. MMP inhibitors (MMPIs) have been extensively investigated as anti-tumor drugs, although the clinical trials thus far have been disappointing. In order to better understand the role of MMPs in cancer growth and metastasis, as well as improve the therapeutic efficacy of MMPIs, there is a need to develop new procedures to assess and/or monitor MMP activity in vivo. In addition to determining whether MMPs are present in tumors, it would be desirable to have an imaging agent that better probes other processes associated with MMP overproduction, including angiogenesis and the establishment of the growth of metastatic lesions in distant organ sites. In this paper we review the studies relating to the recent development of in vivo imaging of MMP expression. One of the purposes of this review is to discuss the current status of imaging MMP expression, which includes the types of tracers being developed and the types of imaging modalities available. Although imaging MMP expression is a relatively new area of research, the progress thus far is highly promising.


Subject(s)
Diagnostic Imaging/methods , Matrix Metalloproteinases/metabolism , Neoplasms/diagnosis , Neoplasms/metabolism , Neovascularization, Pathologic/diagnosis , Neovascularization, Pathologic/metabolism , Radioisotopes/pharmacokinetics , Animals , Humans , Matrix Metalloproteinase Inhibitors , Matrix Metalloproteinases/classification , Neoplasms/blood supply , Spectroscopy, Near-Infrared/methods , Tomography, Emission-Computed/methods
20.
Spinal Cord ; 40(12): 666-8, 2002 Dec.
Article in English | MEDLINE | ID: mdl-12483501

ABSTRACT

STUDY DESIGN: A case report of self-injurious behavior in four children and adolescents with spinal cord injuries (SCI). OBJECTIVES: To report a relatively unusual complication of pediatric-onset SCI, focusing on the potential role that dysesthesia may play in self-injurious behavior. SETTING: A Spinal Cord Injury Program in a Children's Hospital in Chicago that serves children from midwestern and south-central United States of America. METHOD: Case reports and literature review. RESULTS: Case reports are presented of four children or adolescents with SCI who exhibited self-injurious behavior. Two of the subjects had symptoms consistent with dysesthesia. The self-injurious behavior in these two subjects and a very young child responded to treatment with anticonvulsants. The self-injurious behavior in the fourth patient was probably the result of poor technique of using his mouth to move his hands, which responded to conservative management including education, occupational therapy and gloves. CONCLUSION: Self-injurious behavior is a relatively unusual complication of pediatric onset SCI, and may be a manifestation of dysesthesia and be responsive to treatment with anticonvulsants.


Subject(s)
Acetates/therapeutic use , Amines , Anticonvulsants/therapeutic use , Carbamazepine/therapeutic use , Cyclohexanecarboxylic Acids , Self Mutilation/drug therapy , Self Mutilation/etiology , Spinal Cord Injuries/complications , gamma-Aminobutyric Acid , Adolescent , Child , Female , Gabapentin , Humans , Male , Occupational Therapy , Paresthesia/physiopathology , Self Mutilation/therapy , Spinal Cord Injuries/physiopathology
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