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Charles M Krafchak,Hemant Pawar,Sayoko E Moroi,Alan Sugar,Paul R Lichter,David A Mackey,Shahzad Mian,Theresa Nairus,Victor Elner,Miriam T Schteingart,Catherine A Downs,Theresa Guckian Kijek,Jenae M Johnson,Edward H Trager,Frank W Rozsa,Md Nawajes Ali Mandal,Michael P Epstein,Douglas Vollrath,Radha Ayyagari,Michael Boehnke,Julia E Richards Posterior polymorphous corneal dystrophy (PPCD, also known as PPMD) is a rare disease involving metaplasia and overgrowth of corneal endothelial cells. In patients with PPCD, these cells manifest in an epithelial morphology and gene expression pattern, produce an aberrant basement membrane, and, so... ( view more )metimes, spread over the iris and nearby structures in a way that increases the risk for glaucoma. We previously mapped PPCD to a region (PPCD3) on chromosome 10 containing the gene that encodes the two-handed zinc-finger homeodomain transcription factor TCF8. Here, we report a heterozygous frameshift mutation in TCF8 that segregates with PPCD in the family used to map PPCD3 and four different heterozygous nonsense and frameshift mutations in TCF8 in four other PPCD probands. Family reports of inguinal hernia, hydrocele, and possible bone anomalies in affected individuals suggest that individuals with TCF8 mutations should be examined for nonocular anomalies. We detect transcripts of all three identified PPCD genes (VSX1, COL8A2, and TCF8) in the cornea. We show presence of a complex (core plus secondary) binding site for TCF8 in the promoter of Alport syndrome gene COL4A3, which encodes collagen type IV alpha 3, and we present immunohistochemical evidence of ectopic expression of COL4A3 in corneal endothelium of the proband of the original PPCD3 family. Identification of TCF8 as the PPCD3 gene provides a valuable tool for the study of critical gene regulation events in PPCD pathology and suggests a possible role for TCF8 mutations in altered structure and function of cells lining body cavities other than the anterior chamber of the eye. Thus, this study has identified TCF8 as the gene responsible for approximately half of the cases of PPCD, has implicated TCF8 mutations in developmental abnormalities outside the eye, and has presented the TCF8 regulatory target, COL4A3, as a key, shared molecular component of two different diseases, PPCD and Alport syndrome. ( view less ) Kenji Kabashima,Theresa A Banks,K Mark Ansel,Theresa T Lu,Carl F Ware,Jason G Cyster The factors regulating dendritic cell (DC) development and homeostasis are incompletely understood. Here, we demonstrate that DCs express the lymphotoxin (LT)-beta receptor (LT beta R) and that in mice lacking the LT beta R in hematopoietic cells, spleen, and lymph node, CD8- DC numbers are reduced... ( view more ). B cells are a key source of LT alpha 1 beta 2 for splenic DC homeostasis, and transgenic overexpression of LT alpha 1 beta 2 on B cells leads to expansion of the CD8- DC compartment. Furthermore, we find that about 5% of splenic DCs are undergoing cell division, and the number of dividing CD8- DCs is disproportionately reduced in the absence of the LT beta R. In parabiosis experiments, splenic DCs were only partially replaced by circulating precursors over a 6 week period. We conclude that LT alpha 1 bet a2 acts on DCs or DC precursors to promote DC homeostasis, and we suggest that DC proliferation is an important pathway for locally maintaining these cells in the steady state. ( view less ) P Richard Harrigan,Theresa Mo,Brian Wynhoven,Jennifer Hirsch,Zabrina Brumme,Paula McKenna,Theresa Pattery,Johan Vingerhoets,Lee T Bacheler BACKGROUND: The K103N mutation in HIV-1 reverse transcriptase (RT) confers high-level resistance to current non-nucleoside reverse transcriptase inhibitors (NNRTI). The prevalence and resistance profile of HIV-1 with other substitutions at RT codon 103 is less well documented. METHODS: K103 substit... ( view more )utions among over 70,000 clinical samples submitted for routine antiretroviral resistance testing at two independent centres were examined. Phenotypic resistance profiles of isolates harboring rare K103 variants in the absence of known NNRTI-associated resistance mutations were retrieved from Virco's correlative genotype/phenotype database. Genotyped samples with known treatment histories were retrieved from the British Columbia Centre for Excellence in HIV/AIDS database. Site-directed mutants containing K103 variants were constructed and phenotyped. RESULTS: K103N, R and S were observed in 29, 1.8, and 0.9% of Virco isolates and in 16, 1.5 and 0.4% of British Columbia isolates. K103T/Q/H substitutions were observed only rarely (<0.2%). The prevalence of unusual codon 103 substitutions remained stable over 5 years, except K103S, which increased over fourfold in both datasets. K103R/Q-containing clinical isolates remained phenotypically susceptible to NNRTI, whereas K103S/T/H-containing isolates showed over 10-fold decreased NNRTI susceptibility. Among patients with a known treatment history, K103S/T/H were observed primarily in individuals failing NNRTI-containing regimens. Site-directed mutants confirmed decreased susceptibility to NNRTI in K103S/T/H-containing recombinants. CONCLUSION: Variants at HIV RT codon 103 other than K103N are observed relatively rarely in clinical isolates, but K103 S, T and H confer decreased susceptibility to NNRTI. These data are relevant for interpretive genotype algorithms and in the design of assays specific to RT codon 103 mutations. ( view less ) Theresa Mitchell,Steve JonesLeading and co-ordinating a research project is a complex undertaking involving management of people, processes, budget and material resources. The task is even more complex when there are nine nurse researchers collecting and analysing data in the midst of their practice. In this paper, Theresa Mi... ( view more )tchell and Steve Jones describe the experiences of the project lead and co-ordinator responsible for planning and managing such a project in the Oncology Centre in Gloucestershire, UK. Using examples from the project, the authors share some of the potential problems encountered during research activities, and propose strategies and remedies to address them. ( view less ) Don C Des Jarlais,Theresa Diaz,Theresa Perlis,David Vlahov,Carey Maslow,Mary Latka,Russell Rockwell,Vincent Edwards,Samuel R Friedman,Edgar Monterroso,Ian Williams,Richard S Garfein Cohort studies of young (aged 18-30 years) injecting drug users recruited in 1997-1999 in the Harlem and Lower East Side areas of New York City, New York, were used to assess the incidence of human immunodeficiency virus (HIV), hepatitis B virus (HBV), and hepatitis C virus (HCV). The authors found... ( view more ) that HIV incidence was low at both sites: 0.8/100 person-years at the Harlem site and 0/100 person-years at the Lower East Side site. In contrast, HBV incidence was moderate (12.2/100 person-years) at the Harlem site and high (30.7/100 person-years) at the Lower East Side site. Similarly, HCV incidence was moderate (9.3/100 person-years) at the Harlem site and high (34.0/100 person-years) at the Lower East Side site. Results show that high rates of HBV and HCV transmission do not imply high rates of HIV transmission, even within an area of high HIV seroprevalence. ( view less ) Wei Qi,Aming Zhang,Dhara Patel,Sungmun Lee,Jamie L Harrington,Liming Zhao,David Schaefer,Theresa A Good,Erik J Fernandez Increasing evidence indicates that soluble aggregates of amyloid beta protein (Abeta) are neurotoxic. However, difficulty in isolating these unstable, dynamic species impedes studies of Abeta and other aggregating peptides and proteins. In this study, hydrogen-deuterium exchange (HX) detected by ma... ( view more )ss spectrometry (MS) was used to measure Abeta(1-40) aggregate distributions without purification or modification that might alter the aggregate structure or distribution. Different peaks in the mass spectra were assigned to monomer, low molecular weight oligomer, intermediate, and fibril based on HX labeling behavior and complementary assays. After 1 h labeling, the intermediates incorporated approximately ten more deuterons relative to fibrils, indicating a more solvent exposed structure of such intermediates. HX-MS also showed that the intermediate species dissociated much more slowly to monomer than did the very low molecular weight oligomers that were formed at very early times in Abeta aggregation. Atomic force microscopy (AFM) measurements revealed the intermediates were roughly spherical with relatively homogenous diameters of 30-50 nm. Quantitative analysis of the HX mass spectra showed that the amount of intermediate species was correlated with Abeta toxicity patterns reported in a previous study under the same conditions. This study also demonstrates the potential of the HX-MS approach to characterizing complex, multi-component oligomer distributions of aggregating peptides and proteins. ( view less ) Jobiba Chinkhumba,Andrew Tomkins,Theresa Banda,Catherine Mkangama,Pamela Fergusson A prospective cohort study measured mortality during nutritional rehabilitation among HIV-infected and uninfected children, aged 6-59 months, with severe acute malnutrition (SAM). Children were tested for HIV and CD4% on admission to the nutrition rehabilitation unit (NRU). Mortality was assessed b... ( view more )y following children to 4 months post discharge from the NRU or death if earlier. Overall mortality was 14.8% (67/454) and HIV prevalence was 17.4% (79/454). HIV-infected children were significantly more likely to die than uninfected children [35.4% (28/79) vs. 10.4% (39/375), P<0.001], and 85.7% of deaths occurred in children with a CD4% less than 20. Forty percent (18/45) of HIV-infected children with a CD4% <20 died, in contrast to 15% (3/20) of HIV-infected children with a CD4% >20 (P=0.05). Routine testing and treatment for HIV among all malnourished children is necessary to improve quality of care and reduce mortality among children with SAM. ( view less ) Theresa L Cromeans,Xiaoyan Lu,Dean D Erdman,Charles D Humphrey,Vincent R Hill Enteric adenoviruses, important agents of infantile gastroenteritis, are difficult to culture with low titers and limited CPE. Consequently, few plaque assays have been reported and none are used routinely by investigators who may need reproducible quantitative assays for these viruses. CPE in A549... ( view more ) cells (an epithelial lung carcinoma cell line) was induced by isolates of human adenovirus (HAdV) serotypes 40 or 41 that were obtained by prior limited passage in primary cynmolgous monkey kidney (pCMK), human embryonic kidney (HEK), and Graham 293 cells. CPE with HAdV 40 (Dugan strain) and HAdV 41 (Tak strain) inoculated in A549 cells was also observed. Monolayers of A549 cells were inoculated with a low multiplicity of infection (MOI) of the archived stock isolates and harvested at days 10-14 with full CPE. Subsequent passages were harvested in as few as 7 days with 100% CPE to prepare virus stocks for plaque assay. Large individual plaques under agarose overlay were picked prior to staining and clonal stocks prepared. Titers of final stock preparations after six to eight passages in A549 cells were in the range of 5x10(7)-1x10(8)PFU/ml, which provides adequate virus for quantitative recovery studies. The particle to infectivity (P:I) ratios of the early passages of virus stocks were in the range reported previously. The ratio of non-infectious to infectious particles decreased with successive passages of HAdVs 40 and 41 in A549 cells. The specificity of the assay was confirmed by neutralization of plaques with type-specific antisera. Furthermore, sequence analysis of the HAdVs 40 and 41 plaque forming stocks ruled out contamination with any other HAdVs. The plaque assay developed will be useful for evaluation of virus recovery methods from water, food or other environmental matrices, as well as determination of the efficacy of water treatment techniques for inactivation of these viruses. ( view less ) Michael P Carey,Peter A Vanable,Theresa E Senn,Patricia Coury-Doniger,Marguerite A Urban BACKGROUND: Sexually transmitted infection (STI) clinics provide an opportune setting for HIV prevention efforts. This randomized controlled trial evaluated a unique, two-step approach to sexual risk reduction at a publicly-funded STI clinic. METHODS: During an initial visit, patients completed an ... ( view more )audio-computer assisted self-interview (ACASI), were randomized to and received one of two brief interventions, obtained medical care, and completed a post-assessment. Next, two-thirds of the patients were assigned to attend an intensive sexual risk reduction workshop. At 3, 6, and 12 months, patients completed additional ACASIs and provided urine specimens to assess behavior change and incident STIs. RESULTS: During a 28-month interval, 5613 patients were screened, 2691 were eligible, and 1483 consented to participate and were randomized; the modal reason for declining was lack of time (82%). Consenting patients included 688 women and 795 men; 64% of participants were African-American. The sample was low-income, with 57% reporting an annual income of less than $15,000; most participants (62%) had a high school education or less, and 51% were unemployed. Sexual risk behavior was common, as indicated by multiple sexual partners (mean=32.8, lifetime; mean=2.8, past 3 months), unprotected sex (mean=17.3 episodes, past 3 months), and prior STIs (mean=3.3, lifetime; 23% at baseline). Bivariate analyses confirmed our prediction that HIV-related motivation and behavioral skills would be related to current sexual risk behavior. All patients received a brief intervention; patient satisfaction ratings were uniformly high for both interventions (all means >or=3.7 on 4-point scales). Fifty-six percent of invited patients attended the intensive workshop, and attendance did not differ as a function of brief intervention. Patient satisfaction ratings were also uniformly positive for the workshop interventions (all means >or=3.6). Return to follow-up assessments exceeded 70%. CONCLUSIONS: Results demonstrate that implementing an HIV preventive program in a busy, public clinic is feasible and well-accepted by patients. Ongoing evaluation will determine if the interventions reduce sexual risk behavior and lower incident STIs. ( view less ) Eila K Roberts,Vasantha Padmanabhan,Theresa M Lee The process of sexual differentiation leaves genetically female individuals at risk of being masculinized by exogenous androgens. Previous research with sheep indicates that exposure to excess testosterone from Gestational Day (GD) 30 to GD 90 of the 147-day gestation masculinizes and defeminizes b... ( view more )ehavior as well as genitalia. Lower doses and shorter durations produce animals with varying degrees of genital virilization and alterations of the hypothalamic-pituitary-gonadal axis, but to our knowledge, the effects on complex behavior and its prediction by the amount of external virilization have not been explored. Previous research in rodents has suggested that sexual differentiation of the central nervous system and the external genitalia can be dissociated. Therefore, we hypothesized that the extent of virilization of external genitalia would not be predictive of the lack of female-typical, or the presence of male-typical, mating behavior. To test this hypothesis, we compared control females, females exposed to exogenous testosterone from GD 30 to GD 90 (T60 females) that have virilized genitalia, and females exposed to testosterone from GD 60 to GD 90 (T30 females) that have female-typical genitalia. Both natural behavioral estrus in the flock and hormonally controlled behavioral tests were used to explore reproductive behavior. The T60 and T30 females exhibited more masculinized reproductive behavior than the controls; however, the T30 females also exhibited feminine behavior. Neither testosterone-treated group was receptive or was mounted at rates comparable to those of controls. These data illustrate that variation in the timing or duration of exposure to prenatal testosterone during a critical period for masculinization can have variable effects on defeminization and that the effects of testosterone on genitalia are not entirely predictive of behavior. ( view less ) Theresa M Casey,Jonathan Eneman,Abigail Crocker,Jeffrey White,Joseph Tessitore,Mary Stanley,Seth Harlow,Janice Y Bunn,Donald Weaver,Hyman Muss,Karen Plaut Cancer associated fibroblasts (CAFs) are believed to promote tumor growth and progression. Our objective was to measure the effect of TGF-beta1 on fibroblasts isolated from invasive breast cancer patients. Fibroblasts were isolated from tissue obtained at surgery from patients with invasive breast ... ( view more )cancer (CAF; n = 28) or normal reduction mammoplasty patients (normal; n = 10). Myofibroblast activation was measured by counting cells immunostained for smooth muscle alpha actin (ACTA2) in cultures +/- TGF-beta1. Conditioned media (CM) was collected for invasion assays and RNA was isolated from cultures incubated in media +/- TGF-beta1 for 24 h. Q-PCR was used to measure expression of cyclin D1, fibronectin, laminin, collagen I, urokinase, stromelysin-1, and ACTA2 genes. Invasion rate was measured in chambers plated with MDA-MB-231 cells and exposed to CM in the bottom chamber; the number of cells that invaded into the bottom chamber was counted. Wilcox Rank Sum tests were used to evaluate differences in CAFs and normal fibroblasts and the effect of TGF-beta1. There was no difference in percent myofibroblasts or invasion rate between normal and CAF cultures. However, TGF-beta1 significantly increased the percent of myofibroblasts (P < 0.01) and invasion rate (P = 0.02) in CAF cultures. Stromelysin-1 expression was significantly higher in normal versus CAF cultures (P < 0.01). TGF-beta1 significantly increased ACTA2 expression in both normal and CAF cultures (P < 0.01). Expression of fibronectin and laminin was significantly increased by TGF-beta in CAF cultures (P < 0.01). CAFs were measurably different from normal fibroblasts in response to TGF-beta1, suggesting that TGF-beta stimulates changes in CAFs that foster tumor invasion. ( view less ) Nimmi Arora,Theresa Scognamiglio,Baixin Zhu,Thomas J Fahey Iii BACKGROUND: Benign thyroid tumors account for most nodular thyroid disease. Determination of whether a thyroid nodule is benign or malignant is a major clinical dilemma and underlies the decision to proceed to surgery in many patients. Although the accuracy of thyroid nodule fine-needle aspiration ... ( view more )(FNA) has reduced the need for surgery over the years, questions regarding how to follow FNA-designated benign nodules remain unresolved. This is true at least in part because of uncertainty over whether some benign nodules harbor malignant potential. METHODS: An evidence-based review of recent clinical, pathologic, and molecular data is presented. A summary of data and observations from our own experience is also provided. RESULTS: Review of our recent 10-year experience indicates that 2% of thyroid malignancies arise within a preexisting benign thyroid nodule. In addition, both cytologic and molecular tumor markers, including Gal-3, CITED1, HBME-1, Ras, RET/PTC, and PAX8/PPARgamma, have been identified in some histopathologically classified benign nodules. Gene expression profiling suggests that follicular adenomas and Hürthle cell adenomas have similarities to both benign and malignant tumors, suggesting that some of these tumors are premalignant. In addition, 10% of surgically excised follicular tumors are encapsulated follicular lesions with nuclear atypia, which have been termed "well-differentiated tumors of uncertain malignant potential." The data available suggest that these tumors could be precursors to carcinoma. CONCLUSION: Some benign thyroid nodules have malignant potential. Further molecular testing of these tumors can shed light on the pathogenesis of early malignant transformation. ( view less ) Stephan W Glasser,Albert P Senft,Jeffrey A Whitsett,Melissa D Maxfield,Gary F Ross,Theresa R Richardson,Daniel R Prows,Yan Xu,Thomas R Korfhagen To determine the role of surfactant protein C (SP-C) in host defense, SP-C-deficient (Sftpc(-/-)) mice were infected with the pulmonary pathogen Pseudomonas aeruginosa by intratracheal injection. Survival of young, postnatal day 14 Sftpc(-/-) mice was decreased in comparison to Sftpc(+/+) mice. The... ( view more ) sensitivity to Pseudomonas bacteria was specific to the 129S6 strain of Sftpc(-/-) mice, a strain that spontaneously develops interstitial lung disease-like lung pathology with age. Pulmonary bacterial load and leukocyte infiltration were increased in the lungs of Sftpc(-/-) mice 24 h after infection. Early influx of polymorphonuclear leukocytes in the lungs of uninfected newborn Sftpc(-/-) mice relative to Sftpc(+/+) mice indicate that the lack of SP-C promotes proinflammatory responses in the lung. Mucin expression, as indicated by Alcian blue staining, was increased in the airways of Sftpc(-/-) mice following infection. Phagocytic activity of alveolar macrophages from Sftpc(-/-) mice was reduced. The uptake of fluorescent beads in vitro and the number of bacteria phagocytosed by alveolar macrophages in vivo was decreased in the Sftpc(-/-) mice. Alveolar macrophages from Sftpc(-/-) mice expressed markers of alternative activation that are associated with diminished pathogen response and advancing pulmonary fibrosis. These findings implicate SP-C as a modifier of alveolar homeostasis. SP-C plays an important role in innate host defense of the lung, enhancing macrophage-mediated Pseudomonas phagocytosis, clearance and limiting pulmonary inflammatory responses. ( view less ) Theresa A Gannon,Rachael M Collie,Tony Ward,Jo Thakker Whether treatment programs are effective at rehabilitating rapists is yet to be determined empirically. From a scientist-practitioner perspective, treatment should be based on an empirical understanding of rape and rapists, and evidence-based knowledge of treatment outcome with rapists. In this pap... ( view more )er we comprehensively review the characteristics of rapists, etiological features implicated in the commission of rape, and relevant treatment outcome research. We pay particular attention to contemporary knowledge about the core vulnerabilities and features required to understand and treat rapists effectively, and, where possible, highlight similarities and differences between rapists, child molesters and non-sexual violent offenders. We use an epistemological framework to (a) critique the various etiological accounts of rape available and (b) help guide professionals' use of such knowledge in both treatment design and evaluation. Gaps in the understanding of rapists' characteristics and etiological features are highlighted, as are discrepancies between current knowledge and treatment approaches. We conclude by highlighting areas for future research and practice innovation. ( view less ) Kenneth Gundersen,Jan Terje Kvaløy,Jo Kramer-Johansen,Theresa Mariero Olasveengen,Joar Eilevstjønn,Trygve Eftestøl BACKGROUND: Analysis of the electrocardiogram (ECG) can to a certain extent predict if a cardiac arrest patient in ventricular fibrillation will get return of spontaneous circulation (ROSC) if defibrillated. The accuracy of such methods determines how useful it is clinically and for retrospective a... ( view more )nalysis. METHODS AND RESULTS: We have tested the accuracy of a new shock outcome prediction algorithm that is the first to use an updating algorithm capable of learning from previous shocks within a resuscitation effort. The algorithm relies on known predictive features from the pre-shock ECG, but for each delivered shock it re-estimates the patient-dependent relationship between predictive feature value and probability of ROSC by incorporating the information from the already performed shocks. The predictive features mean-slope, median-slope, cardioversion-outcome-predictor and amplitude-spectrum-analysis originally had areas under the receiver operating characteristics curve of 0.843, 0.846, 0.837 and 0.819, respectively. The improvements in areas after applying the algorithm were (bootstrap estimate of mean improvement, 95% confidence interval in parentheses): mean-slope, 0.019 (0.00036, 0.042); median-slope, 0.024 (0.0013, 0.048); cardioversion-outcome-predictor, 0.021 (0.0010, 0.051); amplitude-spectrum-analysis, 0.026 (0.0016, 0.051). The predictions for the first shock to each patient were not included when calculating the areas, as for the first shocks the new algorithm has no previous shocks to learn from and give predictions identical to those of the original features. CONCLUSIONS: It is possible to improve current shock prediction methods by using an updating algorithm capable of learning from previous shocks within a resuscitation effort. ( view less ) Kevin Chatham-Stephens,Theresa Devere,Judith Guzman-Cottrill,Peter KurreSweet's syndrome (SS) is an acute, febrile neutrophilic dermatosis that frequently presents with leukocytosis and erythematous plaques. Lesions show neutrophilic infiltration of the dermis and rarely other organs. We report the case of an adolescent male with chronic pancytopenia secondary to Fanco... ( view more )ni anemia (FA) who presented with acute respiratory distress. Despite an exhaustive and ultimately unrevealing work-up, the diagnosis of pulmonary SS was not made until he developed characteristic cutaneous lesions 4 months later. Comprehensive review of pathological specimens revealed metachronous SS manifestations with infiltrates in lung parenchyma, dermis, and subcutis in this neutropenic patient. ( view less ) Eirik Skogvoll,Trygve Eftestøl,Kenneth Gundersen,Jan Terje Kvaløy,Jo Kramer-Johansen,Theresa Mariero Olasveengen,Petter Andreas Steen BACKGROUND: The state or rhythm during resuscitation, i.e. ventricular fibrillation/tachycardia (VF/VT), asystole (ASY), pulseless electrical activity (PEA), or return of spontaneous circulation (ROSC) determines management. The state is unstable and will change either spontaneously (e.g. PEA-->ASY... ( view more )) or by intervention (e.g. VF-->ASY after DC shock); temporary ROSC may also occur. To gain insight into the dynamics of this process, we analyzed the state transitions over time using real-life data. METHODS: Detailed recordings from 304 episodes of attempted resuscitation from out-of-hospital cardiac arrests of presumed cardiac etiology were obtained from modified Heartstart((R)) 4000 defibrillators. State transitions were visualized and described, and analyzed in terms of a Markov probability model. RESULTS: The median number of state transitions was 5 (range 1-39), and more transitions were observed with VF than PEA or asystole as the initial rhythm. Of 105 patients (35%) who regained ROSC at some point during CPR, only 65 (21%) achieved sustained ROSC; suggesting an unrealized survival potential. A 3-min transition probability matrix was estimated: for example, a patient early in VF has a probability of 31% to be in ASY, 32% of still being in VF, 5% to have temporary ROSC, and 2% to have sustained ROSC after 3min. CONCLUSION: The dynamics of resuscitation can be described in terms of state transitions and a Markov probability model. This framework enables prediction of short-term clinical development, supports informed decisions during CPR, and suggests a novel area for research. ( view less ) John J Osterholzer,Jeffrey L Curtis,Timothy Polak,Theresa Ames,Gwo-Hsiao Chen,Rod McDonald,Gary B Huffnagle,Galen B Toews Pulmonary clearance of the encapsulated yeast Cryptococcus neoformans requires the development of T1-type immunity. CCR2-deficient mice infected with C. neoformans develop a non-protective T2 immune response and persistent infection. The mechanisms responsible for this aberrant response are unknown... ( view more ). The objective of this study was to define the number, phenotype, and microanatomic location of dendritic cells (DC) residing within the lung of CCR2(+/+) or CCR2(-/-) mice throughout a time course following infection with C. neoformans. Results demonstrate the CCR2-mediated recruitment of conventional DC expressing modest amounts of costimulatory molecules. DC recruitment was preceded by the up-regulation in the lung of the CCR2 ligands CCL2 and CCL7. Colocalization of numerous DC and CD4(+) T cells within bronchovascular infiltrates coincided with increased expression of IL-12 and IFN-gamma. By contrast, in the absence of CCR2, DC recruitment was markedly impaired, bronchovascular infiltrates were diminished, and mice developed features of T2 responses, including bronchovascular collagen deposition and IL-4 production. Our results demonstrate that CCR2 is required for the recruitment of large numbers of conventional DC to bronchovascular infiltrates in mice mounting a T1 immune response against a fungal pathogen. These findings shed new insight into the mechanism(s) by which DC recruitment alters T cell polarization in response to an infectious challenge within the lung. ( view less ) Margaret L Hunter,Rajiv Patel,Theresa Loyn,Maria Z Morgan,Ruth Fairchild,Jeremy S Rees BACKGROUND: Only one previous study has investigated the effect which dilution has on the erosive potential of dilutable fruit drinks. Based solely on measurement of neutralizable acidity, the authors concluded that the erosive potential of diluting juices may be reduced substantially by the additi... ( view more )on of water. This has not been verified in an in vitro enamel erosion model. OBJECTIVE: The aim of this study was to examine the effect of dilution on the erosive potential of five dilutable fruit drinks. METHODS: For each product, the erosive potential of three dilutions (1 : 3, 1 : 6 and 1 : 15) was assessed by measuring the initial pH and neutralizable acidity; in vitro enamel erosion was measured by profilometry following immersion for 1 h. RESULTS: For the majority of products, increasing dilution from 1 : 3 to 1 : 15 produced a statistically significant rise in initial pH, though they remained markedly acidic. Increasing the dilution factor consistently produced a statistically significant decrease in neutralizable acidity. Increasing the dilution factor from 1 : 3 to 1 : 15 produced a statistically significant reduction in in vitro enamel erosion in only three of the five products tested. CONCLUSIONS: Increasing the dilution factor of dilutable fruit drinks within a range likely to be acceptable to the consumer may not effectively reduce their erosive potential. ( view less ) John J McAtee,Jason W Dodson,Sarah E Dowdell,Karl Erhard,Gerald R Girard,Krista B Goodman,Mark A Hilfiker,Jian Jin,Clark A Sehon,Deyou Sha,Dongchuan Shi,Feng Wang,Gren Z Wang,Ning Wang,Yonghui Wang,Andrew Q Viet,Catherine C K Yuan,Daohua Zhang,Nambi V Aiyar,David J Behm,Luz H Carballo,Christopher A Evans,Harvey E Fries,Rakesh Nagilla,Theresa J Roethke,Xiaoping Xu,Stephen A Douglas,Michael J NeebLead compound 1 was successfully redesigned to provide compounds with improved pharmacokinetic profiles for this series of human urotensin-II antagonists. Replacement of the 2-pyrrolidinylmethyl-3-phenyl-piperidine core of 1 with a substituted N-methyl-2-(1-pyrrolidinyl)ethanamine core as in compou... ( view more )nd 7 resulted in compounds with improved oral bioavailability in rats. The relationship between stereochemistry and selectivity for hUT over the kappa-opioid receptor was also explored. ( view less ) DeAnna L Adkins,J Edward Hsu,Theresa A Jones Cortical stimulation (CS) as a means to modulate regional activity and excitability in cortex is emerging as a promising approach for facilitating rehabilitative interventions after brain damage, including stroke. In this study, we investigated whether CS-induced functional improvements are linked ... ( view more )with synaptic plasticity in peri-infarct cortex and vary with the severity of impairments. Adult rats that were proficient in skilled reaching received subtotal unilateral ischemic sensorimotor cortex (SMC) lesions and implantation of chronic epidural electrodes over remaining motor cortex. Based on the initial magnitude of reaching deficits, rats were divided into severely and moderately impaired subgroups. Beginning two weeks post-surgery, rats received 100 Hz cathodal CS at 50% of movement thresholds or no-stimulation control procedures (NoCS) during 18 days of rehabilitative training on a reaching task. Stereological electron microscopy methods were used to quantify axodendritic synapse subtypes in motor cortical layer V underlying the electrode. In moderately, but not severely impaired rats, CS significantly enhanced recovery of reaching success. Sensitive movement analyses revealed that CS partially normalized reaching movements in both impairment subgroups compared to NoCS. Additionally, both CS subgroups had significantly greater density of axodendritic synapses and moderately impaired CS rats had increases in presumed efficacious synapse subtypes (perforated and multiple synapses) in stimulated cortex compared to NoCS. Synaptic density was positively correlated with post-rehabilitation reaching success. In addition to providing further support that CS can promote functional recovery, these findings suggest that CS-induced functional improvements may be mediated by synaptic structural plasticity in stimulated cortex. ( view less ) John J McAtee,Jason W Dodson,Sarah E Dowdell,Gerald R Girard,Krista B Goodman,Mark A Hilfiker,Clark A Sehon,Deyou Sha,Gren Z Wang,Ning Wang,Andrew Q Viet,Daohua Zhang,Nambi V Aiyar,David J Behm,Luz H Carballo,Christopher A Evans,Harvey E Fries,Rakesh Nagilla,Theresa J Roethke,Xiaoping Xu,Catherine C K Yuan,Stephen A Douglas,Michael J NeebThis work describes the development of potent and selective human Urotensin-II receptor antagonists starting from lead compound 1, (3,4-dichlorophenyl)methyl{2-oxo-2-[3-phenyl-2-(1-pyrrolidinylmethyl)-1-piperidinyl]ethyl}amine. Several problems relating to oral bioavailability, cytochrome P450 inhi... ( view more )bition, and off-target activity at the kappa opioid receptor and cardiac sodium channel were addressed during lead development. hUT binding affinity relative to compound 1 was improved by more than 40-fold in some analogs, and a structural modification was identified which significantly attenuated both off-target activities. ( view less ) Christoph Bergmann,Laura Strauss,Yun Wang,Miroslaw J Szczepanski,Stephan Lang,Jonas T Johnson,Theresa L Whiteside PURPOSE: Regulatory T cells play a major role in tumor escape from immunosurveillance. T regulatory cells type 1 (Tr1), a subset of regulatory T cells present in the tumor and peripheral circulation of patients with head and neck squamous cell carcinoma (HNSCC), mediate immune suppression and might... ( view more ) contribute to tumor progression. EXPERIMENTAL DESIGN: CD4(+)CD25(-)T cells were isolated from peripheral blood mononuclear cells (PBMC) or tumor-infiltrating lymphocytes (TIL) of 26 HNSCC patients and 10 normal controls. The Tr1 cell phenotype was determined before and after culture in the presence of interleukin (IL)-2, IL-10, and IL-15, each at 10 to 20 IU/mL. Suppression was measured in carboxyfluorescein diacetate succinimidyl ester-based proliferation assays with or without neutralizing anti-IL-10 or anti-transforming growth factor-beta(1) (TGF-beta(1)) monoclonal antibodies in Transwell systems. ELISA was used to define the Tr1 cytokine profile. RESULTS: Tr1 cells originate from CD4(+)CD25(-) precursors present in TIL and PBMC of HNSCC patients. Cytokine-driven ex vivo expansion of Tr1 precursors yielded CD4(+)CD25(-)Foxp3(low)CD132(+)IL-10(+)TGF-beta(1)(+) populations that mediated higher suppression than Tr1 cells of normal controls (P < 0.0001). Tr1 cells suppressed proliferation of autologous responders via IL-10 and TGF-beta(1) secretion. Expression of these cytokines was higher in TIL-derived than PBMC-derived Tr1 cells (P < 0.0001). The Tr1 cell frequency and suppressor function were significantly higher in patients presenting with advanced than early disease stages and in patients "cured" by oncologic therapies than in those with active disease. CONCLUSIONS: In HNSCC, Tr1 cell generation is promoted at the tumor site. Tr1 cells use TGF-beta and IL-10 to mediate suppression. They expand during disease progression and also following cancer therapy in patients with no evident disease. ( view less ) Theresa J Ochoa,Elsa Chea-Woo,Miguel Campos,Iris Pecho,Ana Prada,Robert J McMahon,Thomas G ClearyWe conducted a randomized, double-blind, placebo-controlled trial comparing supplementation with bovine lactoferrin versus placebo for the prevention of diarrhea in children. Comparison of overall diarrhea incidence and prevalence rates found no significant difference between the 2 groups. However,... ( view more ) there was a lower prevalence of colonization with Giardia species and better growth among children in the lactoferrin group. ( view less ) Christine D Palmer,Brenda E Mutch,Theresa H Page,Nicole J Horwood,Brian M J Foxwell Discordant cytokine production is characteristic of chronic inflammatory conditions like rheumatoid arthritis (RA), and anti-cytokine therapeutics are becoming routinely used to treat RA in the clinic. Fibroblasts from rheumatoid synovium have been shown to contribute to cytokine production in infl... ( view more )amed joints; likewise these cells also produce cytokines in response to inflammatory mediators signalling through Toll like receptors (TLRs). Tyrosine kinase activity is essential to LPS-induced cytokine production, and we have previously implicated a role for the Tec kinase, Bmx, in inflammatory cytokine production. Here we show that Bmx kinase activity in RASF is increased following LPS stimulation and that Bmx is involved in the regulation of LPS-induced IL-6 and VEGF production via mRNA stabilisation. This is an important insight into the regulation of VEGF, which is involved in a wide range of different pathologies, and may lead to more effective design of novel anti-inflammatory/angiogenic therapeutics for conditions such as RA. ( view less )
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