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Recent studies have demonstrated conflicting results regarding the value of CDX2 for distinguishing primary ovarian mucinous tumors from metastatic mucinous carcinomas in the ovary. Utility of coordinate expression of cytokeratins 7 and 20 is restricted to distinction of ovarian mucinous tumors from lower gastrointestinal tract metastases and data comparing coordinate expression of all three markers is limited. Immunohistochemical studies were performed to compare expression of CDX2 and cytokeratin 20, both markers of intestinal differentiation, in conjunction with coordinate expression of cytokeratin 7, in 90 mucinous tumors involving the ovary: 42 primary ovarian mucinous tumors (31 atypical proliferative (borderline) mucinous tumors (gastrointestinal type), 11 mucinous carcinomas) and 48 metastatic mucinous carcinomas of upper (pancreaticobiliary tract: 14; stomach: five) and lower (colon and rectum: 25; appendix: four) gastrointestinal tract origin. Primary ovarian tumors expressed CDX2 (40%) less frequently than cytokeratin 20 (83%) (P

Ovarian atypical proliferative (borderline) mucinous tumors of gastrointestinal and seromucinous types are considered subtypes within the mucinous tumor category despite the presence of distinctive clinicopathologic features that seromucinous tumors share with pure serous tumors. Immunophenotypic differences have not been extensively investigated. Immunohistochemical studies were performed to compare the expression patterns of cytokeratins 7 and 20 (CK7, CK 20), estrogen and progesterone receptors (ER, PR), CA-125, mesothelin, and WT-1 in 28 tumors of gastrointestinal type and 12 tumors of seromucinous type. Both gastrointestinal and seromucinous type tumors had a high frequency of CK7 expression (93% and 100%, respectively). The gastrointestinal type tumors were characterized by frequent expression of CK20 (86%) and CDX2 (39%), infrequent expression of CA-125 (11%) and mesothelin (7%), and lack of expression of ER, PR, and WT-1. In contrast, the seromucinous type tumors were characterized by frequent expression of ER (100%), PR (67%), CA-125 (92%), and mesothelin (83%), infrequent expression of WT-1 (8%), and lack of expression of CK20 and CDX2. The gastrointestinal and seromucinous types of atypical proliferative mucinous tumors are immunophenotypically distinctive tumors. The former are characterized by expression of markers of gastrointestinal-type differentiation (CK20 and CDX2), whereas the latter are characterized by expression of "müllerian-type" markers (ER, PR, CA-125, and mesothelin). Expression of the latter markers in the seromucinous tumors, which also are expressed in pure serous tumors, and lack of expression of gastrointestinal-type markers, combined with the clinicopathologic features these tumors share with pure serous tumors, support the concept that this subtype is more closely related to serous than gastrointestinal type mucinous tumors and justify the designation "seromucinous."

Whole-brain irradiation of animals and humans has been reported to lead to late delayed structural (vascular damage, demyelination, white matter necrosis) and functional (cognitive impairment) alterations. However, most of the experimental data on late delayed radiation-induced brain injury have been generated with large single doses or short fractionation schemes that may provide a less accurate indication of the events that occur after clinical whole-brain radiotherapy. The pilot study reported here investigates cerebral vascular pathology in male Fischer 344 rats after whole-brain irradiation with a fractionated total dose of 137Cs gamma rays that is expected to be biologically similar to that given to brain tumor patients. The brains of young adult rats (4 months old) were irradiated with a total dose of 40 Gy, given as eight 5-Gy fractions twice per week for 4 weeks. Brain capillary and arteriole pathology was studied using an alkaline phosphatase enzyme histochemistry method; vessel density and length were quantified using a stereology method with computerized image processing and analysis. Vessel density and length were unchanged 24 h after the last dose, but at 10 weeks postirradiation, both were substantially decreased. After 20 weeks, the rate of decline in the vessel density and length in irradiated rats was similar to that in unirradiated age-matched controls. No gross gliosis or demyelination was observed 12 months postirradiation using conventional histopathology techniques. We suggest that the early (10-week) and persistent vascular damage that occurs after a prolonged whole-brain irradiation fractionation scheme may play an important role in the development of late delayed radiation-induced brain injury.

Introduction: Cephalometric analyses have been used by orthodontists to track growth and monitor treatment effects. Most of these analyses have normative values to which patients are compared, but some "normal" patients vary quite a bit from the normative values. The centrographic analysis is a visual analysis with no angles to measure or normative values to compare. After a reference plane is developed, the relative position of variable landmarks can be seen. Methods: We used the centroid centrographic analysis to study the effects of 1-phase and 2-phase orthodontic treatment. Phase 1 treatment consisted of bionator (n = 66), headgear/biteplane (n = 69), or observation (n = 65) until a Class I molar relationship was achieved or 2 years had elapsed. After 1 year, all subjects underwent full orthodontic treatment with fixed appliances. Results: Centrographic analysis showed that early treatment has effects on the mandible. However, the differences were not apparent by the end of fixed appliance treatment. Conclusions: The skeletal effects of phase 1 treatment disappear by the end of fixed appliance treatment.

Four conformationally flexible benzamide analogs having a high affinity and outstanding selectivity for sigma(2) versus sigma(1) receptors were synthesized and radiolabeled with carbon-11 by reaction with [(11)C]methyl iodide. The four (11)C-labeled radiotracers were evaluated for their potential to image the proliferative status of breast tumors with positron emission tomography (PET). In vivo studies in female BALB/C mice bearing EMT-6 breast tumors showed that one radiotracer, (2-methoxy-(11)C)-N-(4-(3,4-dihydro-6,7-dimethoxy-isoquinolin-2(1H)-yl)butyl)-5-methylbenzamide ([(11)C]2), had a high tumor uptake and suitable tumor/background ratio for imaging purposes. Blocking studies were consistent with the labeling of sigma(2) receptors in vivo. A study comparing the in vivo properties of [(11)C]2 and (18)F-3'-fluoro-3'-deoxy-L-thymidine ([(18)F]FLT) indicated that [(11)C]2 had either similar (lung, fat) or better (blood, muscle) tumor/organ ratios than [(18)F]FLT in the tissues that are important for breast tumor imaging. Consequently, [(11)C]2 is a potential radiotracer for imaging the proliferative status of breast tumors in vivo with PET.

The innate immune system is the oldest mammalian defence against invading micro-organisms and provides the first line of defence against them, however until recently a detailed understanding of its complexity has been lacking. This review describes recent advances that have been made in understanding the components of the innate immune system, including the pathogen sensing mechanisms, receptor and intracellular signalling pathways, linkage to the acquired immune system, and effectors of the innate immune response. These discoveries have created an opportunity for the development of novel drugs through the identification of targets for rational drug design. The opportunity for the development of novel anti-inflammatory and antimicrobial drugs through modulation of pro-inflammatory or antimicrobial signals within the innate immune system, are discussed. A more detailed understanding of the effectors of the innate immune system is providing an opportunity for the design of effector mimetics as novel antimicrobial drugs. The innate immune system is providing the basis for much-needed alternative approaches to controlling infection and inflammation in human medicine.

Regulation of coronary vascular tone is critical for proper perfusion and function of the myocardium. Many disease processes result in compromised regulation of coronary vascular tone and impaired myocardial perfusion. A common result of coronary vascular dysfunction is the development of areas of replacement fibrosis within the myocardium and surrounding the vasculature. Both intravascular processes, such as coronary atherosclerosis and endothelial dysfunction, and extravascular processes, including compromised myocardial metabolism, hormone excesses, and altered local signaling, may result in coronary vascular dysregulation. Coronary occlusion events, in turn, lead to myocardial damage and the activation of inflammatory cells and fibroblasts. The role of fibroblasts in regulating myocardial fibrosis and the contribution of myofibroblasts, cells that have limited contractile potential while retaining many of the extracellular matrix regulating processes of the fibroblast, may also contribute to the development of myocardial disease. In this review we examine the recent literature on myocardial fibrosis and myofibroblast activity, highlighting the effects of several classes of cardiovascular agents on the remodeling process.

A Gaussian random wavefunction that satisfies Dirichlet and Neumann conditions locally on a convex circular boundary is introduced. The average of the square of the wavefunction and its derivatives are computed and their asymptotics studied in the semi-classical limit. The mean nodal line length (L) is calculated and the first order boundary effect shown to be of order log k, where k is the wavenumber. In the limit of vanishing boundary curvature (large boundary radius) these results are shown to approach those for a straight wall boundary.

Health care fraud remains an enormous problem within the United States. In 1993, health care fraud was identified as the number two crime problem in America after violent crime. Though the social and political landscape of this nation has changed much since 1993, health care fraud remains a top priority because of the large financial implications to taxpayers. In 2010, Americans spent $2.6 trillion on health care. The National Health Care Anti-Fraud Association, a nonprofit collaboration of federal and private health care fraud special investigative units, estimates that between 3% and 10% of health care expenditures are lost to fraud, resulting in approximately $78 to $260 billion stolen from taxpayers on an annual basis. Health care fraud consists of several different methods or schemes. In almost every case, health care fraud is focused on the money. In the past, health care fraud was almost solely perpetrated by medical providers. However, recent trends are showing that health care fraud perpetrators are now more likely to have little to no medical experience and tend to have criminal backgrounds or are part of large pharmaceutical organizations that engage in illegal practices. Last year, GlaxoSmithKline (GSK) admitted to off-label marketing of pharmaceuticals and withholding safety data from the Food and Drug Administration. GSK settled the case with the U.S. Government for $3 billion. In May 2013, Ranbaxy USA pleaded guilty to introducing adulterated drugs into U.S. commerce. Other current schemes include billing for services not rendered, substituting inferior products and billing for more expensive ones, performing unnecessary procedures to gain greater reimbursement, engaging in kickbacks, and prescribing narcotics for money rather than medical necessity. Victims of health care fraud may include the patients, health care workers, insurance companies, and, in the case of federally funded medical benefits plans, the taxpayer. The Military Health System (MHS), funded by the Defense Health Program (DHP) appropriation, is not immune to this growing trend. The MHS supports 9.6 million uniformed service members, their spouses and children, retirees and their families worldwide and is the second largest medical benefits program administered by the government. Within MHS, health benefits are offered to beneficiaries through two methods: direct care through the military treatment facilities (MTFs) and purchased care through the TRICARE program. Total annual costs for the MHS are approximately $50 billion. With such a large program comes great risk for fraud, waste, and abuse within the program. Accordingly, MHS leadership has the responsibility to identify and deter fraud to the maximum extent possible to ensure the proper utilization of the government’s resources. Although the national economic climate remains cloudy and medical expenditures continue to grow, the MHS has developed an exceptional tool to battle against fraud and abuse. Specifically, MHS’ TRICARE Management Activity’s (TMA) Program Integrity (PI) Office uses its worldwide authority to combat fraud, waste and abuse within and against the TRICARE program. The office monitors contractor PI activities, coordinates with the Department of Defense (DoD) and external investigative agencies, and initiates administrative remedies as required. In addition, TMI PI

Objective To describe the effectiveness and safety of surgical treatment of adult patients with strabismus, and to review the reported functional benefits and complications of strabismus surgery for adults. Methods A literature search was conducted in September 2001. It was repeated and updated in April 2003, with retrieval of relevant citations. Panel members reviewed the articles and rated them according to their relevance to the topic and methodology. Results The literature search identified 49 reports that describe the surgical treatment of strabismus in adult patients and meet predetermined review criteria. Of these reports, 2 were of randomized controlled trials, and 1 addressed the primary objective of this review. In this randomized study of adults with strabismus, direct comparison of surgical correction with botulinum toxin A chemodenervation indicated that surgical treatment was superior to botulinum toxin A in realigning the eyes (76.9% vs. 29.4%, P = 0.027). Several large case series of adults with strabismus (level III evidence) with successful surgical realignment rates of 68% to 85% have been reported. Functional benefits of surgical treatment are reported in many patients. These include elimination of diplopia, development of binocular fusion, expansion of binocular visual fields, and improvement of head position. Surgical complications, including new, postoperative diplopia (1%–14%) or scleral perforation (0.8%–1.8%), occur in a minority of patients. Unplanned reoperations (subsequent strabismus procedures that were not anticipated as part of a staged treatment) were needed in up to 21% of patients in large case series of comitant strabismus, and in up to 50% of patients with thyroid ophthalmopathy. Conclusions Despite the paucity of level I evidence from randomized controlled trials, the existing literature suggests that surgical treatment of adults with strabismus is safe and effective in improving ocular alignment. In many cases it improves visual function, based largely on level III evidence. Risks include unplanned reoperation, postoperative diplopia, and scleral perforation. Additional level I studies of surgical treatment of adult patients would help to document the effectiveness and substantiate the safety of this treatment.

Genetic defects in the plasma membrane-associated sarcoglycan complex produce cardiomyopathy characterized by focal degeneration. The infarct-like pattern of cardiac degeneration has led to the hypothesis that coronary artery vasospasm underlies cardiomyopathy in this disorder. We evaluated the coronary vasculature of gamma-sarcoglycan mutant mice and found microvascular filling defects consistent with arterial vasospasm. However, the vascular smooth muscle sarcoglycan complex was intact in the coronary arteries of gamma-sarcoglycan hearts with perturbation of the sarcoglycan complex only within the adjacent myocytes. Thus, in this model, coronary artery vasospasm derives from a vascular smooth muscle-cell extrinsic process. To reduce this secondary vasospasm, we treated gamma-sarcoglycan-deficient mice with the calcium channel antagonist verapamil. Verapamil treatment eliminated evidence of vasospasm and ameliorated histological and functional evidence of cardiomyopathic progression. Echocardiography of verapamil-treated, gamma-sarcoglycan-null mice showed an improvement in left ventricular fractional shortening (44.3 +/- 13.3% treated versus 37.4 +/- 15.3% untreated), maximal velocity at the aortic outflow tract (114.9 +/- 27.9 cm/second versus 92.8 +/- 22.7 cm/second), and cardiac index (1.06 +/- 0.30 ml/minute/g versus 0.67 +/- 0.16 ml/minute/g, P0.05). These data indicate that secondary vasospasm contributes to the development of cardiomyopathy and is an important therapeutic target to limit cardiomyopathy progression.

Lumbar spinal stenosis is a common condition seen in patients presenting to physicians who specialize in pain management or perform spine surgery. The designation of "spinal stenosis" without other qualifiers is vague and as such holds little practical value. Classifications have been created in order to more specifically describe the various etiologies as well as the site(s) of narrowing. For this purpose, diagnostic imaging studies are vital. These include myelography, computed tomography (CT), and magnetic resonance imaging (MRI). Each imaging modality has its own inherent advantages and limitations in demonstrating anatomical structures and how they may contribute to the stenotic process. Since proper treatment follows accurate identification of the pathology, it is important for physicians to have a sound understanding of normal and abnormal spinal elements as they are depicted on various imaging studies.

The mechanisms regulating involution of mammary glands after weaning are not clear, but engorgement with milk is a key trigger. Many cell types require to be anchored to an extracellular matrix (ECM) as a prerequisite for survival and this is achieved via intregrins binding to specific motifs and signalling their attachment, intracellularly, via focal adhesion kinase (FAK). We sought to determine firstly, if expression of β1-integrin and FAK is reduced during the first stage of involution. Expression of β1-integrin and FAK was significantly reduced at 6 h after sealing teats and this was accompanied with a decreased abundance of cytochrome C in mitochondria. Secondly, we sought to determine if expression of β1-integrin and FAK was restored during the first, partially reversible stage of involution (at 24 h), but not during the second irreversible stage, which occurs after 72 h. Re-suckling restored full expression of the 80 kDa fragment of FAK, but not of the 125 kDa protein or β1-integrin at 24 h after weaning. Re-suckling did not restore expression of either peptide after 72 h. Changes in expression of cytochrome C and pro-caspase-3 (apoptotic markers) were similar to that of the 80 kDa fragment of FAK. These data suggest that epithelial cells can restore partial contact with their basement membrane during the first, reversible stage, but not during the second irreversible stage of involution. We speculate that decreased contact between epithelial cells and their basement membrane initiates apoptosis in mammary glands at weaning. This process begins within 6 h of pup withdrawal. © 2004 Wiley-Liss, Inc.

Protein restriction in rat pregnancy programmes the development of elevated systolic blood pressure and vascular dysfunction in the offspring. A recent study has shown that hypertension is reversed by maternal glycine supplementation. Whether this protective effect is exerted directly on the embryo and fetus, or indirectly via effects on the mother, is unknown although we have previously shown abnormalities in the maternal vasculature. We tested the hypothesis that dietary glycine repletion would reverse endothelial dysfunction in protein-restricted pregnant rat dams using wire myography. Impaired acetylcholine- (P < 0.01) and isoprenaline-induced (P < 0.05) vasodilatation in isolated mesenteric arteries (MA) from protein-restricted pregnant dams was accompanied by reduced vascular nitric oxide (NO) release (P < 0.05). Dietary glycine supplementation reversed vascular dysfunction in MA (P < 0.05) and improved NO release thus potentially protecting the maternal circulation. The impaired NO release in the MA of low protein diet dams was not accompanied by reduced eNOS mRNA expression, suggesting that eNOS activity was altered. Protein restriction did not alter the vascular function of a conduit artery, the thoracic aorta. These results provide evidence that adequate provision of glycine, a conditionally essential amino acid in pregnancy, may play a role in the vascular adaptations to pregnancy, protecting the fetus from abnormal programming of the cardiovascular system.

Introduction Our previous studies detected focal disruptions in myoepithelial cell layers of several ducts with carcinoma in situ. The cell cluster overlying each of the myoepithelial disruptions showed a marked reduction in or a total loss of immunoreactivity for the estrogen receptor (ER). This is in contrast to the adjacent cells within the same duct, which were strongly immunoreactive for the ER. The current study attempts to confirm and expand previous observations on a larger scale. Methods Paraffin sections from 220 patients with ER-positive intraductal breast tumors were double immunostained with the same protocol previously used. Cross-sections of ducts lined by ≥ 40 epithelial cells were examined for myoepithelial cell layer disruptions and for ER expression. In five selected cases, ER-negative cells overlying the disrupted myoepithelial cell layer and adjacent ER-positive cells within the same duct were separately microdissected and assessed for loss of heterozygosity and microsatellite instability. Results Of the 220 cases with 5698 duct cross-sections examined, 94 showed disrupted myoepithelial cell layers with 405 focal disruptions. Of the 94 cases, 79 (84%) contained only ER-negative cell clusters, nine (9.6%) contained both ER-negative and ER-positive cell clusters, and six (6.4%) contained only ER-positive cell clusters overlying disrupted myoepithelial cell layers. Of the 405 disruptions, 350 (86.4%) were overlain by ER-negative cell clusters and 55 (13.6%) were overlain by ER-positive cell clusters (P < 0.01). Microdissected ER-negative and ER-positive cells within the same duct from all five selected cases displayed a different frequency or pattern of loss of heterozygosity and/or microsatellite instability at 10 of the 15 DNA markers. Conclusions Cells overlying focally disrupted myoepithelial layers and their adjacent counterparts within the same duct displayed different immunohistochemical and molecular features. These features potentially represent an early sign of the formation of a biologically more aggressive cell clone and the myoepithelial cell layer breakdown possibly associated with tumor progression or invasion.

The purpose of this study was to compare the dentoalveolar outcomes after 1-phase and 2-phase orthodontic treatment of Class II malocclusions. Class II subjects (n = 208) were randomized to 1-phase or 2-phase treatment with either bionator or headgear/biteplate. The peer assessment rating (PAR) was calculated from pretreatment, prephase 2, and final study models. Chi-square, Kruskal-Wallis, and Wilcoxon rank sum tests were used to evaluate the differences among treatment groups, sexes, races, pretreatment, mandibular plane angle, severity, and compliance. Spearman rank correlation coefficients were used to examine relationships between PAR at different times. The dropout rate of 24.6% did not adversely affect the ability to detect differences of clinical importance or impact treatment groups disproportionately. There were no significant differences with respect to initial PAR or final PAR among the 3 treatment protocols. The 2 early treatment groups had lower PAR scores than the 1-phase group before phase 2 (P =.0001). Lower PAR scores were achieved at both the beginning and end of phase 2 in girls (P =.03; P =.02, respectively). There were differences in the pre-phase-2 and post-phase-2 PAR scores based on initial severity (P =.0006; P =.02, respectively), with greater improvement in the patients whose malocclusions were less severe initially. Mandibular plane angle had no effect on pre-phase-2 or post-phase-2 PAR scores. These results do not support the hypothesis that different dentoalveolar outcomes are obtained between 2-phase and 1-phase treatment of Class II malocclusions.

Objective To measure local (peritoneal fluid) and systemic (plasma) cytokine profiles in patients with infection-inflammation of the vermiform appendix, a relatively mild, localized inflammatory process. Summary background data The systemic host response to invading microorganisms, often termed the systemic inflammatory response syndrome (SIRS), includes changes in heart rate, respiratory rate, body temperature, and circulating white blood cell numbers. Although these changes can be induced experimentally by administering proinflammatory cytokines, the mediators that appear in the bloodstream during early, localized infection in humans have not been defined. Methods The authors studied 56 patients with pathologically proven appendicitis. Blood was obtained before the induction of anesthesia, when 82% of the patients met the criteria for SIRS. Peritoneal fluid (PF) was obtained by intraoperative lavage. Cytokines were measured by immunoassay. To assess the net impact of the mediators within plasma, the authors studied the ability of patient plasma to augment or suppress bacterial lipopolysaccharide (LPS) stimulation of monocytes in vitro. Results Of the proinflammatory cytokines, tumor necrosis factor-alpha was present in PF but not in plasma, interleukin (IL)-1beta and interferon-gamma were found in low concentrations in both PF and plasma, and IL-12 (p70) was detectable in plasma but not PF. In contrast, IL-6 and IL-1 receptor antagonist (IL-1ra) were the most abundant cytokines in the PF and plasma, and the concentrations of IL-4 and IL-10 were also elevated in both compartments. Patients with more severe appendicitis had higher plasma levels of IL-6 and IL-10 and lower plasma levels of IL-12 and interferon-gamma than did those with uncomplicated disease. Patient plasma inhibited LPS-induced stimulation of a monocyte cell line, and this inhibition was accentuated by complicated disease. Conclusions As judged from the pattern of soluble cytokines in plasma and the effect of the plasma on monocyte activation by LPS, mild, localized infection can induce a systemic response that is predominantly anti-inflammatory.

Saccharomyces cerevisiae , bakers' yeast, is not a pathogen in healthy individuals, but is increasingly isolated from immunocompromised patients. The more frequent isolation of S. cerevisiae clinically raises a number of questions concerning the origin, survival, and virulence of this organism in human hosts. Here we compare the virulence of a human isolate, a strain isolated from decaying fruit, and a common laboratory strain in a mouse infection model. We find that the plant isolate is lethal in mice, whereas the laboratory strain is avirulent. A knockout of the SSD1 gene, which alters the composition and cell wall architecture of the yeast cell surface, causes both the clinical and plant isolates to be more virulent in the mouse model of infection. The hypervirulent ssd1 Δ/ ssd1 Δ yeast strain is a more potent elicitor of proinflammatory cytokines from macrophages in vitro . Our data suggest that the increased virulence of the mutant strains is a consequence of unique surface characteristics that overstimulate the proinflammatory response.

To enhance milk composition and milk processing efficiency by increasing the casein concentration in milk, we have introduced additional copies of the genes encoding bovine beta- and kappa-casein (CSN2 and CSN3, respectively) into female bovine fibroblasts. Nuclear transfer with four independent donor cell lines resulted in the production of 11 transgenic calves. The analysis of hormonally induced milk showed substantial expression and secretion of the transgene-derived caseins into milk. Nine cows, representing two high-expressing lines, produced milk with an 8-20% increase in beta-casein, a twofold increase in kappa-casein levels, and a markedly altered kappa-casein to total casein ratio. These results show that it is feasible to substantially alter a major component of milk in high producing dairy cows by a transgenic approach and thus to improve the functional properties of dairy milk.

The protein synthesis inhibitor cycloheximide (Chx) suppresses prolactin-induced beta-casein gene expression in the mammary epithelial cell line COMMA-D. As the mechanism underlying this effect is unclear, the effects of protein synthesis inhibitors on interactions of transcription factors with the beta-casein promoter were examined. Suppression of prolactin-induced beta-casein gene expression occurred in both COMMA-D cells and primary mammary cell cultures with as little as 2 h protein synthesis inhibition. This was associated with changes in transcription factors interacting at a response element in the proximal region of the rat beta-casein promoter. Inhibition of protein synthesis was associated with NF-kappaB binding at a site immediately 3' to the Stat5-binding site at position 97-89 of the beta-casein promoter, suppression of Stat5 DNA-binding activity, and inhibition of Stat5 tyrosine phosphorylation. Treatment with the NF-kappaB inhibitor parthenolide failed to restore prolactin responsiveness. These results show that protein synthesis inhibition is associated with both blockage of prolactin-Stat5 signaling and NF-kappaB binding to the beta-casein promoter, but that the latter is not necessary for the suppression of beta-casein expression.

There are several public cloud providers that provide service across different cloud models such as IaaS, PaaS, and SaaS. End users require an objective means to assess the performance of the services being offered by the various cloud providers. Benchmarks have typically been used to evaluate the performance of various systems and can play a vital role in assessing performance of the different public cloud platforms in a vendor neutral manner. Amazon's EC2 Service is one of the leading public cloud service providers and offers many different levels of service. The research in this chapter focuses on system level benchmarks and looks into evaluating the memory, CPU, and I/O performance of two different tiers of hardware offered through Amazon's EC2. Using three distinct types of system benchmarks, the performance of the micro spot instance and the M1 small instance are measured and compared. In order to examine the performance and scalability of the hardware, the virtual machines are set up in a cluster formation ranging from two to eight nodes. The results show that the scalability of the cloud is achieved by increasing resources when applicable. This chapter also looks at the economic model and other cloud services offered by Amazon's EC2, Microsoft's Azure, and Google's App Engine.

Background Considerably divergent data have been published from attempts to model the E. coli vs. S. aureus specific immune reaction of the udder using primary cultures of bovine mammary epithelial cells from cows (pbMEC). Some groups reported a swift, strong and transient inflammatory response against challenges with E. coli and only a weak and retarded response against S. aureus, in agreement with the respective reaction of the udder. Others found almost the reverse. Presence or absence of fetal calf serum distinguished the experimental setting between both groups. We examined here if this causes the divergent reaction of the pbMEC towards both pathogen species. We challenged pbMEC with proteins from heat killed E. coli or S. aureus pathogens or purified TLR2 and TLR4 ligands. The stimuli were applied in normal growth medium with (SM10) or without (SM0) 10 % fetal calf serum, or in the basal medium supplemented with 10 mg/ml milk proteins (SM Milk). Results Withdrawal of FCS slowed down and decreased the extent by which E. coli or LPS enhanced the expression of cyto- and chemokine encoding genes through impaired TLR4 signalling but enforced their expression during stimulation with S. aureus. SM Milk strongly quenched the induction of those genes. S. aureus strain specific differences in the reaction of the pbMEC could only be recorded in SM0. NF-κB factors were activated by E. coli in all stimulation media, but only to a small extent by S. aureus, solely in SM0. Purified ligands for TLR2 stimulated expression of those genes and activated NF-κB equally well in SM10 and SM0. The mRNA destabilizing factor tristetraproline was only induced by E. coli in SM10 and by purified PAMPs. Conclusions Our data cross validate the correctness of previously published divergent data on the pathogen-specific induction of key immune genes in pbMEC. The differences are due to the presence of FCS, modulating signalling through TLR4 and TLR-unrelated pathogen receptors. S. aureus does not substantially activate any TLR signalling in MEC. Rather, receptors distinct from TLRs perceive the presence of S. aureus and control the immune response against this pathogen in MEC. Electronic supplementary material The online version of this article (doi:10.1186/s12917-015-0489-3) contains supplementary material, which is available to authorized users.

Saliva influences rumen function in cattle, yet the biochemical role for most of the bovine salivary proteins (BSPs) has yet to be established. Two cDNAs (BSP30a and BSP30b) from bovine parotid salivary gland were cloned and sequenced, each coding for alternate forms of a prominent protein in bovine saliva. The BSP30 cDNAs share 96% sequence identity with each other at the DNA level and 83% at the amino acid level, and appear to arise from separate genes. The predicted BSP30a and BSP30b proteins share 26–36% amino acid identity with parotid secretory protein (PSP) from mouse, rat and human. BSP30 and PSP are in turn more distantly related to a wider group of proteins that includes lung-specific X protein, also known as palate, lung, and nasal epithelium clone (LUNX/PLUNC), von Ebner's minor salivary gland protein (VEMSGP), bactericidal permeability increasing protein (BPI), lipopolysaccharide binding protein (LBP), cholesteryl ester transfer protein (CETP), and the putative olfactory ligand-binding proteins RYA3 and RY2G5. Bovine cDNAs encoding homologs of LUNX/PLUNC and VEMSGP were isolated and sequenced. Northern blot analysis showed that LUNX/PLUNC, BSP30 and VEMSGP are expressed in bovine salivary tissue and airways, and that they have non-identical patterns of expression in these tissues. The expression of both BSP30a and BSP30b is restricted to salivary tissue, but within this tissue they have distinct patterns of expression. The proximity of the human genes coding for the PSP/LBP superfamily on HSA20q11.2, their similar amino acid sequence, and common exon segmentation strongly suggest that these genes evolved from a common ancestral gene. Furthermore, they imply that the BSP30a and BSP30b proteins may have a function in common with other members of this gene family.

Four Frankia-Casuarina endosymbiont strains were tested for their infectivity and effectiveness on C. equisetifolia plants after 1 d, 3 and 6 months of Frankia storage at 7, 28 and 40 degrees C as liquid-cultures and polyacrylamide gel (PAG)-immobilized inocula. At lower temperature the number of nodules was the same or higher than control for liquid inocula except after 6 months of storage. For the PAG-entrapped Frankia lower numbers of nodules were recorded due to the use of high Frankia titers. In general, the results showed comparable plant dry mass, total nitrogen, root to shoot and nodules to plant ratios at lower temperature treatments. Increasing time and temperature was accompanied with reduced infectivity and effectiveness on inoculated plants. No nodulation was scored on plants inoculated with liquid and PAG-entrapped inocula stored at 40 degrees C for 6 months; subsequently, plant growth was inhibited. The variations in results obtained for different strains and treatments lead to variations in plant nitrogen-fixing ability. The use of PAG as a carrier for different Frankia strains is suggested; the recommended storage temperature for PAG-immobilized Frankia in 7-28 degrees C for up to 3 months.

Increases in the vertical dimension can be detrimental in certain facial types or unstable in others. By using cephalograms, this study examined the skeletal and dental changes as well as their permanence in a group of children participating in a randomized clinical trial on the timing of Class II orthodontic treatment. Children (mean age, 9.6 years) were assigned to either a headgear/biteplane (n = 94), bionator (n = 86), or observation (n = 82) group. Cephalometric data were obtained from tracings at baseline, after Class I molar was obtained or 2 years after treatment had elapsed, and after 1 year of retention or observation. In general, the skeletal changes, such as total anterior and posterior face height and mandibular plane angle, observed during the active phase of early treatment persisted until the end of phase I. Phase I treatment also produced dental changes, such as molar eruption and tip. With the exception of the eruption of the mandibular molars in the bionator group, these changes could not be maintained until the end of phase I. Whether the subjects were in retention or not had little impact on any of the studied parameters. Mandibular plane angle, pretreatment, and retention had an affect on the mandibular molar tip over the course of the study.

The dystrophin glycoprotein complex (DGC) is found at the plasma membrane of muscle cells, where it provides a link between the cytoskeleton and the extracellular matrix. A subcomplex within the DGC, the sarcoglycan complex, associates with dystrophin and mediates muscle membrane stability. Mutations in sarcoglycan genes lead to muscular dystrophy and cardiomyopathy in both humans and mice. In invertebrates, there are three sarcoglycan genes, while in mammals there are additional sarcoglycan genes that probably arose from gene duplication events. We identified a novel mammalian sarcoglycan, zeta-sarcoglycan, that is highly related to gamma-sarcoglycan and delta-sarcoglycan. We generated a zeta-sarcoglycan-specific antibody and found that zeta-sarcoglycan associated with other members of the sarcoglycan complex at the plasma membrane. Additionally, zeta-sarcoglycan was reduced at the membrane in muscular dystrophy, consistent with a role in mediating membrane stability. zeta-Sarcoglycan was also found as a component of the vascular smooth muscle sarcoglycan complex. Together, these data demonstrate that zeta-sarcoglycan is an integral component of the sarcoglycan complex and, as such, is important in the pathogenesis of muscular dystrophy.

Activity of acetyl-CoA carboxylase (ACC)-alpha is rate limiting for de novo synthesis of fatty acids. The encoding gene is expressed by three different promoters. We characterized promoter III (PIII) from cow, previously only known from sheep. Quantitation of transcripts by RNAse protection assays and real time PCR revealed that PIII is primarily expressed and strongly induced ( approximately 28-fold) in the lactating mammary gland. PIII transcripts are expressed in mammary epithelial cells (MEC) as shown by in situ hybridization. A 2999 bp segment of the PIII promoter conferred prolactin and dexamethasone inducibility to a luciferase reporter gene in stably transfected mouse MEC cells. Lactogenic induction was abolished if a unique signal transducer and activator of transcription (STAT)-binding site at position -797 was inactivated by two point mutations. An oligonucleotide probe harboring this STAT-site specifically bound nuclear proteins from the lactating mammary gland. Binding was abolished by those two point mutations and super-shift analyses showed that STAT5A factors are present in this complex. Hence, prolactin, acting through STAT5, contributes to the activation of ACC expression in the milk producing cells of the lactating mammary gland. We discuss that STAT5 might be important in determining the milk composition by coordinating fatty acid and protein synthesis during lactation.

When force is applied to a tooth during orthodontic tooth movement, mechanical stress is loaded on the alveolar bone. Alveolar bone and the periodontal ligament (PDL) are compressed on one side, while on the opposite side, the PDL is stretched. The mechanical stress of the stretched PDL induces alveolar bone modeling (surface apposition of bone), while the mechanical compression produces bone remodeling (the turnover of bone in small packets). The steps leading from the application of an orthodontic force to the appearance of a biologic response (mechanotranduction) have not been fully elucidated. The possible mechanisms by which osteoblasts and/or osteocytes can sense a mechanical stimulus include strain-sensitive ion channels, shear stress receptors, adhesin/integrin activation, and cytoskeleton reorganization. Once detected, the signal is internalized and then potentiated in the cytosol by the generation of second messengers and protein kinases. This, in turn, activates transcription of genes into messenger RNA that is then translated into proteins to be used by the cell or exported to modulate the activity of additional enzymes, eventually leading to altered gene expression. In the nucleus, the induction of the so-called immediate early genes, which occur shortly after a cell is stimulated, may also play a critical role in the signal transduction pathway.

Background: A hall mark of hypertrophic cardiomyopathy (HCM) is the heterogeneity of phenotype. Although HCM is often characterized by asymmetrical left ventricular hypertrophy, contemporary studies have focused on global assessment of left ventricular longitudinal strain (LS). We assessed whether

Hypertrophic cardiomyopathy (HCM) is a leading cause of sudden cardiac death among athletes in the United States [(1)][1]. The electrocardiogram (ECG) can be used for screening athletes to identify HCM [(2–4)][2]; however, its routine use remains controversial due to false positives [(5)][3]. We

Early defense against mucosal pathogens consists of both an epithelial barrier and innate immune cells. The immunocompetency of both, and their intercommunication, are paramount for the protection against infections. The interactions of epithelial and innate immune cells with a pathogen are best investigated in vivo, where complex behavior unfolds over time and space. However, existing models do not allow for easy spatio-temporal imaging of the battle with pathogens at the mucosal level. The model developed here creates a mucosal infection by direct injection of the fungal pathogen, Candida albicans, into the swimbladder of juvenile zebrafish. The resulting infection enables high-resolution imaging of epithelial and innate immune cell behavior throughout the development of mucosal disease. The versatility of this method allows for interrogation of the host to probe the detailed sequence of immune events leading to phagocyte recruitment and to examine the roles of particular cell types and molecular pathways in protection. In addition, the behavior of the pathogen as a function of immune attack can be imaged simultaneously by using fluorescent protein-expressing C. albicans. Increased spatial resolution of the host-pathogen interaction is also possible using the described rapid swimbladder dissection technique. The mucosal infection model described here is straightforward and highly reproducible, making it a valuable tool for the study of mucosal candidiasis. This system may also be broadly translatable to other mucosal pathogens such as mycobacterial, bacterial or viral microbes that normally infect through epithelial surfaces.

Background There is controversy regarding Q wave criteria for assessing risk for hypertrophic cardiomyopathy (HCM) in young athletes. Methods The 12-lead ECGs from Preparticipation screening in healthy athletes and patients with HCM were studied retrospectively. All 12 leads were measured using the same automated ECG analysis program. Results There were a total of 225 HCM patients and 1124 athletes with 12-lead electrocardiograms available for analysis. Athletes were on average 20 years of age, 65% were male and 24% were African–American. Patients with HCM were on average 51 years of age, 56% were male and 5.8% were African–American. Q waves by either amplitude, duration or area criteria were more prevalent in males than females, in lateral leads than inferior and in HCM patients than athletes. The most striking difference in Q waves between the groups was in Limb lead I and in the females. Tall, skinny Q waves were rare in athletes and had the highest prevalence of only 3.7% in male HCM patients. Conclusion Q waves are more common in males compared to females and in patients with HCM compared to athletes. Q waves of 30 ms or more in limb lead I appear to offer the greatest discriminatory value for separating patients with HCM from athletes.

The innate immune system is a critical line of defense against pathogenic fungi. Macrophages act at an early stage of infection, detecting and phagocytizing infectious propagules. To avoid killing at this stage, fungal pathogens use diverse strategies ranging from evasion of uptake to intracellular parasitism. This article will discuss five of the most important human fungal pathogens (Candida albicans, Aspergillus fumigatus, Cryptococcus neoformans, Coccidiodes immitis, and Histoplasma capsulatum) and consider the strategies and virulence factors adopted by each to survive and replicate within macrophages.

Prolactin (PRL) is important in the regulation of milk synthesis in mammary epithelial cells (MEC). In cattle, circulating levels of PRL are not limiting, suggesting the possible involvement of other factors that may control the response to PRL at the cellular level. The effects of milking frequency (MF) on milk synthesis are controlled locally within mammary glands and involve PRL signaling. To further investigate this relationship between MF and PRL signaling, udder halves of 17 dairy cows were milked either 4 times a day (4×) or once a day (1×) for 14 d in early lactation. Mammary biopsies were obtained 3 to 5h following milking from both udder halves of 10 cows, and changes in PRL and associated pathways were measured. The abundance of STAT5A mRNA was higher after 4× milking, whereas that of the PRL receptor (PRLR) and STAT3 were lower relative to that after 1× milking. In 4× mammary tissues, the protein levels of STAT5, activated STAT5, and β1-integrin were higher, whereas the those of the long isoform of PRL receptor and activated STAT3 were lower than 1× tissues. The activation of STAT5 correlated strongly with major milk protein mRNA abundance (r=0.86 to 0.94) and β1-integrin protein levels (r=0.91). These results confirm that major milk protein gene expression is associated with STAT5 activation and suggests that the STAT5 and β1-integrin signaling pathways are linked. Modulation of β1-integrin abundance in response to changes in MF may be a mechanism that controls the MEC ability to respond to PRL and therefore its secretory activity.

In dairy cows, short-term changes of milking frequency in early lactation have been shown to produce an immediate and a long-term effect on milk yield in stall-fed cows. The effect is controlled locally within mammary glands and could be a function of either secretory mammary epithelial cell number or activity. To resolve this and determine its applicability in other feed management systems, a unilateral milking frequency experiment was conducted with udder halves of 17 multiparous, pasture-fed dairy cows milked either 4 times (4×) or once a day (1×) for 14d from 5±2d in milk. Mean half-udder milk yield during the treatment period was higher from the 4× compared with 1× udder halves and continued to be higher until 200d in milk once returned to twice a day milking. Mammary biopsies were obtained on d 14 of treatment from both udder halves of 10 cows. Proliferation of mammary cells was higher in 4× udder halves compared with 1×, whereas no difference in apoptosis levels was detected. Abundance of αS1-casein, β-casein, α-lactalbumin, and β-lactoglobulin mRNA was higher in tissue samples from 4× udder halves compared with 1×, whereas lactoferrin mRNA abundance was lower in 4× udder halves. In summary, change in milking frequency during early lactation affects proliferation of mammary cells as well as expression of the major milk protein genes, which both contribute to the observed changes in milk yield during and after unilateral milking frequency treatment.

Regular exercise and a physically active lifestyle have favorable effects on health. Several issues related to this theme are addressed in this report. A comment on the requirements of personalized exercise medicine and in-depth biological profiling along with the opportunities that they offer is presented. This is followed by a brief overview of the evidence for the contributions of genetic differences to the ability to benefit from regular exercise. Subsequently, studies showing that mutations in TP53 influence exercise capacity in mice and humans are succinctly described. The evidence for effects of exercise on endothelial function in health and disease also is covered. Finally, changes in cardiac and skeletal muscle in response to exercise and their implications for patients with cardiac disease are summarized. Innovative research strategies are needed to define the molecular mechanisms involved in adaptation to exercise and to translate them into useful clinical and public health applications.

We conducted a review of patients undergoing heart transplantation (HT) at our institution for amyloid cardiomyopathy (ACM) between 2008 and 2013. Complete follow-up was available for all patients. Nineteen patients with ACM underwent HT during the study period, accounting for 9.4% of all HT performed at our institution during this period. Amyloid subtype was light chain (AL) in 9 patients and transthyretin (ATTR) in 10 (2 wild-type, 7 familial, 1 unknown). Eight of nine patients with AL amyloidosis began chemotherapy prior to HT, six have resumed chemotherapy since HT, and five have undergone autologous stem cell transplantation. Most recent free light chain levels in AL patients decreased by a median of 85% from peak values. Only one patient developed recurrent graft amyloidosis, occurring at 3.5 years post-HT and asymptomatic. After a median follow-up of 380 days, 17 (89.5%) patients are alive. To our knowledge, this is the largest single-center series reported of ACM patients undergoing HT in the modern era. Our results suggest that acceptable outcomes following HT can be achieved in the short-to-intermediate term and that this is a feasible option for end-stage ACM with careful patient selection and aggressive control of amyloidogenic light chains in AL patients.

Purpose To determine whether stereopsis of infants treated for monocular cataracts varies with the type of optical correction used. Design Randomized prospective clinical trial. Methods The Infant Aphakia Treatment Study randomized 114 patients with unilateral cataracts at age 1–7 months to either primary intraocular lens (IOL) or contact lens correction. At 4.5 years of age a masked examiner assessed stereopsis on these patients using 3 different tests: (1) Frisby; (2) Randot Preschool; and (3) Titmus Fly. Results Twenty-eight patients (25%) had a positive response to at least 1 of the stereopsis tests. There was no statistically significant difference in stereopsis between the 2 treatment groups: Frisby (contact lens, 6 [11%]; IOL, 7 [13%]; P = .99), Randot (contact lens, 3 [6%]; IOL, 1 [2%]; P = .62), or Titmus (contact lens, 8 [15%]; IOL, 13 [23%]; P = .34). The median age at surgery for patients with stereopsis was younger than for those without stereopsis (1.2 vs 2.4 months; P = .002). The median visual acuity for patients with stereopsis was better than for those without stereopsis (20/40 vs 20/252; P = .0003). Conclusion The type of optical correction did not influence stereopsis outcomes. However, 2 other factors did: age at surgery and visual acuity in the treated eye at age 4.5 years. Early surgery for unilateral congenital cataract and the presence of visual acuity better than or equal to 20/40 appear to be more important than the type of initial optical correction used for the development of stereopsis.