MR elastography derived shear stiffness--a new imaging biomarker for the assessment of early tumor response to chemotherapy.
The overall goal is to develop magnetic resonance elastography derived shear stiffness as a biomarker for the early identification of chemotherapy response, allowing dose, agent type and treatment regimen to be tailored on a per patient basis, improving therapeutic outcome and minimizing normal tissue toxicity. The specific purpose of this study is to test the feasibility of this novel biomarker to measure the treatment response in a well-known chemotherapy model. Tumors were grown in the right flank of genetically modified mice by subcutaneous injection of DoHH2 (non-Hodgkin's lymphoma) cells. Magnetic resonance elastography was used to quantify tumor stiffness before and after injection of a chemotherapeutic agent or saline. Histological tests were also performed on the tumors. A significant decrease (P < 0.0001) in magnetic resonance elastography-derived tumor shear stiffness was observed within 4 days of chemotherapy treatment, while no appreciable change was observed in saline-treated tumors. No significant change in volume occurred at this early stage, but there were decreased levels of cellular proliferation in chemotherapy-treated tumors. These results demonstrate that magnetic resonance elastography-derived estimates of shear stiffness reflect an initial response to cytotoxic therapy and suggest that this metric could be an early and sensitive biomarker of tumor response to chemotherapy.


Cigarette smoking and subsequent risk of lung cancer in men and women: analysis of a prospective cohort study.
Whether women are more susceptible than men to lung cancer caused by cigarette smoking has been controversial. To address this question, we aimed to compare incidence rates of lung cancer by stratum of smoking use in men and women of the National Institutes of Health (NIH)-AARP cohort. Participants in the NIH-AARP Diet and Health study responded to a postal questionnaire between Oct 13, 1995, and May 6, 1996, and were followed up until Dec 31, 2003. The questionnaire asked participants about their past and current smoking status, demographics, alcohol intake, tobacco smoking, physical activity, and included a food-frequency questionnaire of 124 items. Incident lung cancers were identified by linkage to individual state cancer registries. We present age-standardised incidence rates for cancer and multivariate hazard ratios (HRs) adjusted for potential confounders, with 95% CIs. This study conforms to the STROBE guidelines. 279 214 men and 184 623 women from eight states in the USA aged 50-71 years at study baseline were included in this analysis. During follow-up, lung cancers occurred in 4097 men and 2237 women. Incidence rates were 20.3 (95% CI 16.3-24.3) per 100 000 person-years in men who had never smoked (99 cancers) and 25.3 (21.3-29.3) in women who had never smoked (152 cancers); for this group, the adjusted HR for lung cancer was 1.3 (1.0-1.8) for women compared with men. Smoking was associated with increased risk of lung cancer in men and women. The incidence rate of current smokers who smoked more than two packs per day was 1259.2 (1035.0-1483.3) in men and 1308.9 (924.2-1693.6) in women. In current smokers, in a model adjusted for typical smoking dose, the HR was 0.9 (0.8-0.9) for women compared with men. For former smokers, in a model adjusted for years of cessation and typical smoking dose, the HR was 0.9 (0.9-1.0) for women compared with men. Incidence rates of adenocarcinoma, small-cell carcinoma, and undifferentiated tumours were similar in men and women; incidence rates of squamous tumours in men were about twice that in women. Our findings suggest that women are not more susceptible than men to the carcinogenic effects of cigarette smoking in the lung. In smokers, incidence rates tended to be higher in men than women with comparable smoking histories, but differences were modest; smoking was strongly associated with lung cancer risk in both men and women. Future studies should confirm whether incidence rates are indeed higher in women who have never smoked than in men who have never smoked.


The differential influence of allogeneic tumor cell death via DNA damage on dendritic cell maturation and antigen presentation.
Dendritic cells (DCs) respond to danger signals from tissue injury by amplifying their immune-inducing capacity. In the cancer context, this may lead to in vivo antitumor synergism between DCs and DNA-damaging chemotherapeutic agents. Neither the interaction between DCs and dying tumor cells nor whether different ways of inducing cell injury can deliver danger signals of different strength to DCs nor the potential role of damaged DNA as a danger signal has been studied rigorously. Here we report that coculture of immature DCs with tumor cells treated with the alkylating agents melphalan and chlorambucil leads to enhanced autologous and allogeneic T-cell activation, up-regulation of surface expression of MHC and costimulatory molecules, and increased interleukin (IL)-12 secretion. Exposure of the same DCs to tumor cells killed by cytarabine or by freeze-thaw (primary necrosis) resulted in significantly less T-cell proliferation and IL-12 production, indicating that DCs are able to sense and respond differentially to the mode of cell death. Exposure of DCs to DNA purified from tumor cells treated with alkylating agents also increased their T-cell-stimulating capacity, expression of CD86, and IL-12 secretion, supporting the hypothesis that the activating effects of tumor cells are linked to the nature of the DNA damage. This is the first study that shows that DCs respond differentially to killed tumor cells, depending upon the mechanism of DNA damage and consequent cell death.


Morphological responses of mitochondria-rich cells to hypersaline environment in the Australian mudskipper, Periophthalmus minutus.
A population of the Australian mudskipper, Periophthalmus minutus, was found to inhabit mudflat that remained uncovered by tide for more than 20 days in some neap tides. During these prolonged emersion periods, P. minutus retreated into burrows containing little water, with a highest recorded salinity of 84 ± 7.4 psu (practical salinity unit). To explore the mechanical basis for this salinity tolerance in P. minutus, we determined the densities of mitochondria-rich cells (MRCs) in the inner and outer opercula and the pectoral fin skin, in comparison with P. takita, [corrected] from an adjacent lower intertidal habitat, and studied morphological responses of MRCs to exposure to freshwater (FW), and 100% (34-35 psu) and 200% seawater (SW). Periophthalmus minutus showed a higher density of MRCs in the inner operculum (3365 ± 821 cells mm(-2)) than in the pectoral fin skin (1428 ± 161) or the outer operculum (1100 ± 986), all of which were higher than the MRC densities in p. takita. [corrected]. No mortality occurred in 100% or 200% SW, but half of the fish died within four days in FW. Neither 200% SW nor FW exposure affected MRC density. Transfer to 200% SW doubled MRC size after 9-14 days with no change in the proportion of MRCs with apical pits or plasma sodium concentration. In contrast, transfer to FW resulted in a rapid closing of pits and a significant reduction in plasma sodium concentration. These results suggest that P. minutus has evolved morphological and physiological mechanisms to withstand hypersaline conditions that they may encounter in their habitat.


Are Prophylactic Antibiotic Agents Indicated in Anorectal Surgery for Patients with Human Immunodeficiency Virus?
With increased survival among patients with human immunodeficiency virus (HIV), surgeons have been seeing more cases of anal dysplasia and cancer. There is, however, no data on the incidence of surgical site infections (SSIs) in HIV-positive patients undergoing elective anorectal procedures, nor on the administration of prophylactic antibiotic agents. We reviewed a HIV-positive population that has undergone elective anorectal biopsy of areas of dysplasia observed on office anoscopy to assess the need for antibiotic prophylaxis. A retrospective chart review was performed of all HIV-positive patients seen as outpatients in the Colorectal Surgery Division from 2007-2016. Demographics, dates of surgery and follow-up, antibiotic prophylaxis, and pre-operative CD4 count and HIV viral load were recorded for 229 patients. Post-operative examination notes were reviewed to determine the presence of SSIs. The proportion of patients who received prophylaxis was assessed and the SSI rate was calculated. Surgical site infections occurred in 2 of 237 (0.8%) cases without antibiotic prophylaxis and in none of the 38 cases with prophylaxis. This infection rate was found to be lower than that of the general surgery population, with no statistical difference from hemorrhoidectomy patients without HIV. One SSI occurred in a 51-year-old male with a pre-operative CD4 count of 612 per microliter and viral load of zero. Another occurred in a 57-year-old female with an unknown CD4 count and viral load. A χ2 analysis showed the incidence of SSIs in the groups with and without prophylaxis was not significantly different (p = 0.563). Surgical site infection rates in HIV-positive patients undergoing biopsies for anal dysplasia were similar to patients without HIV undergoing similar minor anorectal procedures, and no difference was noted in the rate of SSI with the administration of prophylactic antibiotic agents. We do not recommend routine use of prophylactic antibiotic agents in this population.


Diagnostic dilemma: osteopetrosis with superimposed rickets causing neonatal hypocalcemia.
Osteopetrosis is a rare genetic condition of reduced osteoclastic bone resorption which causes defective bone remodeling and skeletal sclerosis during growth, having effects on many organs and tissues. Mutation of T-cell immune regulator 1 (TCRG1) gene is the most common genetic defect leading to osteopetrosis, with poor prognosis. The autosomal recessive form presents in the infantile period (also known as malignant infantile osteopetrosis--MIOP), and is characterized by fractures, short stature, hepatosplenomegaly, compressive neuropathies, hypocalcemia and pancytopenia. Being a rare disease with non-specific clinical manifestations, the diagnosis is difficult and usually delayed. Rickets is a characteristic feature of MIOP which results from the defect in osteoclasts to provide a normal Ca/P balance resulting in the poor mineralization of the osteoid. Various treatment options have been suggested for osteopetrosis, but hematopoietic stem cell transplantation still remains the only curative treatment option presently. The authors report the case of a 46-day-old girl with late-onset neonatal hypocalcemia and rickets that was later diagnosed as osteopetrosis. This case report emphasizes that infantile osteopetrosis is an important cause of neonatal hypocalcemia. As irreversible complications develop within the first months of life, immediate diagnosis and early intervention are crucial and may be life-saving.


Managing dyspepsia without alarm signs in primary care: new national guidance for England and Wales.
To report new recommendations for the primary care management of dyspepsia without alarm signs in England and Wales. An independent, representative group of health care professionals, patient representatives and researchers developed the guideline using evidence-based and small group-working principles, and incorporated extensive peer-reviewing and feedback from stakeholder organizations. Referral to investigate dyspepsia should be made for alarm signs and not on the basis of age alone, reflecting the balance of benefit and harm from endoscopy. Empirical management without formal diagnosis is appropriate for most patients: reviewing patient history, lifestyle, over-the-counter medicines, and providing a course of proton-pump inhibitors and/or Helicobacter pylori test and treatment. Patients with ongoing symptoms require at least annual review to discuss symptoms and lifestyle, and as appropriate, encourage stepping down prescribed medication and returning to self-care. A new strategy included in the step down process is the use of therapies 'on-demand'. The guideline provides structured and supported recommendations for both undiagnosed and endoscopically investigated dyspepsia. Some favour increased investigation to detect Barrett's oesophagus and carcinoma. However, there is inconclusive evidence that patients without alarm signs will benefit subsequently from endoscopy, while investigation involves a small but real risk of harm.


Development of a monoclonal antibody-based lateral flow immunoassay for the detection of benzoic acid in liquid food.
To monitor benzoic acid (BA) residues in liquid food samples, a monoclonal antibody (mAb)-based lateral flow immunoassay (LFA) was developed in this study. First, 2-aminobenzoic acid (2-AA), 3-aminobenzoic acid (3-AA), and 4-aminobenzoic acid (4-AA) were conjugated to BSA and used as immunogens. After cell fusion, mAb 6D8 from 4-AA-BSA performed best with an IC50 value of 0.21 mg L-1 using 3-AA-OVA as a heterogeneous antigen, which represented a 3.4-fold improvement compared with the homogeneous antigen 4-AA-BSA. Subsequently, eight kinds of CGNPs with sizes varying from 20.94 nm to 90.00 nm were synthesized for screening the suitable size to develop a sensitive LFA. Finally, a sensitive LFA based on colloidal gold (23.27 nm) nanoparticles was developed for screening BA with a cut-off value of 4 mg L-1, which could meet the requirement of BA detection in milk, Fanta, Sprite, Coca-Cola, and Smart samples.


Growth hormone therapy for adult growth hormone deficiency.
Growth hormone deficiency in adult life is associated with a number of adverse biological changes which include osteopenia, reduced exercise capacity, altered body composition, deleterious alterations in the lipid profile and insulin status, and reduced quality of life. Most of these changes can be reversed by growth hormone replacement therapy, but in an era of health rationing, such therapy is unlikely to be offered to every growth hormone deficient adult. Therefore we have proposed a strategy aimed at delineating which adults with growth hormone deficiency might benefit most from growth hormone therapy.


[Natural resistance of Drosophila melanogaster cells to 6-mercaptopurine and 8-azaguanine].
The growth of cells and the activity of some enzymes in diploid embryonic cells of Drosophila and Mosquito which cultivated for a long time in the presence of 6-mercaptopurine and 8-azaguanine were studied. It is shown on the culture of Drosophila cells that a high level of the resistance to 6-mercaptopurine and 8-azaguanine correlates with the absence of hypoxanthine-guanine phyosphorybosyltransferase (HGPRT) in them. HGPRT activity was not also revealed in extracts obtained from flies of different lines, which draws to the conclusion on the natural resistance of Drosophila cells to hypoxanthine and quanine analogues. Similar resistance was found out in Mosquito cell lines Mos 20 A. The second enzyme of such a type, adenine phosphoribosyltransferase, is present in the cells of all investigated lines. This fact explaines the retention of the sensitivity to adenine analogues in these cells.


Preclinical Development of EBI-005: An IL-1 Receptor-1 Inhibitor for the Topical Ocular Treatment of Ocular Surface Inflammatory Diseases.
Topical interleukin (IL)-1 receptor (R)1 blockade is therapeutically active in reducing signs and symptoms of dry eye disease. Herein, we describe in vitro and in vivo nonclinical Investigational New Drug (IND)-enabling studies of EBI-005, a novel protein chimera of IL-1β and IL-1 receptor antagonist (IL-1Ra or anakinra) that potently binds IL-1R1 and blocks signaling. These studies provide an assessment of receptor affinity, drug bioavailability, immunogenic response, safety, and tolerability in mice and rabbits. In vitro and in silico along with Good Laboratory Practices (GLP) and non-GLP in vivo studies in mice and rabbits assessed the topical ocular and systemic immunogenicity and toxicology of EBI-005. Animals were treated with EBI-005 once daily subcutaneously or four times daily by topical ocular administration for up to 6 weeks (with 2-week recovery phase). EBI-005 has 500 times higher affinity than anakinra to IL-1R1. Predictive immunogenicity testing suggested that EBI-005 is not more immunogenic. Systemic bioavailability of EBI-005 is low (1.4% in mice and 0.2% in rabbits) after topical ocular administration. EBI-005 penetrated into the anterior ocular tissues within 15 min of topical ocular administration. However, it is low or undetectable after 4 hr and does not form a depot after repeated topical ocular administration. EBI-005 was safe and well tolerated, and exposure to drug was maintained despite an antidrug antibody response after systemic administration, based on IND-enabling toxicology and safety pharmacology studies. Ocular doses of EBI-005 at 50 mg/mL in mice and rabbits totaling 0.15 mg/eye in mice and 1.5 mg/eye in rabbits, administered 4 times daily, did not produce adverse effects, and demonstrated excellent bioavailability in target tissues with low systemic exposure. In addition, immunogenic response to the drug did not cause adverse effects or diminish the drug's activity in most cases. The results support drug administration of the highest anticipated human clinical study dose of a 20 mg/mL solution (40 μL 3 times daily in each eye).


The ethical contours of research in crisis settings: five practical considerations for academic institutional review boards and researchers.
The number of research studies in the humanitarian field is rising. It is imperative, therefore, that institutional review boards (IRBs) consider carefully the additional risks present in crisis contexts to ensure that the highest ethical standards are upheld. Ethical guidelines should represent better the specific issues inherent to research among populations grappling with armed conflict, disasters triggered by natural hazards, or health-related emergencies. This paper seeks to describe five issues particular to humanitarian settings that IRBs should deliberate and on which they should provide recommendations to overcome associated challenges: staged reviews of protocols in acute emergencies; flexible reviews of modification requests; addressing violence and the traumatic experiences of participants; difficulties in attaining meaningful informed consent among populations dependent on aid; and ensuring reviews are knowledgeable of populations' needs. Considering these matters when reviewing protocols will yield more ethically sound research in humanitarian settings and hold researchers accountable to appropriate ethical standards.


[Severe hypertriglyceridemia--an important cause of pancreatitis].
Moderate hypertriglyceridaemia is a risk factor for cardiovascular disease and serious hypertriglyceridaemia, with triglyceride values above 10 mmol/L, increases the risk of pancreatitis. Gallstones and alcohol abuse are regarded as the two most important causes of acute pancreatitis, but the considerable risk posed by hypertriglyceridaemia has probably been underrated. It is therefore crucial to acquire updated knowledge and awareness of the fact that high levels of triglycerides can cause pancreatitis. This article is based on current literature retrieved though a search on the topic and clinical experience. Serious hypertriglyceridaemia is a relatively rare condition and its usual cause is genetic predisposition combined with obesity, diabetes or alcohol abuse. Certain types of medication, as well as pregnancy, are also well known causes. Current literature suggests that hypertriglyceridaemia is the cause of pancreatitis in 1-38% of the cases--a substantial variation. The condition is often accompanied by low amylase values and may therefore be underrated as a cause. Our case reports illustrate that the etiology is complex. Plasmapheresis or LDL-apheresis may be indicated when conservative treatment proves insufficient.


Alcoholism and other drug addictions: a primary disease entity, 1991 update.
Intensive work needs to continue on the genetic characteristics of the X-Y chromosome. There needs to be further clarification of the allele penetrants and the oncogenes, as well as determination of the specific chromosomes. It is now apparent that the neurotransmitters, particularly dopamine, seratonin, and norepinephrine, as related to the MFB receptor sites, are a determining factor in the presence or absence of this disease. However, the exact manner in which these neurotransmitters and receptor sites act, particularly in terms of the feed-back systems, remains obscure. New receptor sites are also being discovered and additional neurotransmitter systems, such as the second messenger system, uncovered. It would be naive at this point to attempt to delineate specific receptor sites or neurotransmitter systems as the absolute determinant of the disease. However, the basic heuristic model accepted by the Georgia Impaired Health Professionals Program remains valid. The heuristic model proposed in the past 15 years remains basically intact and valid. In this model, it is apparent that the essential disease criteria are fulfilled by alcohol and other drug addictions. Chemical dependency is a primary, psychosocial, biogenetic disease. The American culture, however, often fails to appreciate the difference between controlled abusive use and the uncontrolled addictive user. They do not understand that controlled abusive use can lead to severe physical changes, even death.(ABSTRACT TRUNCATED AT 250 WORDS)


A Prospective Observational Study of the Epidemiology, Management, and Outcomes of Skin and Soft Tissue Infections Due to Carbapenem-Resistant Enterobacteriaceae.
This study was performed to characterize the epidemiology, management, and outcomes of skin and soft tissue infection (SSTI) and colonization due to carbapenem-resistant Enterobacteriaceae (CRE). Patients from the Consortium on Resistance Against Carbapenem in Klebsiella and Other Enterobacteriaceae (CRACKLE-1) from December 24, 2011 to October 1, 2014 with wound cultures positive for CRE were included in the study. Predictors of surgical intervention were analyzed. Molecular typing of isolates was performed using repetitive extragenic palindromic polymerase chain reaction (PCR). Carbapenemase genes were detected using PCR. One hundred forty-two patients were included: 62 had SSTI (44%) and 56% were colonized. Mean age was 61 years, and 48% were male: median Charlson score was 3 (interquartile range, 1-5). Forty-eight percent of patients were admitted from long-term care facilities (LTCFs), and 31% were from the community. Two strain types (ST258A and ST258B) were identified (73% of 45 tested). Carbapenemase genes were detected in 40 of 45 isolates (blaKPC-3 [47%], blaKPC-2 [42%]). Sixty-eight patients (48%) underwent surgical intervention, 63% of whom had SSTI. Patients admitted from LTCFs were less likely to undergo surgical intervention (odds ratio [OR], 0.36; 95% confidence interval [CI], 0.18-0.71). In multivariable analysis, among patients with SSTI, those admitted from LTCFs were less likely to undergo debridement (OR, 0.18; 95% CI, 0.04-0.93). Patients admitted from LTCFs with CRE SSTI were less likely to undergo surgical intervention. Sixteen percent of the patients died, and approximately 50% of survivors required more intensive care upon discharge. These findings suggest a unique, impactful syndrome within the CRE infection spectrum. Further studies are needed to assess the role of surgical debridement in management of CRE-SSTI, particularly among LTCF residents.


Modifiable risk factors for non-adherence to immunosuppressants in renal transplant recipients: a cross-sectional study.
Non-adherence to immunosuppressants is a major cause of renal transplant failure. Interventions to improve adherence need to target modifiable risk factors. Adherence was measured using the 'gold standard' measure of electronic monitoring in 58 adult renal transplant recipients from a UK transplant unit. Subjects were identified from a stratified random sample of 153 recipients recruited to a larger cross-sectional study comparing different measures of adherence. Inclusion criteria included age over 18 years and a functioning renal transplant, transplanted 6-63 months previously. Exclusion criteria included residence outside the region served by the unit and inability to give informed consent. Health beliefs, depression and functional status were measured using standardized questionnaires (Beliefs about Medicines Questionnaire, Illness Perception Questionnaire, Revised Clinical Interview Schedule and SF-36) and semi-structured interview. Transplant and demographic details were collected from the notes. Seven [12%, 95% confidence interval (CI) 4-20%] subjects missed at least 20% of days medication and 15 (26%, 15-37%) missed at least 10% of days. Lower belief in the need for medication and having a transplant from a live donor were the major factors associated with non-adherence. Depression was common, although not strongly associated with non-adherence. Beliefs about medication are a promising target for interventions designed to improve adherence. The lower adherence in recipients of transplants from live donors needs confirming but may be clinically important in light of the drive to increase live donation.


Production of transgenic Xenopus laevis by restriction enzyme mediated integration and nuclear transplantation.
Stable integration of cloned gene products into the Xenopus genome is necessary to control the time and place of expression, to express genes at later stages of embryonic development, and to define how enhancers and promoters regulate gene expression within the embryo. The protocol demonstrated here can be used to efficiently produce transgenic Xenopus laevis embryos. This transgenesis approach involves three parts: 1. Sperm nuclei are isolated from adult X. laevis testis by treatment with lysolecithin, which permeabilizes the sperm plasma membrane. 2. Egg extract is prepared by low speed centrifugation, addition of calcium to cause the extract to progress to interphase of the cell cycle, and a high-speed centrifugation to isolate interphase cytosol. 3. Nuclear transplantation: the nuclei and extract are combined with the linearized plasmid DNA to be introduced as the transgene and a small amount of restriction enzyme. During a short reaction, egg extract partially decondenses the sperm chromatin and the restriction enzyme generates chromosomal breaks that promote recombination of the transgene into the genome. The treated sperm nuclei are then transplanted into unfertilized eggs. Integration of the transgene usually occurs prior to the first embryonic cleavage such that the resulting embryos are not chimeric. These embryos can be analyzed without any need to breed to the next generation, allowing for efficient and rapid generation of transgenic embryos for analyses of promoter and gene function. Adult X. laevis resulting from this procedure also propagate the transgene through the germline and can be used to generate lines of transgenic animals for multiple purposes.


High frequency of IgE receptor Fc epsilonRIbeta variant (Leu181/Leu183) in Kuwaiti Arabs and its association with asthma.
The presence of IgE receptor Fc epsilonRIbeta polymorphism (181/183) was investigated in Kuwaiti asthmatic patients and controls using an allele refractory mutation screening (ARMS) test. The variant sequence (Leu181/Leu183) was detected in 72% (320/442) chromosomes analysed. Homozygous LL genotype was detected in 48% (46/96) asthmatic subjects compared to 31% (39/125) in non-asthmatics. In 11/52 families mothers of the asthmatic children were themselves asthmatic and 3/11 asthmatic mothers had homozygous LL genotype. 80% of the homozygous LL asthmatics showed a positive skin prick test (SPT) compared with 28% of non-asthmatics with the same genotype. In heterozygous NL asthmatics a positive SPT was found in 60% cases compared to 17% in non asthmatics with the same genotype. The incidence of hay fever (HF) and eczema (E) was also found to be higher in homozygous LL asthmatics compared with the non-asthmatics with the same genotype. This study reports a high prevalence of IgE receptor Fc epsilonRIbeta variants in Kuwaiti Arabs compared with British, Australian and Austrian populations studied before. The association of Leu181/Leu183 variant with asthma in Kuwaitis underlines its significance as a risk factor in manifesting the clinical phenotype in this population.


Atypical Teratoid/ Rhabdoid Tumor of Brain: a Clinicopathologic Study of Eleven Patients and Review of Literature
Background: Atypical teratoid/ rhabdoid tumor (AT/RT) is a rare aggressive embryonal central nervous system (CNS) tumor of infancy and early childhood. Majority of the cases arise in the posterior fossa, and remaining in the cerebrum. Aims: To analyze the clinicopathologic features of AT/RT on a cohort of cases. Materials and methods: All reported cases of AT/RT at the Department of Pathology and Laboratory Medicine, Aga Khan University Hospital (AKUH) from 2007 to 2016 were reviewed for clinical and pathological features. Immunohistochemical stain for INI-1 was performed in all 11 cases. Follow up was obtained. Results: A total of 11 cases were identified. Seven patients were males and 4 were females. The ages ranged from 1 month to 48 months (mean 26.6 months). Six tumors were located in the cerebrum and 3 in the posterior fossa. Exact Location was not known in 2 cases. Histologically, rhabdoid cells were present in sheets in variable proportions in five cases, Medulloblastoma and PNET like areas were seen in 2 cases each. Immunohistochemical stains EMA (10/10), vimentin (7/7), CKAE1/AE3 (8/9), and CD99 (3/4), GFAP (6/10), ASMA (3/4) and synaptophysin (3/4) were positive in varying proportions while desmin was negative in all 6 cases in which it was performed. All 11 tumors lacked immunoreactivity for INI-1 protein. Four patients died of disease with a follow up ranging from 5 to 24 months. Conclusions: AT/RT is a rare highly aggressive embryonal tumor of CNS. A male predominance was noted in our series. We report the first and largest series from Pakistan.


Cross-cultural adaptation, reliability and validity of the Spanish version of the Upper Limb Functional Index.
The Upper Limb Functional Index (ULFI) is an internationally widely used outcome measure with robust, valid psychometric properties. The purpose of study is to develop and validate a ULFI Spanish-version (ULFI-Sp). A two stage observational study was conducted. The ULFI was cross-culturally adapted to Spanish through double forward and backward translations, the psychometric properties were then validated. Participants (n = 126) with various upper limb conditions of >12 weeks duration completed the ULFI-Sp, QuickDASH and the Euroqol Health Questionnaire 5 Dimensions (EQ-5D-3 L). The full sample determined internal consistency, concurrent criterion validity, construct validity and factor structure; a subgroup (n = 35) determined reliability at seven days. The ULFI-Sp demonstrated high internal consistency (α = 0.94) and reliability (r = 0.93). Factor structure was one-dimensional and supported construct validity. Criterion validity with the EQ-5D-3 L was fair and inversely correlated (r = -0.59). The QuickDASH data was unavailable for analysis due to excessive missing responses. The ULFI-Sp is a valid upper limb outcome measure with similar psychometric properties to the English language version.


Neurobehavioral toxicity study of dibutyl phthalate on rats following in utero and lactational exposure.
To investigate the neurobehavioral effects of dibutyl phthalate (DBP), an important endocrine disruptor known for reproductive toxicity, on rodent offspring following in utero and lactational exposure, pregnant Wistar rats were treated with DBP (0, 0.037, 0.111, 0.333 and 1% in the diet) from gestation day (GD) 6 to postnatal day (PND) 28, and selected developmental and neurobehavioral parameters of the offspring were measured. There were no significant effects of DBP on body weight gain of the dams during GD 6-20 or on the pups' ages of pinna detachment, incisor eruption or eye opening. Exposure to 1% DBP prolonged gestation period, decreased body weight in both male and female pups, depressed surface righting (PND 7) in male pups, shortened forepaw grip time (PND 10), enhanced spatial learning and reference memory (PND 35) in male pups. Exposure to 0.037% DBP also shortened forepaw grip time (PND 10), but inhibited spatial learning and reference memory in male pups. Sex x treatment effects were found in forepaw grip time (PND 10), spatial learning and reference memory, and the male pups appeared to be more susceptible than the females. However, all levels of DBP exposure did not significantly alter surface righting (PND 4), air righting (PND 16), negative geotaxis (PND 4 or 7), cliff avoidance (PND 7) or open field behavior (PND 28) in either sex. Overall, the dose level of DBP in the present study produced a few adverse effects on the neurobehavioral parameters, and it may alter cognitive abilities of the male rodent.


Directional asymmetries in the optokinetic response of larval zebrafish (Danio rerio).
Photographic images of the optokinetic response (OKR) of larval zebrafish permitted the calculation of the amplitude and velocity of the response to gratings of various spatial frequencies rotating at different speeds. At low spatial frequencies, the amplitude of the OKR did not vary significantly for drum speeds ranging from 24 to 108 degrees/sec. Plotting the velocity of the OKR as a function of drum speed gave rise to a bell-shaped curve, with a maximum at about 48 degrees/sec. Interestingly, both eyes exhibited an asymmetric response to the rotating drum, that is, they were more responsive to temporal-to-nasal rotation than to nasal-to-temporal motion. Although this asymmetry persisted over the entire range of drum speeds tested, the situation was reversed when tested with gratings of higher spatial frequency (i.e., the eyes became more responsive to rotation in the nasal-to-temporal direction). The amplitude of the OKR for both eyes exhibited an inverse relation to increasing spatial frequency of the stimulus, whereas the velocity of the OKR showed a steep decline within the range of 0.08 to 0.14 cycles/degree. The data indicate that zebrafish are more responsive to objects with low spatial frequencies moving from behind the animal's head toward the frontal plane, and to high spatial frequencies of objects moving across the frontal plane (perpendicular to the anterior-posterior axis of the eye).


Approaching the genomics of risk-taking behavior.
Individual animals differ in their propensity to engage in dangerous situations, or in their risk-taking behavior. There is a heritable basis to some of this variation, but the environment plays an important role in shaping individuals' risk-taking propensity as well. This chapter describes some of the challenges in studying the genetic basis of individual differences in risk-taking behavior, arguing new insights will emerge from studies which take a whole-genome approach and which simultaneously consider both genetic and environmental influences on the behavior. The availability of genomic tools for three-spined stickleback, a small fish renowned for its variable behavior, opens up new possibilities for studying the genetic basis of natural, adaptive variation in risk-taking behavior. After introducing the general biology of sticklebacks, the chapter summarizes the existing literature on the genetic and environmental influences on risk-taking behavior, and describes the overall strategy that our group is taking to identify inherited and environmentally responsive genes related to risk-taking behavior in this species. Insights gleaned from such studies will be relevant to our understanding of similar behaviors in other organisms, including ourselves.


Classification and prediction of pilot weather encounters: A discriminant function analysis.
Flight into adverse weather continues to be a significant hazard for General Aviation (GA) pilots. Weather-related crashes have a significantly higher fatality rate than other GA crashes. Previous research has identified lack of situational awareness, risk perception, and risk tolerance as possible explanations for why pilots would continue into adverse weather. However, very little is known about the nature of these encounters or the differences between pilots who avoid adverse weather and those who do not. Visitors to a web site described an experience with adverse weather and completed a range of measures of personal characteristics. The resulting data from 364 pilots were carefully screened and subject to a discriminant function analysis. Two significant functions were found. The first, accounting for 69% of the variance, reflected measures of risk awareness and pilot judgment while the second differentiated pilots in terms of their experience levels. The variables measured in this study enabled us to correctly discriminate between the three groups of pilots considerably better (53% correct classifications) than would have been possible by chance (33% correct classifications). The implications of these findings for targeting safety interventions are discussed.


CpG island methylation in premalignant stages of gastric carcinoma.
There are limited reports on methylation analysis of the premalignant lesions of gastric carcinoma thus far. This is despite the fact that gastric carcinoma is one of the tumors with a high frequency of CpG island hypermethylation. To determine the frequency and timing of hypermethylation during multistep gastric carcinogenesis, non-neoplastic gastric mucosa (n = 118), adenomas (n = 61), and carcinomas (n = 64) were analyzed for their p16, human Mut L homologue 1 (hMLH1), death-associated protein (DAP)-kinase, thromobospondin-1 (THBS1), and tissue inhibitor of metalloproteinase 3 (TIMP-3) methylation status using methylation-specific PCR. Three different classes of methylation behaviors were found in the five tested genes. DAP-kinase was methylated at a similar frequency in all four stages, whereas hMLH1 and p16 were methylated in cancer samples (20.3% and 42.2%, respectively) more frequently than in intestinal metaplasia (6.3% and 2.1%, respectively) or adenomas (9.8% and 11.5%, respectively). However, hMLH1 and p16 were not methylated in chronic gastritis. THBS-1 and TIMP-3 were methylated in all stages but showed a marked increase in hypermethylation frequency from chronic gastritis (10.1% and 14.5%, respectively) to intestinal metaplasia (34.7% and 36.7%, respectively; P < 0.05) and from adenomas (28.3% and 26.7%, respectively) to carcinomas (48.4% and 57.4%, respectively: P < 0.05). The hMLH1, THBS1, and TIMP-3 hypermethylation frequencies were similar in both intestinal metaplasia and adenomas, but the p16 hypermethylation frequency tended to be higher in adenomas (11.5%) than in intestinal metaplasia (2.1%; P = 0.073). The average number of methylated genes was 0.6, 1.1, 1.1, and 2.0 per five genes per sample in chronic gastritis, intestinal metaplasia, adenomas, and carcinomas, respectively. This shows a marked increase in methylated genes from non-metaplastic mucosa to intestinal metaplasia (P = 0.001) as well as from premalignant lesions to carcinomas (P = 0.002). These results suggest that CpG island hypermethylation occur early in multistep gastric carcinogenesis and tend to accumulate along the multistep carcinogenesis.


Carrier frequency of the GJB2 mutations that cause hereditary hearing loss in the Japanese population.
Hearing impairment is one of the most common sensory disorders that affect ~1 in 1000 children, and half of them are considered to be hereditary. Information about the carrier frequencies of mutations that underlie autosomal recessive disorders is indispensable for accurate genetic counseling to predict the probability of patients' children's disease. However, there have been few reports specific to the Japanese population. GJB2 mutations are reported to be the most frequent cause of hereditary hearing loss, and the mutation spectrum and frequency of GJB2 mutations were reported to vary among different ethnic groups. In this study, we investigated the carrier frequency of GJB2 mutations and the mutation spectrum in 509 individuals randomly selected from the general Japanese population. We show that the carrier frequencies of the two most common pathogenic mutations are 1.57% (8/509) for c.235delC and 1.77% (9/509) for p.Val37Ile. In addition to these mutations, we found two pathogenic variants (p.[Gly45Glu;Tyr136*] and p.Arg143Trp), and the total carrier frequency was estimated to be around 3.73% (19/509). We also detected six unclassified variants, including two novel variants (p.Cys60Tyr and p.Phe106Leu), with the former predicted to be pathogenic. These findings will provide indispensable information for genetic counseling in the Japanese population.


Duration of Hyperthyroidism and Lack of Sufficient Treatment Are Associated with Increased Cardiovascular Risk.
Cardiovascular disease remains the most prevalent cause of death in hyperthyroidism. However, the impact on cardiovascular events of varying thyroid status and of treatment remains unclarified. The aims of this study were to investigate the association between hyperthyroidism and cardiovascular events in treated and untreated hyperthyroid individuals, as well as exploring the impact of cumulative periods of hyperthyroidism as a proxy for undertreatment on cardiovascular events. This was a case-control study nested within a population-based cohort of individuals attending health services in Funen County, Denmark, in the period from 1995 to 2011. Data on comorbidities and mortality were collected from The Danish National Patient Register and The Danish Register of Causes of Death. Participants were 275,467 individuals with at least one serum thyrotropin (TSH) measurement in the study period. Hyperthyroidism was defined as at least two measurements of decreased serum TSH within six months, separated by at least 14 days. Incident cases of cardiovascular disease (myocardial infarction, atrial fibrillation, heart failure, stroke, and cardiovascular death) were matched with controls. Conditional logistic regression analyses were performed to calculate odds ratios (OR) for exposure to hyperthyroidism, adjusting for preexisting comorbidities. A total of 20,651 individuals experienced a cardiovascular event (9.5% incidence rate 13.2/1000 person-years [confidence interval (CI) 13.0-13.4]) compared to euthyroid individuals, conditional logistic regression showed increased cardiovascular risk in untreated hyperthyroid patients (OR = 1.25 [CI 1.06-1.48], p = 0.007) but not in treated hyperthyroid patients (OR = 1.04 [CI 0.90-1.22], p = 0.57)]. The OR for cardiovascular events per six months of decreased TSH was 1.09 ([CI 1.05-1.14], p < 0.001) in treated hyperthyroid individuals, and 1.10 ([CI 1.05-1.15], p < 0.001) in untreated hyperthyroid individuals. The risk of cardiovascular disease was found to be increased in untreated hyperthyroid patients, and the duration of decreased TSH associated with increasing risk of cardiovascular outcomes in both treated and untreated hyperthyroid individuals. This suggests that increased cardiovascular risk is driven not only by lack of treatment but also by insufficient therapy. The results support timely treatment and careful monitoring of hyperthyroid patients in order to reduce cardiovascular risk.


Monoclonal Antibodies Reveal Dynamic Plasticity Between Lgr5- and Bmi1-Expressing Intestinal Cell Populations.
Continual renewal of the intestinal epithelium is dependent on active- and slow-cycling stem cells that are confined to the crypt base. Tight regulation of these stem cell populations maintains homeostasis by balancing proliferation and differentiation to support critical intestinal functions. The hierarchical relation of discrete stem cell populations in homeostasis or during regenerative epithelial repair remains controversial. Although recent studies have supported a model for the active-cycling leucine-rich repeat-containing G-protein-coupled receptor 5 (Lgr5)+ intestinal stem cell (ISC) functioning upstream of the slow-cycling B lymphoma Mo-MLV insertion region 1 homolog (Bmi1)-expressing cell, other studies have reported the opposite relation. Tools that facilitate simultaneous analyses of these populations are required to evaluate their coordinated function. We used novel monoclonal antibodies (mAbs) raised against murine intestinal epithelial cells in conjunction with ISC-green fluorescent protein (GFP) reporter mice to analyze relations between ISC populations by microscopy. Ex vivo 3-dimensional cultures, flow cytometry, and quantitative reverse-transcription polymerase chain reaction analyses were performed. Two novel mAbs recognized distinct subpopulations of the intestinal epithelium and when used in combination permitted isolation of discrete Lgr5GFP and Bmi1GFP-enriched populations with stem activity. Growth from singly isolated Lgr5GFP ISCs gave rise to small spheroids. Spheroids did not express Lgr5GFP and instead up-regulated Bmi1GFP expression. Conversely, Bmi1-derived spheroids initiated Lgr5GFP expression as crypt domains were established. These data showed the functional utility of murine mAbs in the isolation and investigation of Lgr5GFP and Bmi1GFP ISC-enriched populations. Ex vivo analyses showed hierarchical plasticity between different ISC-expressing states; specifically Lgr5GFP ISCs gave rise to Bmi1GFP cells, and vice versa. These data highlight the impact of temporal and physiological context on unappreciated interactions between Lgr5GFP and Bmi1GFP cells during crypt formation.


Novel treatments for bipolar disorder.
Treatments other than lithium have recently emerged as equally important in the management of bipolar disorder. The spectrum of efficacy of newer treatments differs from lithium and among the novel drug treatments valproate, generally used as the better tolerated divalproex form, principally benefits manic symptomatology both acutely and in prophylaxis. Atypical antipsychotic drugs have demonstrated efficacy in reducing acute manic symptoms. No controlled evidence of efficacy in prophylaxis has been published. Lamotrigine has demonstrated efficacy in both acute bipolar depression and maintenance efficacy in rapid cycling bipolar patients, especially those patients with bipolar II disorder, which is principally manifested as depression. Randomised, double-blind, placebo- controlled studies provide good evidence that regimens of risperidone or olanzapine in combination with lithium or valproate provide greater improvement in acute mania than the mood stabilisers alone. Similarly, valproate combined with antipsychotics provided greater improvement in mania than antipsychotic medication alone and resulted in lower dosage of the antipsychotic medication. A positive but unclear placebo-controlled study of omega-3 fatty acids added to lithium in bipolar disorder needs confirmation in standard clinical trial paradigms. Several other drugs that were reported as beneficial in various facets of bipolar disorder in open trials have not differed from placebo when studied in randomised, placebo-controlled trials.


Function of two ficolin-like proteins in innate immune defense of the oriental river prawn, Macrobrachium nipponense.
Ficolins have crucial functions in recognizing and eliminating pathogens in innate immunity. In this study, we identified two ficolin-like genes from the oriental river prawn, Macrobrachium nipponense. These genes were designated as MnFico1 and MnFico2. MnFico1 cDNA has 1600 bp, whereas MnFico2 has 1486 bp. In addition to a coiled-coil region or a low complexity region, the two ficolins both contained a signal peptide and a fibrinogen-related domain. qRT-PCR results showed that the highest expression level of MnFico1 expression was in the gills, whereas that of MnFico2 was in the heart. The transcripts of MnFico1 and MnFico2 could both respond to bacteria challenge. The transcription of antilipopolysaccharide factors ALFs (MnALF1 and MnALF2) and crustin genes (MnCru4, MnCru5, MnCru6, and MnCru7) was inhibited in the gills of MnFico1 or MnFico2 knockdown prawns at 24 h Vibrio parahaemolyticus challenge. Recombinant proteins of rMnFico1 and rMnFico2 could bind toward diverse bacteria and agglutinate Gram-negative and Gram-positive bacteria with the presence of calcium (Ca2+). rMnFico1 and rMnFico2 proteins also have lipopolysaccharide and peptidoglycan binding activity. Both recombinant ficolin proteins could help the prawn to facilitate the clearance of V. parahaemolyticus in vivo. Our results suggested that MnFico1 and MnFico2 might serve as pattern recognition receptors in M. nipponense.


Down-regulation of MTA1 protein leads to the inhibition of migration, invasion, and angiogenesis of non-small-cell lung cancer cell line.
Metastasis-associated protein 1 (MTA1) high expression has been detected in a wide variety of human aggressive tumors and plays important roles in the malignant biological behaviors such as invasion, metastasis, and angiogenesis. However, the specific roles and mechanisms of MTA1 protein in regulating the malignant behaviors of non-small-cell lung cancer (NSCLC) cells still remain unclear. To elucidate the detailed functions of MTA1 protein, we down-regulated the MTA1 protein expression in NSCLC cell line by RNA interference (RNAi) in vitro, and found that down-regulation of MTA1 protein significantly inhibited the migration and invasion potentials of 95D cells. Further research revealed that down-regulation of MTA1 protein significantly decreased the activity of matrix metalloproteinase-9, which could be the mechanism responsible for the inhibition of 95D cells migration and invasion. In addition, the tube formation assay demonstrated that the number of complete tubes induced by the conditioned medium of MTA1-siRNA 95D cells was significantly smaller than that of 95D cells. These findings demonstrate that MTA1 protein plays important roles in regulating the migration, invasion, and angiogenesis potentials of 95D cells, suggesting that MTA1 protein down-regulation by RNAi might be a novel therapeutic approach to inhibit the progression of NSCLC.


Photoemission evidence for a remnant fermi surface and a d-wave-like dispersion in insulating Ca2CuO2Cl2
An angle-resolved photoemission study is reported on Ca2CuO2Cl2, a parent compound of high-Tc superconductors. Analysis of the electron occupation probability, n(k), from the spectra shows a steep drop in spectral intensity across a contour that is close to the Fermi surface predicted by the band calculation. This analysis reveals a Fermi surface remnant, even though Ca2CuO2Cl2 is a Mott insulator. The lowest energy peak exhibits a dispersion with approximately the &cjs3539;coskxa - coskya&cjs3539; form along this remnant Fermi surface. Together with the data from Dy-doped Bi2Sr2CaCu2O8+delta, these results suggest that this d-wave-like dispersion of the insulator is the underlying reason for the pseudo gap in the underdoped regime.


Lentiviral vectors pseudotyped with minimal filovirus envelopes increased gene transfer in murine lung.
A human immunodeficiency virus (HIV)-based vector pseudotyped with the Ebola Zaire (EboZ) viral envelope glycoprotein (GP) was recently shown to transduce murine airway epithelia cells in vivo. In this study, the vector was further redesigned to improve gene transfer and also to increase safety. We used mutant EboZ envelopes for pseudotyping, which resulted in higher titers and increased transduction of airway cells in vivo compared to vectors pseudotyped with wild-type EboZ GP. As these envelopes lack regions associated with toxicity of the wild-type EboZ GP, they should also be safer to use for pseudotyping of lentiviral vectors. In addition, lentiviral vectors were created based on feline immunodeficiency virus and shown to have similar efficiency of transduction compared to HIV-based vectors. The creation of lentiviral vectors with highly engineered EboZ envelopes improved the performance of the system and should also increase its safety since only minimal regions of the EboZ envelope, which lack the toxic domain, are used.


[Protein concentrate of African oil palm (Elaesis guineensis, Jacquin), extraction process and functional properties].
A study was carried out for the purpose of obtaining and characterizing a protein concentrate obtained from defatted oil palm cake using alkaline extraction, and compare it with a commercial soy meal. The oil palm cake came from a national industry as a subproduct of the oil extraction of the palm kernels. The moisture, protein, fat, crude fiber and ash content of the oil palm was then determined. The optimum conditions for extraction and precipitation of the proteins were selected. These were the following: extraction at pH, 11.4; adding NaOH 0.06 M solvent; a meal/solvent relation of 1:20 g/ml and extraction time, 20 minutes with magnetic agitation, and precipitation at pH 5.3. The protein concentrate obtained contained: 66.50% protein; 0.07% fat, 0.90% crude fiber, and 3.20% ashes. Then the following functional properties were analyzed: solubility, according to the pH; water absorption (250); oil absorption (175); emulsion activity (27.2), and stability (13.6). The author concludes that the protein concentrate has good water and oil absorption when compared to soymeal; the emulsion, however, was found to be unstable to heat.


Near-infrared fluorescence imaging of microcalcification in an animal model of breast cancer.
At present, there is no animal model of breast cancer that forms reproducible microcalcification. The aim of this study was to develop a straightforward, reproducible model system that could be used to develop multimodality contrast agents for the identification of breast cancer microcalcification. The R3230 mammary adenocarcinoma cell line was implanted in the mammary fat pad of female Fischer 344 rats (two rats with two implanted tumors and two rats with a single implanted tumor). After growth to 1-2 cm in diameter, tumors were implanted with 100 microm hydroxyapatite crystals (positive control) or calcium oxalate crystals (negative control). Twenty-four hours after crystal implantation, rats were injected intravenously with a previously described near-infrared fluorescent bisphosphonate derivative known as Pam78, and the tumors were imaged using a reflectance optical imaging system. Tumors implanted with hydroxyapatite displayed bright, focal, near-infrared fluorescence in the area of crystal implantation. Control tumors, grown in the same animal and implanted with calcium oxalate, did not display any near-infrared fluorescence, even along the needle track used for crystal implantation. A simple and rapid animal model of focal calcification in breast cancer tumors has been developed and validated. The model used Pam78, a near-infrared fluorescent contrast agent specific for hydroxyapatite. The potential usefulness of the model for developing similar contrast agents for magnetic resonance and other imaging modalities is discussed.


Multispectral texture synthesis using fractal concepts.
Natural textures are synthesized across several spectral bands using no more than a single second-order descriptor for each band. The Fractal model for monochrome textures is used as a null-hypothesis. The eye's sensitivity to Fractal dimension under specific conditions is determined. Multispectral camouflage is generated to imitate a range of natural textures.


The Role of Audiology in an Outpatient Interdisciplinary Post-Trauma Clinic.
Patients in the process of recovering from severe bodily injury will encounter several barriers to effective treatment. When present, untreated hearing loss can create additional obstacles in a process that is already difficult. This article describes an outpatient post-trauma clinic associated with a tertiary care hospital trauma unit that consolidates rehabilitation resources 2 weeks after inpatient discharge to help these patients on their path to recovery. The role of audiology in the interdisciplinary clinic is described and data related to services are presented. Some practical tips for implementation of audiologic services in this type of environment are provided.


Determination of residues responsible for substrate and product specificity of Solanum habrochaites short-chain cis-prenyltransferases.
Isoprenoids are diverse compounds that have their biosynthetic origin in the initial condensation of isopentenyl diphosphate and dimethylallyl diphosphate to form C10 prenyl diphosphates that can be elongated by the addition of subsequent isopentenyl diphosphate units. These reactions are catalyzed by either cis-prenyltransferases (CPTs) or trans-prenyltransferases. The synthesis of volatile terpenes in plants typically proceeds through either geranyl diphosphate (C10) or trans-farnesyl diphosphate (C15), to yield monoterpenes and sesquiterpenes, respectively. However, terpene biosynthesis in glandular trichomes of tomato (Solanum lycopersicum) and related wild relatives also occurs via the cis-substrates neryl diphosphate (NPP) and 2Z,6Z-farnesyl diphosphate (Z,Z-FPP). NPP and Z,Z-FPP are synthesized by neryl diphosphate synthase1 (NDPS1) and Z,Z-farnesyl diphosphate synthase (zFPS), which are encoded by the orthologous CPT1 locus in tomato and Solanum habrochaites, respectively. In this study, comparative sequence analysis of NDPS1 and zFPS enzymes from S. habrochaites accessions that synthesize either monoterpenes or sesquiterpenes was performed to identify amino acid residues that correlate with the ability to synthesize NPP or Z,Z-FPP. Subsequent structural modeling, coupled with site-directed mutagenesis, highlighted the importance of four amino acids located within conserved domain II of CPT enzymes that form part of the second α-helix, for determining substrate and product specificity of these enzymes. In particular, the relative positioning of aromatic amino acid residues at positions 100 and 107 determines the ability of these enzymes to synthesize NPP or Z,Z-FPP. This study provides insight into the biochemical evolution of terpene biosynthesis in the glandular trichomes of Solanum species.


Body mass estimates of an exceptionally complete Stegosaurus (Ornithischia: Thyreophora): comparing volumetric and linear bivariate mass estimation methods.
Body mass is a key biological variable, but difficult to assess from fossils. Various techniques exist for estimating body mass from skeletal parameters, but few studies have compared outputs from different methods. Here, we apply several mass estimation methods to an exceptionally complete skeleton of the dinosaur Stegosaurus. Applying a volumetric convex-hulling technique to a digital model of Stegosaurus, we estimate a mass of 1560 kg (95% prediction interval 1082-2256 kg) for this individual. By contrast, bivariate equations based on limb dimensions predict values between 2355 and 3751 kg and require implausible amounts of soft tissue and/or high body densities. When corrected for ontogenetic scaling, however, volumetric and linear equations are brought into close agreement. Our results raise concerns regarding the application of predictive equations to extinct taxa with no living analogues in terms of overall morphology and highlight the sensitivity of bivariate predictive equations to the ontogenetic status of the specimen. We emphasize the significance of rare, complete fossil skeletons in validating widely applied mass estimation equations based on incomplete skeletal material and stress the importance of accurately determining specimen age prior to further analyses.


Effects of earthworms on surface clogging characteristics of intermittent sand filters.
Intermittent sand filters (ISFs) are effective and economical in treating wastewater, but they are easy to clog up. To explore a feasible and simple method to alleviate clogging, two pilot-scale ISFs were constructed, one of which contained earthworms and the other did not. During the operation, the effects of earthworms on the hydraulic behaviour of ISFs were investigated. The results showed that both ISFs exhibited good performance in wastewater treatment. However, they showed different hydraulic characteristics although operated at the same organic loading rate (approximately 300 g m(-2) d(-1)). The ISF without earthworms clogged only after 53 d operation, and was partially recovered after 7 d resting, but after that, clogging occurred again, and more rapidly than the initial clogging event (40 d). However, water on the medium surface of the ISF with earthworms was not observed during the whole experiments. In addition, 11-13% of effective porosity and 0.015-0.026 cm s(-1) of infiltration rate were measured in the upper 20 cm of the ISF at the end of the experiments. The facts demonstrated that earthworms played a positive role in alleviating clogging and earthworms fed filter could alleviate surface clogging effectively.


Effect of aerosolized prostacyclin and inhaled nitric oxide on experimental hypoxic pulmonary hypertension.
To compare the effect of different concentrations of inhaled nitric oxide and doses of nebulized prostacyclin on hypoxia-induced pulmonary hypertension in pigs. Prospective, controlled animal study. Animal research facilities of an university hospital. After reducing the fraction of inspired oxygen (FIO(2)) from 1.0 to 0.1, two groups of five pigs each were submitted to inhalation of three concentrations of nitric oxide (5, 10 and 20 ppm) or three doses of prostacyclin (2.5, 5, 10 ng x kg(-1) x min(-1)). All doses of prostacyclin and concentrations of nitric oxide resulted in a decrease in mean pulmonary arterial pressure and pulmonary vascular resistance when compared to hypoxic ventilation (p < 0.001) which was independent of the dose or concentration of either drug used. While inhalation of nitric oxide caused a reduction in mean pulmonary arterial pressure back to values obtained during ventilation with FIO(2) 1.0, values achieved with prostacyclin were still significantly higher when compared to measurements prior to the initiation of hypoxic ventilation. However, direct comparison of the effect of 20 ppm nitric oxide and 10 ng x kg(-1) x min(-1) prostacyclin on mean pulmonary arterial pressure revealed no differences between the drugs. All other hemodynamic and gas exchange parameters remained stable throughout the study. Inhalation of clinically used concentrations of nitric oxide and doses of prostacyclin can decrease elevated pulmonary arterial pressure in an animal model of hypoxic pulmonary vasoconstriction without impairing systemic hemodynamics or gas exchange.


An ultraviolet floral polymorphism associated with life history drives pollinator discrimination in Mimulus guttatus.
• Ultraviolet (UV) floral patterns are common in angiosperms and mediate pollinator attraction, efficiency, and constancy. UV patterns may vary within species, yet are cryptic to human observers. Thus, few studies have explicitly described the distribution or ecological significance of intraspecific variation in UV floral patterning. Here, we describe the geographic distribution and pattern of inheritance of a UV polymorphism in the model plant species Mimulus guttatus (Phrymaceae). We then test whether naturally occurring UV phenotypes influence pollinator interactions within M. guttatus.• We document UV patterns in 18 annual and 19 perennial populations and test whether UV pattern is associated with life history. To examine the pattern of inheritance, we conducted crosses within and between UV phenotypes. Finally, we tested whether bee pollinators discriminate among naturally occurring UV phenotypes in two settings: wild bee communities and captive Bombus impatiens.• Within M. guttatus, perennial populations exhibit a small bulls-eye pattern, whereas a bilaterally symmetric runway pattern occurs mainly in annual populations. Inheritance of UV patterning is consistent with a single-locus Mendelian model in which the runway phenotype is dominant. Bee pollinators discriminate against unfamiliar UV patterns in both natural and controlled settings.• We describe a widespread UV polymorphism associated with life history divergence within Mimulus guttatus. UV pattern influences pollinator visitation and should be considered when estimating reproductive barriers between life history ecotypes. This work develops a new system to investigate the ecology and evolution of UV floral patterning in a species with extensive genomic resources.


ROS/Autophagy/Nrf2 Pathway Mediated Low-Dose Radiation Induced Radio-Resistance in Human Lung Adenocarcinoma A549 Cell.
Low-dose ionizing radiation (LDIR) can induce radio-resistance to following high dose radiation in various mammalian cells. The protective role of LDIR has been thought to be associated with the overall outcomes of cancer radiotherapy. NF-E2 related factor 2 (Nrf2) is a transcription factor that plays pivotal roles in maintaining cellular oxidative equilibrium. Since oxidative stress has been indicated to be a mediator of LDIR induced radio-resistance, the role of Nrf2 in this process was investigated in this research. Our results showed that in human lung adenocarcinoma A549 cell, 5cGy alpha particle induced radio-resistance to following 75cGy alpha particle radiation. The expression level of Nrf2 and its target Heme Oxygenase-1(HO-1) increased after 5cGy radiation. Both the shRNA of Nrf2 and the chemical inhibitor of HO-1 suppressed the induced radio-resistance, indicating the involvement of Nrf2 antioxidant pathway in this process. Further, we found 5cGy radiation stimulated autophagy process in A549. Inhibition of the autophagy process resulted in suppression of the radio-resistance and the induced expression of Nrf2 and HO-1. ROS scavenger N-acetyl-L-cysteine (NAC) blocked the autophagy process induced by 5cGy alpha particle, the upregulation of Nrf2 and HO-1, as well as the induced radio-resistance. In conclusion, ROS elevation caused by LDIR promoted Autophagy/Nrf2-HO-1 and conferred radio-resistance in A549.


Blood pressure and vascular calcification.
The aim of this study was to determine the associations between the presence and extent of calcified atherosclerosis in multiple vascular beds and systolic blood pressure, diastolic blood pressure, pulse pressure, mean arterial pressure, isolated systolic hypertension, and hypertension. A total of 9510 patients (42.5% women) underwent electron beam computed tomography scanning as part of a routine health maintenance screening. At the same visit, blood pressure was measured with the participant in the seated position using a mercury sphygmomanometer. Mean age was 58+/-11.4 years, and body mass index was 27.1+/-4.5. The prevalences of any calcification in the carotids, coronaries, subclavians, thoracic aorta, abdominal aorta, and iliacs were 31.9%, 57.2%, 31.7%, 37.0%, 54.3%, and 48.8%, respectively. In separate multivariable logistic models containing traditional cardiovascular disease risk factors, pulse pressure and systolic blood pressure were significantly associated with the presence of calcification in all of the vascular beds except the iliacs and subclavians, respectively, with pulse pressure having stronger magnitudes of the associations for most of the vascular beds. Age-stratified analyses indicated that these associations were stronger in those >60 years of age compared with subjects <60 years of age, and sex-stratified analyses demonstrated that men had a greater association compared with women. Also, the magnitudes of the associations for isolated systolic hypertension were, in general, larger than those for hypertension. Pulse pressure and isolated systolic hypertension are robust and important correlates for calcified atherosclerosis in different vascular beds. Isolated systolic hypertension may be clinically relevant in diagnosing or preventing calcified atherosclerosis.


The dual role of GoPGF reveals that the pigment glands are synthetic sites of gossypol in aerial parts of cotton.
Gossypol and the related terpenoids are stored in the pigment gland to protect cotton plants from biotic stresses, but little is known about the synthetic sites of these metabolites. Here, we showed that GoPGF, a key gene regulating gland formation, was expressed in gland cells and roots. The chromatin immunoprecipitation sequencing (ChIP-seq) analysis demonstrated that GoPGF targets GhJUB1 to regulate gland morphogenesis. RNA-sequencing (RNA-seq) showed high accumulation of gossypol biosynthetic genes in gland cells. Moreover, integrated analysis of the ChIP-seq and RNA-seq data revealed that GoPGF binds to the promoter of several gossypol biosynthetic genes. The cotton callus overexpressing GoPGF had dramatically increased the gossypol levels, indicating that GoPGF can directly activate the biosynthesis of gossypol. In addition, the gopgf mutant analysis revealed the existence of both GoPGF-dependent and -independent regulation of gossypol production in cotton roots. Our study revealed that the pigment glands are synthetic sites of gossypol in aerial parts of cotton and that GoPGF plays a dual role in regulating gland morphogenesis and gossypol biosynthesis. The study provides new insights for exploring the complex relationship between glands and the metabolites they store in cotton and other plant species.


[Effect and side effects of apraclonidine].
Apraclonidine is a alpha 2 adrenergic agonic which is a para-amino derivative of clonidine. This molecular alteration helps prevent apraclonidine from crossing the blood-brain barrier so it minimally influences the heart rate and blood pressure. Apraclonidine reduces the intraocular pressure primarily by an aqueous suppressant mechanism. Its ocular hypotensive efficacy as monotherapy is similar to timolol maleate 0.5% up to eight hours post dosing, but is less than timolol maleate by 12 hours following instillation. As an adjunctive agent apraclonidine decreases the intraocular pressure approximately 16% over that of timolol maleate done. These clinical effects are stable out of three months after the initiation of therapy. Apraclonidine is also additive to maximally tolerated medical therapy. As many as 60% of these patients do not require additional therapy after adding apraclonidine for as long as five months. Apraclonidine also is useful to prevent transiently increased intraocular pressure following anterior segment laser procedures and intraocular surgery. Apraclonidine is well tolerated systemically with no serious adverse event attributed previously to this medicine. The most common ocular side effect is anterior segment intolerance in between 8-36% of patients. Symptoms of itching, as well as burning, and signs of conjunctival inflammation may appear. The intolerance quickly resolves upon discontinuation of the medicine. In summary, apraclonidine is useful in chronic therapy for chronic open-angle glaucoma and helping to prevent acute increased intraocular pressure following anterior segment laser procedures.


Defining desmosomal plakophilin-3 interactions.
Plakophilin 3 (PKP3) is a recently described armadillo protein of the desmosomal plaque, which is synthesized in simple and stratified epithelia. We investigated the localization pattern of endogenous and exogenous PKP3 and fragments thereof. The desmosomal binding properties of PKP3 were determined using yeast two-hybrid, coimmunoprecipitation and colocalization experiments. To this end, novel mouse anti-PKP3 mAbs were generated. We found that PKP3 binds all three desmogleins, desmocollin (Dsc) 3a and -3b, and possibly also Dsc1a and -2a. As such, this is the first protein interaction ever observed with a Dsc-b isoform. Moreover, we determined that PKP3 interacts with plakoglobin, desmoplakin (DP) and the epithelial keratin 18. Evidence was found for the presence of at least two DP-PKP3 interaction sites. This finding might explain how lateral DP-PKP interactions are established in the upper layers of stratified epithelia, increasing the size of the desmosome and the number of anchoring points available for keratins. Together, these results show that PKP3, whose epithelial and epidermal desmosomal expression pattern and protein interaction repertoire are broader than those of PKP1 and -2, is a unique multiprotein binding element in the basic architecture of a vast majority of epithelial desmosomes.


Effects of ethylene glycol monomethyl ether on various parameters of testicular function in the F344 rat.
These studies were designed to define the target cell for ethylene glycol monomethyl ether (EGME) action in the rodent testis, and to compare fluid production and protein secretory patterns in the testes of treated and control animals. Adult F344 rats were treated with 150 mg/kg/day, 5 day/week, and sacrificed 1, 2, 4, 7 or 10 days after the first dose. In the testes of treated animals, the most sensitive cells were the premeiotic and meiotic spermatocytes. As length of treatment continued, the younger transitional spermatocytes, and more pachytene spermatocytes, were affected. Early and late stage spermatids did not appear affected, nor was the visual appearance of spermatogonia changed by exposure to EGME. Fluid production, androgen-binding protein secretion and electrophoreograms of proteins found in the ligated rete testis were also unchanged by EGME, showing that these indices of Sertoli cell function were not affected by EGME. The histologic and biochemical data suggest that both early and late spermatocytes are targets for EGME in the testis, and that the Sertoli cells are relatively unaffected.


Analytic solution of ray equations in cylindrically inhomogeneous guiding media. Part 2: skew rays.
Analytic solutions are found for skew ray paths in inhomogeneous cylindrical systems such as GRIN rods and gas lenses, which focus rays by virtue of a gradual radial decrease in refractive index. The solutions are obtained by employing digital computers to carry out analytic, as opposed to numerical, calculations.


Rapid and selective cerebral hypothermia achieved using a cooling helmet.
Hypothermia is by far the most potent neuroprotectant. Nevertheless, timely and safe delivery of hypothermia remains a clinical challenge. To maximize neuroprotection yet minimize systemic complications, ultra-early delivery of selective cerebral hypothermia by Emergency Medical Service (EMS) personnel in the field would be advantageous. The authors (W.E. and H.W.) have developed a cooling helmet by using National Aeronautics and Space Administration spinoff technology. In this study its effectiveness in lowering brain temperature in patients with severe stroke or head injury is examined. Patients were randomly assigned to groups receiving either the cooling helmet or no cooling, and brain temperatures (0.8 cm below the cortical surface) were continuously monitored for a mean of 48 to 72 hours with a Neurotrend sensor and then compared with the patients' core temperatures. There were eight patients in the study group and six in the control group. The mean change in temperature (brain-body temperature) calculated from 277 data hours in the study group was -1.6 degrees C compared with a mean change in temperature of +0.22 degrees C calculated from 309 data hours in the control group. This was statistically significant (p < 0.0001). On average, 1.84 degrees C of brain temperature reduction (range 0.9-2.4 degrees C) was observed within 1 hour of helmet application. It took a mean of 3.4 hours (range 2-6 hours) to achieve a brain temperature lower than 34 degrees C and 6.67 hours (range 1-12 hours) before systemic hypothermia (< 36 degrees C) occurred. Use of the helmet resulted in no significant complications. There was, however, one episode of asymptomatic bradycardia (heart rate < 40) that responded to a 0.5 degrees C body temperature increase. This helmet delivers initial rapid and selective brain cooling and maintains a significant temperature gradient between the core and brain temperatures throughout the hypothermic period to provide sufficient regional hypothermia yet minimize systemic complications. It results in delayed systemic hypothermia, creating a safe window for possible ultra-early delivery of regional hypothermia by EMS personnel in the field.


Intramedullary nailing of femoral shaft fractures. Part III: Long-term effects of static interlocking fixation.
The results of treatment of fractures of the femoral shaft with static interlocking nailing were reviewed retrospectively to determine the clinical importance of any stress-riser or stress-shielding properties of the nail. These properties, if relevant, would have been manifested by refracture of the femur, either through a hole used for a locking screw or through the original site of fracture after extraction of the device. Two hundred and fourteen fractures that had been treated with static interlocking nailing and that had healed without conversion to dynamic intramedullary fixation were divided into two groups. In Group I, which comprised 111 fractures, the static interlocking-fixation device was retained and in Group II, which comprised 103 fractures, the static interlocking-fixation device was removed during one operative procedure at an average of fourteen months after the injury. The average duration of follow-up was thirty months from the time of the original fixation in both groups. All patients in Group II were followed for a minimum of six months after removal of the nail. No femur in Group I, in which the static interlocked nail remained in situ, refractured. No femur in either group fractured through the proximal or the distal holes used for the locking screws. No locking screws or nails broke. One patient (1 per cent) in Group II had a refracture of the femoral shaft through the site of the original fracture six weeks after removal of the nail.(ABSTRACT TRUNCATED AT 250 WORDS)


Air-breathing during activity in the fishes amia calva and lepisosteus oculatus
Many osteichthyan fishes obtain oxygen from both air, using a lung, and water, using gills. Although it is commonly thought that fishes air-breathe to survive hypoxic aquatic habitats, other reasons may be more important in many species. This study was undertaken to determine the significance of air-breathing in two fish species while exercising in oxygen-rich water. Oxygen consumption from air and water was measured during mild activity in bowfin (Amia calva) and spotted gar (Lepisosteus oculatus) by sealing a fish in an acrylic flume that contained an air-hole. At 19-23 degreesC, the rate of oxygen consumption from air in both species was modest at rest. During low-level exercise, more than 50 % of the oxygen consumed by both species was from the air (53.0+/-22.9 % L. oculatus; 66.4+/-8.3 % A. calva). <P>


Impact of MRI timing on tumor volume and anatomic displacement for brain metastases undergoing stereotactic radiosurgery.
The objective of this study was to evaluate the impact of the time interval between planning imaging and stereotactic radiosurgery (SRS) delivery on tumor volumes and spatial anatomic displacements of brain metastases (BM). Consecutive patients diagnosed with BM treated with SRS over a 3-year period were evaluated. Only patients who underwent an institutionally standardized diagnostic MRI (MRI-1) and a treatment planning MRI (MRI-2) were included. The impact of histology, inter-scan time interval, lesion location, tumor volume, and diameter were evaluated on final lesion diameter, volume, anatomic displacement, and ultimate need for change in management (ie, expanding margins, rescanning). 101 patients (531 lesions) with a median inter-scan time interval of 8 days (range: 1-42 days) met the inclusion criteria. The median percentage increase in BM diameter and volume were 9.5% (IQR: 2.25%-24.0%) and 20% (IQR: 0.7%-66.7%). Overall, 147 lesions (27.7%) in 57 patients (56.4%) required a change in management. There was a statistically significant relationship between initial tumor diameter (cm) and change in management (OR: 2.69, 95% CI: 1.93-3.75; P < .001). Each day between MRI-1 and MRI-2 was associated with a change in management with an OR of 1.05 (95% CI: 1.03-1.07; P < .001). Changes in tumor diameter, volume, and spatial position occur as a function of time. Planning imaging for SRS is recommended to occur in close temporal proximity to treatment; for those with delays, a larger setup margin may need to be used to ensure tumor coverage and account for positional changes.


Association between parathyroid hormone (PTH)/PTH-related peptide receptor gene polymorphism and the extent of bone mass reduction in primary hyperparathyroidism.
Genetic contributions to bone mineral density (BMD) and bone turnover are well known. In the present study, we analyzed the relationship between polymorphism of parathyroid hormone (PTH)/PTH-related peptide (PTHrP) receptor gene existing in exon M7 and the clinical characteristics of primary hyperparathyroidism (pHPT). PTH/PTHrP receptor genotypes were analyzed in 92 pHPT patients by direct sequence to determine whether nucleotide 1417 of the cDNA was C or T. BMD levels at the lumbar spine and at the radius before and one year after parathyroidectomy, as well as serum levels of calcium, phosphorus, alkaline phosphatase (ALP) and intact PTH were measured. Although there were no significant differences in serum levels of calcium, phosphorus and intact PTH, ALP was significantly lower in the CT genotype compared with the TT genotype. BMD level at the radius was significantly higher in the CT genotype than in the CC genotype. Moreover, an increase in radial BMD one year after parathyroidectomy was significantly less in CT genotype than two other genotypes (CC, TT). The present study is the first to indicate that the polymorphism of PTH/PTHrP receptor gene is closely related to the extent of bone mass reduction in pHPT and that this polymorphism would be one of the genetic factors responsible for the severity of the pathological state of pHPT.


Expression of adipocyte-derived leucine aminopeptidase in endometrial cancer. Association with tumor grade and CA-125.
Adipocyte-derived leucine aminopeptidase (A-LAP) is a novel zinc-metallopeptidase involved in angiotensin II (AngII) metabolism, cell migration and antigen presentation. These functions are implicated in the progression of cancer, whereas A-LAP expression and involvement have not been studied in any type of cancer. We investigated the expression of A-LAP in endometrial cancer as well as its association with angiogenesis and clinicopathological features. Immunohistochemical staining of 58 endometrial endometrioid adenocarcinoma specimens revealed that 37 were A-LAP immunoreactive. We also found that A-LAP staining correlated with histological tumor grade in a significant and reverse manner. In addition, serum CA-125 levels in patients with A-LAP positive cancers were significantly higher. However, contrary to our hypothesis that A-LAP suppresses angiogenic activity via AngII metabolism, A-LAP expression was not associated with the microvessel count determined by CD34 immunostaining. Our results suggest that A-LAP is involved in endometrial cancer cell growth and differentiation. However, further studies, especially of the biological roles of A-LAP, are required to confirm this notion.


Self-assembling, amphiphilic polymer-gemcitabine conjugate shows enhanced antitumor efficacy against human pancreatic adenocarcinoma.
The therapeutic efficacy of gemcitabine is severely compromised due to its rapid plasma metabolism. Moreover, its hydrophilicity poses a challenge for its efficient entrapment in nanosized delivery systems and to provide a sustained release profile. In this study, gemcitabine was covalently conjugated to poly(ethylene glycol)-block-poly(2-methyl-2-carboxyl-propylene carbonate) (PEG-PCC) which could self-assemble into micelles of 23.6 nm. These micelles afforded protection to gemcitabine from plasma metabolism as evident by negligible amount of gemcitabine and its metabolite dFdU detected in the plasma after 24 h. A controlled release of gemcitabine from the micelles was observed with 53.89% drug release in 10 days in the presence of protease enzyme Cathepsin B. Gemcitabine conjugated micelles were cytotoxic, showed internalization, and induced cell apoptosis in MIA PaCa-2 and L3.6pl pancreatic cancer cell lines. These micelles efficiently inhibited tumor growth when injected intravenously into MIA PaCa-2 cell derived xenograft tumor bearing NSG mice at a dose of 40 mg/kg in terms of reduced tumor volume and tumor weight (0.38 g vs 0.58 g). TUNEL assay revealed that gemcitabine conjugated micelles induced a much higher extent of apoptosis in the tumor tissues compared to free gemcitabine. In conclusion, gemcitabine conjugated micelles were able to enhance the drug payload, protect it from rapid plasma metabolism, and provide a sustained release and showed enhanced antitumor activity, and thus have the potential to provide a better therapeutic alternative for treating pancreatic cancer.


Loss of ventricular preexcitation during noninvasive testing does not exclude high-risk accessory pathways: A multicenter study of WPW in children.
Abrupt loss of ventricular preexcitation on noninvasive evaluation, or nonpersistent preexcitation, in Wolff-Parkinson-White syndrome (WPW) is thought to indicate a low risk of life-threatening events. The purpose of this study was to compare accessory pathway (AP) characteristics and occurrences of sudden cardiac arrest (SCA) and rapidly conducted preexcited atrial fibrillation (RC-AF) in patients with nonpersistent and persistent preexcitation. Patients 21 years or younger with WPW and invasive electrophysiology study (EPS) data, SCA, or RC-AF were identified from multicenter databases. Nonpersistent preexcitation was defined as absence/sudden loss of preexcitation on electrocardiogram, Holter monitoring, or exercise stress test. RC-AF was defined as clinical preexcited atrial fibrillation with shortest preexcited R-R interval (SPERRI) ≤ 250 ms. AP effective refractory period (APERP), SPERRI at EPS , and shortest preexcited paced cycle length (SPPCL) were collected. High-risk APs were defined as APERP, SPERRI, or SPPCL ≤ 250 ms. Of 1589 patients, 244 (15%) had nonpersistent preexcitation and 1345 (85%) had persistent preexcitation. There were no differences in sex (58% vs 60% male; P=.49) or age (13.3±3.6 years vs 13.1±3.9 years; P=.43) between groups. Although APERP (344±76 ms vs 312±61 ms; P<.001) and SPPCL (394±123 ms vs 317±82 ms; P<.001) were longer in nonpersistent vs persistent preexcitation, there was no difference in SPERRI at EPS (331±71 ms vs 316±73 ms; P=.15). Nonpersistent preexcitation was associated with fewer high-risk APs (13% vs 23%; P<.001) than persistent preexcitation. Of 61 patients with SCA or RC-AF, 6 (10%) had nonpersistent preexcitation (3 SCA, 3 RC-AF). Nonpersistent preexcitation was associated with fewer high-risk APs, though it did not exclude the risk of SCA or RC-AF in children with WPW.


[Meningeal carcinomatosis as first manifestation of carcinoma of the bladder: report of 2 cases].
Meningeal carcinomatosis may occur in 0.8-8% of patients with solid tumors. The most common tumors associated with that condition are breast and small cell lung cancer. Meningeal carcinomatosis from urothelial cancer is rare, and it appears described in advanced stages of disease, generally, after chemotherapy. Two cases of meningeal carcinomatosis as the first manifestation of bladder cancer were reported. In the first case, a 46-year-old man presented signs and symptoms indicative of involvement of the spinal roots, subsequently neurologic dysfunction of the brain and cranial nerves appeared. In the second case, a 68-year-old man was admitted to our hospital with a history compatible with panhypopituitarism and pulmonary lymphangitic carcinomatosis from cancer of unknown primary site. Follow-up revealed a transitional cell carcinoma of the bladder and hydrocephalus due to cerebrospinal fluid outflow obstruction. We emphasize in the polymorphic presentation of meningeal carcinomatosis and the necessity to consider the bladder as primary tumor localization.


Serum Immune Profiling for Early Detection of Cervical Disease.
The most recent (2012) worldwide estimates from International Agency for Research on Cancer indicate that approximately 528,000 new cases and 270,000 deaths per year are attributed to cervical cancer worldwide. The disease is preventable with HPV vaccination and with early detection and treatment of pre-invasive cervical intraepithelial neoplasia, CIN. Antibodies (Abs) to HPV proteins are under investigation as potential biomarkers for early detection. To detect circulating HPV-specific IgG Abs, we developed programmable protein arrays (NAPPA) that display the proteomes of two low-risk HPV types (HPV6 and 11) and ten oncogenic high-risk HPV types (HPV16, 18, 31, 33, 35, 39, 45, 51, 52 and 58). Arrays were probed with sera from women with CIN 0/I (n=78), CIN II/III (n=84), or invasive cervical cancer (ICC, n=83). Abs to any early (E) HPV protein were detected less frequently in women with CIN 0/I (23.7%) than women with CIN II/III (39.0%) and ICC (46.1%, p<0.04). Of the E Abs, anti-E7 Abs were the most frequently detected (6.6%, 19.5%, and 30.3%, respectively). The least frequently detected Abs were E1 and E2-Abs in CIN 0/I (1.3%) and E1-Abs in CIN II/III (1.2%) and ICC (7.9%). HPV16-specific Abs correlated with HPV16 DNA detected in the cervix in 0% of CIN 0/I, 21.2% of CIN II/III, and 45.5% of ICC. A significant number (29 - 73%) of E4, E7, L1, and L2 Abs had cross-reactivity between HPV types. HPV protein arrays provide a valuable high-throughput tool for measuring the breadth, specificity, and heterogeneity of the serologic response to HPV in cervical disease.


Mechanisms involved in the anti-inflammatory action of inhaled tea tree oil in mice.
Tea tree oil (TTO) is well known as an antimicrobial and immunomodulatory agent. In the present study we confirmed the anti-inflammatory properties of TTO and investigated the involvement of the hypothalamic-pituitary-adrenal (HPA) axis in the immunomodulatory action of TTO administered by inhalation. Sexually mature, 6-8-week-old, C(57)BI(10) x CBA/H (F(1)) male mice were used. One group of animals was injected intra-peritoneally (ip) with Zymosan to elicit peritoneal inflammation and was then submitted to four sessions of TTO inhalation (15 mins each). Some of the mice were simultaneously injected ip with Antalarmin, a CRH-1 receptor antagonist, to block HPA axis functions. Twenty-four hours after the injections the mice were killed by CO(2) asphyxia, and peritoneal leukocytes (PTLs) were isolated and counted. Levels of reactive oxygen species (ROS) and cyclooxygenase (COX) activity in PTLs were assessed by fluorimetric and colorimetric assays, respectively. The results obtained show that sessions of TTO inhalation exert a strong anti-inflammatory influence on the immune system stimulated by Zymosan injection, while having no influence on PTL number, ROS level, and COX activity in mice without inflammation. The HPA axis was shown to mediate the anti-inflammatory effect of TTO; Antalarmin abolished the influence of inhaled TTO on PTL number and their ROS production in mice with experimental peritonitis, but it had no effect on these parameters in mice without inflammation.


Polyphenols from brown alga, Padina boergesenii (Allendar & Kraft) decelerates renal cancer growth involving cell cycle arrest and induction of apoptosis in renal carcinoma cells.
In our previous work, we assessed the ameliorative effect of crude extract from Padina boergesenii. In this report, we have extended these study by fractionating the methanol extract with methanol:ethylacetate (1:3,v/v) (MME-F) and assessed the cytotoxic effect of MME-F fraction in human renal carcinoma cell lines (A498 and ACHN). The fraction had time-and dose-dependent inhibition of cancer cell proliferation, migration with deceleration of cancer growth at EC50 -22.73 μg in A498 and 26.43 μg in ACHN cells. Cells treated at EC50 value 25 μg displayed twofold greater ability to induce early and late stage of apoptosis. The cells treated with polyphenolic fraction (MME-F) induced cell cycle arrest at G2/M phase. HPLC/DAD chromatographic procedures quantified polyphenols from active fraction (MME-F). These data revealed the functional activity of polyphenols from brown alga, P. boergesenii as a potent inhibitor of cancer proliferation with induction of apoptosis, it suggest their applicability in preventing cancer metastasis.


Comparison Between Doppler-Echocardiography and Uncalibrated Pulse Contour Method for Cardiac Output Measurement: A Multicenter Observational Study.
Echocardiography and pulse contour methods allow, respectively, noninvasive and less invasive cardiac output estimation. The aim of the present study was to compare Doppler echocardiography with the pulse contour method MostCare for cardiac output estimation in a large and nonselected critically ill population. A prospective multicenter observational comparison study. The study was conducted in 15 European medicosurgical ICUs. We assessed cardiac output in 400 patients in whom an echocardiographic evaluation was performed as a routine need or for cardiocirculatory assessment. None. One echocardiographic cardiac output measurement was compared with the corresponding MostCare cardiac output value per patient, considering different ICU admission categories and clinical conditions. For statistical analysis, we used Bland-Altman and linear regression analyses. To assess heterogeneity in results of individual centers, Cochran Q, and the I statistics were applied. A total of 400 paired echocardiographic cardiac output and MostCare cardiac output measures were compared. MostCare cardiac output values ranged from 1.95 to 9.90 L/min, and echocardiographic cardiac output ranged from 1.82 to 9.75 L/min. A significant correlation was found between echocardiographic cardiac output and MostCare cardiac output (r = 0.85; p < 0.0001). Among the different ICUs, the mean bias between echocardiographic cardiac output and MostCare cardiac output ranged from -0.40 to 0.45 L/min, and the percentage error ranged from 13.2% to 47.2%. Overall, the mean bias was -0.03 L/min, with 95% limits of agreement of -1.54 to 1.47 L/min and a relative percentage error of 30.1%. The percentage error was 24% in the sepsis category, 26% in the trauma category, 30% in the surgical category, and 33% in the medical admission category. The final overall percentage error was 27.3% with a 95% CI of 22.2-32.4%. Our results suggest that MostCare could be an alternative to echocardiography to assess cardiac output in ICU patients with a large spectrum of clinical conditions.


Zinc treatment for 5 or 10 days is equally efficacious in preventing diarrhea in the subsequent 3 months among Bangladeshi children.
We conducted a randomized, double-blind placebo controlled, community trial in rural Bangladesh in children 4-59 mo of age to compare the efficacy of a 5- and 10-d course of zinc therapy on the incidence and duration of diarrhea over the subsequent 90-d follow-up after initial treatment for an acute childhood diarrheal (ACD) episode. Children (n = 1622) with ACD were randomly allocated to either 5 or 10 d of zinc treatment. Female field workers visited each child daily, supervised the administration of zinc, recorded the duration of current episode, and the occurrence and duration of diarrhea over the subsequent 3 mo. The incidence of diarrhea over the 90 d of follow-up did not differ between the 5-d (1.08 ± 1.38 episodes) and 10-d (1.02 ± 1.35 episodes) groups (P = 0.35). Children in both groups experienced a comparable duration of diarrheal episodes (3.1 ± 5.6 d vs. 2.9 ± 5.6 d, 5-d vs. 10-d, respectively; P = 0.64) with a mean difference between groups within the defined range of equivalence. Time to onset of the first episode and the proportion children experiencing diarrhea during the 90-d follow-up also did not differ between groups. These findings suggest that among Bangladeshi children, a 5-d zinc treatment for ACD is as efficacious as 10 d in preventing diarrhea in the subsequent 3 mo.


Tidal expiratory flow patterns in airflow obstruction.
Tidal expiratory flow pattern was analysed in 99 subjects with a view to assessing it as a quantitative measurement of airflow obstruction. Fifteen normal volunteers, nine patients with dyspnoea referred for investigation in whom airway resistance was within normal limits, 24 patients with restrictive lung disorders, and 51 patients with airway obstruction were studied. The expiratory flow pattern against time had a quadrilateral configuration in airway obstruction, which differed from the more sinusoidal form that is seen in subjects without airflow obstruction. The rapid rise to tidal peak flow was analysed in two ways, percentage of volume expired at tidal peak flow (delta V/V) and percentage of expiratory time to tidal peak flow (delta t/t). Both these indices correlated significantly with conventional measurements of airway obstruction. The pattern of expiratory flow in airflow obstruction during quiet breathing resembles that of a forced expiratory maneuver at similar lung volumes. In some cases this may be caused by dynamic compression occurring during tidal breathing. In others, the pattern may result from the static recoil of the lung being permitted to drive flow freely in expiration, rather than being braked by postinspiratory contraction of inspiratory musculature.