Influence of irradiation time on subsurface degree of conversion and microhardness of high-viscosity bulk-fill resin composites.
To evaluate the influence of irradiation time on degree of conversion (DC) and microhardness of high-viscosity bulk-fill resin composites in depths up to 6 mm. Four bulk-fill materials (Tetric EvoCeram Bulk Fill--TECBF; x-tra fil--XF; QuixFil--QF; SonicFill-SF) and one conventional nano-hybrid resin composite (Tetric EvoCeram--TEC) were irradiated for 10, 20, or 30 s at 1,170 mW/cm(2). DC and Knoop microhardness (KHN) were recorded after 24-h dark storage at five depths: 0.1, 2, 4, 5, and 6 mm. Data were statistically analyzed using ANOVA and Bonferroni's post-hoc test (α = 0.05). With increasing bulk thickness, DC and KHN significantly decreased for TEC. TECBF and SF showed a significant decrease in DC and KHN at 4-mm depth after 10-s irradiation, but no decrease in DC after 30-s irradiation (p > 0.05). XF and QF demonstrated no significant DC decrease at depths up to 6 mm after irradiation of at least 20 s. At 4-mm depth, all materials tested achieved at least 80 % of their maximum DC value, irrespective of irradiation time. However, at the same depth (4 mm), only XF and QF irradiated for 30 s achieved at least 80 % of their maximum KHN value. Regarding DC, the tested bulk-fill resin composites can be safely used up to at least 4-mm incremental thickness. However, with respect to hardness, only XF and QF achieved acceptable results at 4-mm depth with 30 s of irradiation. Minimum irradiation times stated by the manufacturers cannot be recommended for placement of high-viscosity bulk-fill materials in 4-mm increments.


Separation of sanguinarine and chelerythrine in Macleaya cordata (Willd) R. Br. based on methyl acrylate-co-divinylbenzene macroporous adsorbents.
Sanguinarine (SAN) and chelerythrine (CHE) are known as major effective components in the quaternary benzo[c]phenanthridine isoquinoline alkaloids (QBA) fraction of Macleaya cordata (Willd) R. Br. but possess different biological activities. In this study, a method for the separation of SAN and CHE based on methyl acrylate-co-divinylbenzene (MA-co-DVB) macroporous adsorbents was established. The relationship between the polarities of the adsorbents and their adsorption-desorption behaviors towards SAN and CHE was investigated. The results showed that, among three selected commercial adsorbents and seven synthesized macroporous polymeric adsorbents with different MA content, the adsorbent No. 5 with 50% MA content provided the best separation power, and the two alkaloids were separated successfully in a gradient eluent process with 60% (v/v) ethanol aqueous and 80% ethanol aqueous contained 8% acetic acid. Dynamic adsorption and desorption tests had been performed in the column packed with the adsorbent No. 5 for optimizing the process parameters. Under the optimized conditions, the ratio of SAN and CHE transformed from 2:1 in the QBA fraction of M. cordata to 1:13 and 25:1 in the products obtained from the two-step gradient elution, and the recoveries of both SAN and CHE were nearly 90%.


Combination of interleukin-12 gene therapy, metronomic cyclophosphamide and DNA cancer vaccination directs all arms of the immune system towards tumor eradication.
In this work a combination therapy that acts upon the immune suppressive, the innate and specific arms of the immune system is proposed. This combination therapy, which consists of intratumoral interleukin-12 (IL-12) gene therapy, human tyrosinase (hTyr) DNA vaccination and metronomic cyclophosphamide (CPX), was evaluated in a B16-F10 mouse model. The following groups were compared: (1) no treatment, (2) control vector, (3) intratumoral IL-12 gene therapy, (4) intratumoral IL-12 gene therapy+metronomic CPX, (5) intratumoral IL-12 gene therapy+metronomic CPX+hTyr DNA vaccination. Next to clinical efficacy and safety, we characterized acute effects of IL-12 and anti-tumor immune response after a second tumor challenge. All treatment groups showed increased survival and higher cure rates than control groups. Survival of non-cured mice was increased when metronomic CPX was combined with IL-12 gene therapy. Furthermore, mice that received metronomic CPX had significantly lower percentages of regulatory T cells. Addition of the hTyr DNA vaccine increased cure rate and resulted in increased survival compared to other treatment groups. We also demonstrated that the manifest necrosis within days after IL-12 gene therapy is at least partly due to IL-12 mediated activation of NK cells. All cured mice were resistant to a second challenge. A humoral memory response against the tumor cells was observed in all groups that received IL-12 gene therapy, while a cellular memory response was observed only in the vaccinated mice. In conclusion, every component of this combination treatment contributed a unique immunologic trait with associated clinical benefits.


Effect of horticultural therapy on wellbeing among dementia day care programme participants: A mixed-methods study (Innovative Practice).
Fourteen people attending an adult day programme were recruited to a structured horticultural therapy programme which took place over 10 weeks. The effects were assessed using Dementia Care Mapping and questionnaires completed by family carers. High levels of wellbeing were observed while the participants were engaged in horticultural therapy, and these were sustained once the programme was completed. This study adds to the growing evidence on the benefits of horticultural therapy for people with dementia who have enjoyed gardening in the past.


Preliminary experience with power Doppler imaging of solid breast masses.
The purpose of our study was to assess the potential of power Doppler imaging (PDI) to differentiate benign from malignant solid breast masses. Sixty-nine biopsy-proven solid breast masses were evaluated with PDI using 7- to 10-MHz transducers optimized for low-volume flow sensitivity. The extent of flow on PDI was estimated as a percentage of the lesion area on multiple longitudinal and transverse static sonographic images. Flow was categorized as avascular; less than 10%; 10-25%; 25.1-50%; and greater than 50%. Of the 69 lesions evaluated, 33 were malignant and 36 were benign. Of the avascular lesions, nine were malignant and eight were benign. Significant overlap was seen in the vascularity of the other 52 lesions: both malignant and benign lesions revealed a similar range of vascular patterns. Preliminary experience with PDI suggests that both malignant and benign lesions can be avascular and that the presence of color within a solid breast mass is a nonspecific finding. Assessing the extent of vascularity with PDI appears to be of limited value in the diagnostic evaluation of solid breast masses.


Sharp emergence of feature-selective sustained activity along the dorsal visual pathway.
Sustained activity encoding visual working memory representations has been observed in several cortical areas of primates. Where along the visual pathways this activity emerges remains unknown. Here we show in macaques that sustained spiking activity encoding memorized visual motion directions is absent in direction-selective neurons in early visual area middle temporal (MT). However, it is robustly present immediately downstream, in multimodal association area medial superior temporal (MST), as well as and in the lateral prefrontal cortex (LPFC). This sharp emergence of sustained activity along the dorsal visual pathway suggests a functional boundary between early visual areas, which encode sensory inputs, and downstream association areas, which additionally encode mnemonic representations. Moreover, local field potential oscillations in MT encoded the memorized directions and, in the low frequencies, were phase-coherent with LPFC spikes. This suggests that LPFC sustained activity modulates synaptic activity in MT, a putative top-down mechanism by which memory signals influence stimulus processing in early visual cortex.


Management of colon cancer at a tertiary referral center in India - Patterns of presentation, treatment, and survival outcomes.
To define the patterns of disease presentation, treatment strategies, and outcomes for patients with colon cancer at a tertiary referral center in India over 1 year period. This is a retrospective analysis of a prospectively maintained database. All consecutive patients with proven or suspected colonic adenocarcinoma between July 2013 and July 2014 were evaluated in a dedicated analysed multidisciplinary clinic at the Tata Memorial Hospital, Mumbai. The demography, treatment plan, pathology, stage, and survival data were examined. The median age of presentation was 49 years with 60.1% male patients. In total, 151 cases (57.4%) underwent treatment with curative intent consisting of surgery with adjuvant chemotherapy as indicated. The rest were offered either palliative chemotherapy (36.9%) or best supportive care (5.7%). Approximately, 70% patients had advanced stage disease (Stage III/IV) at presentation and 41.8% presented with metastatic disease with the liver being the most common site of disease dissemination. With a median follow-up of 29 months, the estimated 3-year disease free survival for patients treated with curative intent was 67.1%. The median progression free survival was 12.3 months for patients treated with palliative intent. The estimated 3-year overall survival was 89.7%, 65.5%, and 22.8% for Stage I/II, Stage III, and Stage IV, respectively. Indian patients with colon cancer, at a tertiary referral center, tend to present at more advanced stages of the disease as compared to the West. However, curative treatment with surgery and chemotherapy offers similar survival outcomes when compared stage for stage.


Effects of LPS injection on the hypothalamic and testicular mRNA expression levels of reproductive factors in male rats.
In the hypothalamus, kisspeptin and RFamide-related peptide (RFRP) regulate gonadotropin-releasing hormone expression. Kisspeptin and RFRP are also found in the testes and might play roles in steroidogenesis and spermatogenesis. The present study demonstrated that the hypothalamic mRNA expression level of the kisspeptin receptor was decreased by the injection of lipopolysaccharide (LPS) (500 μg/kg) in male rats, and it was suggested that such changes might contribute to reductions in serum luteinizing hormone levels. Contrary to our expectations, hypothalamic RFRP and testicular GPR147 (the RFRP receptor) mRNA expression were also decreased by LPS injection. We speculate that changes in hypothalamic RFRP expression might represent a protective response aimed at attenuating LPS-induced anorectic responses.


Human embryonic stem cells and their differentiated derivatives are less susceptible to immune rejection than adult cells.
Differentiated cell types derived from human embryonic stem cells (hESCs) may serve in the future to treat various human diseases. A crucial step toward their successful clinical application is to examine the immune response that might be launched against them after transplantation. We used two experimental platforms to examine the in vivo leukocyte response toward hESCs. First, immunocompetent and immunodeficient mouse strains were used to identify T cells as the major component that causes xenorejection of hESCs. Second, mice that were conditioned to carry peripheral blood leukocytes from human origin were used to test the human leukocyte alloresponse toward undifferentiated and differentiated hESCs. Using this model, we have detected only a minute immune response toward undifferentiated as well as differentiated hESCs over the course of 1 month, although control adult grafts were repeatedly infiltrated with lymphocytes and destroyed. Our data show that the cells evade immune destruction due to a low immunostimulatory potential. Nevertheless, a human cytotoxic T lymphocyte clone that was specifically prepared to recognize two hESC lines could lyse the cells after major histocompatibility complex class I (MHC-I) induction. Although MHC-I levels in hESCs are sufficient for rejection by cytotoxic T cells, our data suggest that the immunostimulatory capacity of the cells is very low. Thus, immunosuppressive regimens for hESC-based therapeutics could be highly reduced compared with conventional organ transplantation because direct allorejection processes of hESCs and their derivatives are considerably weaker.


Single-cell microinjection of cytochrome c can result in gap junction-mediated apoptotic cell death of bystander cells in head and neck cancer.
Gap junction intercellular channels are required for metabolic cooperation between cells and regulate normal tissue homeostasis by means of the transfer of small molecules between contacting cells. Not surprisingly, the gap junction phenotype is frequently lost during carcinogenesis in human tissues (including those of the upper aerodigestive tract), freeing individual cancer cells from the growth control signals of normal surrounding tissues and less aggressive adjacent cancer cells. We hypothesized that gap junctional intercellular communication (GJIC) could mediate a bystander effect (apoptotic cell death) in squamous cell carcinoma of the head and neck (SCCHN) cells adjacent to individually targeted SCCHN cells. Single-cell microinjection of cytochrome c was used to induce apoptosis in target SCCHN cells with endogenous GJIC activity and in an SCCHN cell line with exogenously introduced GJIC activity. Apoptosis was followed in target and surrounding bystander cells through light and time course microscopic characterization. All of the preceding experiments were carried out in the absence and presence of 18-beta-glycerretinic acid, a pharmacologic inhibitor of GJIC. When cytochrome c was introduced into SCCHN cells with endogenous GJIC activity through single-cell microinjection, bystander effects (apoptosis of nontarget cells) were observed. When GJIC activity was blocked with the specific pharmacologic inhibitor of gap junctions, 18-beta-glycerretinic acid, a bystander effect was never seen in GJIC active SCCHN cell lines. Gap junction intercellular channels can mediate a bystander effect in SCCHN. Inconsistencies in our data will be discussed in the context of recent advances in this field, as well as our future research directions.


The clinical features of immunoglobulin light-chain (AL) amyloidosis with heart involvement.
We reviewed clinical presentation, investigations, therapy, prognosis and outcome of 232 patients with primary (AL) cardiac amyloidosis. There were 142 men and 90 women. Median age at presentation was 59 years (range 29-85). AL heart disease was unusual both in patients under the age of 40 (3.0%) and in non-Caucasians (6.5%). Fatigue and weakness were the commonest presenting symptoms. Hallmark features of periorbital ecchymoses and macroglossia were present in 12.5% and 27.2%, respectively. AL cardiac amyloidosis was unusual in isolation (3.9%), and most frequently patients had features of multiorgan dysfunction; heavy proteinuria and features of malabsorption predominating in this respect. Heart involvement represents the worst prognostic indicator, with a median survival from diagnosis of 1.08 years, falling to 0.75 years with the onset of heart failure. Current therapeutic procedures appear to prolong survival, with left ventricular wall thickness, mass and ejection fraction on echocardiography and late potentials on signal averaged electrocardiography of use in prognostic stratification. Cardiac involvement from AL amyloidosis is rapidly fatal. It should be suspected in all patients with heart failure who have wall thickening on echo, normal chamber sizes, low EKG voltages and evidence suggesting a multisystem disease.


Regulation of muscarinic receptor expression by changes in mRNA stability.
Regulation of muscarinic acetylcholine receptor (mAChR) subtype mRNAs was investigated in the human neuroblastoma cell line IMR-32 and in transfected CHO cells. IMR-32 cells express both m1 and m3 subtypes of mAChR. Exposure of IMR-32 cells to the muscarinic agonist, carbamylcholine (CBC) leads to a time dependent down-regulation of mAChRs which was maximal by 9 hours. mAChR activation resulted in a differential regulation of mAChR subtype mRNAs. m1 mAChR mRNA was down-regulated following 12 hours of agonist treatment and was associated with a decreased stability of the receptor transcript. In contrast, the m3 mAChR mRNA was resistant to agonist treatment for up to 24 hours. Using transfected CHO cells, we identified sequence elements within the 3'-untranslated region (3'-UTR) of the m1 mAChR gene which dictate agonist-induced destabilization of the m1 mAChR mRNA. Removal of these sequences abolished the ability of chronic agonist exposure to destabilize m1 mAChR mRNA. These findings suggest that sequence specific differences between m1 and m3 mAChR subtypes, which both preferentially couple to hydrolysis of phosphoinositides, may be responsible for differences in the regulation of mAChR gene expression.


Photochemical activation of SF6 by N-heterocyclic carbenes to provide a deoxyfluorinating reagent.
The activation of the greenhouse gas SF6 using electron-rich N-heterocyclic carbenes gave 2,2-difluoroimidazolines or 2,2-difluoroimidazolidines as well as 2-thio derivatives of the carbene precursors. The N-heterocyclic carbenes can also convert SF4 to give similar products. The difluoroimidazolidine derivatives were applied in deoxyfluorination reactions. Furthermore, the activation of SF6 and the fluorination can be coupled in a one-pot process to convert 1-octanol into 1-fluorooctane.


Characterization of LE3 and LE4, the only lytic phages known to infect the spirochete Leptospira.
Leptospira is a phylogenetically unique group of bacteria, and includes the causative agents of leptospirosis, the most globally prevalent zoonosis. Bacteriophages in Leptospira are largely unexplored. To date, a genomic sequence is available for only one temperate leptophage called LE1. Here, we sequenced and analysed the first genomes of the lytic phages LE3 and LE4 that can infect the saprophyte Leptospira biflexa using the lipopolysaccharide O-antigen as receptor. Bioinformatics analysis showed that the 48-kb LE3 and LE4 genomes are similar and contain 62% genes whose function cannot be predicted. Mass spectrometry led to the identification of 21 and 23 phage proteins in LE3 and LE4, respectively. However we did not identify significant similarities with other phage genomes. A search for prophages close to LE4 in the Leptospira genomes allowed for the identification of a related plasmid in L. interrogans and a prophage-like region in the draft genome of a clinical isolate of L. mayottensis. Long-read whole genome sequencing of the L. mayottensis revealed that the genome contained a LE4 phage-like circular plasmid. Further isolation and genomic comparison of leptophages should reveal their role in the genetic evolution of Leptospira.


Is a new sonic toothbrush more effective in plaque removal than a manual toothbrush?
Powered or manual toothbrushes are daily-used instrument in the Western area for the control and removal of bacterial biofilm. Among powered-toothbrushes, sonic technology has shown to produce fluid turbulent activity that might assist in plaque removal; however, limited knowledge is available in-vivo. The objectives of this study were to compare the plaque removal efficacy of two different toothbrushes in a population not familiar with sonic technology, and to collect and analyse data regarding oral hygiene habits. The null-hypothesis was that a sonic toothbrush is able to remove a superior amount of plaque compared to the manual type. Forty young adult patients were enrolled in the study. A single-cohort crossover clinical trial was designed. For each patient, three appointments were scheduled: the first (T0) was used for oral care education and explanations of toothbrushes techniques, for a preliminary professional hygiene session, and for delivery of a questionnaire; at one week (T1), plaque evaluation was performed (Turesky modification of the Quigley and Hein index) at baseline and after asking patients to brush with the randomly selected manual or sonic device. At the last appointment (week 3, T2), the same plaque evaluations of T1 were repeated asking patients to brush with the other toothbrush. Entire mouth indexes were calculated and mean reductions in whole mouth plaque scores were obtained (pre-brushing minus post-brushing values) for the two tested toothbrushes. Multiple ANOVA tests (p = 0.05 ) were used 1) to compare plaque levels between male and female subjects at baseline and post-brushing, regardless the type of toothbrush, and 2) to differentiate between mean reductions in whole mouth plaque scores according to the type of toothbrush (manual versus sonic). The study population was subjected to descriptive statistical analysis; potential relationships between socio-demographic variables and obtained plaque scores were evaluated (Mann-Whitney and Kruskal-Wallis tests). Full-mouth plaque levels were reduced at post-brushing sessions, regardless the device, by approximately 62% (p<0.0001). Mean plaque index reductions for manual and sonic toothbrush were of 1.05 ± 0.22 and 1.19 ± 0.37, respectively. A statistically significant difference was found between the two devices (p = 0.0342). The powered sonic toothbrush removed about 10% more plaque than the manual type. From the collected questionnaire financial data, willingness to pay (WTP) values expressing economic efforts of patients for the purchase of toothbrushes were of Euros 4.83 ± 3.86 and of Euros 54.75 ± 36, for the manual and sonic devices, respectively. Within the limitations of the study, in subjects without any previous experience of a similar technology, the single use of the sonic toothbrush showed a significantly greater plaque reduction compared to the manual traditional toothbrush (null-hypothesis accepted).


Interleukin-1 promoter region polymorphism role in rheumatoid arthritis: a meta-analysis of IL-1B-511A/G variant reveals association with rheumatoid arthritis.
IL-1 has a central role mediating inflammation and joint destruction in RA. Single nucleotide polymorphisms (SNPs) and haplotype structure in the promoter region can modulate IL-1 function. This study examined the effects of four common promoter SNPs in the IL-1 region on susceptibility and clinical characteristics of RA in British Caucasian patients and assessed the risk of RA by meta-analysis of published studies. Using PCR-based methods, 756 RA patients and 625 healthy controls (HCs) were genotyped for IL-1A (-889 C/A, rs17561), IL-1B (-511 A/G, rs16944), IL-1B (-1464 C/G, rs1143623) and IL-1B (-3737 G/A, rs4848306) SNPs. Further meta-analysis was performed for IL-1B (-511 A/G) incorporating 3712 RA patients and 2331 HC from six association studies. The IL-1B (-1464 C/G) G allele was found to be less common in the RA group [P = 0.01; odds ratio (OR) 1.24; 95% CI 1.04, 1.48]. There was no association between IL-1 SNPs and the presence of HLA-DRB1 shared epitope, RF or clinical characteristics. Meta-analysis revealed statistically significant association between IL-1B (-511 A/G) and RA (P = 0.02; pooled OR 1.13; 95% CI 1.02, 1.26). There may be a protective effect in RA from the IL-1B (-1464 C/G) G variant. No direct association between the polymorphisms studied and clinical severity characteristics were observed. Further meta-analysis revealed IL-1B (-511 A/G) to be associated with increased susceptibility to RA.


Chemical Leslie effect in Langmuir monolayers: a complete experimental characterization.
We propose a complete characterization of the chemical Leslie effect in a Langmuir monolayer of a chiral liquid crystal. To reach this goal, we developed new experimental techniques using an electric field and a humidifier to prepare large monodomains in which the molecules can freely rotate. We also designed six independent experiments to precisely measure the four material constants involved in the dynamics of the monolayer, namely the Leslie coefficient, the rotational viscosity, the curvature elasticity constant and the surface polarization. The relevance of the inverse Leslie effect is also discussed.


Distinctive properties of mammalian DNA polymerases.
DNA polymerase-alpha and -beta can be distinguished from one another by the differential effects of N-ethylmaleimide, KCl, ara-CTP and temperature, as well as on the basis of sedimentation. The sensitivity of DNA polymerase-beta to elevated temperatures as compared to DNA polymerase-alpha provides a new means of distinguishing between these two enzymes even in crude extracts and a possible probe for determining their function. DNA polymerase-alpha and -beta share several properties in common, including the ability to readily incorporate dUTP in place of dTTP. The Km for dUTP varies from 10 to 30 micron with different preparations of DNA polymerase-alpha and -beta. Thus, in mammalian cells, dUMP could be incorporated into DNA, and if excised by an endonuclease, would lead to discontinuities. Initial analyses of fidelity in direct comparative studies indicate that beta-class DNA polymerases are highly accurate in base selection when copying poly[d(A-T)]. Less than one molecule of dGMP is incorporated for every 12 000-45 000 molecules of dAMP and dTMP polymerized. DNA polymerase-alpha is somewhat less accurate, making one mistake for every 4000-10 000 correct nucleotides incorporated. Since both polymerases lack an exonucleolytic activity, this accuracy must be the result of selectivity for the complementary nucleotide by the polymerase.


PDK4 Deficiency Suppresses Hepatic Glucagon Signaling by Decreasing cAMP Levels.
In fasting or diabetes, gluconeogenic genes are transcriptionally activated by glucagon stimulation of the cAMP-protein kinase A (PKA)-CREB signaling pathway. Previous work showed pyruvate dehydrogenase kinase (PDK) inhibition in skeletal muscle increases pyruvate oxidation, which limits the availability of gluconeogenic substrates in the liver. However, this study found upregulation of hepatic PDK4 promoted glucagon-mediated expression of gluconeogenic genes, whereas knockdown or inhibition of hepatic PDK4 caused the opposite effect on gluconeogenic gene expression and decreased hepatic glucose production. Mechanistically, PDK4 deficiency decreased ATP levels, thus increasing phosphorylated AMPK (p-AMPK), which increased p-AMPK-sensitive phosphorylation of cyclic nucleotide phosphodiesterase 4B (p-PDE4B). This reduced cAMP levels and consequently p-CREB. Metabolic flux analysis showed that the reduction in ATP was a consequence of a diminished rate of fatty acid oxidation (FAO). However, overexpression of PDK4 increased FAO and increased ATP levels, which decreased p-AMPK and p-PDE4B and allowed greater accumulation of cAMP and p-CREB. The latter were abrogated by the FAO inhibitor etomoxir, suggesting a critical role for PDK4 in FAO stimulation and the regulation of cAMP levels. This finding strengthens the possibility of PDK4 as a target against diabetes.


PATCHED and p53 gene alterations in sporadic and hereditary basal cell cancer.
It is widely accepted that disruption of the hedgehog-patched pathway is a key event in development of basal cell cancer. In addition to patched gene alterations, p53 gene mutations are also frequent in basal cell cancer. We determined loss of heterozygosity in the patched and p53 loci as well as sequencing the p53 gene in tumors both from sporadic and hereditary cases. A total of 70 microdissected samples from tumor and adjacent skin were subjected to PCR followed by fragment analysis and DNA sequencing. We found allelic loss in the patched locus in 6/8 sporadic basal cell cancer and 17/19 hereditary tumors. All sporadic and 7/20 hereditary tumors showed p53 gene mutations. Loss of heterozygosity in the p53 locus was rare in both groups. The p53 mutations detected in hereditary tumors included rare single nucleotide deletions and unusual double-base substitutions compared to the typical ultraviolet light induced missense mutations found in sporadic tumors. Careful microdissection of individual tumors revealed genetically linked subclones with different p53 and/or patched genotype providing an insight on time sequence of genetic events. The high frequency and co-existence of genetic alterations in the patched and p53 genes suggest that both these genes are important in the development of basal cell cancer.


How do top cable news websites portray cognition as an aging issue?
We examined messages that the websites of the top cable news companies (MSNBC, FOX, and CNN) conveyed about cognition between January 2007 and March 2010. Drawing on agenda-setting theory, this work assessed the frequency, prominence, and attributes of cognitive topics in messages targeting an aging audience. We used quantitative content analysis to examine the frequency and prominence of cognitive topics and cognitive goals, as well as how the cognitive discussions were framed. Chi-square analyses were conducted to compare cognitive health information discussed in news items that did and did not target an "aging audience." Qualitative analysis of the aging audience subgroup was used to further examine age-associated cognitive messages. Within the 229 cognitive items identified, we found significantly more coverage of cognitive functioning and unspecified dementia and significantly less coverage of cognitive disease not dementia, specified dementia, and accidents or injury for the aging audience. Our qualitative analysis of news items aimed at an aging audience documented a focus on maintaining functioning and avoiding decline through various individual lifestyle behaviors. However, contextual information about level of cognition to be maintained, particular cognitive functions targeted, specific norms about cognitive aging, and how cognitive function is determined was lacking. Our research points to a communication gap in the delivery of academic research findings to a lay audience through online journalism. We suggest more clarity by researchers in defining cognitive concepts and measurement of cognitive function for journalistic translation and public consumption.


EGFR inhibition promotes an aggressive invasion pattern mediated by mesenchymal-like tumor cells within squamous cell carcinomas.
Squamous cell carcinomas (SCC) with an infiltrative invasion pattern carry a higher risk of treatment failure. Such infiltrative invasion may be mediated by a mesenchymal-like subpopulation of malignant cells that we have previously shown to arise from epithelial-mesenchymal transition (EMT) and resist epidermal growth factor receptor (EGFR) targeting. Here, we show that SCCs with infiltrative, high-risk invasion patterns contain abundant mesenchymal-like cells, which are rare in tumors with low-risk patterns. This cellular heterogeneity was modeled accurately in three-dimensional culture using collagen-embedded SCC spheroids, which revealed distinct invasive fronts created by collective migration of E-cadherin-positive cells versus infiltrative migration of individual mesenchymal-like cells. Because EGFR expression by mesenchymal-like cells was diminished in the spheroid model and in human SCCs, we hypothesized that SCCs shift toward infiltrative invasion mediated by this subpopulation during anti-EGFR therapy. Anti-EGFR treatment of spheroids using erlotinib or cetuximab enhanced infiltrative invasion by targeting collective migration by E-cadherin-positive cells while sparing mesenchymal-like cells; by contrast, spheroid invasion in absence of mesenchymal-like cells was abrogated by erlotinib. Similarly, cetuximab treatment of xenografts containing mesenchymal-like cells created an infiltrative invasive front composed of this subpopulation, whereas no such shift was observed upon treating xenografts lacking these cells. These results implicate mesenchymal-like SCC cells as key mediators of the infiltrative invasion seen in tumors with locally aggressive behavior. They further show that EGFR inhibition can promote an infiltrative invasion front composed of mesenchymal-like cells preferentially in tumors where they are abundant before therapy.


Cellular dynamic simulator: an event driven molecular simulation environment for cellular physiology.
In this paper, we present the Cellular Dynamic Simulator (CDS) for simulating diffusion and chemical reactions within crowded molecular environments. CDS is based on a novel event driven algorithm specifically designed for precise calculation of the timing of collisions, reactions and other events for each individual molecule in the environment. Generic mesh based compartments allow the creation / importation of very simple or detailed cellular structures that exist in a 3D environment. Multiple levels of compartments and static obstacles can be used to create a dense environment to mimic cellular boundaries and the intracellular space. The CDS algorithm takes into account volume exclusion and molecular crowding that may impact signaling cascades in small sub-cellular compartments such as dendritic spines. With the CDS, we can simulate simple enzyme reactions; aggregation, channel transport, as well as highly complicated chemical reaction networks of both freely diffusing and membrane bound multi-protein complexes. Components of the CDS are generally defined such that the simulator can be applied to a wide range of environments in terms of scale and level of detail. Through an initialization GUI, a simple simulation environment can be created and populated within minutes yet is powerful enough to design complex 3D cellular architecture. The initialization tool allows visual confirmation of the environment construction prior to execution by the simulator. This paper describes the CDS algorithm, design implementation, and provides an overview of the types of features available and the utility of those features are highlighted in demonstrations.


Hospital at home: home-based end-of-life care.
The policy in a number of countries is to provide people with a terminal illness the choice of dying at home. This policy is supported by surveys indicating that the general public and people with a terminal illness would prefer to receive end-of-life care at home. This is the fourth update of the original review. To determine if providing home-based end-of-life care reduces the likelihood of dying in hospital and what effect this has on patients' symptoms, quality of life, health service costs, and caregivers, compared with inpatient hospital or hospice care. We searched the following databases until April 2015: Cochrane Central Register of Controlled Trials (CENTRAL) (the Cochrane Library), Ovid MEDLINE(R) (from 1950), EMBASE (from 1980), CINAHL (from 1982), and EconLit (from 1969). We checked the reference lists of potentially relevant articles identified and handsearched palliative care publications, clinical trials registries, and a database of systematic reviews for related trials (PDQ-Evidence 2015). Randomised controlled trials, interrupted time series, or controlled before and after studies evaluating the effectiveness of home-based end-of-life care with inpatient hospital or hospice care for people aged 18 years and older. Two review authors independently extracted data and assessed study quality. We combined the published data for dichotomous outcomes using fixed-effect Mantel-Haenszel meta-analysis. When combining outcome data was not possible, we reported the results from individual studies. We included four trials in this review and did not identify new studies from the search in April 2015. Home-based end-of-life care increased the likelihood of dying at home compared with usual care (risk ratio (RR) 1.33, 95% confidence interval (CI) 1.14 to 1.55, P = 0.0002; Chi(2) = 1.72, df = 2, P = 0.42, I(2) = 0%; 3 trials; N = 652; high quality evidence). Admission to hospital while receiving home-based end-of-life care varied between trials, and this was reflected by a high level of statistical heterogeneity in this analysis (range RR 0.62 to RR 2.61; 4 trials; N = 823; moderate quality evidence). Home-based end-of-life care may slightly improve patient satisfaction at one-month follow-up and reduce it at six-month follow-up (2 trials; low quality evidence). The effect on caregivers is uncertain (2 trials; low quality evidence). The intervention may slightly reduce healthcare costs (2 trials, low quality evidence). No trial reported costs to patients and caregivers. The evidence included in this review supports the use of home-based end-of-life care programmes for increasing the number of people who will die at home, although the numbers of people admitted to hospital while receiving end-of-life care should be monitored. Future research should systematically assess the impact of home-based end-of-life care on caregivers.


Membrane fusion.
The fusion of biological membranes results in two bilayer-based membranes merging into a single membrane. In this process the lipids have to undergo considerable rearrangement. The nature of the intermediates that are formed during this rearrangement has been investigated. Certain fusion proteins facilitate this process. In many cases short segments of these fusion proteins have a particularly important role in accelerating the fusion process. Studies of the interaction of model peptides with membranes have allowed for increased understanding at the molecular level of the mechanism of the promotion of membrane fusion by fusion proteins. There is an increased appreciation of the roles of several independent segments of fusion proteins in promoting the fusion process. Many of the studies of the fusion of biological membranes have been done with the fusion of enveloped viruses with other membranes. One reason for this is that the number of proteins involved in viral fusion is relatively simple, often requiring only a single protein. For many enveloped viruses, the structure of their fusion proteins has certain common elements, suggesting that they all promote fusion by an analogous mechanism. Some aspects of this mechanism also appears to be common to intracellular fusion, although several proteins are involved in that process which is more complex and regulated than is viral fusion.


Breast cancer stromal elastosis is associated with mammography screening detection, low Ki67 expression and favourable prognosis in a population-based study.
Mammography screen-detected breast cancers have a better prognosis than predicted from established prognostic markers. A search for additional features that are characteristic for these tumours and their prognosis is needed to reduce overtreatment, a recognized challenge in breast cancer patient management today. Here, we have investigated the occurrence and importance of tumour elastosis. We performed a population based retrospective study of breast cancers detected in the Norwegian Breast Cancer Screening Programme in Vestfold County during 2004-2009. In total, 197 invasive screen-detected cancers and 75 interval cancers in patients aged 50-69 years were compared with regard to standard clinico-pathological parameters and tumour shape, as well as ER, PR, HER2 and Ki67 expression. In particular, the presence of elastotic material in tumours was graded on a 4-tiered scale (score 0-3). Screen-detected cancers had a significantly higher content of stromal elastosis than interval cancers (p < 0.001). High content of elastosis (score 3) correlated strongly with stellate tumour shape, low histological grade, and ER+/HER2- status. Further, high elastosis score was significantly associated with lower Ki67 expression. In survival analyses, cases with high elastosis demonstrated increased recurrence free (p = 0.03) and disease-specific survival (p = 0.11) compared to cases with low elastosis. There is a strong correlation between the presence of tumour elastosis, stellate tumour shape and mammography detection of breast cancers. To our knowledge, this is the first time elastosis has been studied in relation to breast cancer detection method. Presence of elastosis is associated with low tumour cell proliferation (Ki67) and a good prognosis. The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/13000_2014_230.


The impact of a cadaver-based airway lab on critical care fellows' direct laryngoscopy skills.
This study sought to determine the impact of a cadaver-based airway lab on critical care medicine fellows' direct laryngoscopy skills and hypothesised that fellows can improve their self-reported percentage of glottic opening (POGO) scores in cadaver models to achieve POGO scores similar to that of expert faculty. Nineteen fellows attended an airway management skills laboratory utilising five modified cadavers. Initial fellow POGO (POGOi) visualised was recorded at the onset with direct laryngoscopy. Maximum fellow POGO score with optimised direct laryngoscopy was recorded after two additional hours of teaching and also during a testing phase several days later. Data was assessed for significant differences between trainee POGO scores at each time interval and between trainee POGO scores and expert scores. A departmental procedural database was utilised to examine success rates of intubation as a clinical correlation. Fellows' mean POGOi scores, averaged across all five specimens, were significantly lower than both their maximum POGO scores and their testing phase POGO scores. Mean POGOi scores for fellows, averaged over all five cadavers, were lower than the instructors' POGOi scores. There was no difference between fellows' and instructors' mean maximum POGO or mean testing phase POGO scores. Clinical success rates of intubation were over 98%. A short training session using modified cadavers can be utilised to teach new critical care medicine fellows additional techniques for airway management and assist them in obtaining higher POGO scores, similar to those of expert instructors. Success rates of clinical intubations were favourable in the wake of this training.


Fourteen-week toxicity study of green tea extract in rats and mice.
The toxicity of green tea extract (GTE) was evaluated in 14-week gavage studies in male and female F344/NTac rats and B6C3F1 mice at doses up to 1,000 mg/kg. In the rats, no treatment-related mortality was noted. In the mice, treatment-related mortality occurred in male and female mice in the 1,000 mg/kg dose groups. The cause of early deaths was likely related to liver necrosis. Treatment-related histopathological changes were seen in both species in the liver, nose, mesenteric lymph nodes, and thymus. In addition, in mice, changes were seen in the Peyer's patches, spleen, and mandibular lymph nodes. The no adverse effect level (NOAEL) for the liver in both species was 500 mg/kg. In the nose of rats, the NOAEL in males was 62.5 mg/kg, and in females no NOAEL was found. No NOAEL was found in the nose of female or male mice. The changes in the liver and nose were considered primary toxic effects of GTE, while the changes in other organs were considered to be secondary effects. The nose and liver are organs with high metabolic enzyme activity. The increased susceptibility of the nose and liver suggests a role for GTE metabolites in toxicity induction.


Reactions of the oxidized organic cofactor in copper-depleted bovine serum amine oxidase.
A novel copper-depleted bovine serum amine oxidase (BSAO), in which about half the molecules contained the organic cofactor in the oxidized form, was prepared by adding a reductant in anaerobic conditions to the cyanide-reacted protein. The CuI-semiquinone formed in these conditions reoxidizes after the removal of copper. The inactive derivative was reduced by benzylamine at approx. 1/1000 the rate of BSAO. The pseudo-first-order reaction was preceded by the formation of a protein-benzylamine complex with dissociation constant, Kd, of 4.9+/-0.5 mM, similar to the Km of BSAO (2.2 mM). Also the reactions with phenylhydrazine and benzohydrazide were considerably slower than in holo-BSAO, whereas the reactions with p-pyridine-2-ylphenylacetohydrazide, containing a longer aromatic tail, and semicarbazide, lacking an aromatic moiety, were less severely affected. Removal of copper had no effect on the optical spectra of BSAO and of most adducts, containing the cofactor in quinol form, showing that copper is bound to neither the oxidized nor the reduced cofactor. Benzylhydrazine did not produce optical effects but was tightly bound, as inferred from its inhibitory effect on reaction with other molecules. Substrate and inhibitors might bind a hydrophobic pocket at some distance from the quinone, probably near the protein surface, with their affinity depending on the hydrophobic character and pKa. The binding, which is not greatly influenced by copper removal, probably induces a copper-dependent change of conformation, 'opening' a pathway to the active site buried in the protein interior.


Development and quantitative performance evaluation of a noninvasive EMG computer interface.
This paper describes a noninvasive electromyography (EMG) signal-based computer interface and a performance evaluation method based on Fitts' law. The EMG signals induced by volitional wrist movements were acquired from four sites in the lower arm to extract users' intentions, and six classes of wrist movements were distinguished using an artificial neural network. Using the developed interface, a user can move the cursor, click buttons, and type text on a computer. The test setup was built to evaluate the developed interface, and the mouse was tested by five volunteers with intact limbs. The performance of the developed computer interface and the mouse was tested at 1.299 and 7.733 b/s, respectively, and these results were compared with the performance of a commercial noninvasive brain signal interface (0.386 b/s). The results show that the developed interface performed better than the commercial interface, but less satisfactorily than a computer mouse. Although some issues remain to be resolved, the developed EMG interface has the potential to help people with motor disabilities to access computers and Internet environments in a natural and intuitive manner.


2015 American College of Rheumatology Guideline for the Treatment of Rheumatoid Arthritis.
To develop a new evidence-based, pharmacologic treatment guideline for rheumatoid arthritis (RA). We conducted systematic reviews to synthesize the evidence for the benefits and harms of various treatment options. We used the Grading of Recommendations Assessment, Development and Evaluation (GRADE) methodology to rate the quality of evidence. We employed a group consensus process to grade the strength of recommendations (either strong or conditional). A strong recommendation indicates that clinicians are certain that the benefits of an intervention far outweigh the harms (or vice versa). A conditional recommendation denotes uncertainty over the balance of benefits and harms and/or more significant variability in patient values and preferences. The guideline covers the use of traditional disease-modifying antirheumatic drugs (DMARDs), biologic agents, tofacitinib, and glucocorticoids in early (<6 months) and established (≥6 months) RA. In addition, it provides recommendations on using a treat-to-target approach, tapering and discontinuing medications, and the use of biologic agents and DMARDs in patients with hepatitis, congestive heart failure, malignancy, and serious infections. The guideline addresses the use of vaccines in patients starting/receiving DMARDs or biologic agents, screening for tuberculosis in patients starting/receiving biologic agents or tofacitinib, and laboratory monitoring for traditional DMARDs. The guideline includes 74 recommendations: 23% are strong and 77% are conditional. This RA guideline should serve as a tool for clinicians and patients (our two target audiences) for pharmacologic treatment decisions in commonly encountered clinical situations. These recommendations are not prescriptive, and the treatment decisions should be made by physicians and patients through a shared decision-making process taking into account patients' values, preferences, and comorbidities. These recommendations should not be used to limit or deny access to therapies.


Rasch-built Overall Disability Scale (R-ODS) for immune-mediated peripheral neuropathies.
To develop a patient-based, linearly weighted scale that captures activity and social participation limitations in patients with Guillain-Barré syndrome (GBS), chronic inflammatory demyelinating polyradiculoneuropathy (CIDP), and gammopathy-related polyneuropathy (MGUSP). A preliminary Rasch-built Overall Disability Scale (R-ODS) containing 146 activity and participation items was constructed, based on the WHO International Classification of Functioning, Disability and Health, literature search, and patient interviews. The preliminary R-ODS was assessed twice (interval: 2-4 weeks; test-retest reliability studies) in 294 patients who experienced GBS in the past (n = 174) or currently have stable CIDP (n = 80) or MGUSP (n = 40). Data were analyzed using the Rasch unidimensional measurement model (RUMM2020). The preliminary R-ODS did not meet the Rasch model expectations. Based on disordered thresholds, misfit statistics, item bias, and local dependency, items were systematically removed to improve the model fit, regularly controlling the class intervals and model statistics. Finally, we succeeded in constructing a 24-item scale that fulfilled all Rasch requirements. "Reading a newspaper/book" and "eating" were the 2 easiest items; "standing for hours" and "running" were the most difficult ones. Good validity and reliability were obtained. The R-ODS is a linearly weighted scale that specifically captures activity and social participation limitations in patients with GBS, CIDP, and MGUSP. Compared to the Overall Disability Sum Score, the R-ODS represents a wider range of item difficulties, thereby better targeting patients with different ability levels. If responsive, the R-ODS will be valuable for future clinical trials and follow-up studies in these conditions.


Determination of α-Keggin structure of [GeW11O39Ru(III)(H2O)]5-. Reaction of [GeW11O39Ru(III)(H2O)]5- with dimethyl sulfoxide to form [GeW11O39Ru(III)(dmso)]5- and their structural characterization.
Reaction of ruthenium((III))-substituted Keggin-type germanotungstate [GeW(11)O(39)Ru(III)(H(2)O)](5-) (1) with a dimethyl sulfoxide (dmso) produced a dmso-coordinating derivative [GeW(11)O(39)Ru(III)(dmso)](5-) (2). Structural characterization of compound 2 by using cyclic voltammetry, UV-Vis spectroscopy, IR spectroscopy, elemental analysis, (1)H-NMR spectroscopy and single crystal structure analysis (monoclinic, P2(1)/c, a = 13.461(1), b = 20.198(1), c = 18.078(1), β = 90.426(1), Z = 4) revealed that Ru(III) was incorporated in the α-Keggin-type framework and coordinated by dmso through an Ru-S bond. The IR spectra of 1 and 2 were very similar, indicating that 1 also has an α-Keggin structure. Cyclic voltammetry indicated that the incorporated Ru(III)-dmso was reversibly reduced to the Ru(II)-dmso derivative and oxidized to the Ru(IV)-dmso derivative. The redox potential in [α-XW(11)O(39)Ru(III/II)(dmso)](n-) (X = P, Ge, Si) decreased in the order of P > Ge > Si. Reaction of the complex 2 with ascorbic acid produced a one-electron-reduced compound [α-GeW(11)O(39)Ru(II)(dmso)](6-).


Network cosmology.
Prediction and control of the dynamics of complex networks is a central problem in network science. Structural and dynamical similarities of different real networks suggest that some universal laws might accurately describe the dynamics of these networks, albeit the nature and common origin of such laws remain elusive. Here we show that the causal network representing the large-scale structure of spacetime in our accelerating universe is a power-law graph with strong clustering, similar to many complex networks such as the Internet, social, or biological networks. We prove that this structural similarity is a consequence of the asymptotic equivalence between the large-scale growth dynamics of complex networks and causal networks. This equivalence suggests that unexpectedly similar laws govern the dynamics of complex networks and spacetime in the universe, with implications to network science and cosmology.


Relationship between encounter time and candidate performance on COMLEX-USA Level 2-PE.
The National Board of Osteopathic Medical Examiners uses a standardized timing format for all osteopathic medical candidates across all stations in the Comprehensive Osteopathic Medical Licensing Examination-USA Level 2-Performance Evaluation (COMLEX-USA Level 2-PE). Examination standardization, and therefore validity, is weakened if time administered affects student performance. To investigate the relationship between the amount of time students spent with standardized patients (SPs) and their clinical skill component scores. Data were analyzed from 249 candidates in 21 test sessions from the 2007-2008 testing cycle. Trained SPs recorded the time each examinee entered and exited each encounter. Outcome measures of the examination included pass (1 if pass; 0 if fail) performance outcome, pass by domain (biomedical/biomechanical and humanistic), and pass by component scores (data gathering, osteopathic manipulative treatment, and patient note scores). Analysis of variance (ANOVA), followed by pairwise comparisons, was used to compare encounter times across cases. A total of 2988 SP encounters were analyzed. Examinees spent an average 12.1 minutes in each patient encounter. Candidates spent less time with cases classified as health promotion and disease prevention than with cases that primarily involved a problem visit, but amount of time spent with cases representing various content areas was not significantly different (P>.05). On average, candidates spent the least amount of time (11.95 minutes) with gastrointestinal cases and the most time (12.26 minutes) with patients who had complaints relating to the neuromusculoskeletal system. A trivial relationship was shown between the continuous variable encounter time as the average amount of time a student spends in the encounter and the dichotomous clinical skill outcome of the examination (r=.03). The relationship between these 2 variables was consistent when examined separately for the humanistic domain (r=-.10) and the biomedical/biomechanical domain (r=.12). The strongest correlation was between encounter time and the data-gathering component score (r=.28). Time spent in the encounter was not influential in overall candidate performance in the COMLEX-USA Level 2-PE. Candidates spent a similar amount of time with cases of different content areas. This finding suggests that 14 minutes is sufficient for this osteopathic high-stakes performance evaluation.


Clinicopathological characterization of so-called "cholangiocarcinoma with intraductal papillary growth" with respect to "intraductal papillary neoplasm of bile duct (IPNB)".
Cholangiocarcinoma (CC) of the biliary tract occasionally presents a predominant intraductal papillary growth in the bile ducts, called as biliary tract carcinoma (BTC) of papillary growth (PG) and intrahepatic CC (ICC) of intraductal growth (IG) type. Recently, intraductal papillary neoplasm of bile duct (IPNB) has been proposed as a pre-invasive biliary neoplasm. This study was performed to characterize pathologically BTC of PG type and ICC of IG type with respect to IPNB. It was found that 126 of such 154 CCs (81.8%) fulfilled the criteria of IPNB, while the remaining 28 cases showed different histologies, such as tubular adenocarcinoma and carcinosarcoma. These IPNBs occurred in old aged patients with a male predominance, and the left lobe was rather frequently affected in the liver. A majority of these cases were high grade IPNB (43 cases) and invasive IPNB (77 cases), while low grade IPNB was rare (6 cases). Pancreatobiliary type was predominant (48 cases) followed by gastric (30 cases), intestinal (29 cases) and oncocytic (19 cases) types. Mucus hypersecretion was found in 45 cases, and this was frequent in IPNB at the intrahepatic large bile duct and hilar bile ducts but rare at the extrahepatic bile ducts. Interestingly, 36 cases of high grade and invasive IPNBs contained foci of moderately differentiated adenocacinoma within the intraductal papillary tumor. In conclusion, a majority of ICC of IG type and BTC of PG type could be regarded as a IPNB lineage, and clinically detectable IPNBs were already a malignant papillary lesion.


A codon substitution model that incorporates the effect of the GC contents, the gene density and the density of CpG islands of human chromosomes.
Developing a model for codon substitutions is essential for the analyses of protein sequences. Recent studies on the mutation rates in the non-coding regions have shown that CpG mutation rates in the human genome are negatively correlated to the local GC content and to the densities of functional elements. This study aimed at understanding the effect of genomic features, namely, GC content, gene density, and frequency of CpG islands, on the rates of codon substitution in human chromosomes. Codon substitution rates of CpG to TpG mutations, TpG to CpG mutations, and non-CpG transitions and transversions in humans were estimated by comparing the coding regions of thousands of human and chimpanzee genes and inferring their ancestral sequences by using macaque genes as the outgroup. Since the genomic features are depending on each other, partial regression coefficients of these features were obtained. The substitution rates of codons depend on gene densities of the chromosomes. Transcription-associated mutation is one such pressure. On the basis of these results, a model of codon substitutions that incorporates the effect of genomic features on codon substitution in human chromosomes was developed.


Antiapoptotic effects of vitamins C and E against cypermethrin-induced oxidative stress and spermatogonial germ cell apoptosis.
Toxicological studies have demonstrated the relation between use of agrochemicals and fertility issues within males. Thus, the present study aimed to elucidate the propensity of cypermethrin (CYP) in bringing testicular germ cell apoptosis and effective attenuation by vitamins C and E in caprines. Reproductive toxicity of CYP was evaluated using histomorphological, cytological, and biochemical changes in the testicular germ cells in dose-dependent (1, 5, 10 μg/mL) and time-dependent (4, 6, 8 h) manner. Histological and ethidium bromide/acridine orange fluorescence staining exhibited that vitamins C and E (0.5 and 1.0 mM) successfully diminished the CYP-induced testicular germ cells apoptosis. CYP exposure along with vitamins C and E supplementation also resulted in significantly increased ferric reducing antioxidant power activity along with the antioxidant enzymes, namely catalase, superoxide dismutase, and glutathione-s-transferase, and decreased lipid peroxidation in testicular germ cells. Thus, vitamins C and E ameliorated CYP-induced testicular germ cell apoptosis, thereby preventing spermatogonial cells degeneration and male infertility.


Peste des Petits Ruminants virus: an emerging threat to goat farming in Pakistan.
Pakistan at present is having more than 60 million head of goats, which consist of about 37 well-recognized breeds found in different regions of the country. Although the goat farming on commercial level is escalating in Pakistan, there are threats, which result this initiative into a loss. Among these threats, Peste des Petits Ruminants (PPR) outbreaks are causing huge economic damages. In this study, three outbreaks of PPR were confirmed at three different commercial farms in various regions of Punjab province and their economic impact on small ruminants farming was calculated. The disease started after 1-2 months of the establishment of these farms as the animals were purchased from different livestock markets. Disease started with sudden onset of respiratory and enteric clinical signs and spreads quickly. Disease caused mortality and morbidity of 10-15% and 20-40%, respectively, within a time period of 01-03 weeks. At these three farms, 116 of 365 animals exhibited the clinical disease, with an overall morbidity rate of 31.78%. A total of 43 animals died with mortality rate of 11.78% (43/365) causing a direct financial loss of $4300 (Pakistan Rupees 430,000/-), while the indirect cost due to treatment, loss of animal body condition, reduction in market value, increase veterinary services and labour was $7911 (Pak Rs. 791,100/-). Taken together, the results demonstrate that there is an urgent need to assess the economic impact of the disease throughout the country and to give proper emphasis for controlling PPR in sensitive regions where it is discouraging the investment in goat farming.


Effect of cranberry juice deacidification on its antibacterial activity against periodontal pathogens and its anti-inflammatory properties in an oral epithelial cell model.
Cranberries are widely recognized as a functional food that can promote oral health. However, the high concentration of organic acids in cranberry juice can cause tooth enamel erosion. Electrodialysis with bipolar membrane (EDBM) is a process used for the deacidification of cranberry juice. The present study investigated whether the removal of organic acids (0%, 19%, 42%, 60%, and 79%) from cranberry juice by EDBM affects its antibacterial activity against major periodontopathogens as well as its anti-inflammatory properties in an oral epithelial cell model. A deacidification rate ≥60% attenuated the bactericidal effect against planktonic and biofilm-embedded Aggregatibacter actinomycetemcomitans but had no impact on Porphyromonas gingivalis and Fusobacterium nucleatum. Cranberry juice increased the adherence of A. actinomycetemcomitans and P. gingivalis to oral epithelial cells, but reduced the adherence of F. nucleatum by half regardless of the deacidification rate. F. nucleatum produced more hydrogen sulfide when it was exposed to deacidified cranberry juice with a deacidification rate ≥42% compared to the raw beverage. Interestingly, the removal of organic acids from cranberry juice lowered the cytotoxicity of the beverage for oral epithelial cells. Deacidification attenuated the anti-inflammatory effect of cranberry juice in an in vitro oral epithelial cell model. The secretion of IL-6 by lipopolysaccharide (LPS)-stimulated oral epithelial cells exposed to cranberry juice increased proportionally with the deacidification rate. No such effect was observed with respect to the production of IL-8. This study provided evidence that organic acids, just like phenolic compounds, might contribute to the health benefits of cranberry juice against periodontitis.


Capsaicinoids-induced changes of plasma glucose, free fatty acid and glycerol concentrations in rats.
Red peppers are used as a spice for enhancing the palatability of foods. Two major capsaicinoids, dihydrocapsaicin (DHC) and capsaicin (CAP) are responsible for up to 90% of the total pungency of pepper fruits. These capsaicinoids are known to enhance energy metabolism and thermogenesis. However, there is a little information on the effects of capsaicinoids on the lipolysis and carbohydrate metabolism. We studied the effects of DHC and CAP on plasma glucose, free fatty acid (FFA) and glycerol concentrations in rats. Male six-week-old Sprague Dawley rats were divided into the DHC, CAP and control groups. Each capsaicinoid (dose = 3 mg/kg BW/day) was subcutaneously administered to rats for 10 days. DHC increased markedly plasma glucose, FFA and glycerol concentrations on day 1-10 by 14-35%, 61-103% and 108-174%, respectively, as compared with those of the control group. CAP increased relatively plasma glucose concentrations on day 1-3 by 15-17%, as compared with the control group. However, there were no significant differences in plasma glucose concentrations on day 7-10 among three groups. On the contrary, CAP did not change plasma FFA and glycerol concentrations on day 1-3. However, CAP increased markedly plasma FFA and glycerol concentrations on day 7-10 by 54-89% and 92-98%, respectively, as compared with the control group. DHC and CAP did not change the weights of white (perirenal and periepididymal) and brown (interscapular) adipose tissues. In conclusion, the effects of capsaicinoids on plasma glucose, FFA and glycerol concentrations were relatively higher in the DHC than in the CAP, and capsaicinoids did not change the weight of white and brown adipose tissues.


Certification and licensing of clinical medical physicists in AFOMP countries.
In many countries the education and training of medical physicists has changed over the last few decades from being rather ad hoc to becoming structured with residency programs becoming quite common. These are often followed by formal certification by independent bodies, and medical physics professional organizations (MPPOs) affirming the quality of certified physicists and recognizing their competence and ability to practice independently. Policies on the training and education of medical physicists have been developed by the International Organization for Medical Physics (IOMP) and by the Asia-Oceania Federation of Organizations for Medical Physics (AFOMP). Currently nine countries in Asia-Oceania operate systems for the certification of clinical medical physicists and four others are planning or implementing such systems. The existing systems, which are described in this paper, generally conform to the policies. Licensing of medical physicists, which is primarily the responsibility of government bodies, is almost unknown in that region.


Prospective, Randomized Ex Vivo Trial to Assess the Ideal Stapling Site for Endoscopic Fundoplication with Medigus Ultrasonic Surgical Endostapler.
Background and Aims. Endoscopic fundoplication is an emerging technique for the treatment of gastroesophageal reflux disease (GERD). The aim of this study is to determine the ideal position of the staples in relation to gastroesophageal junction (GEJ). Methods. Ten endoscopic fundoplication procedures were performed in each group using fresh ex vivo porcine stomachs: Group A: 2 staples each at 3 cm above the GEJ and 180° apart; Group B: 2 staples at 3 cm and 90° apart; Group C: 2 staples at 4 cm and 180° apart; Group D: 3 staples at 3 cm with 90° between each staple (180° total). After the procedure, the stomach was gradually filled with water. Gastric yield pressure (GYP) was determined by detection of reflux of the water in esophagus or by rupture of staples. Results. Mean increase of GYPs (±SD) after the procedure was as follows: Group A: 16.9 ± 8.7; Group B: 8.1 ± 7.9; Group C: 12.2 ± 9.4; Group D: 22.7 ± 13.3. GYP in Group A and Group D was higher than Group B (p = 0.03 and p = 0.01, resp.). Conclusions. We recommend the placement of 3 staples at 3 cm distance from the GEJ, which resulted in the highest increase of GYP.


A randomised, double blind, placebo controlled trial of the effect of theophylline in prevention of vasomotor nephropathy in very preterm neonates with respiratory distress syndrome.
Vasomotor nephropathy is a common renal dysfunction in very preterm neonates. To determine whether theophylline could prevent vasomotor nephropathy in very preterm infants with respiratory distress syndrome. A randomised, double blind, placebo controlled trial of 50 preterm infants of gestational age < or = 32 weeks needing assisted ventilation. Infants received an intravenous dose of theophylline (1 mg/kg) or placebo for three days. The 24 hour urine volume was measured daily. On days 2, 5, and 11, blood samples and 12 hour urine collections were analysed for electrolytes, creatinine, and urea. On day 1, urine output was significantly higher in the theophylline (2.4 (0.9) ml/kg/h) than the placebo (1.6 (1.0) ml/kg/h; p = 0.023) group (values are mean (SD)). The incidence of oligoanuria was significantly lower in the theophylline treated (5%) than the placebo (33%) group. Twenty four hours after the first administration of theophylline/placebo, serum creatinine concentration was significantly lower in the theophylline (0.76 (0.23) mg/dl) than the placebo (1.0 (0.41) mg/dl; p = 0.025) group. On day 5 an increase in serum creatinine was observed in both groups. On day 11 a significant reduction in serum creatinine was observed, compared with day 5, with no difference between the two groups. The results suggest that, in very preterm infants with respiratory distress syndrome, early theophylline administration improves renal function during the first two days of life.


Parents' perspectives on physician-parent communication near the time of a child's death in the pediatric intensive care unit.
Communicating bad news about a child's illness is a difficult task commonly faced by intensive care physicians. Greater understanding of parents' scope of experiences with bad news during their child's hospitalization will help physicians communicate more effectively. Our objective is to describe parents' perceptions of their conversations with physicians regarding their child's terminal illness and death in the pediatric intensive care unit (PICU). A secondary analysis of a qualitative interview study. Six children's hospitals in the National Institute of Child Health and Human Development Collaborative Pediatric Critical Care Research Network. Fifty-six parents of 48 children who died in the PICU 3-12 months before the study. Parents participated in audio recorded semistructured telephone interviews. Interviews were analyzed using established qualitative methods. Of the 56 parents interviewed, 40 (71%) wanted to provide feedback on the way information about their child's terminal illness and death was communicated by PICU physicians. The most common communication issue identified by parents was the physicians' availability and attentiveness to their informational needs. Other communication issues included honesty and comprehensiveness of information, affect with which information was provided, withholding of information, provision of false hope, complexity of vocabulary, pace of providing information, contradictory information, and physicians' body language. The way bad news is discussed by physicians is extremely important to most parents. Parents want physicians to be accessible and to provide honest and complete information with a caring affect, using lay language, and at a pace in accordance with their ability to comprehend. Withholding prognostic information from parents often leads to false hopes and feelings of anger, betrayal, and distrust. Future research is needed to investigate whether the way bad news is discussed influences psychological adjustment and family functioning among bereaved parents.


Clinicians' Views on Management and Terminology for Papillary Thyroid Microcarcinoma: A Qualitative Study.
There is growing acceptance that the increase in thyroid cancer incidence is in part a result of overdiagnosis of small low-risk papillary microcarcinomas (PMCs) with indolent clinical course. Although surgery is the cornerstone treatment for patients with PMCs, recent management guidelines are shifting toward inclusion of more conservative treatments such as active surveillance. There is little evidence on clinicians' experience in managing PMC patients and their attitudes toward treatment options, including their willingness to accept a nonsurgical option. The aim of this study was to understand how clinicians perceive a diagnosis of PMC, potential changes to terminology, and the treatment options available to patients. This was a qualitative study using semi-structured interviews conducted between November 2015 and May 2016 with 22 clinicians (seven endocrinologists and 15 thyroid surgeons). Transcribed audio-recordings were thematically coded, and a framework method was used to analyze the data. Across a sample of clinicians who manage thyroid cancer patients, awareness of overdiagnosis and overtreatment of PMC was common. However, there was little acceptance of active surveillance to manage these patients. Clinicians did not feel comfortable recommending this management approach, as they were worried about the risk of metastases, did not feel that evidence to support this approach was strong enough, and also believed that patients currently have a high preference for surgery. The majority of clinicians did not believe that changing the terminology of this diagnosis was a viable strategy to reduce patients' anxiety and their perceived preference for more aggressive treatments. However, most clinicians felt that thyroid nodules <1 cm should not be biopsied, which could help minimize the risk of overdiagnosis of PMC. This study, based on a non-representative sample of 22 clinicians, which remains an important limitation, provides revealing insight into clinicians' management preferences and decision making for small low-risk thyroid cancers at a time when management guidelines and practices are evolving. It suggests that clinicians may not be ready to accept nonsurgical options, or changes in terminology, until evidence to support these options and changes is stronger.


Bleeding in acute coronary syndromes and percutaneous coronary interventions: position paper by the Working Group on Thrombosis of the European Society of Cardiology.
Bleeding has recently emerged as an important outcome in the management of acute coronary syndromes (ACS), which is relatively frequent compared with ischaemic outcomes and has important implications in terms of prognosis, outcomes, and costs. In particular, there is evidence that patients experiencing major bleeding in the acute phase are at higher risk for death in the following months, although the causal nature of this relation is still debated. This position paper aims to summarize current knowledge regarding the epidemiology of bleeding in ACS and percutaneous coronary intervention, including measurement and definitions of bleeding, with emphasis on the recent consensus Bleeding Academic Research Consortium (BARC) definitions. It also provides an European perspective on management strategies to minimize the rate, extent, and consequences of bleeding. Finally, the research implications of bleeding (measuring and reporting bleeding in trials, the importance of bleeding as an outcome measure, and bleeding as a subject for future research) are also discussed.


Microfabricated Device for High-Resolution Imaging of Preimplantation Embryos.
The mouse preimplantation embryo is an excellent system for studying how mammalian cells organize dynamically into increasingly complex structures. Accessible to experimental and genetic manipulations, its normal or perturbed development can be scrutinized ex vivo by real-time imaging from fertilization to late blastocyst stage. High-resolution imaging of multiple embryos at the same time can be compromised by embryos displacement during imaging. We have developed an inexpensive and easy-to-produce imaging device that facilitates greatly the imaging of preimplantation embryo. In this chapter, we describe the different steps of production and storage of the imaging device as well as its use for live imaging of mouse preimplantation embryos expressing fluorescent reporters from genetically modified alleles or after in vitro transcribed mRNA transfer by microinjection or electroporation.


Meteterakis saotomensis n. sp. (Nematoda: Heterakidae) from Schistometopum thomense (Bocage) (Gymnophiona: Dermophiidae) on São Tomé Island.
Meteterakis saotomensis n. sp. is described from Schistometopum thomense (Bocage), a gymnophionan endemic to the oceanic island of São Tomé in the Gulf of Guinea. The specimens were assigned to Meteterakis Karve, 1930, based on the possession of a head with three rounded lips, not set-off from the body, the absence of interlabia and cordons, females with a long vagina and males with a preanal sucker, surrounded by a cuticularised rim and caudal alae that are supported by fleshy papillae. The new species is characterised by: body length 4.2-4.5 mm (males) and 5.1-6.4 mm (females); total length of oesophagus, including pharyngeal portion and oesophageal bulb, 820-856 µm (males) and 898-1,070 µm (females); length of pharynx 57-58 µm (males) and 65-68 µm (females); spicules equal, 410-521 µm long, with tessellated ornamentation throughout their length and alae, and with bevelled tip; gubernaculum or 'gubernacular mass' absent; tail length 164-176 µm (males) and 214-239 µm (females), with elongated tip; vulva at 2.3-2.8 mm from anterior end, with anterior lip forming small flap. This is the second species of Meteterakis reported from gymnophionan hosts and the first from the Afrotropical region. Selected comparative morphological data for Meteterakis spp. are presented, and data on host range and geographic distribution are updated. The name M. striaturus Oshmarin & Demshin, 1972 is corrected to M. striatura to reflect the female gender of the genus name.


Proteasome inhibition can induce an autophagy-dependent apical activation of caspase-8.
Antiapoptotic Bcl-2 family proteins are often highly expressed in chemotherapy-resistant cancers and impair mitochondrial outer membrane permeabilisation (MOMP), an important requirement for caspase activation via the intrinsic apoptosis pathway. Interestingly, although Bcl-2 overexpression in HeLa cervical cancer cells abrogated caspase processing in response to intrinsic apoptosis induction by staurosporine, tunicamycin or etoposide, residual caspase processing was observed following proteasome inhibition by bortezomib ([(1R)-3-methyl-1-({(2S)-3-phenyl-2-[(pyrazin-2-ylcarbonyl)amino]propanoyl}amino)butyl]boronic acid), epoxomicin (N-acetyl-N-methyl-lisoleucyl-L-isoleucyl-N-[(1S)-3-methyl-1-[[(2R)-2-methyloxiranyl]carbonyl]butyl]-L-threoninamide) or MG-132 (N-(benzyloxycarbonyl)leucinylleucinylleucinal). Similar responses were found in Bcl-2-overexpressing H460 NSCLC cells and Bax/Bak-deficient mouse embyronic fibroblasts. Mild caspase processing resulted in low DEVDase activities, which were MOMP independent and persisted for long periods without evoking immediate cell death. Surprisingly, depletion of caspase-3 and experiments in caspase-7-depleted MCF-7-Bcl-2 cells indicated that the DEVDase activity did not originate from effector caspases. Instead, Fas-associated death domain (FADD)-dependent caspase-8 activation was the major contributor to the slow, incomplete substrate cleavage. Caspase-8 activation was independent of death ligands, but required the induction of autophagy and the presence of Atg5. Depletion of XIAP or addition of XIAP-antagonising peptides resulted in a switch towards efficient apoptosis execution, suggesting that the requirement for MOMP was bypassed by activating the caspase-8/caspase-3 axis. Combination treatments of proteasome inhibitors and XIAP antagonists therefore represent a promising strategy to eliminate highly resistant cancer cells, which overexpress antiapoptotic Bcl-2 family members.


Inhibitory effect of caffeic acid phenethyl ester on bleomycine-induced lung fibrosis in rats.
Pulmonary fibrosis (PF) induced by anticancerogenic bleomycin (BLM) is one of the more common side effects encountered during cancer treatment. It has been suggested in the last decades that the main responsible agent in PF is reactive oxygen species which were generated also in normal physiological conditions in the human body. In this experimental study, we investigated the preventive or attenuating effects of caffeic acid phenethyl ester (CAPE) that has been demonstrated to have anti-inflammatory, cytocytatic, anticancerogenic, antiprolipherative and antioxidant effects on BLM-induced PF. Thirty-six Sprague-Dawley rats were divided randomly into four groups as sham operation, BLM, BLM + vitamin E (vit E), and BLM + CAPE groups. BLM (7.5 mg/kg, single dose) was applied intratracheally, and CAPE and vit E intraperitoneally in the appropriate groups. At the end of the fibrosis processes, lung tissues were removed and the levels of tissues hydroxyproline (OH-proline), malondialdehyde (MDA) and NO as well as the activities of superoxide dismutase (SOD), catalase (CAT) and myeloperoxidase (MPO) were determined. Also, the weights of the rats were recorded at 7th and 14th days of the experiments. BLM application to the rats resulted in a significant increase in the OH-proline level as compared to the controls. Administration of CAPE and vit E led to the remarkable reduction of total lung OH-proline levels compared to the rats treated with BLM alone (p < 0.0001). There were a decreases in antioxidant enzyme (SOD and CAT) activities while an increase in MPO activity in BLM group was found vs. the control group (p < 0.0001). CAPE had a regulator effect on these parameters: the increase in CAT and SOD activities and the decrease in MPO activity were seen after CAPE application. NO, MDA and OH-proline levels were increased in BLM group vs. the control group. CAPE was more effective in decreasing the tissue levels of NO, MDA and OH-proline than vit E. MPO activity, as a good marker of neutrophil sequestration to the tissues, in the BLM group was decreased by CAPE approximately to the control group. We suggest that CAPE is more effective on the prevention of BLM-induced fibrosis via antioxidant and free radical scavenger properties than vit E at the doses used in the present study. CAPE has some attenuating effects on BLM-induced PF affecting both oxidant and antioxidant systems as well as neutrophils sequestration.


Detection of pyrrolizidine alkaloids in commercial honey using liquid chromatography-ion trap mass spectrometry.
Pyrrolizidine alkaloids (PAs) are known secondary plant metabolites which can cause hepatotoxicity in both humans and livestock. PAs can be consumed through the use of plants for food, medicinal purposes and as contaminants of agricultural crops and food. PA contaminated grain has posed the largest health risk, although any PA contamination in our food chain should be recognised as a potential health threat. For this purpose, retail honeys were tested by LC-MS/MS. The method allows for specific identification of toxic retronecine and otonecine-type PAs by comparison to reference compounds via a spectral library. In total, 50 honey samples were matched to the reference spectra within a set of tolerance parameters. Accurate data analysis and quick detection of positive samples was possible. Positive samples contained an average PA concentration of 1260 μg kg(-1) of honey. Good linear calibrations were obtained (R(2)>0.991). LOD and LOQ ranged from 0.0134 to 0.0305 and 0.0446 to 0.1018 μg mL(-1), respectively.


How prepared are we for cross-border outbreaks? An exploratory analysis of cross-border response networks for outbreaks of multidrug resistant microorganisms in the Netherlands and Germany.
The emergence and spread of multidrug resistant microorganisms is a serious threat to transnational public health. Therefore, it is vital that cross-border outbreak response systems are constantly prepared for fast, rigorous, and efficient response. This research aims to improve transnational collaboration by identifying, visualizing, and exploring two cross-border response networks that are likely to unfold during outbreaks involving the Netherlands and Germany. Quantitative methods were used to explore response networks during a cross-border outbreak of carbapenem resistant Enterobacteriaceae in healthcare settings. Eighty-six Dutch and German health professionals reflected on a fictive but realistic outbreak scenario (response rate ≈ 70%). Data were collected regarding collaborative relationships between stakeholders during outbreak response, prior working relationships, and trust in the networks. Network analysis techniques were used to analyze the networks on the network level (density, centralization, clique structures, and similarity of tie constellations between two networks) and node level (brokerage measures and degree centrality). Although stakeholders mainly collaborate with stakeholders belonging to the same country, transnational collaboration is present in a centralized manner. Integration of the network is reached, since several actors are beneficially positioned to coordinate transnational collaboration. However, levels of trust are moderately low and prior-existing cross-border working relationships are sparse. Given the explored network characteristics, we conclude that the system has a promising basis to achieve effective coordination. However, future research has to determine what kind of network governance form might be most effective and efficient in coordinating the necessary cross-border response activity. Furthermore, networks identified in this study are not only crucial in times of outbreak containment, but should also be fostered in times of non-crisis.


Preparation, Optimization and Toxicity Evaluation of (SPION-PLGA) ±PEG Nanoparticles Loaded with Gemcitabine as a Multifunctional Nanoparticle for Therapeutic and Diagnostic Applications.
The aim of this study was to develop a novel multifunctional nanoparticle, which encapsulates SPION and Gemcitabine in PLGA ± PEG to form multifunctional drug delivery system. For this aim, super paramagnetic iron oxide nanoparticles (SPIONs) were simultaneously synthesized and encapsulated with Gemcitabine (Gem) in PLGA ± PEG copolymers via W/O/W double emulsification method. Optimum size and encapsulation efficiency for radiosensitization, hyperthermia and diagnostic applications were considered and the preparation parameters systematically were investigated and physicochemical characteristics of optimized nanoparticle were studied. Then SPION-PLGA and PLGA-Gem nanoparticles were prepared with the same optimized parameters and the toxicity of these nanoparticles was compared with Gemcitabine in human breast cancer cell line (MCF-7). The optimum preparation parameters were obtained with Gem/polymer equal to 0.04, SPION/polymer equal to 0.8 and 1% sucrose per 20 mg of polymer. The hydrodynamic diameters of all nanoparticles were under 200 nm. Encapsulation efficiency was adjusted between 13.2% to 16.1% for Gemcitabine and 48.2% to 50.1% for SPION. In-vitro Gemcitabine release kinetics had controlled behavior. Enhancement ratios for PLGA-Gem and SPION-PLGA-Gem at concentration of nanoparticles equal to IC50 of Gemcitabine were 1.53 and 1.89 respectively. The statistical difference was significant (p-value = 0.006 for SPION-PLGA-Gem and p-value = 0.015 for PLGA-Gem compared with Gemcitabine). In conclusion, we have successfully developed a Gemcitabine loaded super paramagnetic PLGA-Iron Oxide multifunctional drag delivery system. Future work includes in-vitro and in-vivo investigation of radiosensitization and other application of these nanoparticles.


Kaposi's sarcoma in patients with and without human immunodeficiency virus infection, in a tertiary referral centre in Kenya.
The clinical features of Kaposi's sarcoma (KS), in patients with and without HIV infection, were investigated in a tertiary referral centre in Kenya between 1997 and 1999. Although 186 cases were identified prospectively, the data analysis was restricted to the 91 (49%) cases who had pathological confirmation of Kaposi's sarcoma and documented HIV serostatus. Among these 91 subjects (58% of whom were male), the age-group holding the largest number of KS cases was that of individuals aged 31-40 years; most of the paediatric cases were aged 6-10 years. The ratio of HIV-seropositives to HIV-seronegatives was 8.5:1 for the adult cases and 0.9:1 for the paediatric. Of the signs and symptoms of Kaposi's sarcoma seen at presentation, only peripheral lympadenopathy was found to be significantly associated with underlying HIV infection (P = 0.05). The median survival was 104 days. It is apparent that, as the HIV epidemic advances in regions of the world with endemic KS, the clinical presentation and natural history of the endemic KS are blending with those of the epidemic or AIDS-associated disease, leading to a reduction in the mean age of the cases and a nearly identical incidence in men and women. In regions of the world where patients have ready access to such chemotherapy, the impact of treatment with highly active antiretroviral drugs on the incidence and natural history of KS has been dramatic. It will be important to monitor the clinico-pathological features of KS in the developing world, as more active antiretroviral regimens become available in clinical practice there.


Genetic variation and mating success in managed american plains bison.
The American plains bison (Bison bison) was pushed to the brink of extinction in the late 1800s but has since rebounded. Less than 5% of animals currently exist in conservation herds that are critical for maintaining genetic variability. Here, we use 25 microsatellite loci to assess genetic diversity and patterns of mating success over a 3-year period in a managed conservation herd at Konza Prairie Biological Station, Kansas (total number of individuals genotyped = 587). Heterozygosity was comparable to and allelic diversity higher than that in 11 other wild and managed herds for which similar estimates are available. Parentage analyses revealed that males within the oldest age classes (5-7 years) sired >90% of calves over the study period, consistent with a polygynous breeding system. Asymmetries in siring success also were observed within age classes, with the same males enjoying high siring success over multiple seasons. Empirical results of paternity will facilitate future modeling and empirical efforts to determine how demographic factors, population size, and variation in siring success interact to determine the retention (or loss) of genetic diversity in natural and managed herds, thus allowing informed recommendations for management practices and conservation efforts of this symbolic North American species.


Soluble epoxide hydrolase inhibitor, AUDA, prevents early salt-sensitive hypertension.
In stroke-prone spontaneously hypertensive rats (SHRSP) end-organ damage is markedly accelerated by high-salt (HS) intake. Since epoxyeicosatrienoic acids (EETs) possess vasodepressor and natriuretic activities, we examined whether a soluble epoxide hydrolase (sEH) inhibitor, 12-(3-adamantan-1-yl-ureido)-dodecanoic acid (AUDA), to inhibit the metabolism of EETs, would protect against pathologic changes in SHRSP. Seven-week-old male SHRSP were treated as follows: normal salt (NS), NS + AUDA, HS and HS + AUDA. Systolic blood pressure (SBP) (205 +/- 4 v 187 +/- 7 mmHg) and proteinuria (3.7 +/- 0.2 v 2.6 +/- 0.2 mg/6 h), but not plasma EETs (11.0 +/- 0.9 v 9.7 +/- 1.1 ng/ml), were significantly increased at 9 weeks of age in HS v NS SHRSP. HS was associated with fibrinoid degeneration and hypertrophy of arterioles in the kidney and perivascular fibrosis and contraction band necrosis in the heart. AUDA ameliorated these early salt-dependent changes in saline-drinking SHRSP and increased plasma levels of EETs but did not affect water and electrolyte excretion. sEH inhibition may provide a therapeutic strategy for treating salt-sensitive hypertension and its sequelae.


Uranium 238U/235U isotope ratios as indicators of reduction: results from an in situ biostimulation experiment at Rifle, Colorado, U.S.A.
The attenuation of groundwater contamination via chemical reaction is traditionally evaluated by monitoring contaminant concentration through time. However, this method can be confounded by common transport processes (e.g., dilution, sorption). Isotopic techniques bypass the limits of concentration methods, and so may provide improved accuracy in determining the extent of reaction. We apply measurements of 238U/235U to a U bioremediation field experiment at the Rifle Integrated Field Research Challenge Site in Rifle, Colorado. An array of monitoring and injection wells was installed on a 100 m2 plot where U(VI) contamination was present in the groundwater. Acetate-amended groundwater was injected along an up-gradient gallery to encourage the growth of dissimilatory metal reducing bacteria (e.g., Geobacter species). During amendment, U concentration dropped by an order of magnitude in the experiment plot. We measured 238U/235U in samples from one monitoring well by MC-ICP-MS using a double isotope tracer method. A significant approximately 1.00 per thousand decrease in 238U/235U occurred in the groundwater as U(VI) concentration decreased. The relationship between 238U/235U and concentration corresponds approximately to a Rayleigh distillation curve with an effective fractionation factor (alpha) of 1.00046. We attribute the observed U isotope fractionation to a nuclear field shift effect during enzymatic reduction of U(VI)(aq) to U(IV)(s).


Astonishing 35S rDNA diversity in the gymnosperm species Cycas revoluta Thunb.
In all eukaryotes, the highly repeated 35S ribosomal DNA (rDNA) sequences encoding 18S-5.8S-26S ribosomal RNA (rRNA) typically show high levels of intragenomic uniformity due to homogenisation processes, leading to concerted evolution of 35S rDNA repeats. Here, we compared 35S rDNA divergence in several seed plants using next generation sequencing and a range of molecular and cytogenetic approaches. Most species showed similar 35S rDNA homogeneity indicating concerted evolution. However, Cycas revoluta exhibits an extraordinary diversity of rDNA repeats (nucleotide sequence divergence of different copies averaging 12 %), influencing both the coding and non-coding rDNA regions nearly equally. In contrast, its rRNA transcriptome was highly homogeneous suggesting that only a minority of genes (<20 %) encode functional rRNA. The most common SNPs were C > T substitutions located in symmetrical CG and CHG contexts which were also highly methylated. Both functional genes and pseudogenes appear to cluster on chromosomes. The extraordinary high levels of 35S rDNA diversity in C. revoluta, and probably other species of cycads, indicate that the frequency of repeat homogenisation has been much lower in this lineage, compared with all other land plant lineages studied. This has led to the accumulation of methylation-driven mutations and pseudogenisation. Potentially, the reduced homology between paralogs prevented their elimination by homologous recombination, resulting in long-term retention of rDNA pseudogenes in the genome.


Transcription of the Staphylococcus aureus cid and lrg murein hydrolase regulators is affected by sigma factor B.
The Staphylococcus aureus lrg and cid loci are homologous operons that have been shown to regulate murein hydrolase activity and affect sensitivity to penicillin. Although the mode of action of these operons has not been demonstrated, a model based on the similarities of the lrgA and cidA gene products to the bacteriophage holin family of proteins has been proposed. In this study, the transcription organization and regulation of these operons were examined by Northern blot analyses. Unexpectedly, cidB and a gene located immediately downstream, designated cidC, were found to be cotranscribed on a 2.7-kb transcript. Maximal cidBC transcription occurred during early exponential growth, and high-level transcription of cidBC was dependent on the rsbU-mediated activation of the alternative sigma factor B (sigmaB). In contrast, lrgAB transcription in stationary phase was negatively regulated by sigmaB. Although cidABC transcription was not detected by Northern blot analysis, reverse transcriptase PCR revealed that these genes are also cotranscribed as a single RNA message in early exponential growth. Primer extension analysis revealed the presence of two cidBC transcription start sites, but no apparent sigmaB-dependent promoter consensus sequence was identified in these regions. The rsbU gene was also shown to have a positive impact on murein hydrolase activity but a negligible effect on sensitivity to penicillin-induced killing. These results suggest that the lrgAB and cidBC genes may be part of the S. aureus sigmaB-controlled stress regulon.


Activation of gamma delta T cells for production of IFN-gamma is mediated by bacteria via macrophage-derived cytokines IL-1 and IL-12.
Gamma delta T cells are found at sites of microbial infection and have been reported to proliferate in response to bacterial Ags. We show here that although the response by Listeria-elicited peritoneal gamma delta T cells to heat-killed bacteria in the presence of macrophage accessory cells may be partially mediated via the TCR, it is predominantly mediated via cytokines produced by the macrophages. Macrophage cytokines IL-12 and IL-1 synergize to induce some proliferation and considerable IFN-gamma production by peritoneal gamma delta T cells. This cytokine synergy pattern differs from that reported for NK cells, in which IL-12 in combination with either IL-2 or TNF-alpha induces NK cells to produce IFN-gamma. The combination of IL-12 and IL-1 provides a strong stimulus for IFN-gamma production by gamma delta T cells, but a relatively weak signal for proliferation. This is in contrast to the strong proliferative signal from the combination of IL-7 and IL-1 and the relatively weak stimulation of IFN-gamma production by the IL-7/IL-1 combination. Thus, there is differential regulation of NK and gamma delta T cells by cytokines and differential regulation of activation functions within the gamma delta T cell population by combinations of cytokines. These data provide evidence for a potentially important pathway for augmentation of IFN-gamma secretion at sites of infection where gamma delta T cells are found in abundance and where IFN-gamma may play a major role in the control of the infection.