Sedation protocols to reduce duration of mechanical ventilation in the ICU: a Cochrane Systematic Review.
Assess the effects of protocol-directed sedation management on the duration of mechanical ventilation and other relevant patient outcomes in mechanically ventilated intensive care unit patients. Sedation is a core component of critical care. Sub-optimal sedation management incorporates both under- and over-sedation and has been linked to poorer patient outcomes. Cochrane systematic review of randomized controlled trials. Cochrane Central Register of Controlled trials, MEDLINE, EMBASE, CINAHL, Database of Abstracts of Reviews of Effects, LILACS, Current Controlled Trials and US National Institutes of Health Clinical Research Studies (1990-November 2013) and reference lists of articles were used. Randomized controlled trials conducted in intensive care units comparing management with and without protocol-directed sedation were included. Two authors screened titles, abstracts and full-text reports. Potential risk of bias was assessed. Clinical, methodological and statistical heterogeneity were examined and the random-effects model used for meta-analysis where appropriate. Mean difference for duration of mechanical ventilation and risk ratio for mortality, with 95% confidence intervals, were calculated. Two eligible studies with 633 participants comparing protocol-directed sedation delivered by nurses vs. usual care were identified. There was no evidence of differences in duration of mechanical ventilation or hospital mortality. There was statistically significant heterogeneity between studies for duration of mechanical ventilation. There is insufficient evidence to evaluate the effectiveness of protocol-directed sedation as results from the two randomized controlled trials were conflicting.


How do plant viruses induce disease? Interactions and interference with host components.
Plant viruses are biotrophic pathogens that need living tissue for their multiplication and thus, in the infection-defence equilibrium, they do not normally cause plant death. In some instances virus infection may have no apparent pathological effect or may even provide a selective advantage to the host, but in many cases it causes the symptomatic phenotypes of disease. These pathological phenotypes are the result of interference and/or competition for a substantial amount of host resources, which can disrupt host physiology to cause disease. This interference/competition affects a number of genes, which seems to be greater the more severe the symptoms that they cause. Induced or repressed genes belong to a broad range of cellular processes, such as hormonal regulation, cell cycle control and endogenous transport of macromolecules, among others. In addition, recent evidence indicates the existence of interplay between plant development and antiviral defence processes, and that interference among the common points of their signalling pathways can trigger pathological manifestations. This review provides an update on the latest advances in understanding how viruses affect substantial cellular processes, and how plant antiviral defences contribute to pathological phenotypes.


Neighborhood level risk factors for type 1 diabetes in youth: the SEARCH case-control study.
European ecologic studies suggest higher socioeconomic status is associated with higher incidence of type 1 diabetes. Using data from a case-control study of diabetes among racially/ethnically diverse youth in the United States (U.S.), we aimed to evaluate the independent impact of neighborhood characteristics on type 1 diabetes risk. Data were available for 507 youth with type 1 diabetes and 208 healthy controls aged 10-22 years recruited in South Carolina and Colorado in 2003-2006. Home addresses were used to identify Census tracts of residence. Neighborhood-level variables were obtained from 2000 U.S. Census. Multivariate generalized linear mixed models were applied. Controlling for individual risk factors (age, gender, race/ethnicity, infant feeding, birth weight, maternal age, number of household residents, parental education, income, state), higher neighborhood household income (p = 0.005), proportion of population in managerial jobs (p = 0.02), with at least high school education (p = 0.005), working outside the county (p = 0.04) and vehicle ownership (p = 0.03) were each independently associated with increased odds of type 1 diabetes. Conversely, higher percent minority population (p = 0.0003), income from social security (p = 0.002), proportion of crowded households (0.0497) and poverty (p = 0.008) were associated with a decreased odds. Our study suggests that neighborhood characteristics related to greater affluence, occupation, and education are associated with higher type 1 diabetes risk. Further research is needed to understand mechanisms underlying the influence of neighborhood context.


Indoor air pollution and exposure assessment of the gulf cooperation council countries: A critical review.
Indoor air pollution is one of the human health threat problems in the Gulf Cooperation Council (GCC) countries. In these countries, due to unfavorable meteorological conditions, such as elevated ambient temperature, high relative humidity, and natural events such as dust storms, people spend a substantial amount of their time in indoor environments. In addition, production of physical and biological aerosols from air conditioners, cooking activities, burning of Arabian incense, and overcrowding due to pilgrimage programs are common causes of low quality indoor air in this region. Thus, due to infiltration of outdoor sources as well as various indoor sources, people living in the GCC countries are highly exposed to indoor air pollutants. Inhalation of indoor air pollutants causes mortalities and morbidities attributed to cardiorespiratory, pulmonary, and lung cancer diseases. Hence, the aim of this review study is to provide a summary of the major findings of indoor air pollution studies in different microenvironments in six GCC countries. These include characterization of detected indoor air pollutants, exposure concentration levels, source identifications, sustainable building designs and ventilation systems, and the mitigation strategies. To do so, >130 relevant indoor air pollution studies across the GCC countries were critically reviewed. Particulate matters (PM10 and PM2.5), total volatile organic compounds (TVOCs), carbon dioxide (CO2), sulfur dioxide (SO2), nitrogen dioxide (NO2), and heavy metals were identified as the reported indoor air pollutants. Apart from them, indoor Radon and bioaerosols were studied only in specific GCC countries. Thus, future studies should also focus on the investigation of emerging indoor air pollutants, such as ultrafine and nanoparticles and their associated health effects. Furthermore, studies on the mitigation of indoor air pollution through the development of advanced air purification and ventilation systems could improve the indoor air quality (IAQ) in the GCC region.


Alpha Radiation-Induced Luminescence by Am-241 in Aqueous Nitric Acid Solution.
When exposed to air, alpha particles cause the production of light by exciting the molecules surrounding them. This light, the radioluminescence, is indicative of the presence of alpha radiation, thus allowing for the optical sensing of alpha radiation from distances larger than the few centimeters an alpha particle can travel in air. While the mechanics of radioluminescence in air and other gas compositions is relatively well understood, the same cannot be said about the radioluminescence properties of liquids. Better understanding of the radioluminescence properties of liquids is essential to design methods for the detection of radioactively contaminated liquids by optical means. In this article, we provide radioluminescence images of Am-241 dissolved in aqueous nitric acid ( HNO 3 ) solution and present the recorded radioluminescence spectrum with a maximum between 350 nm and 400 nm , and a steep decrease at the short wavelength side of the maximum. The shape of the spectrum resembles a luminescence process rather than Cerenkov light, bremsstrahlung, or other mechanisms with broadband emission. We show that the amount of light produced is about 150 times smaller compared to that of the same amount of Am-241 in air. The light production in the liquid is evenly distributed throughout the sample volume with a slight increase on the surface of the liquid. The radioluminescence intensity is shown to scale linearly with the Am-241 concentration and not be affected by the HNO 3 concentration.


Trump's "America first" energy policy, contingency and the reconfiguration of the global energy order.
The global energy system is in transition to a new energy order characterized by the emergence of the United States as a net oil exporter, the shale revolution and the gradual shift towards low-carbon sources and renewables. The shale boom in the US was a game changer, as was the election of Donald Trump as US president. Trump pushed an ambitious "America first" agenda aimed at transforming the US into a global energy superpower. The purpose of this article is two-fold: first, it outlines the key pillars of the emerging global energy order. Second, it underscores the role of contingent events, a factor neglected by some previous studies because of their reliance on what A.O.Hirschman (1970) termed "paradigmatic thinking". The recent transition in the international energy order is an outcome of two paramount, yet largely unanticipated events: the shale revolution in the US and Trump's neomercantilist and unilateralist economic policies. While contingencies are an inherent feature of social reality, the scenario approach can be a useful heuristic for dealing with uncertainties. The article concludes by discussing the implications of these developments for global energy governance.


Corticomotor excitability during precision motor tasks.
The aim of this preliminary study was to investigate motor cortex (cortical) excitability between a similar fine visuomotor task of varying difficulty. Ten healthy adults (three female, seven male; 20-45 years of age) participated in the study. Participants were instructed to perform a fine visuomotor task by statically abducting their first index finger against a force transducer which displayed the level of force (represented as a marker) on a computer monitor. This marker was to be maintained between two stationary bars, also displayed on the computer monitor. The level of difficulty was increased by amplifying the position of the marker, making the task more difficult to control. Cortical measures of motor evoked potential (MEP) and silent period (SP) duration in first dorsal interosseous (FDI) muscle were obtained using transcranial magnetic stimulation (TMS) while the participant maintained the "easy" or "difficult" static task. An 11.8% increase in MEP amplitude was observed when subjects undertook the "difficult" task, but no differences in MEP latency or SP duration. The results from this preliminary study suggest that cortical excitability increases reflect the demand required to perform tasks requiring greater precision with suggestions for further research discussed.


Rapid induction of apoptosis by deregulated uptake of polyamine analogues.
Treatment of Chinese hamster ovary cells with alpha-difluoromethylornithine for 3 days, followed by exposure to cycloheximide, led to an unregulated, rapid and massive accumulation of polyamine analogues. This accumulation led to cell death by apoptosis within a few hours. Clear evidence of DNA fragmentation was seen in response to both N-terminally ethylated polyamines and to polyamines containing methyl groups on the terminal carbon atoms. Programmed cell death was induced within 2-4 h of exposure to 1 microM or higher concentrations of N1,N11-bis(ethyl)norspermine. The presence of cycloheximide increased the uptake of the polyamine analogues and therefore led to cell death at lower analogue concentrations, but it was not essential for the induction of apoptosis, since similar effects were seen when the protein synthesis inhibitor was omitted and the concentration of N1, N11-bis(ethyl)norspermine was increased to 5 microM or more. The induction of apoptosis was blocked both by the addition of the caspase inhibitor N-benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone, or by the addition of the polyamine oxidase inhibitor N1-methyl-N2-(2,3-butadienyl)butane-1,4-diamine (MDL 72,527). These experiments provide evidence to support the concepts that: (1) polyamines or their oxidation products may be initiators of programmed cell death; (2) regulation of polyamine biosynthesis and uptake prevents the accumulation of toxic levels of polyamines; and (3) the anti-neoplastic effects of bis(ethyl) polyamine analogues may be due to the induction of apoptosis in sensitive tumour cells.


Oceanic fronts in the Sargasso Sea control the early life and drift of Atlantic eels.
Anguillid freshwater eels show remarkable life histories. In the Atlantic, the European eel (Anguilla anguilla) and American eel (Anguilla rostrata) undertake extensive migrations to spawn in the oceanic Sargasso Sea, and subsequently the offspring drift to foraging areas in Europe and North America, first as leaf-like leptocephali larvae that later metamorphose into glass eels. Since recruitment of European and American glass eels has declined drastically during past decades, there is a strong demand for further understanding of the early, oceanic phase of their life cycle. Consequently, during a field expedition to the eel spawning sites in the Sargasso Sea, we carried out a wide range of dedicated bio-physical studies across areas of eel larval distribution. Our findings suggest a key role of oceanic frontal processes, retaining eel larvae within a zone of enhanced feeding conditions and steering their drift. The majority of the more westerly distributed American eel larvae are likely to follow a westerly/northerly drift route entrained in the Antilles/Florida Currents. European eel larvae are generally believed to initially follow the same route, but their more easterly distribution close to the eastward flowing Subtropical Counter Current indicates that these larvae could follow a shorter, eastward route towards the Azores and Europe. The findings emphasize the significance of oceanic physical-biological linkages in the life-cycle completion of Atlantic eels.


Therapist and client emotional expression and psychotherapy outcomes: A meta-analysis.
Although emotion has long been considered important to psychotherapeutic process, empirical assessment of its impact has emerged only recently. The present study applied two meta-analyses to explore the association between therapist expression of emotion and psychotherapy outcome, and client expression of emotion and psychotherapy outcome. Overall, 66 studies (13 for the therapist meta-analysis and 43 for the client meta-analysis) were included. A significant medium effect size was found between the therapist's emotional expression and outcomes (d = 0.56) and a significant medium-to-large effect size between the client's emotional expression and outcomes (d = 0.85). Third-party rating of emotional expression emerged as a significant moderator of outcomes. Limitations of the research, diversity considerations, and therapeutic practices that conclude the article are then presented. (PsycINFO Database Record (c) 2018 APA, all rights reserved).


Creutzfeldt-Jakob disease with unusually extensive neuropathology in a child treated with native human growth hormone.
We report a case of iatrogenic Creutzfeldt-Jakob disease(iCJD) in a child with a neonatal growth hormone (GH) deficiency that was treated with native human growth hormone (hGH) between the ages of 9 months and 7 years. Three years after the end of treatment a progressive neurological syndrome consistent with Creutzfeldt-Jakob disease (CJD) developed, leading to death within a year, at age 11. Neuropathological examination showed an unusual widespread form of CJD, notably characterized by (i) involvement of the cerebellar white matter, (ii) cortico-spinal degeneration and (iii) ballooned neurons. A transitional form of the disease between common iatrogenic and panencephalopathic CJD is suggested.


Characterizing the composition and evolution of homoeologous genomes in hexaploid wheat through BAC-end sequencing on chromosome 3B.
Bread wheat (Triticum aestivum) is one of the most important crops worldwide. However, because of its large, hexaploid, highly repetitive genome it is a challenge to develop efficient means for molecular analysis and genetic improvement in wheat. To better understand the composition and molecular evolution of the hexaploid wheat homoeologous genomes and to evaluate the potential of BAC-end sequences (BES) for marker development, we have followed a chromosome-specific strategy and generated 11 Mb of random BES from chromosome 3B, the largest chromosome of bread wheat. The sequence consisted of about 86% of repetitive elements, 1.2% of coding regions, and 13% remained unknown. With 1.2% of the sequence length corresponding to coding sequences, 6000 genes were estimated for chromosome 3B. New repetitive sequences were identified, including a Triticineae-specific tandem repeat (Fat) that represents 0.6% of the B-genome and has been differentially amplified in the homoeologous genomes before polyploidization. About 10% of the BES contained junctions between nested transposable elements that were used to develop chromosome-specific markers for physical and genetic mapping. Finally, sequence comparison with 2.9 Mb of random sequences from the D-genome of Aegilops tauschii suggested that the larger size of the B-genome is due to a higher content in repetitive elements. It also indicated which families of transposable elements are mostly responsible for differential expansion of the homoeologous wheat genomes during evolution. Our data demonstrate that BAC-end sequencing from flow-sorted chromosomes is a powerful tool for analysing the structure and evolution of polyploid and highly repetitive genomes.


Author Correction: Large-scale transcriptome-wide association study identifies new prostate cancer risk regions.
The original version of this Article contained an error in the spelling of a member of the PRACTICAL Consortium, Manuela Gago-Dominguez, which was incorrectly given as Manuela Gago Dominguez. This has now been corrected in both the PDF and HTML versions of the Article. Furthermore, In the original HTML version of this Article, the order of authors within the author list was incorrect. The consortium PRACTICAL consortium was incorrectly listed after Bogdan Pasaniuc and should have been listed after Kathryn L. Penney. This error has been corrected in the HTML version of the Article; the PDF version was correct at the time of publication.


Chromosome mapping of the sweet potato little leaf phytoplasma reveals genome heterogeneity within the phytoplasmas.
To further understand the genomic diversity and genetic architecture of phytoplasmas, a physical and genetic map of the sweet potato little leaf (SPLL) strain V4 phytoplasma chromosome was determined. PFGE was used to determine the size of the SPLL-V4 genome, which was estimated to be 622 kb. A physical map was prepared by two-dimensional reciprocal digestions using the restriction endonucleases BssHII, Smal, Eagl and I-Ceul. Sixteen cleavage sites were located on the map. Southern hybridizations of digested SPLL-V4 chromosomal DNA were done using random clones and PCR-amplified genes as probes. This confirmed fragment positions and located the two rRNA operons and the linked fus/tuf genes encoding elongation factors G and Tu, respectively, on the physical map. An inversion of one of the rRNA operons was observed from hybridization data. Sequence analysis of one of the random clones identified a gid gene encoding a glucose-inhibited division protein. Digestions of the tomato big bud (TBB) phytoplasma chromosome with the same four enzymes revealed genome heterogeneity when compared to the closely related SPLL-V4, and a preliminary chromosome size for the TBB phytoplasma of 662 kb was estimated. This mapping information has revealed that significant genome diversity exists within the phytoplasmas.


Wide geographic variations of sensitivity to MOTT sensitins in Greece.
Several studies have suggested that large bodies of water are a main source of infection with mycobacteria other than tuberculosis (MOTT). If this is correct, there should be a gradient in the infection rate with MOTT between mountainous and seaside areas. To test this hypothesis, we performed skin testing with tuberculin and sensitins in 19,470 Greek Armed Forces recruits. Initially, several MOTT sensitins were used, but when it became clear that the Mycobacterium scrofulaceum sensitin was the most appropriate, the study was continued with it alone in 17,403 recruits. Finally, in order to evaluate the geographical distribution of sensitivity to sensitins, we studied the results of 8,507 of these recruits living in or near their birthplace. They were divided into three geophysical areas: seaside 3,389 recruits; mountains 2,692 recruits; and inland plains 2,426 recruits. MOTT sensitivity rates were 4.1% in mountainous areas and 7.1% in seaside areas. All small Aegean islands had high MOTT rates (above 8%). In inland plains, high MOTT rates (above 8%) were observed among those living near big rivers. This geographical distribution of MOTT sensitivity supports the theory that large bodies of water are a main source of infection with MOTT.


Behaviour of motor units of human arm muscles: differences between slow isometric contraction and relaxation.
The behaviour of motor units in the m. biceps brachii (long head), in the m. brachialis and in the m. supinator during slow isometric contraction and relaxation was studied when subjects were performing different motor tasks. These tasks were: flexion of the elbow joint, supination of the forearm and exorotation of the humerus. Motor unit activity was recorded by means of bipolar fine wire electrodes. In the long head of the biceps, motor unit activity was recorded at medial, central and lateral sites. When the subject relaxed from flexion, the firing rate of motor units located in the biceps and the brachialis was always found to be lower than that at the corresponding level of flexion force during contraction. The firing rate during relaxation decreased slowly and almost linearly with force. However, during relaxation from supination or exorotation, the firing rate of motor units at medial and central locations in the biceps was more or less constant until decruitment. The firing rate of motor units of the supinator during relaxation from supination decreased slowly and was lower than during contraction. Motor units located medially and centrally in the biceps had decruitment thresholds for flexion that were lower than their recruitment thresholds. Motor units on the lateral side of the biceps did not show such a difference. In the brachialis decruitment thresholds for flexion were usually higher than the recruitment thresholds. Differences between decruitment and recruitment thresholds for motor units in the biceps were much more pronounced for supination and exorotation than for flexion. For motor units in the supinator the decruitment threshold during relaxation from supination was higher than the recruitment threshold. The time that had passed after the onset of firing of a motor unit did not influence its decruitment threshold. If, after complete relaxation, the exerted force was increased again, it appeared that the recruitment threshold was changed. It took about 4 s to reach the original recruitment threshold. It is concluded that the relation between the firing rate of a motor unit and total exerted force depends on the phase of contraction. This relation varies within a muscle and between muscles. Furthermore, the results indicate an interchange of activity within the motoneurone pools of the synergists involved in isometrical motor tasks.


Fast joint estimation of silhouettes and dense 3D geometry from multiple images.
We propose a probabilistic formulation of joint silhouette extraction and 3D reconstruction given a series of calibrated 2D images. Instead of segmenting each image separately in order to construct a 3D surface consistent with the estimated silhouettes, we compute the most probable 3D shape that gives rise to the observed color information. The probabilistic framework, based on Bayesian inference, enables robust 3D reconstruction by optimally taking into account the contribution of all views. We solve the arising maximum a posteriori shape inference in a globally optimal manner by convex relaxation techniques in a spatially continuous representation. For an interactively provided user input in the form of scribbles specifying foreground and background regions, we build corresponding color distributions as multivariate Gaussians and find a volume occupancy that best fits to this data in a variational sense. Compared to classical methods for silhouette-based multiview reconstruction, the proposed approach does not depend on initialization and enjoys significant resilience to violations of the model assumptions due to background clutter, specular reflections, and camera sensor perturbations. In experiments on several real-world data sets, we show that exploiting a silhouette coherency criterion in a multiview setting allows for dramatic improvements of silhouette quality over independent 2D segmentations without any significant increase of computational efforts. This results in more accurate visual hull estimation, needed by a multitude of image-based modeling approaches. We made use of recent advances in parallel computing with a GPU implementation of the proposed method generating reconstructions on volume grids of more than 20 million voxels in up to 4.41 seconds.


Effects of chronic blockade of angiotensin II receptor on the maintenance of hypertension and vascular changes in spontaneously hypertensive rats.
Previous studies have shown that chronic treatment of spontaneously hypertensive rats (SHR) with angiotensin converting enzyme inhibitors is effective in causing the regression of hypertension and vascular structural changes in adult SHR. The purpose of this study was to determine the involvement of angiotensin type 1 receptors in the maintenance of hypertension and vascular changes in adult SHR. Three groups of 15-week-old male SHR were treated with three different doses of the angiotensin type 1 receptor antagonist L-158,809. After 12 weeks of treatment, average systolic blood pressure was reduced in a dose-dependent manner from 206 +/- 2 mmHg (1 mmHg = 133.3 Pa) in the control SHR to 154 +/- 7 mmHg in the 0.6 mg/kg group, 172 +/- 3 mmHg in the 0.3 mg/kg group, and 196 +/- 8 mmHg in the 0.1 mg/kg group. Withdrawal of the treatment caused a rapid rise in blood pressure within 1 week, and 14-16 weeks after treatment had been withdrawn, blood pressure of SHR treated with the highest dose had increased to 169 +/- 8 mmHg. Blood pressure of the SHR treated with the two lower doses had increased to the level of the untreated SHR. Mean blood pressure, diastolic pressure, and pulse pressure were significantly reduced in the SHR treated with 0.6 mg/kg L-158,809. The arterial media cross-sectional area and media to lumen ratio were reduced by L-158,809 treatment (0.6 mg/kg dose). Treatment did not affect body weight or heart rate. Contractile response of the mesenteric arteries to nerve or norepinephrine stimulation was not affected by the treatment. However, arteries from treated SHR did not show an angiotensin II potentiation of nerve-stimulated response, as was observed in the control SHR. We conclude that persistent control of blood pressure by treatment with L-158,809 is associated with its effects on the structure and function of the arteries in the SHR.


Severe brief pressure-controlled hemorrhagic shock after traumatic brain injury exacerbates functional deficits and long-term neuropathological damage in mice.
Hypotension after traumatic brain injury (TBI) worsens outcome. We published the first report of TBI plus hemorrhagic shock (HS) in mice using a volume-controlled approach and noted increased neuronal death. To rigorously control blood pressure during HS, a pressure-controlled HS model is required. Our hypothesis was that a brief, severe period of pressure-controlled HS after TBI in mice will exacerbate functional deficits and neuropathology versus TBI or HS alone. C57BL6 male mice were randomized into four groups (n=10/group): sham, HS, controlled cortical impact (CCI), and CCI+HS. We used a pressure-controlled shock phase (mean arterial pressure [MAP]=25-27 mm Hg for 35 min) and its treatment after mild to moderate CCI including, a 90 min pre-hospital phase, during which lactated Ringer's solution was given to maintain MAP >70 mm Hg, and a hospital phase, when the shed blood was re-infused. On days 14-20, the mice were evaluated in the Morris water maze (MWM, hidden platform paradigm). On day 21, the lesion and hemispheric volumes were quantified. Neuropathology and hippocampal neuron counts (hematoxylin and eosin [H&E], Fluoro-Jade B, and NeuN) were evaluated in the mice (n=60) at 24 h, 7 days, or 21 days (n=5/group/time point). HS reduced MAP during the shock phase in the HS and CCI+HS groups (p<0.05). Fluid requirements during the pre-hospital phase were greatest in the CCI+HS group (p<0.05), and were increased in HS versus sham and CCI animals (p<0.05). MWM latency was increased on days 14 and 15 after CCI+HS (p<0.05). Swim speed and visible platform latency were impaired in the CCI+HS group (p<0.05). CCI+HS animals had increased contusion volume versus the CCI group (p<0.05). Hemispheric volume loss was increased 33.3% in the CCI+HS versus CCI group (p<0.05). CA1 cell loss was seen in CCI+HS and CCI animals at 24 h and 7 days (p<0.05). CA3 cell loss was seen after CCI+HS (p<0.05 at 24 h and 7 days). CA1 cell loss at 21 days was seen only in CCI+HS animals (p<0.05). Brief, severe, pressure-controlled HS after CCI produces robust functional deficits and exacerbates neuropathology versus CCI or HS alone.


Antisense oligonucleotides to Cux-1, a Cut-related homeobox gene, cause increased apoptosis in mouse embryonic kidney cultures.
Cux-1 is a murine homeobox gene that is highly and transiently expressed in the developing kidney. To further evaluate the role of Cux-1 in mammalian kidney development, organotypic cultures of embryonic mouse kidney were incubated with phosphorothioate-coupled antisense Cux-1 oligonucleotides (ODNs) in the presence of cationic liposomes. Inhibition of Cux-1 expression by antisense ODNs was verified by reverse transcription-PCR. Metanephroi that were incubated with antisense Cux-1 ODNs were 23% smaller than metanephroi that were incubated with sense Cux-1 ODNs. Morphologic analysis of metanephroi that were treated with antisense Cux-1 ODNs revealed that ureteric buds and induced epithelial structures were present. However, extensive areas of cell death containing shrunken cells with pyknotic nuclei were also evident. The presence of increased apoptosis was verified by ultrastructural and terminal transferase-mediated dUTP nick end labeling analyses. Two different antisense Cux-1 ODNs targeting either the translation start codon or the homeobox produced increased apoptosis. In contrast, metanephroi incubated with sense ODNs exhibited only occasional apoptotic cells. We conclude that the presence of antisense Cux-1 ODNs does not block nephron induction, but results instead in increased apoptosis. Proper regulation of Cux-1 expression may be necessary for normal kidney development.


Towards global consensus on core outcomes for hidradenitis suppurativa research: an update from the HISTORIC consensus meetings I and II.
A core outcomes set (COS) is an agreed minimum set of outcomes that should be measured and reported in all clinical trials for a specific condition. Hidradenitis suppurativa (HS) has no agreed-upon COS. A central aspect in the COS development process is to identify a set of candidate outcome domains from a long list of items. Our long list had been developed from patient interviews, a systematic review of the literature and a healthcare professional survey, and initial votes had been cast in two e-Delphi surveys. In this manuscript, we describe two in-person consensus meetings of Delphi participants designed to ensure an inclusive approach to generation of domains from related items. To consider which items from a long list of candidate items to exclude and which to cluster into outcome domains. The study used an international and multistakeholder approach, involving patients, dermatologists, surgeons, the pharmaceutical industry and medical regulators. The study format was a combination of formal presentations, small group work based on nominal group theory and a subsequent online confirmation survey. Forty-one individuals from 13 countries and four continents participated. Nine items were excluded and there was consensus to propose seven domains: disease course, physical signs, HS-specific quality of life, satisfaction, symptoms, pain and global assessments. The HISTORIC consensus meetings I and II will be followed by further e-Delphi rounds to finalize the core domain set, building on the work of the in-person consensus meetings.


Frequency of anatomical variation of the sacroiliac joint in asymptomatic young adults and its relationship with sacroiliac joint degeneration.
To evaluate the variations of the sacroiliac joint (SIJ) in asymptomatic young adults and their relationship to SIJ degeneration. A total of 860 SIJs were retrospectively evaluated in 430 abdominal computed tomographies (CT) of abdominal pain cases. The study population consisted of 430 patients (176 female, 254 male) aged 20-45 years with abdominal pain. The anatomical variations of the SIJ were classified as (1) accessory sacroiliac joint, (2) iliosacral complex, (3) bipartite iliac bony plate, (4) semicircular defects, (5) crescent-like iliac bony plate, and (6) ossification center. The CT findings of SIJ degeneration included joint space narrowing, sclerosis, subcortical cysts, vacuum phenomenon, and osteophytes. Chi-square and cross tabulation tests were used for statistical analysis. The iliosacral complex was the most common (bilateral) anatomical variation (7.7%, n = 66). A total of 71 SIJ degenerations were detected in cases with anatomical variations. They were determined in 22 (2.6%) iliosacral complex, 19 (2.2%) bipartite iliac bony plate, 15 (1.7%) accessory sacroiliac joint, 6 (0.7%) semicircular defect, 5 (0.6%) crescent-like iliac bony plate, and 4 (0.5%) ossification center cases. We found that iliosacral complex was the most common SIJ variation in asymptomatic young adults and also that there was significantly more SIJ degeneration among subjects with anatomical variations of the SIJ than those without. SIJ degeneration was most common in subjects with the iliosacral complex variation.


Design considerations for polarization-sensitive optical coherence tomography with a single input polarization state.
Using a generalized design for a polarization-sensitive optical coherence tomography (PS-OCT) system with a single input polarization state (SIPS), we prove the existence of an infinitely large design space over which it is possible to develop simple PS-OCT systems that yield closed form expressions for birefringence. Through simulation and experiment, we validate this analysis by demonstrating new configurations for PS-OCT systems, and present guidelines for the general design of such systems in light of their inherent inaccuracies. After accounting for systemic errors, alternative designs exhibit similar performance on average to the traditional SIPS PS-OCT system. This analysis could be extended to systems with multiple input polarization states and could usher in a new generation of PS-OCT systems optimally designed to probe specific birefringent samples with high accuracy.


Surface-enhanced coherent anti-Stokes Raman imaging of lipids.
This work describes in detail a wide-field surface-enhanced coherent anti-Stokes Raman scattering (CARS) microscope, which enables enhanced detection of sample structures in close proximity (∼100 nm) of the substrate interface. Unlike conventional CARS microscopy, where the sample is illuminated with freely propagating light, the current implementation uses evanescent fields to drive Raman coherences across the entire object plane. By coupling the pump and Stokes excitation beams to the surface plasmon-polariton mode at the interface of a 30 nm thick gold film, we obtained strong CARS signals from cholesteryl oleate droplets adhered to the surface. The surface-enhanced CARS imaging system visualizes lipid structures with vibrational selectivity using illumination doses per unit area that are more than four orders of magnitude lower than in point-scanning CARS microscopy.


Robotic Mitral Valve Repair for Simple and Complex Degenerative Disease: Midterm Clinical and Echocardiographic Quality Outcomes.
Severe primary (degenerative) mitral regurgitation (MR) is repaired with durable results when simple single-scallop disease is addressed. The midterm quality outcomes of minimally invasive repair for complex disease are unknown, however. From January 2008 to January 2015, 487 patients (56±11 years, 360 men, ejection fraction 65±6%, 98.8% complete follow-up) underwent robotic mitral valve repair for severe nonischemic degenerative MR. Simple pathology was addressed in 289 of 487 (59%) patients, and complex repair (all others) was performed in 198 of 487 (41%). Four patients died during follow-up with a 5-year survival rate 99.5% (99.4% simple; 99.5% complex; hazard ratio, 0.48; 95% confidence interval, 0.05-4.59); and New York Heart Association functional class I/II was documented in 97.9% (477/487). Eight patients had recurrence of moderate-to-severe MR (4 simple, 4 complex), with a 5-year freedom from MR of 94.6% (96.2% simple; 92.7%, complex; P=0.67; hazard ratio, 1.36; 95% confidence interval, 0.34-5.43). Seven patients (2 simple, 5 complex), underwent mitral reoperation, with a 5-year freedom from reoperation of 97.7% (99.1% simple; 95.7% complex; P=0.13; hazard ratio, 3.35; 95% confidence interval, 0.65-17.32). At a large tertiary care referral center, midterm quality outcomes after robotic correction of degenerative MR are excellent, with very high survival, infrequent complications, and a low likelihood of MR recurrence, regardless of mitral valve repair complexity. Awareness of these improvements in outcome is important to inform contemporary decisions regarding high-quality alternatives to conventional and percutaneous mitral repair.


Micronutrient treatment for children with emotional and behavioral dysregulation: a case series.
In clinical studies of adults and children, broad-spectrum micronutrients (minerals and vitamins) have proven beneficial for improving mood regulation and attention. We report here pilot work whose primary objective was to evaluate the feasibility of studying micronutrient treatment in school-aged children with emotional and behavioral problems. Issues examined included feasibility of participant recruitment from a culturally diverse population, probability of sample retention for a 12-week trial, acceptability of the outcome measures, supplement adherence, as well as trends in treatment benefit. The families of two boys (ages 5 and 6) and one girl (age 14) were invited to participate in a 12-week pilot trial of micronutrients carried out during the summer months. All children were enrolled in the private school at which future research was being considered. During the previous school year, all three had been extremely difficult to educate due to their inability to pay attention and learn, as well as their behavior problems. Although the two younger children had not been formally diagnosed, parents and teachers provided reports of hyperactivity and inability to focus on education in the classroom. The oldest child was often aggressive, and had been diagnosed with bipolar disorder, attention deficit hyperactivity disorder, and oppositional defiant disorder. All three children were Hispanic and spoke both Spanish and English. For 12 weeks, after signing consent forms, the children's parents provided weekly ratings on the parent-report Child Mania Rating Scale; the children consumed the micronutrient formula daily and provided a daily rating of how they felt. The parent ratings revealed significantly improved behavior, p = .002. Children's ratings approached the ideal level of 7, indicating "happy" self-reports. Parent interviews confirmed the weekly scores. Several feasibility questions were answered: all three children completed the 12-week trial, all scores were completed by parents and children, adherence to the protocol was excellent, and no adverse reactions emerged. Family physicians and pediatricians are often confronted with the challenge of improving the lives of families whose children experience school crises due to emotional and behavioral dysregulation. Three children, who participated in pilot work to determine the feasibility of further investigations, experienced impressive changes that clearly warrant both research and clinical exploration.


Current strategies against COVID-19.
Coronavirus disease (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) recently was declared a pandemic by world health organization (WHO) Due to sudden outbreaks, currently, no completely effective vaccine or drug is clinically approved. Several therapeutic strategies can be envisaged to prevent further mortality and morbidity. Based on the past contribution of traditional Chinese medicines (TCM) and immune-based therapies as a treatment option in crucial pathogen outbreaks, we aimed to summarize potential therapeutic strategies that could be helpful to stop further spread of SARS-CoV-2 by effecting its structural components or modulation of immune responses. Several TCM with or without modification could be effective against the structural protein, enzymes, and nucleic acid should be tested from available libraries or to identify their immune-stimulatory activities to enhance several antiviral biological agents for effective elimination of SARS-CoV-2 from the host. TCM is not only effective in the direct inhibition of virus attachment and internalization in a cell but can also prevent their replication and can also help to boost up host immune response. Immune-modulatory effects of TCMs may lead to new medications and can guide us for the scientific validity of drug development. Besides, we also summarized the effective therapies in clinical for controlling inflammation. This review will be not only helpful for the current situation of COVID-19, but can also play a major role in such epidemics in the future.


PrPC is sorted to the basolateral membrane of epithelial cells independently of its association with rafts.
PrP(C) is a glycosylphosphatidylinositol-anchored protein expressed in neurons as well as in the cells of several peripheral tissues. Although the normal function of PrP(C) remains unknown, a conformational isoform called PrP(Sc) (scrapie) has been proposed to be the infectious agent of transmissible spongiform encephalopathies in animals and humans. Where and how the PrP(C) to PrP(Sc) conversion occurs in the cells is not yet known. Therefore, dissecting the intracellular trafficking of the wild-type prion protein, as well as of the scrapie isoform, can be of major relevance to the pathogenesis of the diseases. In this report we have analyzed the exocytic pathway of transfected mouse PrP(C) in thyroid and kidney polarized epithelial cells. In contrast to the majority of glycosylphosphatidylinositol-anchored proteins, we found that PrP(C) is localized mainly on the basolateral domain of the plasma membrane of both cell lines. This is reminiscent of the predominant somatodendritic localization found in neurons. However, similarly to apical glycosylphosphatidylinositol-proteins, PrP(C) associates with detergent-resistant microdomains, which have been suggested to have a role in apical sorting of glycosylphosphatidylinositol-proteins, as well as in the conversion process of PrP(C) to PrP(Sc). In order to discriminate whether detergent-resistant microdomains have a direct role in PrP(Sc) conversion, or whether they are involved in the transport of the protein to the site of its conversion, we have examined the effect of disruption of detergent-resistant microdomain association on PrP(C) intracellular traffic. Consistent with the unusual basolateral localization of this glycosylphosphatidylinositol-linked protein, our data exclude a classical role for detergent-resistant microdomains in the post-trans-Golgi network sorting and transport of PrP(C) to the plasma membrane.


Micro-biomechanics of the Kebara 2 hyoid and its implications for speech in Neanderthals.
The description of a Neanderthal hyoid from Kebara Cave (Israel) in 1989 fuelled scientific debate on the evolution of speech and complex language. Gross anatomy of the Kebara 2 hyoid differs little from that of modern humans. However, whether Homo neanderthalensis could use speech or complex language remains controversial. Similarity in overall shape does not necessarily demonstrate that the Kebara 2 hyoid was used in the same way as that of Homo sapiens. The mechanical performance of whole bones is partly controlled by internal trabecular geometries, regulated by bone-remodelling in response to the forces applied. Here we show that the Neanderthal and modern human hyoids also present very similar internal architectures and micro-biomechanical behaviours. Our study incorporates detailed analysis of histology, meticulous reconstruction of musculature, and computational biomechanical analysis with models incorporating internal micro-geometry. Because internal architecture reflects the loadings to which a bone is routinely subjected, our findings are consistent with a capacity for speech in the Neanderthals.


p53-dependent apoptosis suppresses radiation-induced teratogenesis.
About half of human conceptions are estimated not to be implanted in the uterus, resulting in unrecognizable spontaneous abortions, and about 5% of human births have a recognizable malformation. In order to find clues to the mechanisms of malformation and abortion, we compared the incidences of radiation-induced malformations and abortions in p53 null (p53-/-) and wild-type (p53+/+) mice. After X-irradiation with 2 Gy on day 9.5 of gestation, p53-/- mice showed a 70% incidence of anomalies and a 7% incidence of deaths, whereas p53+/+ mice had a 20% incidence of anomalies and a 60% incidence of deaths. Similar results were obtained after irradiation on day 3.5 of gestation. This reciprocal relationship of radiosensitivity to anomalies and to embryonic or fetal lethality supports the notion that embryonic or fetal tissues have a p53-dependent "guardian" of the tissue that aborts cells bearing radiation-induced teratogenic DNA damage. In fact, after X-irradiation, the number of cells with apoptotic DNA fragments was greatly increased in tissues of the p53+/+ fetuses but not in those of the p53-/- fetuses.


Dental Pulp Defence and Repair Mechanisms in Dental Caries.
Dental caries is a chronic infectious disease resulting from the penetration of oral bacteria into the enamel and dentin. Microorganisms subsequently trigger inflammatory responses in the dental pulp. These events can lead to pulp healing if the infection is not too severe following the removal of diseased enamel and dentin tissues and clinical restoration of the tooth. However, chronic inflammation often persists in the pulp despite treatment, inducing permanent loss of normal tissue and reducing innate repair capacities. For complete tooth healing the formation of a reactionary/reparative dentin barrier to distance and protect the pulp from infectious agents and restorative materials is required. Clinical and in vitro experimental data clearly indicate that dentin barrier formation only occurs when pulp inflammation and infection are minimised, thus enabling reestablishment of tissue homeostasis and health. Therefore, promoting the resolution of pulp inflammation may provide a valuable therapeutic opportunity to ensure the sustainability of dental treatments. This paper focusses on key cellular and molecular mechanisms involved in pulp responses to bacteria and in the pulpal transition between caries-induced inflammation and dentinogenic-based repair. We report, using selected examples, different strategies potentially used by odontoblasts and specialized immune cells to combat dentin-invading bacteria in vivo.


FGF/EGF signaling regulates the renewal of early nephron progenitors during embryonic development.
Recent studies indicate that nephron progenitor cells of the embryonic kidney are arranged in a series of compartments of an increasing state of differentiation. The earliest progenitor compartment, distinguished by expression of CITED1, possesses greater capacity for renewal and differentiation than later compartments. Signaling events governing progression of nephron progenitor cells through stages of increasing differentiation are poorly understood, and their elucidation will provide key insights into normal and dysregulated nephrogenesis, as well as into regenerative processes that follow kidney injury. In this study, we found that the mouse CITED1(+) progenitor compartment is maintained in response to receptor tyrosine kinase (RTK) ligands that activate both FGF and EGF receptors. This RTK signaling function is dependent on RAS and PI3K signaling but not ERK. In vivo, RAS inactivation by expression of sprouty 1 (Spry1) in CITED1(+) nephron progenitors results in loss of characteristic molecular marker expression and in increased death of progenitor cells. Lineage tracing shows that surviving Spry1-expressing progenitor cells are impaired in their subsequent epithelial differentiation, infrequently contributing to epithelial structures. These findings demonstrate that the survival and developmental potential of cells in the earliest embryonic nephron progenitor cell compartment are dependent on FGF/EGF signaling through RAS.


High dietary diversity is associated with obesity in Sri Lankan adults: an evaluation of three dietary scores.
Dietary diversity is recognized as a key element of a high quality diet. However, diets that offer a greater variety of energy-dense foods could increase food intake and body weight. The aim of this study was to explore association of diet diversity with obesity in Sri Lankan adults. Six hundred adults aged > 18 years were randomly selected by using multi-stage stratified sample. Dietary intake assessment was undertaken by a 24 hour dietary recall. Three dietary scores, Dietary Diversity Score (DDS), Dietary Diversity Score with Portions (DDSP) and Food Variety Score (FVS) were calculated. Body mass index (BMI) ≥ 25 kg.m⁻² is defined as obese and Asian waist circumference cut-offs were used diagnosed abdominal obesity. Mean of DDS for men and women were 6.23 and 6.50 (p=0.06), while DDSP was 3.26 and 3.17 respectively (p=0.24). FVS values were significantly different between men and women 9.55 and 10.24 (p=0.002). Dietary diversity among Sri Lankan adults was significantly associated with gender, residency, ethnicity, education level but not with diabetes status. As dietary scores increased, the percentage consumption was increased in most of food groups except starches. Obese and abdominal obese adults had the highest DDS compared to non obese groups (p<0.05). With increased dietary diversity the level of BMI, waist circumference and energy consumption was significantly increased in this population. Our data suggests that dietary diversity is positively associated with several socio-demographic characteristics and obesity among Sri Lankan adults. Although high dietary diversity is widely recommended, public health messages should emphasize to improve dietary diversity in selective food items.


IFN Regulatory Factor 3 Balances Th1 and T Follicular Helper Immunity during Nonlethal Blood-Stage Plasmodium Infection.
Differentiation of CD4+ Th cells is critical for immunity to malaria. Several innate immune signaling pathways have been implicated in the detection of blood-stage Plasmodium parasites, yet their influence over Th cell immunity remains unclear. In this study, we used Plasmodium-reactive TCR transgenic CD4+ T cells, termed PbTII cells, during nonlethal P. chabaudi chabaudi AS and P. yoelii 17XNL infection in mice, to examine Th cell development in vivo. We found no role for caspase1/11, stimulator of IFN genes, or mitochondrial antiviral-signaling protein, and only modest roles for MyD88 and TRIF-dependent signaling in controlling PbTII cell expansion. In contrast, IFN regulatory factor 3 (IRF3) was important for supporting PbTII expansion, promoting Th1 over T follicular helper (Tfh) differentiation, and controlling parasites during the first week of infection. IRF3 was not required for early priming by conventional dendritic cells, but was essential for promoting CXCL9 and MHC class II expression by inflammatory monocytes that supported PbTII responses in the spleen. Thereafter, IRF3-deficiency boosted Tfh responses, germinal center B cell and memory B cell development, parasite-specific Ab production, and resolution of infection. We also noted a B cell-intrinsic role for IRF3 in regulating humoral immune responses. Thus, we revealed roles for IRF3 in balancing Th1- and Tfh-dependent immunity during nonlethal infection with blood-stage Plasmodium parasites.


Impact-absorbing properties of the human knee.
A biomechanical study has been carried out on 20 cadaveric knees to investigate their load-absorbing mechanism. The impact load was applied using a weight falling onto the transected proximal femur and the force transmitted through the knee was measured at the transected distal tibia using a load transducer. The peak force transmitted increased as, sequentially, meniscus, articular cartilage and subchondral bone were damaged or removed. The most striking result was found in an implanted knee replacement where the transmitted force reached 180% of that in the intact knee. The results show that the joint has an impact-absorbing property in each segment and that in the osteoarthritic knee there is less absorption of shock than in the normal knee. The high impact force in an implanted knee suggests that microfractures of the cancellous bone might be expected and may produce loosening.


Contribution of NAADP to Glutamate-Evoked Changes in Ca2+ Homeostasis in Mouse Hippocampal Neurons.
Nicotinic acid adenine dinucleotide phosphate (NAADP) is a second messenger that evokes calcium release from intracellular organelles by the engagement of calcium release channels, including members of the Transient Receptor Potential (TRP) family, such as TRPML1, the (structurally) related Two Pore Channel type 1 (TPC1) and TPC2 channels as well as Ryanodine Receptors type 1 (RYR1; Guse, 2012). NAADP evokes calcium release from acidic calcium stores of many cell types (Guse, 2012), and NAADP-sensitive Ca2+ stores have been described in hippocampal neurons of the rat (Bak et al., 1999; McGuinness et al., 2007). Glutamate triggers Ca2+-mediated neuronal excitotoxicity in inflammation-induced neurodegenerative pathologies such as Multiple Sclerosis (MS; Friese et al., 2014), and when applied extracellularly to neurons glutamate can elevate NAADP levels in these cells. Accordingly, glutamate-evoked Ca2+ signals from intracellular organelles were inhibited by preventing organelle acidification (Pandey et al., 2009). Analysis of reported RNA sequencing experiments of cultured hippocampal neurons revealed the abundance of Mcoln1 (encoding TRPML1), Tpcn1, and Tpcn2 (encoding TPC1 and TPC2, respectively) as potential NAADP target channels in these cells. Transcripts encoding Ryr1 were not found in contrast to Ryr2 and Ryr3. To study the contribution of NAADP signaling to glutamate-evoked calcium transients in murine hippocampal neurons we used the NAADP antagonists Ned-19 (Naylor et al., 2009) and BZ194 (Dammermann et al., 2009). Our results show that both NAADP antagonists significantly reduce glutamate-evoked calcium transients. In addition to extracellular glutamate application, we studied synchronized calcium oscillations in the cells of the neuronal cultures evoked by addition of the GABAA receptor antagonist bicuculline. Pretreatment with Ned-19 (50 μM) or BZ194 (100 μM) led to an increase in the frequency of bicuculline-induced calcium oscillations at the cost of calcium transient amplitudes. Interestingly, Ned-19 triggered a rise in intracellular calcium concentrations 25 min after bicuculline stimulation, leading to the question whether NAADP acts as a neuroprotective messenger in hippocampal neurons. Taken together, our results are in agreement with the concept that NAADP signaling significantly contributes to glutamate evoked Ca2+ rise in hippocampal neurons and to the amplitude and frequency of synchronized Ca2+ oscillations triggered by spontaneous glutamate release events.


Sudden infant death syndrome among twins.
Sudden infant death syndrome (SIDS) is a major contributor to infant mortality. Previous studies have suggested that infants born of twin pregnancies are at greater risk for SIDS and that a twin who survives after a co-twin dies is at increased risk for SIDS. To attempt to confirm the increased risk of SIDS among and within twin pairs through the use of US vital statistics data. We analyzed data from the US-linked birth and infant death certificate tapes for the years 1987 through 1991 to determine the risk of SIDS in twin births compared with singleton births and to describe the characteristics of twin pairs in whom SIDS occurred. The analysis was limited to live births with weights of 500 g or more and gestational ages of 24 weeks or more. We used an algorithm to match co-twins (infants within a twin pair) to measure sex and birth weight concordancy, to identify twin pairs, in which one or both twins died of SIDS; and to examine, when both twins died, whether they died on the same day. There were 23464 singleton SIDS deaths and 1056 twin SIDS deaths during the 5-year period. The crude relative risk for SIDS among twins compared with singleton births was 2.06 (95% confidence interval, 1.94-2.19). The adjusted relative risk independent of birth weight and sociodemographic variables was 1.13 (95% confidence interval, 0.97-1.31). We successfully matched the co-twins of 172029 twin pregnancies. Of these, 767 were twin pregnancies in which one or both twins died of SIDS. Among the 767 twin pregnancies in which one or both twins experienced SIDS, there were only 7 in which both twins died of SIDS (rate ratio, 4.0 per 100000 twin pregnancies). In only 1 of these 7 did both twins die on the same day (rate ratio, 0.58 per 100000 twin pregnancies). The relative risk for a second twin dying of SIDS was 8.17 (90% confidence interval, 1.18-56.67). Independent of birth weight, twins do not appear to be at greater risk for SIDS compared with singleton births. In addition, the occurrence of both twins dying of SIDS is uncommon, and the occurrence of both twins dying on the same day is extremely uncommon.


Neurodegeneration by activation of the microglial complement-phagosome pathway.
Systemic inflammatory reactions have been postulated to exacerbate neurodegenerative diseases via microglial activation. We now demonstrate in vivo that repeated systemic challenge of mice over four consecutive days with bacterial LPS maintained an elevated microglial inflammatory phenotype and induced loss of dopaminergic neurons in the substantia nigra. The same total cumulative LPS dose given within a single application did not induce neurodegeneration. Whole-genome transcriptome analysis of the brain demonstrated that repeated systemic LPS application induced an activation pattern involving the classical complement system and its associated phagosome pathway. Loss of dopaminergic neurons induced by repeated systemic LPS application was rescued in complement C3-deficient mice, confirming the involvement of the complement system in neurodegeneration. Our data demonstrate that a phagosomal inflammatory response of microglia is leading to complement-mediated loss of dopaminergic neurons.


Molecular Genetics of Emericella nidulans Sexual Development.
Many aspergilli that belongs to ascomycetes have sexuality. In a homothallic or self-fertile fungus, a number of fruiting bodies or cleistothecia are formed in a thallus grown from a single haploid conidia or ascospores. Genome-sequencing project revealed that two mating genes (MAT) encoding the regulatory proteins that are necessary for controlling partner recognition in heterothallic fungi were conserved in most aspergilli. The MAT gene products in some self-fertile species were not required for recognition of mating partner at pheromone-signaling stage but required at later stages of sexual development. Various environmental factors such as nutritional status, culture conditions and several stresses, influence the decision or progression of sexual reproduction. A large number of genes are expected to be involved in sexual development of Emericella nidulans (anamorph: Aspergillus nidulans), a genetic and biological model organism in aspergilli. The sexual development process can be grouped into several development stages, including the decision of sexual reproductive cycle, mating process, growth of fruiting body, karyogamy followed by meiosis, and sporulation process. Complicated regulatory networks, such as signal transduction pathways and gene expression controls, may work in each stage and stage-to-stage linkages. In this review, the components joining in the regulatory pathways of sexual development, although they constitute only a small part of the whole regulatory networks, are briefly mentioned. Some of them control sexual development positively and some do negatively. Regarding the difficulties for studying sexual differentiation compare to asexual one, recent progresses in molecular genetics of E. nidulans enlarge the boundaries of understanding sexual development in the non-fertile species as well as in fertile fungi.


Conformational dynamics of the nucleotide binding domains and the power stroke of a heterodimeric ABC transporter.
Multidrug ATP binding cassette (ABC) exporters are ubiquitous ABC transporters that extrude cytotoxic molecules across cell membranes. Despite recent progress in structure determination of these transporters, the conformational motion that transduces the energy of ATP hydrolysis to the work of substrate translocation remains undefined. Here, we have investigated the conformational cycle of BmrCD, a representative of the heterodimer family of ABC exporters that have an intrinsically impaired nucleotide binding site. We measured distances between pairs of spin labels monitoring the movement of the nucleotide binding (NBD) and transmembrane domains (TMD). The results expose previously unobserved structural intermediates of the NBDs arising from asymmetric configuration of catalytically inequivalent nucleotide binding sites. The two-state transition of the TMD, from an inward- to an outward-facing conformation, is driven exclusively by ATP hydrolysis. These findings provide direct evidence of divergence in the mechanism of ABC exporters.DOI: http://dx.doi.org/10.7554/eLife.02740.001.


An MRI-based parcellation method for the temporal lobe.
The temporal lobe has long been a focus of attention with regard to the underlying pathology of several major psychiatric illnesses. Previous postmortem and imaging studies describing regional volume reductions or perfusion defects in temporal subregions have shown inconsistent findings, which are in part due to differences in the definition of the subregions and the methodology of measurement. The development of precise reproducible parcellation systems on magnetic resonance images may help improve uniformity of results in volumetric MR studies and unravel the complex activation patterns seen in functional neuroimaging studies. The present study describes detailed guidelines for the parcellation of the temporal neocortex. It parcels the entire temporal neocortex into 16 subregions: temporal pole, heschl's gyrus, planum temporale, planum polare, superior temporal gyrus (rostral and caudal), middle temporal gyrus (rostral, intermediate, and caudal), inferior temporal gyrus (rostral, intermediate, and caudal), occipitotemporal gyrus (rostral and caudal), and parahippocampal gyrus (rostral and caudal). Based upon topographic landmarks of individual sulci, every subregion was consecutively traced on a set of serial coronal slices. In spite of the huge variability of sulcal topography, the sulcal landmarks could be identified reliably due to the simultaneous display of three orthogonal (transaxial, coronal, and sagittal) planes, triangulated gray matter isosurface, and a 3-D-rendered image. The reliability study showed that the temporal neocortex could be parceled successfully and reliably; intraclass correlation coefficient for each subregion ranged from 0.62 to 0.99. Ultimately, this method will permit us to detect subtle morphometric impairments or to find abnormal patterns of functional activation in the temporal subregions that might reflect underlying neuropathological processes in psychiatric illnesses such as schizophrenia.


Barriers to Eating Traditional Foods Vary by Age Group in Ecuador With Biodiversity Loss as a Key Issue.
To document the perceptions of indigenous peoples for the sustainable management of natural resources against malnutrition. Initially 4 and then 12 interviews were conducted with 4 different age groups. Eight rural villages in Guasaganda, central Ecuador, were studied in 2011-2012. A total of 75 people (22 children, 18 adolescents, 20 adults, and 15 elders). Benefits, severity, susceptibility, barriers, cues to action, and self-efficacy of eating traditional foods. Qualitative content analysis was completed using NVivo software. Initial analysis was inductive, followed by a content analysis directed by the Health Belief Model. Coding was completed independently by 2 researchers and kappa statistics (κ ≥ 0.65) were used to evaluate agreement. Healthy perceptions toward traditional foods existed and differed by age. Local young people ate traditional foods for their health benefits and good taste; adults cultivated traditional foods that had an economic benefit. Traditional knowledge used for consumption and cultivation of traditional foods was present but needs to be disseminated. Nutrition education in schools is needed that supports traditional knowledge in younger groups and prevents dietary changes toward unhealthy eating. Increased production of traditional food is needed to address current economic realities.


Direction-specific motion blindness induced by focal stimulation of human extrastriate cortex.
Motion blindness (MB) or akinetopsia is the selective disturbance of visual motion perception while other features of the visual scene such as colour and shape are normally perceived. Chronic and transient forms of MB are characterized by a global deficit of direction discrimination (pandirectional), which is generally assumed to result from damage to, or interference with, the motion complex MT+/V5. However, the most characteristic feature of primate MT-neurons is not their motion specificity, but their preference for one direction of motion (direction specificity). Here, we report that focal electrical stimulation in the human posterior temporal lobe selectively impaired the perception of motion in one direction while the perception of motion in other directions was completely normal (unidirectional MB). In addition, the direction of MB was found to depend on the brain area stimulated. It is argued that direction specificity for visual motion is not only represented at the single neuron level, but also in much larger cortical units.


The search for organizing principles as a cure against reductionism in systems medicine.
Biological complexity has forced scientists to develop highly reductive approaches, with an ever-increasing degree of specialization. As a consequence, research projects have become fragmented, and their results strongly dependent on the experimental context. The general research question, that originally motivated these projects, appears to have been forgotten in many highly specialized research programmes. We here investigate the prospects for use of an old regulative ideal from systems theory to describe the organization of cellular systems 'in general' by identifying key concepts, challenges and strategies to pursue the search for organizing principles. We argue that there is no tension between the complexity of biological systems and the search for organizing principles. On the contrary, it is the complexity of organisms and the current level of techniques and knowledge that urge us to renew the search for organizing principles in order to meet the challenges that are arise from reductive approaches in systems medicine. Reductive approaches, as important and inevitable as they are, should be complemented by an integrative strategy that de-contextualizes through abstractions, and thereby generalizes results.


The applicability of the international classification of functioning, disability, and health to study lifestyle and quality of life of colorectal cancer survivors.
Well-designed studies on lifestyle and health-related quality of life (HRQoL) in colorectal cancer survivors based on a biopsychosocial instead of a traditional biomedical approach are warranted. We report on the applicability of the International Classification of Functioning, Disability, and Health (ICF) as useful biopsychosocial framework to improve research on how lifestyle influences colorectal cancer survivors' HRQoL, using the Energy for life after ColoRectal cancer (EnCoRe) study as an example. The ICF was used to develop a conceptual model for studying lifestyle and colorectal cancer survivors' HRQoL, by identifying relevant factors from literature and mapping them within the ICF. Subsequently, this model was used for selection of measurement instruments and biomarkers. By linking meaningful concepts within selected measures to the ICF, we could assess the ICF coverage of our developed conceptual model. Within selected measures, 450 meaningful concepts were identified, of which 88% were linked to the ICF. The linking process resulted in 132 distinctive ICF categories assigned (38% within "Body Functions," 2% within "Body Structures," 46% within "Activities and Participation," and 14% within "Environmental Factors"). The selected EnCoRe study measures broadly cover ICF domains relevant to colorectal cancer survivors, stressing the relevance of using a biopsychosocial approach for studying this population's HRQoL. The developed conceptual model will guide data analyses and interpretation, and facilitate early transfer of results for development, evaluation, and implementation of personalized multidisciplinary lifestyle interventions. We recommend the ICF as an invaluable framework for improving the quality and scope of HRQoL studies.


Identification of a rat glomerular mesangial cell mitogenic 5-HT2A receptor.
Previous studies have demonstrated the presence of mitogenic serotonin [i.e., 5-hydroxytryptamine (5-HT)] receptors on glomerular mesangial cells and have linked those receptors to a complicated array of intracellular and autocrine/paracrine signaling pathways [T. Knauss and H. E. Abboud. Am. J. Physiol. 251 (Renal Fluid Electrolyte Physiol. 20): F844-F850, 1986; and N. Takuwa, M. Ganz, Y. Takuwa, R. B. Sterzel, and H. Rasmussen. Am. J. Physiol. 257 (Renal Fluid Electrolyte Physiol. 26): F431-F439, 1989]. Those studies suggested that the mesangial subtype of 5-HT receptor is a member of the 5-HT2 receptor family, which consists of three known members, designated as subtypes A, B, and C. The purpose of the current study was to identify the subtype of 5-HT2 receptor present on mesangial cells. Northern blot showed detectable mRNA for a putative 5-HT2A receptor, but not for 5-HT1A or 5-HT2C receptors. Reverse transcription-polymerase chain reaction (RT-PCR) with degenerate oligonucleotides derived from the putative third and sixth transmembrane domains of cloned 5-HT2 receptors yielded a 580-nucleotide (nt) fragment. RT-PCR with primers highly specific for the 5-HT2A receptor and designed to amplify > 95% of its coding block yielded a product of 1,320 nt. Nested PCR reactions yielded products of the predicted sizes for the 5-HT2A receptor. Partial sequence information was obtained, and the sequence corresponded exactly (627/627 nt) to that published for the cloned rat brain 5-HT2A receptor. These studies identify the mesangial cell mitogenic 5-HT receptor as a 5-HT2A receptor subtype.


Postoperative Outcomes in Ustekinumab-Treated Patients Undergoing Abdominal Operations for Crohn's Disease.
Ustekinumab, a monoclonal antibody targeting interleukins-12 and -23 is used to treat adults with Crohn's disease [CD]. We determined the 30-day postoperative infectious complication rate among CD patients who received ustekinumab within the 12 weeks prior to an abdominal operation as compared with patients who received anti-tumor necrosis factor [TNF] agents. A retrospective chart review of adults with CD who underwent an abdominal operation between January 1, 2015 and May 1, 2017 was performed across six sites. Surgical site infection [SSI] was defined as superficial skin and soft tissue infection, intra-abdominal abscess, anastomotic leak, and mucocutaneous separation of the stoma. Forty-four patients received ustekinumab and 169 patients received anti-TNF therapy within the 12 weeks prior to surgery. The two groups were similar, except anti-TNF patients were more likely to have received combination therapy with an immunomodulator [P = 0.006]. There were no significant differences in postoperative SSI [13% in ustekinumab versus 20% in anti TNF-treated patients, p = 0.61] or hospital readmission rates [18% versus 10%, respectively, p = 0.14], but ustekinumab-treated patients had a higher rate of return to the operating room [16% versus 5%; P = 0.01]. There were no significant predictors identified on multivariable analysis. Of the 44 patients with CD who received ustekinumab within the 12 weeks prior to a major abdominal operation, 13% experienced a 30-day postoperative SSI, not statistically different from the 20% found in the anti-TNF cohort. Ustekinumab treatment within 12 weeks of surgery does not appear to increase the risk of postoperative SSI above that of CD patients treated with anti-TNF medications.


Lack of C9ORF72 coding mutations supports a gain of function for repeat expansions in amyotrophic lateral sclerosis.
Hexanucleotide repeat expansions in C9ORF72 are a common cause of familial and apparently sporadic amyotrophic lateral sclerosis (ALS) and frontal temporal dementia (FTD). The mechanism by which expansions cause neurodegeneration is unknown, but current evidence supports both loss-of-function and gain-of-function mechanisms. We used pooled next-generation sequencing of the C9ORF72 gene in 389 ALS patients to look for traditional loss-of-function mutations. Although rare variants were identified, none were likely to be pathogenic, suggesting that mutations other than the repeat expansion are not a common cause of ALS, and providing supportive evidence for a gain-of-function mechanism. We also show by repeat-primed PCR genotyping that the C9ORF72 expansion frequency varies by geographical region within the United States, with an unexpectedly high frequency in the Mid-West. Finally we also show evidence of somatic instability of the expansion size by Southern blot, with the largest expansions occurring in brain tissue.


Earliest land plants created modern levels of atmospheric oxygen.
The progressive oxygenation of the Earth's atmosphere was pivotal to the evolution of life, but the puzzle of when and how atmospheric oxygen (O2) first approached modern levels (∼21%) remains unresolved. Redox proxy data indicate the deep oceans were oxygenated during 435-392 Ma, and the appearance of fossil charcoal indicates O2 >15-17% by 420-400 Ma. However, existing models have failed to predict oxygenation at this time. Here we show that the earliest plants, which colonized the land surface from ∼470 Ma onward, were responsible for this mid-Paleozoic oxygenation event, through greatly increasing global organic carbon burial-the net long-term source of O2 We use a trait-based ecophysiological model to predict that cryptogamic vegetation cover could have achieved ∼30% of today's global terrestrial net primary productivity by ∼445 Ma. Data from modern bryophytes suggests this plentiful early plant material had a much higher molar C:P ratio (∼2,000) than marine biomass (∼100), such that a given weathering flux of phosphorus could support more organic carbon burial. Furthermore, recent experiments suggest that early plants selectively increased the flux of phosphorus (relative to alkalinity) weathered from rocks. Combining these effects in a model of long-term biogeochemical cycling, we reproduce a sustained +2‰ increase in the carbonate carbon isotope (δ(13)C) record by ∼445 Ma, and predict a corresponding rise in O2 to present levels by 420-400 Ma, consistent with geochemical data. This oxygen rise represents a permanent shift in regulatory regime to one where fire-mediated negative feedbacks stabilize high O2 levels.


The Eye Book of Master Peter of Spain--a glimpse of diagnosis and treatment of eye disease in the Middle Ages.
Peter Hispanus, who became Pope John XXI, wrote a book about eye diseases and their treatment in the 13th century. De Oculis was not a scholarly treatise about eye diseases, but a manual intended for general physicians. Nevertheless, Latin copies were made into the 16th century, indicating its influence and importance. We present the first published English translation of De Oculis, based on a synthesis of the medieval Latin manuscripts and a German translation from the 19th century. The sources of Peter's understanding of the causes and treatment of eye diseases were the more scholarly and complete treatises by Greek and Arab writers. Many of the diseases described then cannot be correlated precisely with ophthalmic disorders known to us today because of the limited understanding of anatomy, function and pathophysiology that existed. However, De Oculis provides us with a glimpse of the practice of medicine in the Middle Ages.


The MRC/BHF Heart Protection Study: preliminary results.
The Heart Protection Study (HPS), with over 20,500 subjects, is the largest trial of statin therapy ever conducted. It provides important and definitive new information on women, the elderly, diabetics, and people with low baseline cholesterol pre-treatment and those with prior occlusive non-coronary vascular disease. It is a prospective double blind randomised controlled trial with a 2 x 2 factorial design investigating prolonged use (>5 years) of simvastatin 40 mg and a cocktail of antioxidant vitamins (650 mg vitamin E, 250 mg vitamin C and 20 mg beta-carotene). The HPS specifically included patients with high risk for coronary heart disease (CHD) but characteristics that excluded them from participation in previous statin trials. Simvastatin 40 mg treatment showed benefit across all patient groups regardless of age, gender or baseline cholesterol value and proved safe and well tolerated. Results show a 12% reduction in total mortality, a 17% reduction in vascular mortality, a 24% reduction in CHD events, a 27% reduction in all strokes and a 16% reduction in non-coronary revascularisations. Among high-risk patients in this western population (with a minimum total cholesterol [TC] > or = 3.5 mmol/l at entry) there appears to be no threshold cholesterol value below which statin therapy is not associated with benefit; even among those with pre-treatment cholesterol levels below current national recommended targets. Over the 5.5 year study period patients and their doctors were encouraged to add an active non-study statin to the study regimen if they wished to do so. Thus the trial eventually had only two-thirds complying with the original intention-to-treat design. Nevertheless, results were highly significant for the study statin--simvastatin 40 mg once daily. Preliminary results of the HPS are negative for the antioxidant vitamin cocktail but provide reassurance that vitamins do no harm.


Targeting mitochondrial biogenesis to overcome drug resistance to MAPK inhibitors.
Targeting multiple components of the MAPK pathway can prolong the survival of patients with BRAFV600E melanoma. This approach is not curative, as some BRAF-mutated melanoma cells are intrinsically resistant to MAPK inhibitors (MAPKi). At the systemic level, our knowledge of how signaling pathways underlie drug resistance needs to be further expanded. Here, we have shown that intrinsically resistant BRAF-mutated melanoma cells with a low basal level of mitochondrial biogenesis depend on this process to survive MAPKi. Intrinsically resistant cells exploited an integrated stress response, exhibited an increase in mitochondrial DNA content, and required oxidative phosphorylation to meet their bioenergetic needs. We determined that intrinsically resistant cells rely on the genes encoding TFAM, which controls mitochondrial genome replication and transcription, and TRAP1, which regulates mitochondrial protein folding. Therefore, we targeted mitochondrial biogenesis with a mitochondrium-targeted, small-molecule HSP90 inhibitor (Gamitrinib), which eradicated intrinsically resistant cells and augmented the efficacy of MAPKi by inducing mitochondrial dysfunction and inhibiting tumor bioenergetics. A subset of tumor biopsies from patients with disease progression despite MAPKi treatment showed increased mitochondrial biogenesis and tumor bioenergetics. A subset of acquired drug-resistant melanoma cell lines was sensitive to Gamitrinib. Our study establishes mitochondrial biogenesis, coupled with aberrant tumor bioenergetics, as a potential therapy escape mechanism and paves the way for a rationale-based combinatorial strategy to improve the efficacy of MAPKi.


Ecological performance of construction materials subject to ocean climate change.
Artificial structures will be increasingly utilized to protect coastal infrastructure from sea-level rise and storms associated with climate change. Although it is well documented that the materials comprising artificial structures influence the composition of organisms that use them as habitat, little is known about how these materials may chemically react with changing seawater conditions, and what effects this will have on associated biota. We investigated the effects of ocean warming, acidification, and type of coastal infrastructure material on algal turfs. Seawater acidification resulted in greater covers of turf, though this effect was counteracted by elevated temperatures. Concrete supported a greater cover of turf than granite or high-density polyethylene (HDPE) under all temperature and pH treatments, with the greatest covers occurring under simulated ocean acidification. Furthermore, photosynthetic efficiency under acidification was greater on concrete substratum compared to all other materials and treatment combinations. These results demonstrate the capacity to maximise ecological benefits whilst still meeting local management objectives when engineering coastal defense structures by selecting materials that are appropriate in an ocean change context. Therefore, mitigation efforts to offset impacts from sea-level rise and storms can also be engineered to alter, or even reduce, the effects of climatic change on biological assemblages.


Environmental stimulation influence the cognition of developing mice by inducing changes in oxidative and apoptosis status.
Environment condition has been shown to play an important role in brain development. The present study examined the effects of enriched and impoverished environment on both spatial and emotional learning and memory of young mice and explored the underlying mechanisms. 3-week-old mice were housed in enriched environment (n=10, 10 mice in a large cage with toys and a running wheel), or standard environment (n=10, 10 mice in a large cage without objects), or impoverished environment (n=10, single mice in a small cage without objects) for 6weeks. Then, the spatial and emotional cognition of mice were evaluated by the water maze and step-down inhibitory avoidance test, respectively. To explore the underlying mechanisms, oxidation measurement in hippocampus and medial-temporal lobe cortex (MTLC) and apoptosis examination in hippocampus were performed. Results showed that compared with standard environment group, enriched and impoverished mice exhibited high and low performance levels in behavior tests, respectively. The oxidative status of hippocampus and MTLC were decreased in enriched group but increased in impoverished group. Moreover, changes in apoptosis of hippocampus in these two groups showed the same tendency with oxidative status. These results suggest that environment condition can simultaneously influence spatial and emotional learning and memory, which may result from inducing changes in the oxidative and apoptosis status in associated brain regions. Here, we firstly report using young mice to examine the oxidative status as a primary and direct factor to explore the mechanism of effects of different environment on both spatial and emotional cognition.


Single-molecule force-clamp experiments reveal kinetics of mechanically activated silyl ester hydrolysis.
We have investigated the strength of silyl ester bonds formed between carboxymethylated amylose (CMA) molecules and silane-functionalized silicon oxide surfaces using AFM-based single-molecule force spectroscopy in the force-clamp mode. Single tethered CMA molecules were picked up, and bond lifetimes were determined at constant clamp forces of 0.8, 1.0, and 1.2 nN at seven temperatures between 295 and 320 K at pH 2.0. The results reveal biexponential rupture kinetics. To obtain the reaction rate constants for each force and temperature individually, the results were analyzed with a biexponential kinetic model using the maximum likelihood estimation (MLE) method. The force-independent kinetic and structural parameters of the underlying bond rupture mechanisms were extracted by fitting the entire data set with a parallel MLE fit procedure using the Zhurkov/Bell model and, alternatively, an Arrhenius kinetics model combined with a Morse potential as an analytic representation of the binding potential. With activation energies between 37 and 40 kJ mol(-1), and with Arrhenius prefactors between 5 × 10(4) and 2 × 10(6) s(-1), the results point to the hydrolysis of the silyl ester bond.


Effects of ethanol on gastric epithelial cell phospholipid dynamics and cellular function.
The lipid profile of isolated gastric superficial epithelial cells (SEC) was evaluated by proton nuclear magnetic resonance spectroscopy (1H-NMR). The most conspicuous resonance band in SEC spectra was due to the protons of +N(CH3)3 groups of phosphatidylcholine and, to a lesser degree, other phospholipid derivatives, on the basis of their chemical shift and addition of purified phospholipids. NMR of cell lysates and phospholipid extracts of SEC in deutero-chloroform provided further spectral resolution of these components. Phospholipase or ethanol treatments of SEC produced membrane disorganization reflected as increased peak intensity of the phospholipid signals. In addition, ethanol, in a dose-dependent manner, attenuated paranitrophenyl phosphatase activity, which correlated with inhibition of total and ouabain-sensitive 86Rubidium chloride uptake by SEC. This study suggests that NMR used in conjunction with other biochemical techniques can monitor SEC membrane structure-function relationships. NMR is a potentially powerful noninvasive probe to show changes in lipid membrane organization induced by low concentrations of ethanol (1%) and may indicate an early sign of "cytotoxicity" in intact SEC.


The Cell Adhesion Molecule Necl-4/CADM4 Serves as a Novel Regulator for Contact Inhibition of Cell Movement and Proliferation.
Contact inhibition of cell movement and proliferation is critical for proper organogenesis and tissue remodeling. We show here a novel regulatory mechanism for this contact inhibition using cultured vascular endothelial cells. When the cells were confluently cultured, Necl-4 was up-regulated and localized at cell-cell contact sites where it cis-interacted with the vascular endothelial growth factor (VEGF) receptor. This interaction inhibited the tyrosine-phosphorylation of the VEGF receptor through protein-tyrosine phosphatase, non-receptor type 13 (PTPN13), eventually reducing cell movement and proliferation. When the cells were sparsely cultured, Necl-4 was down-regulated but accumulated at leading edges where it inhibited the activation of Rho-associated protein kinase through PTPN13, eventually facilitating the VEGF-induced activation of Rac1 and enhancing cell movement. Necl-4 further facilitated the activation of extracellular signal-regulated kinase 1/2, eventually enhancing cell proliferation. Thus, Necl-4 serves as a novel regulator for contact inhibition of cell movement and proliferation cooperatively with the VEGF receptor and PTPN13.


Conditional Allele Mouse Planner (CAMP): software to facilitate the planning and design of breeding strategies involving mice with conditional alleles.
Transgenic and conditional knockout mouse models play an important role in biomedical research and their use has grown exponentially in the last 5-10 years. Generating conditional knockouts often requires breeding multiple alleles onto the background of a single mouse or group of mice. Breeding these mice depends on parental genotype, litter size, transmission frequency, and the number of breeding rounds. Therefore, a well planned breeding strategy is critical for keeping costs to a minimum. However, designing a viable breeding strategy can be challenging. With so many different variables this would be an ideal task for a computer program. To facilitate this process, we created a Java-based program called Conditional Allele Mouse Planner (CAMP). CAMP is designed to provide an estimate of the number of breeders, amount of time, and costs associated with generating mice of a particular genotype. We provide a description of CAMP, how to use it, and offer it freely as an application.


Effect of coingestion of fat and protein with carbohydrate feedings on muscle glycogen storage.
Dietary guidelines for achieving optimal muscle glycogen storage after prolonged exercise have been given in terms of absolute carbohydrate (CHO) intake (8-10 g.kg-1.day-1). However, it is of further interest to determine whether the addition of fat and protein to carbohydrate feedings affects muscle glycogen storage. Eight well-trained triathletes [23.1 +/- 2.0 (SE) yr; 74.0 +/- 3.4 kg; peak O2 consumption = 4.7 +/- 0.4 l/min] undertook an exercise trial (2 h at 75% peak O2 consumption, followed by four 30-s sprints) on three occasions, each 1 wk apart. For 24 h after each trial, the subjects rested and were assigned to the following diets in randomized order: control (C) diet (high glycemic index CHO foods; CHO = 7 g.kg-1.day-1), added fat and protein (FP) diet (C diet + 1.6 g.kg-1.day-1 fat + 1.2 g.kg-1.day-1 protein), and matched-energy diet [C diet + 4.8 g.kg-1.day-1 additional CHO (Polycose) to match the additional energy in the FP diet]. Meals were eaten at t = 0, 4, 8, and 21 h of recovery. The total postprandial incremental plasma glucose area was significantly reduced after the FP diet (P < 0.05). Serum free fatty acid and plasma triglyceride responses were significantly elevated during the FP trial (P < 0.05). There were no differences between trials in muscle glycogen storage over 24 h (C, 85.8 +/- 2.7 mmol/kg wet wt; FP, 80.5 +/- 8.2 mmol/kg wet wt; matched-energy, 87.9 +/- 7.0 mmol/kg wet wt).(ABSTRACT TRUNCATED AT 250 WORDS)


Correlates of sexual risk for HIV infection in female members of heterosexual California Latino couples: an application of a Bernoulli process model.
Individual HIV risk estimates were generated from reported sexual behavior for 1,146 California Latino Couples Study participants (573 couples). These Bernoulli process model-based estimates proved strongly associated with individual sexually transmitted disease history. Mean estimated background risk from sexual contacts other than with their primary partner was substantially lower for the females than the males (1.4 vs. 7.4 per 10,000). After including their chance of infection from each other, mean net estimated risk was higher for the females than the males (9.2 vs. 8.6 per 10,000). Individual background risk was predicted by individual demographic and psychosocial characteristics (females: coefficient of concordance C = 0.84 predicting any (nonzero) risk; adjusted R(2) = 36% predicting level of risk, given any risk; males: C = 0.78; R(2) = 24%). Characteristics of women with higher risk primary partners were also identifiable (C = 0.65; R(2) = 13%). There was no significant negative association between the male partner's background sexual risk and the aggregate infectivity of the woman from him (taking into account the total number of their condom-protected and unprotected acts of different types).


Nuclear and cytoplasmic survivin: molecular mechanism, prognostic, and therapeutic potential.
Survivin's proposed dual role as an apoptosis inhibitor and a mitotic effector positioned it in the front line of cancer research. Notably, survivin is detected as a cytoplasmic and nuclear protein in cancer patients, which stimulated numerous studies to investigate and to speculate on the functional and prognostic significance of its dynamic localization. Recent evidence shows that the direct interaction of survivin with the nuclear export receptor Crm1 is critically involved in its intracellular localization and cancer-relevant functions. Here, we review our current understanding of the Crm1/survivin interface and discuss its potential prognostic and therapeutic relevance.


The Effect and Mechanisms of Proliferative Inhibition of Crocin on Human Leukaemia Jurkat Cells.
Targeted therapy is a potentially useful approach for the treatment of T-lineage acute lymphoblastic leukaemia. This study aimed to find a highly effective, low toxic anti-tumour drug and further investigate its mechanisms. Jurkat cells were used as the object and were stimulated by different concentrations of crocin. By cell count, growth curve, MTT method for the detection of cell proliferation, annexin V/propidium iodide (PI) method for the apoptosis rates, and reverse transcription-polymerase chain reaction (RT-PCR) for bcl-2 and bax gene expression, the effect and mechanisms of proliferative inhibition of crocin on Jurkat cells were further explored. Crocin promoted Jurkat cell apoptosis and inhibited cell growth, in a dose-time-dependent manner. The mechanism might be related to the inhibition of bcl-2 gene expression and the promotion of bax gene expression. These results suggest that crocin can be used as a suitable clinical agent for the treatment of T-lineage acute lymphoblastic leukaemia.


Identification of a Kavain Analog with Efficient Anti-inflammatory Effects.
Kavain, a compound derived from Piper methysticum, has demonstrated anti-inflammatory properties. To optimize its drug properties, identification and development of new kavain-derived compounds was undertaken. A focused library of analogs was synthesized and their effects on Porphyromonas gingivalis (P. gingivalis) elicited inflammation were evaluated in vitro and in vivo. The library contained cyclohexenones (5,5-dimethyl substituted cyclohexenones) substituted with a benzoate derivative at the 3-position of the cyclohexanone. The most promising analog identifed was a methylated derivative of kavain, Kava-205Me (5,5-dimethyl-3-oxocyclohex-1-en-1-yl 4-methylbenzoate.) In an in vitro assay of anti-inflammatory effects, murine macrophages (BMM) and THP-1 cells were infected with P. gingivalis (MOI = 20:1) and a panel of cytokines were measured. Both cell types treated with Kava-205Me (10 to 200 μg/ml) showed significantly and dose-dependently reduced TNF-α secretion induced by P. gingivalis. In BMM, Kava-205Me also reduced secretion of other cytokines involved in the early phase of inflammation, including IL-12, eotaxin, RANTES, IL-10 and interferon-γ (p < 0.05). In vivo, in an acute model of P. gingivalis-induced calvarial destruction, administration of Kava-205Me significantly improved the rate of healing associated with reduced soft tissue inflammation and osteoclast activation. In an infective arthritis murine model induced by injection of collagen-antibody (ArthriomAb) + P. gingivalis, administration of Kava-205Me was able to reduce efficiently paw swelling and joint destruction. These results highlight the strong anti-inflammatory properties of Kava-205Me and strengthen the interest of testing such compounds in the management of P. gingivalis elicited inflammation, especially in the management of periodontitis.