Organ-specific genetic interactions between paralogues of the PXY and ER receptor kinases enforce radial patterning in Arabidopsis vascular tissue.
In plants, cells do not migrate. Tissues are frequently arranged in concentric rings; thus, expansion of inner layers is coordinated with cell division and/or expansion of cells in outer layers. In Arabidopsis stems, receptor kinases, PXY and ER, genetically interact to coordinate vascular proliferation and organisation via inter-tissue signalling. The contribution of PXY and ER paralogues to stem patterning is not known, nor is their function understood in hypocotyls, which undergo considerable radial expansion. Here, we show that removal of all PXY and ER gene-family members results in profound cell division and organisation defects. In hypocotyls, these plants failed to transition to true radial growth. Gene expression analysis suggested that PXY and ER cross- and inter-family transcriptional regulation occurs, but it differs between stem and hypocotyl. Thus, PXY and ER signalling interact to coordinate development in a distinct manner in different organs. We anticipate that such specialised local regulatory relationships, where tissue growth is controlled via signals moving across tissue layers, may coordinate tissue layer expansion throughout the plant body.


Effect of Intracorporeal Human Adipose-Derived Stem Cells (hADSCs) on Corpora Cavernosa Transforming Growth Factor β1 (TGFβ1) and Collagen Type I Concentration in Wistar Rat Priapism Model.
The ischemic process in priapism can lead to displacement of normal tissue with fibrotic tissue, due to collagen deposition, and eventually leads to erectile dysfunction. Many studies have identified that the supernatant of adipose tissue-derived stem cells (ADSCs) significantly ameliorates fibrosis of different tissue, but limited attention has been paid to its efficacy on fibrosis of the corpora cavernosa. A total of 22 Wistar rats divided into five groups, with two groups each consisting of five male wistar rats with priapism without human ADSC (hADSC) therapy (group I) and two other groups consisting of five rats with priapism, were given 106 cells' intracorporeal hADSC injection after 12 hours of penile clamping (group II) were euthanized after 2 and 4 weeks of observation. The last group consisted of two rats without any treatment or model (group III). Following euthanasia, penises were harvested for TGFβ1 and collagen type I measurement using ELISA. Statistical analysis using independent-sample t-tests was done with SPSS 21.0. Penile TGFβ1 concentration in the treatment group was significantly lower in the second and fourth weeks of observation (p 2=0.004, p 4=0.003), and collagen type I was significantly lower in the second and fourth weeks (p 2=0.003, p 4=0.011). Intracorporeal hADSC injection limited the fibrosis process in a priapism model. Although the mechanism was unclear, it may be related to the potential of hADSCs to produce various growth factors that could limit TGFβ1 and collagen production.


Proteomic identification of urinary biomarkers of diabetic nephropathy.
Diabetic nephropathy is a serious complication of both type 1 and type 2 diabetes, and, unless arrested, leads to end-stage renal disease. Current diagnosis consists of urine assays of microalbuminuria, which have inadequate specificity and sensitivity. We used proteomic analyses to identify novel biomarkers of nephropathy in urine from type 2 diabetic patients with demonstrated normo-, micro-, or macroalbuminuria. Samples were analyzed by fluorescence two-dimensional (2-D) differential in-gel electrophoresis (DIGE), and protein identification was performed by liquid chromatography-tandem mass spectrometry. 2-D DIGE analysis of the urinary proteome in diabetes with nephropathy identified 195 protein spots representing 62 unique proteins. These proteins belonged to several functional groups, i.e., cell development, cell organization, defense response, metabolism, and signal transduction. Comparisons between control and diabetic subjects with different stages of renal dysfunction revealed the differential expression of several proteins. Spot volume quantification identified 7 proteins that were progressively upregulated with increasing albuminuria and 4 proteins that exhibited progressive downregulation. The majority of these potential candidate biomarkers were glycoproteins. These data demonstrate the ability of proteomic analyses to reveal potential biomarkers for diabetic nephropathy in urine, an important step forward in advancing accurate diagnosis and our understanding of disease mechanisms.


Developing a scaffold for urease inhibition based on benzothiazoles: Synthesis, docking analysis, and therapeutic potential.
The synthesis, in silico molecular docking, and in vitro urease inhibition studies of a novel series of benzothiazole derivatives are reported. The title compounds in the two series, namely, 2-({5-[(benzothiazol-2-ylthio)methyl]-1,3,4-oxadiazol-2-yl}thio)-1-(4-substituted-phenyl)ethan-1-one and 2-(benzothiazol-2-ylthio)-1-(4-substituted-phenyl)ethan-1-one oxime, were synthesized by the reaction of benzo[d]thiazole-2-thiol with different kinds of intermediates in several steps using both conventional and microwave techniques. All compounds were found to have an excellent degree of urease-inhibitory potential ranging between 16.16 ± 0.54 and 105.32 ± 2.10 µM when compared with the standard inhibitor acetohydroxamic acid with IC50 = 320.70 ± 4.24 µM. The structure-activity relationship was established in detail. The binding interactions of the compounds with the enzyme were confirmed through molecular docking. Further, 100 -ns molecular dynamics simulations were performed to investigate the stability and structural perturbations experienced by the most potent compound over the urease active site.


The Carcinogenic Agent Diethylnitrosamine Induces Early Oxidative Stress, Inflammation and Proliferation in Rat Liver, Stomach and Colon: Protective Effect of Ginger Extract.
Background: Diethylnitrosamine (DENA), a well-known dietary carcinogen, related to cancer initiation of various organs. The present study investigated the deleterious mechanisms involved in the early destructive changes of DENA in different organs namely, liver, stomach and colon and the potential protective effect of GE against these mechanisms. Methods: Adult male albino rats were assigned into four groups. A normal control group received the vehicle, another group was injected with a single necrogenic dose of DENA (200 mg/kg, i.p) on day 21. Two groups received oral GE (108 or 216 mg/kg) daily for 28 days. Sera, liver, stomach and colon were obtained 7 days after DENA injection. Serum aspartate transaminase and alanine transaminase were detected as well as reduced glutathione (GSH), malondialdehyde, nitric oxide metabolites, interleukin 1β, tumor necrosis factor (TNF-α), alpha-fetoprotein (AFP) and nuclear factorerythroid 2-related factor2 (Nrf2) in liver, stomach and colon. Histopathological studies and immunohistochemical examination of cyclooxygenase-2 (COX2) were conducted. Results: DENA induced elevation in liver function enzymes with significant increase in oxidation and inflammation biomarkers and AFP while decreased levels of Nrf2 in liver, stomach and colon were detected. Histologically, DENA showed degenerative changes in hepatocytes and inflammatory foci. Inflammatory foci displayed increased expression of COX2 in immunohistochemical staining. GE-pretreatment improved liver function and restored normal GSH with significant mitigation of oxidative stress and inflammatory biomarkers compared to DENA-treated group. AFP was reduced by GE in both doses, while Nrf2 increased significantly. Histology and immunostaining of hepatic COX-2 were remarkably improved in GE-treated groups in a dose dependent manner. Conclusion: GE exerted a potential anti-proliferative activity against DENA in liver, stomach and colon via Nrf2 activation, whilst suppression of oxidation and inflammation.


Benefits of Combining Molecular Biology and Controlled Human Infection Model Methodologies in Advancing Vaccine Development.
Infectious diseases continue to account for a significant portion of global deaths despite the use of vaccines for several centuries. Immunization programs around the world are a testament to the great success of multiple vaccines, yet there are still diseases without vaccines and others that require safer more effective ones. Addressing uncontrolled and emerging disease threats is restrained by the limitations and bottlenecks encountered with traditional vaccine development paradigms. Recent advances in modern molecular biology technologies have enhanced the interrogation of host pathogen interaction and deciphered complex pathways, thereby uncovering the myriad interplay of biological events that generate immune protection against foreign agents. Consequent to insights into the immune system, modern biology has been instrumental in the development and production of next generation 21st century vaccines. As these biological tools, commonly and collectively referred to as 'omics, became readily available, there has been a renewed consideration of Controlled Human Infection Models (CHIMs). Successful and reproducible CHIMs can complement modern molecular biology for the study of infectious diseases and development of effective vaccines in a regulated process that mitigates risk, cost, and time, with capacity to discern immune correlates of protection.


Electrochemical and Photophysical Properties of Ruthenium(II) Complexes Equipped with Sulfurated Bipyridine Ligands.
The development of new solar-to-fuel scenarios is of great importance, but the construction of molecular systems that convert sunlight into chemical energy represents a challenge. One specific issue is that the molecular systems have to be able to accumulate redox equivalents to mediate the photodriven transformation of relevant small molecules, which mostly involves the orchestrated transfer of multiple electrons and protons. Disulfide/dithiol interconversions are prominent 2e-/2H+ couples and can play an important role for redox control and charge storage. With this background in mind, a new photosensitizer [Ru(S-Sbpy)(bpy)2]2+ (12+) equipped with a disulfide functionalized bpy ligand (S-Sbpy, bpy = 2,2'-bipyridine) was synthesized and has been comprehensively studied, including structural characterization by X-ray diffraction. In-depth electrochemical studies show that the S-Sbpy ligand in 12+ can be reduced twice at moderate potentials (around -1.1 V vs Fc+/0), and simulation of the cyclic voltammetry (CV) traces revealed potential inversion (E2 > E1) and allowed to derive kinetic parameters for the sequential electron-transfer processes. However, reduction at room temperature also triggers the ejection of one sulfur atom from 12+, leading to the formation of [Ru(Sbpy)(bpy)2]2+(22+). This chemical reaction can be suppressed by decreasing the temperature from 298 to 248 K. Compared to the archetypical photosensitizer [Ru(bpy)3]2+, 12+ features an additional low energy optical excitation in the MLCT region, originating from charge transfer from the metal center to the S-Sbpy ligand (aka MSCT) according to time-dependent density functional theory (TD-DFT) calculations. Analysis of the excited states of 12+ on the basis of ground-state Wigner sampling and using charge-transfer descriptors has shown that bpy modification with a peripheral disulfide moiety leads to an energy splitting between charge-transfer excitations to the S-Sbpy and the bpy ligands, offering the possibility of selective charge transfer from the metal to either type of ligands. Compound 12+ is photostable and shows an emission from a 3MLCT state in deoxygenated acetonitrile with a lifetime of 109 ns. This work demonstrates a rationally designed system that enables future studies of photoinduced multielectron, multiproton PCET chemistry.


Psychological Disorders among Iranian Infertile Couples Undergoing Assisted Reproductive Technology (ART).
Worldwide, infertility affects 10%-15% of couples and most of them seek medical help including Assisted Reproductive Technology (ART) treatments. Undergoing ART treatments create many physical and emotional burdens. This study examined the psychological consequences of infertility in Iranian infertile males and females as well as their spouses, unlike previous studies that examined mainly females with infertility. Subjects in this descriptive analytical design were recruited from the IVF Department of Mirza Koochak Khan Hospital and the Rouyesh Infertility Treatment Center of Tehran, Iran between Aug 2014 and Sep 2015. Overall, 256 couples (64% response rate), consisting of 78 infertile male and their spouses and 50 infertile female and their spouses, were included in this research. The psychological disorders were measured by the Persian version of Symptoms Checklist-90-Revised and Cattle Inventory. Psychological disorders of infertile couples are significantly associated with increasing age, higher education, longer duration of infertility and unemployment (P<0.05). Prevalence of anxiety, depression, hypochondriasis and paranoia in infertile females and spouses of infertile males were significantly higher than husbands of infertile females (P<0.05). Obsession was more sever in infertile females was significantly greater than infertile males (P=0.01). Depression was significantly lower in infertile males than their spouses (P=0.016). Iranian infertile females and spouses of infertile males experienced more psychological disorders than infertile males and spouses of infertile females. These results may be due to the impact of cultural beliefs and gender roles in Iranian society. Anxiety, depression, obsession, paranoia and hypochondriasis should be addressed before any ART treatments.


Comparison of contemporary troponin assays with the novel biomarkers, heart fatty acid binding protein and copeptin, for the early confirmation or exclusion of myocardial infarction in patients presenting to the emergency department with chest pain.
To examine the diagnostic accuracy of novel biomarkers of myocardial injury and troponin assays for diagnosis of myocardial infarction. 850 patients randomised to the point-of-care testing arm of the Randomised Assessment of Panel Assay of Cardiac markers (RATPAC) study in six emergency departments of low-risk patients presenting with chest pain were studied. Blood samples were obtained on admission and 90 min from admission. Myocardial infarction was defined by the universal definition of myocardial infarction. The following diagnostic strategies were compared by receiver operator characteristic curve analysis and comparison of area under the curve: individual marker values and the combination of presentation heart fatty acid binding protein (HFABP) and copeptin with troponin. 68 patients had a final diagnosis of myocardial infarction. Admission samples were available from 838/1132 patients enrolled in the study. Areas under the curve were as follows (CIs in parentheses): cardiac troponin I (cTnI) Stratus CS 0.94 (0.90 to 0.98), cTnI Beckmann 0.92 (0.88 to 0.96), cTnI Siemens ultra 0.90 (0.85 to 0.95), cardiac troponin T high sensitivity 0.92 (0.88 to 0.96), HFABP 1 0.84 (0.77 to 0.90) copeptin 0.62 (0.57 to 0.68). HFABP and copeptin were diagnostically inferior to troponin. The combination of HFABP (at the 95th percentile) and troponin (at the 99th percentile) increased diagnostic sensitivity. High-sensitivity cardiac troponin is the best single marker. Addition of HFABP to high-sensitivity troponin increased diagnostic sensitivity. Additional measurement of copeptin is not useful in the chest pain population.


Real price and affordability as challenges for effective tobacco control policies: an analysis for Argentina.
To describe the evolution of cigarettes' real price and affordability during the last decade in Argentina. To analyze the real price of cigarettes, the weighted average monthly price of a pack of 20 cigarettes was divided by the consumer price index (CPI) from 2004 to 2014. The relative income price (RIP) was evaluated for the same period, defining RIP as the percentage of the income required to buy 100 packs of 20-per-pack cigarettes. The RIP was calculated for first-quartile, median, and third-quartile income groups. The lower the RIP, the higher the affordability. The nominal price of a pack of 20 cigarettes sold in Argentina increased from AR$ 2.24 in March 2004 to AR$ 14.36 in June 2014 (nominal price increase of about 19.7% per year). The real price fell from AR$ 2.24 in March 2004 to AR$ 2.11 in June 2014 (real price drop of about 0.6% per year). Between June 2004 and June 2014, the RIP decreased about 39% for the 3rd quartile income group (from 31.3% to 19.2%), about 42% for the median (from 55.7% to 32.0%), and about 50% for the 1st quartile (from 104.4% to 51.8%). In Argentina, inflation and rising income were greater than growth in cigarette prices. Cigarette affordability increased for each income group, with the highest shifts occurring among the poorest and most vulnerable income earners. The increased affordability of cigarettes might reduce the impact of implemented tobacco control policies.


Epilepsy phenotype and its reproducibility after lateral fluid percussion-induced traumatic brain injury in rats: Multicenter EpiBioS4Rx study project 1.
This study was undertaken to assess reproducibility of the epilepsy outcome and phenotype in a lateral fluid percussion model of posttraumatic epilepsy (PTE) across three study sites. A total of 525 adult male Sprague Dawley rats were randomized to lateral fluid percussion-induced brain injury (FPI) or sham operation. Of these, 264 were assigned to magnetic resonance imaging (MRI cohort, 43 sham, 221 traumatic brain injury [TBI]) and 261 to electrophysiological follow-up (EEG cohort, 41 sham, 220 TBI). A major effort was made to harmonize the rats, materials, equipment, procedures, and monitoring systems. On the 7th post-TBI month, rats were video-EEG monitored for epilepsy diagnosis. A total of 245 rats were video-EEG phenotyped for epilepsy on the 7th postinjury month (121 in MRI cohort, 124 in EEG cohort). In the whole cohort (n = 245), the prevalence of PTE in rats with TBI was 22%, being 27% in the MRI and 18% in the EEG cohort (p > .05). Prevalence of PTE did not differ between the three study sites (p > .05). The average seizure frequency was .317 ± .725 seizures/day at University of Eastern Finland (UEF; Finland), .085 ± .067 at Monash University (Monash; Australia), and .299 ± .266 at University of California, Los Angeles (UCLA; USA; p < .01 as compared to Monash). The average seizure duration did not differ between UEF (104 ± 48 s), Monash (90 ± 33 s), and UCLA (105 ± 473 s; p > .05). Of the 219 seizures, 53% occurred as part of a seizure cluster (≥3 seizures/24 h; p >.05 between the study sites). Of the 209 seizures, 56% occurred during lights-on period and 44% during lights-off period (p > .05 between the study sites). The PTE phenotype induced by lateral FPI is reproducible in a multicenter design. Our study supports the feasibility of performing preclinical multicenter trials in PTE to increase statistical power and experimental rigor to produce clinically translatable data to combat epileptogenesis after TBI.


Multiple Doris "kerguelenensis" (Nudibranchia) species span the Antarctic Polar Front.
Despite strong historical biogeographical links between benthic faunal assemblages of the Magellan region of South America and the Antarctic Peninsula, very few studies have documented contemporary movement and gene flow in or out of the Southern Ocean, especially across the Antarctic Polar Front (APF). In fact, oceanographic barriers such as the APF and Antarctica's long geologic isolation have substantially separated the continents and facilitated the evolution of endemic marine taxa found within the Antarctic region. The Southern Ocean benthic sea slug complex, Doris "kerguelenensis," are a group of direct-developing, simultaneous hermaphrodites that lack a dispersive larval stage. To date, there are 59 highly divergent species known within this complex. Here, we provide evidence to show intraspecific genetic connectivity occurs across the APF for multiple species within the D. "kerguelenensis" nudibranch species complex. We addressed questions of genetic connectivity by examining the phylogeographic structure of the three best-sampled D. "kerguelenensis" species and another three trans-APF species using the protein coding mtDNA gene, cytochrome oxidase I. We also highlight alternative refugia uses among species with the same life history traits (i.e., benthic and direct developers) and for some species, extremely large distributions are established (e.g., circumpolarity). By improving our sampling of these nudibranchs, we gain better insight into the population structure and connectivity of the Antarctic region. This work also demonstrates how difficult it is to make generalizations across Antarctic marine species, even among ecologically-similar, closely related species.


Transcriptome analysis of postharvest blueberries (Vaccinium corymbosum 'Duke') in response to cold stress.
Blueberry (Vaccinium spp.) is a small berry with high economic value. Although cold storage can extend the storage time of blueberry to more than 60 days, it leads to chilling injury (CI) displaying as pedicle pits; and the samples of 0 °C-30 days was the critical point of CI. However, little is known about the mechanism and the molecular basis response to cold stress in blueberry have not been explained definitely. To comprehensively reveal the CI mechanisms in response to cold stress, we performed high-throughput RNA Seq analysis to investigate the gene regulation network in 0d (control) and 30d chilled blueberry. At the same time, the pitting and decay rate, electrolyte leakage (EL), malondialdehyde (MDA) proline content and GSH content were measured. Two cDNA libraries from 0d (control) and 30d chilled samples were constructed and sequenced, generating a total of 35,060 unigenes with an N50 length of 1348 bp. Of these, 1852 were differentially expressed, with 1167 upregulated and 685 downregulated. Forty-five cold-induced transcription factor (TF) families containing 1023 TFs were identified. The DEGs indicated biological processes such as stress responses; cell wall metabolism; abscisic acid, gibberellin, membrane lipid, energy metabolism, cellular components, and molecular functions were significantly responsed to cold storage. The transcriptional level of 40 DEGs were verified by qRT-PCR. The postharvest cold storage leads serious CI in blueberry, which substantially decreases the quality, storability and consumer acceptance. The MDA content, proline content, EL increased and the GSH content decreased in this chilled process. The biological processes such as stress responses, hormone metabolic processes were significantly affected by CI. Overall, the results obtained here are valuable for preventing CI under cold storage and could help to perfect the lack of the genetic information of non-model plant species.


Case management and patient reactions: a study of STD care in a province in Zambia.
Sexually transmitted disease (STD) case management was evaluated through observation and interviews at 2 urban and 4 rural health centres and 2 district hospital STD clinics in one urban and 2 rural districts in Central Province, Zambia. The analysis was limited to 59 patients (42 men and 17 women) who paid first visits for their disease and were managed by a clinical officer. The evaluation suffered from the lack of a standard for case management. Results showed that the patients engaged in risky sexual behaviour without being aware of the risks. At the health institutions, few patients were informed about condoms, the risk of HIV, and abstinence from sex during treatment and few were asked to notify their partners. Clinical officers with special STD training performed better than others but sill informed only one-fifth of the patients. Few clinical officers managed patients according to the syndromic approach recommended by the STD control programme.


Self-assembly of magnetic nanoparticles in evaporating solution.
When deposited from an evaporating solution onto a substrate, even nondescript nanoparticles can organize into intricate spatial patterns. Here we show that a simple but long-ranged anisotropy in nanoparticles' interactions can greatly enrich this scenario. In experiments with colloidal Co nanocrystals, which bear a substantial magnetic dipole, we observe assemblies quite distinct from those formed by nonmagnetic particles. Reflecting the strongly nonequilibrium nature of this process, nanocrystal aggregates also differ substantially from expected low-energy arrangements. Using coarse-grained computer simulations of dipolar nanoparticles, we have identified several dynamical mechanisms from which such unusual morphologies can arise. For particles with modest dipole moments, transient connections between growing domains frustrate phase separation into sparse and dense regions on the substrate. Characteristic length scales of the resulting cellular networks depend non-monotonically on the depth of quenches we use to mimic the effects of solvent evaporation. For particles with strong dipole moments, chain-like aggregates formed at early times serve as the agents of assembly at larger scales. Their effective interactions drive the formation of layered loop structures similar to those observed in experiments.


Evaluating the stability of three oxidative stress biomarkers under sewer conditions and potential impact for use in wastewater-based epidemiology.
Wastewater contains a wealth of information about the population who contribute to it including biological and chemical markers of human activity and exposures. F2-isoprostanes have been proposed as oxidative stress biomarkers that can be measured in wastewater to provide a measure of oxidative stress at the population level. While an association between tobacco use and their level in wastewater has been demonstrated, an in-sewer stability assessment has not been conducted to support their use as oxidative stress biomarkers for wastewater-based epidemiology studies. In this study we investigated the stability of 8-iso-prostaglandin F2α (PGF2α), its metabolite dinor-11β-Prostaglandin F2α (dnPGF2α) and Prostaglandin E2 (PGE2) (representative of other classes of prostaglandins) in laboratory-scale sewer reactors simulating real sewers. PGF2α, dnPGF2α and PGE2 were all found to be sufficiently stable under typical sewer conditions therefore satisfying the stability requirement of wastewater-based epidemiology population health biomarkers.


Stage III non-small cell lung cancer and metachronous brain metastases.
This study was undertaken to identify management strategies that maximize survival of patients with stage III non-small cell lung cancer and metachronous brain metastases and to determine whether any apparent improved survival was due to treatment or simply to patient selection. Treatment evaluations of both primary non-small cell lung cancer and brain metastases were performed in 91 patients. Optimal treatment was identified by multivariable analysis. Propensity scoring and multivariable analysis were used to separate treatment benefit from patient selection. Risk-unadjusted median, 12-, and 24-month survivals were 5.2 months, 22%, and 10%, respectively. Younger age (P =.006), good performance status (P =.003), stage IIIA (P =.001), lung resection (P =.02), no other systemic metastases at time of diagnosis of brain metastases (P =.02), and either metastasectomy (P <.001) or stereotactic radiosurgery (P <.001) predicted best survival. However, metastasectomy or stereotactic radiosurgery was more common after lung resection (P =.02) and in patients with good performance status (P =.006), no other systemic metastases at time of diagnosis of brain metastases (P =.01), and fewer brain metastases (P <.001), suggesting that the patients with the best risk profile were selected for aggressive therapy of both lung primary and brain metastases. Despite this selection, analysis of propensity-matched patients demonstrated the benefit of lung resection and metastasectomy or stereotactic radiosurgery (P <.001). Younger patients with resected stage IIIA non-small cell lung cancer who have isolated metachronous brain metastases and good performance status do best when treated with metastasectomy or stereotactic radiosurgery. This survival benefit is a brain treatment effect, not the result of selecting the best patients for aggressive therapy.


Superimposition of metabolic syndrome magnifies post-stenotic kidney injury in dyslipidemic pigs.
Dyslipidemia aggravates kidney injury distal to atherosclerotic renal artery stenosis (ARAS). Besides dyslipidemia, metabolic syndrome (MetS) also involves development of obesity and insulin-resistance (IR). We hypothesized that concurrent obesity and IR magnify swine stenotic-kidney damage beyond dyslipidemia. Pigs with unilateral RAS were studied after 16 weeks of atherogenic diets without (ARAS) or with (MetS + RAS) development of obesity/IR (n=6 each). Additional pigs on normal diet served as normal or non-dyslipidemic RAS controls (n=6 each). Stenotic-kidney renal blood flow (RBF), glomerular filtration rate (GFR), and microvascular architecture were studied using CT, and oxygenation was studied using blood oxygen level-dependent magnetic-resonance-imaging. We further compared kidney adiposity, oxidative stress, inflammation, apoptosis, fibrosis, and systemic levels of oxidative and inflammatory cytokines. ARAS and MetS + RAS developed hypertension and dyslipidemia, and MetS + RAS also developed obesity and IR. RBF and GFR were similarly decreased in all post-stenotic pig kidneys compared to normal pig kidneys, yet MetS + RAS aggravated and expanded medullary hypoxia and microvascular loss. RAS and ARAS increased systemic levels of tumor necrosis factor (TNF)-α, which were further elevated in MetS + RAS. Renal oxidative stress and TNF-α expression increased in ARAS and further in MetS + RAS, which also upregulated expression of anti-angiogenic angiostatin, and magnified apoptosis, tubular injury, and fibrosis. Beyond dyslipidemia, obesity and insulin-resistance aggravate damage in the post-stenotic kidney in MetS, despite relative hyperfiltration-related preservation of renal function. These observations underscore the need to control systemic metabolic disturbances in order to curb renal damage in subjects with ischemic kidney disease.


Human Infection Challenge Experiments: Then and Now.
In the midst of the Covid-19 pandemic, ethicists, researchers, and journalists have recommended studies that deliberately infect healthy volunteers with the coronavirus as a scientific means of expediting vaccine development. In this essay, we trace the history of infection challenge experiments and reflect on the Nuremberg Code of 1947, issued in response to brutal human experiments conducted by Nazi investigators in concentration camps. We argue that the Code continues to offer valuable guidance for assessing the ethics of this controversial form of research, with respect particularly to the acceptable limits to research risks and the social value of research necessary to justify exposing human participants to these risks.


Comparison of viscoelastic characteristics in triceps surae between Black and White athletes.
The purpose of the present study was to investigate race differences in viscoelastic characteristics of triceps surae muscle group. Black and white college sprint type athletes (n=44) participated in this study. Viscoelastic properties were assessed using the free vibration technique: subjects sat with their forefeet on the edge of a force-plate (Kistler, Switzerland) and support a frame loaded with weights (0-40 kg) on the knees. Oscillations of the triceps surae and Achilles tendon system were initiated with a hand-held hammer by tapping the weight load. Oscillations occur at frequencies of 3-6 Hz and were slightly damped. The damped oscillations in conjunction with the equation of motion of a damped mass-spring model were used to calculate the viscosity of muscle (b), and the elasticity of muscle fibres (k(d)) and tendon (k(t)) in each subject. There were little significant differences in most of physical characteristic variables between black and white athletes. Black athletes have significantly greater muscle viscosity and elasticity than white athletes while tendon elasticity is equivalent. Thus, muscle stiffness is greater among black athletes. Greater muscle stiffness could contribute to greater sprint/jump performance among black athletes, compared with white athletes, through alteration of foot/ground contact and take-off phases during sprinting/jumping.


Integrin-using rotaviruses bind alpha2beta1 integrin alpha2 I domain via VP4 DGE sequence and recognize alphaXbeta2 and alphaVbeta3 by using VP7 during cell entry.
Integrins alpha2beta1, alphaXbeta2, and alphaVbeta3 have been implicated in rotavirus cell attachment and entry. The virus spike protein VP4 contains the alpha2beta1 ligand sequence DGE at amino acid positions 308 to 310, and the outer capsid protein VP7 contains the alphaXbeta2 ligand sequence GPR. To determine the viral proteins and sequences involved and to define the roles of alpha2beta1, alphaXbeta2, and alphaVbeta3, we analyzed the ability of rotaviruses and their reassortants to use these integrins for cell binding and infection and the effect of peptides DGEA and GPRP on these events. Many laboratory-adapted human, monkey, and bovine viruses used integrins, whereas all porcine viruses were integrin independent. The integrin-using rotavirus strains each interacted with all three integrins. Integrin usage related to VP4 serotype independently of sialic acid usage. Analysis of rotavirus reassortants and assays of virus binding and infectivity in integrin-transfected cells showed that VP4 bound alpha2beta1, and VP7 interacted with alphaXbeta2 and alphaVbeta3 at a postbinding stage. DGEA inhibited rotavirus binding to alpha2beta1 and infectivity, whereas GPRP binding to alphaXbeta2 inhibited infectivity but not binding. The truncated VP5* subunit of VP4, expressed as a glutathione S-transferase fusion protein, bound the expressed alpha2 I domain. Alanine mutagenesis of D308 and G309 in VP5* eliminated VP5* binding to the alpha2 I domain. In a novel process, integrin-using viruses bind the alpha2 I domain of alpha2beta1 via DGE in VP4 and interact with alphaXbeta2 (via GPR) and alphaVbeta3 by using VP7 to facilitate cell entry and infection.


Cross-sex testosterone therapy in ovariectomized mice: addition of low-dose estrogen preserves bone architecture.
Cross-sex hormone therapy (XHT) is widely used by transgender people to alter secondary sex characteristics to match their desired gender presentation. Here, we investigate the long-term effects of XHT on bone health using a murine model. Female mice underwent ovariectomy at either 6 or 10 wk and began weekly testosterone or vehicle injections. Dual-energy X-ray absorptiometry (DXA) was performed (20 wk) to measure bone mineral density (BMD), and microcomputed tomography was performed to compare femoral cortical and trabecular bone architecture. The 6-wk testosterone group had comparable BMD with controls by DXA but reduced bone volume fraction, trabecular number, and cortical area fraction and increased trabecular separation by microcomputed tomography. Ten-week ovariectomy/XHT maintained microarchitecture, suggesting that estrogen is critical for bone acquisition during adolescence and that late, but not early, estrogen loss can be sufficiently replaced by testosterone alone. Given these findings, we then compared effects of testosterone with effects of weekly estrogen or combined testosterone/low-dose estrogen treatment after a 6-wk ovariectomy. Estrogen treatment increased spine BMD and microarchitecture, including bone volume fraction, trabecular number, trabecular thickness, and connectivity density, and decreased trabecular separation. Combined testosterone-estrogen therapy caused similar increases in femur and spine BMD and improved architecture (increased bone volume fraction, trabecular number, trabecular thickness, and connectivity density) to estrogen therapy and were superior compared with mice treated with testosterone only. These results demonstrate estradiol is critical for bone acquisition and suggest a new cross-sex hormone therapy adding estrogens to testosterone treatments with potential future clinical implications for treating transgender youth or men with estrogen deficiency.


Scaffolding the attention-deficit/hyperactivity disorder brain using transcranial direct current and random noise stimulation: A randomized controlled trial.
Improving symptomology and cognitive deficits in neurodevelopmental disorders is a crucial challenge. We examined whether neurostimulation protocols, which have been shown to yield long-term effects when combined with cognitive training, could benefit children with Attention-deficit/hyperactivity-disorder (ADHD), the most common neurodevelopmental disorder in childhood. We used a randomized double-blind active-controlled crossover study of 19 unmedicated children with ADHD, who received either anodal transcranial direct current stimulation (tDCS) over the left dorsolateral prefrontal cortex (dlPFC) or random noise stimulation (tRNS) over the bilateral dlPFC, while completing executive functions training. For our primary outcome, tRNS yielded a clinical improvement as indicated by the reduced ADHD rating-scale score from baseline, and in comparison to the changes observed in tDCS. The effect of brain stimulation one week after completion of treatment yielded further improvement, suggesting a neuroplasticity-related effect. Finally, tRNS improved working memory compared to tDCS, and a larger tRNS effect on ADHD rating-scale was predicted for those who showed the greatest improvement in working memory. We found that our intervention can have a lasting effect, rather than a merely immediate effect as was shown for in previous medical interventions in ADHD. Our results provide a promising direction toward a novel intervention in ADHD.


Experimental and clinical observations on hepatic cryosurgery for colorectal metastases.
Cryosurgery, using liquid nitrogen at -196 degrees C, was explored for treating colorectal liver metastases in an experimental study of rat colon cancer and in a clinical investigation of patients with unresectable liver tumors. The viability of rat colon cancer isografts showed that while two or three freeze-thaw cycles were 100% effective in controlling established isografts and preventing isograft take, one freeze-thaw cycle was suboptimal. In these animals cryosurgery was as effective as surgical resection in controlling established experimental liver metastases. Cryosurgery by operative liver exposure and intraoperative ultrasound monitoring were used to treat liver metastases from colorectal cancer in 24 patients. At a median follow-up of 2 years (range, 5 months to 5 years), seven patients (29%) are disease free, eight (33.5%) are alive with recurrent tumors, and nine (37.5%) have died. The patterns of failure were: remaining liver and extrahepatic sites, ten patients (59%); remaining liver only, six patients (35%); and extrahepatic only, one patient (6%). These data demonstrate that cryosurgery is a useful modality for treating unresectable primary and metastatic liver cancers. Addition of systemic adjuvant therapy may improve the tertiary failure following the control of liver metastases.


Identification of the gene PaEMT1 for biosynthesis of mannosylerythritol lipids in the basidiomycetous yeast Pseudozyma antarctica.
The yeast Pseudozyma antarctica produces a large amount of glycolipid biosurfactants known as mannosylerythritol lipids (MELs), which show not only excellent surface-active properties but also versatile biochemical actions. To investigate the biosynthesis of MELs in the yeast, we recently reported expressed sequence tag (EST) analysis and estimated genes expressing under MEL production conditions. Among the genes, a contiguous sequence of 938 bp, PA_004, showed high sequence identity to the gene emt1, encoding an erythritol/mannose transferase of Ustilago maydis, which is essential for MEL biosynthesis. The predicted translation product of the extended PA_004 containing the two introns and a stop codon was aligned with Emt1 of U. maydis. The predicted amino acid sequence shared high identity (72%) with Emt1 of U. maydis, although the amino-terminal was incomplete. To identify the gene as PaEMT1 encoding an erythritol/mannose transferase of P. antarctica, the gene-disrupted strain was developed by the method for targeted gene disruption, using hygromycin B resistance as the selection marker. The obtained ΔPaEMT1 strain failed to produce MELs, while its growth was the same as that of the parental strain. The additional mannosylerythritol into culture allowed ΔPaEMT1 strain to form MELs regardless of the carbon source supplied, indicating a defect of the erythritol/mannose transferase activity. Furthermore, we found that MEL formation is associated with the morphology and low-temperature tolerance of the yeast.


Gynecological cancer survivors and community support services: referral, awareness, utilization and satisfaction.
To quantify gynecological cancer survivors' referral to, awareness of, utilization of and satisfaction with community support services, as well as the factors associated with service use. In 2004, 802 gynecological cancer survivors, 3 months-5 years post-diagnosis, completed a postal questionnaire (56% response rate). Descriptive statistics summarized outcome prevalences. Logistic regression models identified correlates of service utilization. Substantial proportions of women were aware of the main cancer support organization, Cancer Council Queensland (72%), and of information booklets (74%), helplines (66%), support groups (56%) and internet information (50%). Less than half were aware of other services. The most commonly used resources and services were information booklets (37%), the internet (23%), and helplines (20%). More broadly, 43% utilized information/internet support, 30% utilized psychosocial services and 27% utilized functional/practical services. Approximately one-fifth (19%) used more than one support types. Having a health-care provider referral, being diagnosed with lymphedema or living in northern Queensland were associated with higher odds of service use in all three of the support types. While most (86%) of those referred used a service, only a few women received referrals. Among users, satisfaction with services was high. While gynecological cancer survivors accessed a variety of support, there is a need to ensure women are aware of services. Given the low prevalence of referrals and that referral was a key influence on service use, clinician education may be necessary to improve service referral. Organizations should also consider strategies to keep services high on clinicians' radars.


Polymeric nano-assemblies as emerging delivery carriers for therapeutic applications: a review of recent patents.
Multi-morphologic polymer nano-assemblies, such as micelles and vesicles, have been intensively studied recently by scientists in the multidisciplinary fields for their promising applications in bioengineering, biomedicine and pharmaceutics. With the success of several micellar formulations in clinical trials, more and more therapeutics based on polymer assemblies are on the pipeline for clinical applications. The current review summarizes some recent patents on the polymer nano-assemblies including micelles and vesicles, with the focus on drug delivery and gene therapy. For the lack of updated patents, the selected progress made most recently in this field has been presented based on the newly published articles. The future development in this active and exciting field has been discussed as well. Because of the huge literature of scientific papers on delivery systems based on polymer assemblies in recent years, this review attempts to limit the specific examples to those that are currently in clinical trials. Accordingly, it is impossible to properly credit all the scientists in this field. The author apologizes in advance for all omissions.


The effect of heat stress on ovarian function of laying hens.
Reproductive failure associated with heat stress is a well-known phenomenon. The mechanism involved in this failure is not clearly understood. In order to test a possible direct effect of heat stress on ovarian function, 36 White Leghorn laying hens were housed in individual cages in 2 temperature- and light-controlled rooms (n = 18). At 31 wk of age, one group was exposed daily for 12 h to high temperature (42 +/- 3 degrees C), and the second group was maintained under thermoneutral conditions (24 to 26 degrees C) and served as control. Body temperature, feed intake, egg production, and egg weight were recorded daily; heparinized blood samples were drawn every 3 d for plasma hormonal level of luteinizing hormone, follicular stimulating hormone, progesterone, 17beta-estradiol, and testosterone. Six days after exposure half of the birds in each group were killed, and the ovary and oviduct were weighed and preovulatory follicles removed and extracted for mRNA of Cytochrome P 450 aromatase, 17-alpha hydroxylase. The same procedure was repeated 9 d later with the rest of the birds. Short and long heat exposure caused significant hyperthermia and reduction of egg production, egg weight, ovarian weight, and the number of large follicles. In addition, a significant reduction in plasma progesterone and testosterone was detected 2 d after exposing the birds to heat stress, and plasma 17beta-estradiol was significantly reduced 14 d after initiation of heat stress. Short exposure to heat stress caused significant reduction in mRNA expression of cytochrome P450 17-alpha hydroxylase, exposing the birds to long-term heat stress caused significant reduction in expression of mRNA of both steroidogenic enzymes. No significant change was found in plasma luteinizing hormone and follicular stimulating hormone levels during the entire experimental period. We suggest a possible direct effect of heat stress on ovarian function.


You can't keep a good parasite down: evolution of a male-killer suppressor uncovers cytoplasmic incompatibility.
Maternally inherited parasites are known to impose a wide variety of reproductive manipulations upon their host. These often produce strong selection on the host to suppress the parasite, resulting in a reduction in the frequency of the parasite. However, in the butterfly Hypolimnas bolina, infected with a Wolbachia bacterium, field data demonstrate that suppression of the male-killing phenotype does not depress parasite frequency. Here we test and verify one hypothesis to explain this apparent paradox-Wolbachia induces a second phenotype, Cytoplasmic Incompatibility (CI), in populations where host suppression has evolved. We further demonstrate that the capacity to induce CI has not evolved de novo, but instead is instantaneously expressed upon the survival of infected males. The significance of these results is threefold: (1) multiple phenotypes can provide Wolbachia with the means to maintain itself in a host following suppression of a single manipulative phenotype; (2) the ability to induce CI can remain hidden in systems in which male-killing is observed, just as the ability to induce male-killing may be obscured in strains exhibiting CI; (3) the evolutionary maintenance of CI in a system in which it is not expressed suggests a functional link with male-killing or other traits under selection.


MALDI in-source decay of high mass protein isoforms: application to alpha- and beta-tubulin variants.
Tubulin is one of the major targets in cancer chemotherapy and the target of more than twenty percent of the cancer chemotherapic agents. The modulation of isoform content has been hypothesized as being a cause of resistance to treatment. Isoform differences lie mostly in the C-terminus part of the protein. Extensive characterization of this polypeptide region is therefore of critical importance. MALDI-TOF fragmentation of tubulin C-terminal domains was tested using synthetic peptides. Then, isotypes from HeLa cells were successfully characterized for the first time by in-source decay (ISD) fragmentation of their C-terminus coupled to a pseudo MS(3) technique named T(3)-sequencing. The fragmentation occurred in-source, preferentially generating y(n)-series ions. This approach required guanidination for the characterization of the beta(III)-tubulin C-terminus peptide. This study is, to our knowledge, the first example of reflectron in-source decay (reISD) of the C-terminus of a 50 kDa protein. This potentially occurs via a CID-like mechanism occurring in the MALDI plume. There are now new avenues for top-down characterization of important clinical biomarkers such as beta(III)-tubulin isotypes, a potential marker of drug resistance and tumor progression. This paper raises the challenge of protein isotypes characterization for early cancer detection and treatment monitoring.


[Enhancement of ionizing radiation effects on transplantable cerebral gliomas using short-term hyperglycemia].
Experiments were staged on rats with transplantable anaplastic cerebral right hemisphere astrocytomas. Hyperglycemia was induced by means of intraperitoneal fractional administration of 40% glucose solution. Single local x-ray tumor irradiation followed on the 4-5th, 7-8th and 10-12th days after strain transplantation at the doses of 30 and 40 Gy without hyperglycemia and in combination with it. The irradiation effect was assessed by the mean survival time of the animals. Hyperglycemia was shown to increase significantly radiation injury of highly resistant experimental malignant cerebral glial tumors.


The prognostic value of non-linear analysis of heart rate variability in patients with congestive heart failure--a pilot study of multiscale entropy.
The influences of nonstationarity and nonlinearity on heart rate time series can be mathematically qualified or quantified by multiscale entropy (MSE). The aim of this study is to investigate the prognostic value of parameters derived from MSE in the patients with systolic heart failure. Patients with systolic heart failure were enrolled in this study. One month after clinical condition being stable, 24-hour Holter electrocardiogram was recording. MSE as well as other standard parameters of heart rate variability (HRV) and detrended fluctuation analysis (DFA) were assessed. A total of 40 heart failure patients with a mea age of 56±16 years were enrolled and followed-up for 684±441 days. There were 25 patients receiving β-blockers treatment. During follow-up period, 6 patients died or received urgent heart transplantation. The short-term exponent of DFA and the slope of MSE between scale 1 to 5 were significantly different between patients with or without β-blockers (p = 0.014 and p = 0.028). Only the area under the MSE curve for scale 6 to 20 (Area(6-20)) showed the strongest predictive power between survival (n = 34) and mortality (n = 6) groups among all the parameters. The value of Area(6-20)21.2 served as a significant predictor of mortality or heart transplant (p = 0.0014). The area under the MSE curve for scale 6 to 20 is not relevant to β-blockers and could further warrant independent risk stratification for the prognosis of CHF patients.


Interleukin-1 beta, tumor necrosis factor, and prostaglandin E2 response after incubation of peritoneal cells and peripheral blood mononuclear cells with lipopolysaccharide.
The magnitude of response to lipopolysaccharide (LPS) by cells isolated from peritoneal dialysate (PDC) (n = 29) was compared to the response of monocytes (M) from hemodialysis (HD) patients (n = 8) and from normals (n = 9). Further studies compared monocyte (M) and PDC from 8 peritoneal dialysis (PD) patients, with adherence purification of PDC. The cells were cultured at 5 x 10(5)/ml with varying LPS concentrations for 1, 2, 8, 20 and 48 hrs. TNF was measured by L929 assay, PGE2 by RIA and Il-1 beta by ELISA. The data demonstrate a defect in the stimulated response of TNF and Il-1 beta by M from both HD and PD patients. A similar defect is also present in PDC. By contrast PGE2 response to stimulation with LPS does not differ in M from HD and normals. For TNF and PGE2 the depressed response of the PDC may be explained by the lower percentage of macrophages (M phi) if adherence purification has not been carried out. However Il-1 beta production by adherence purified M phi is significantly lower than monocytes, although greater than non-adherent cells. We describe alterations in uremic M and M phi function which may contribute to the clinically observed immune defect in uremia.


Efficient incorporation of large (>2 kb) heterologies into heteroduplex DNA: Pms1/Msh2-dependent and -independent large loop mismatch repair in Saccharomyces cerevisiae.
DNA double-strand break (DSB) repair in yeast is effected primarily by gene conversion. Conversion can conceivably result from gap repair or from mismatch repair of heteroduplex DNA (hDNA) in recombination intermediates. Mismatch repair is normally very efficient, but unrepaired mismatches segregate in the next cell division, producing sectored colonies. Conversion of small heterologies (single-base differences or insertions <15 bp) in meiosis and mitosis involves mismatch repair of hDNA. The repair of larger loop mismatches in plasmid substrates or arising by replication slippage is inefficient and/or independent of Pms1p/Msh2p-dependent mismatch repair. However, large insertions convert readily (without sectoring) during meiotic recombination, raising the question of whether large insertions convert by repair of large loop mismatches or by gap repair. We show that insertions of 2.2 and 2.6 kbp convert efficiently during DSB-induced mitotic recombination, primarily by Msh2p- and Pms1p-dependent repair of large loop mismatches. These results support models in which Rad51p readily incorporates large heterologies into hDNA. We also show that large heterologies convert more frequently than small heterologies located the same distance from an initiating DSB and propose that this reflects Msh2-independent large loop-specific mismatch repair biased toward loop loss.


Primary fibrosarcoma of the scalp: a review of a ten-year record from Lagos, Nigeria.
Primary fibrosarcoma of the scalp is very rare and, although the tumour is slow growing, it often results in major cosmetic and other problems. Repeated surgical excisions for recurrent disease are often necessary. We report our experience of the management of this rare tumour at the Lagos University Teaching Hospital, Nigeria: a national referral centre for malignant tumours requiring radiotherapy. Five patients were seen over a 10-year period. An 80% recurrence-free interval was obtained at 2 years with surgery and radiotherapy.


Maternal immune activation causes age- and region-specific changes in brain cytokines in offspring throughout development.
Maternal infection is a risk factor for autism spectrum disorder (ASD) and schizophrenia (SZ). Indeed, modeling this risk factor in mice through maternal immune activation (MIA) causes ASD- and SZ-like neuropathologies and behaviors in the offspring. Although MIA upregulates pro-inflammatory cytokines in the fetal brain, whether MIA leads to long-lasting changes in brain cytokines during postnatal development remains unknown. Here, we tested this possibility by measuring protein levels of 23 cytokines in the blood and three brain regions from offspring of poly(I:C)- and saline-injected mice at five postnatal ages using multiplex arrays. Most cytokines examined are present in sera and brains throughout development. MIA induces changes in the levels of many cytokines in the brains and sera of offspring in a region- and age-specific manner. These MIA-induced changes follow a few, unexpected and distinct patterns. In frontal and cingulate cortices, several, mostly pro-inflammatory, cytokines are elevated at birth, followed by decreases during periods of synaptogenesis and plasticity, and increases again in the adult. Cytokines are also altered in postnatal hippocampus, but in a pattern distinct from the other regions. The MIA-induced changes in brain cytokines do not correlate with changes in serum cytokines from the same animals. Finally, these MIA-induced cytokine changes are not accompanied by breaches in the blood-brain barrier, immune cell infiltration or increases in microglial density. Together, these data indicate that MIA leads to long-lasting, region-specific changes in brain cytokines in offspring-similar to those reported for ASD and SZ-that may alter CNS development and behavior.


Improvement of Attention-Deficit/Hyperactivity Disorder Symptoms in School-Aged Children, Adolescents, and Young Adults With Autism via a Digital Smartglasses-Based Socioemotional Coaching Aid: Short-Term, Uncontrolled Pilot Study.
People with autism spectrum disorder (ASD) commonly experience symptoms related to attention-deficit/hyperactivity disorder (ADHD), including hyperactivity, inattention, and impulsivity. One-third of ASD cases may be complicated by the presence of ADHD. Individuals with dual diagnoses face greater barriers to accessing treatment for ADHD and respond less positively to primary pharmacologic interventions. Nonpharmacologic technology-aided tools for hyperactivity and inattention in people with ASD are being developed, although research into their efficacy and safety remains limited. The objective of this preliminary study was to describe the changes in ADHD-related symptoms in children, adolescents, and young adults with ASD immediately after use of the Empowered Brain system, a behavioral and social communication aid for ASD running on augmented reality smartglasses. We recruited 8 children, adolescents, and young adults with ASD (male to female ratio of 7:1, mean age 15 years, range 11.7-20.5 years) through a Web-based research signup form. The baseline score on the hyperactivity subscale of the Aberrant Behavioral Checklist (ABC-H), a measure of hyperactivity, inattention, and impulsivity, determined their classification into a high ADHD-related symptom group (n=4, ABC-H≥13) and a low ADHD-related symptom group (n=4, ABC-H<13). All participants received an intervention with Empowered Brain, where they used smartglasses-based social communication and behavioral modules while interacting with their caregiver. We then calculated caregiver-reported ABC-H scores at 24 and 48 hours after the session. All 8 participants were able to complete the intervention session. Postintervention ABC-H scores were lower for most participants at 24 hours (n=6, 75%) and for all participants at 48 hours (n=8, 100%). At 24 hours after the session, average participant ABC-H scores decreased by 54.9% in the high ADHD symptom group and by 20% in the low ADHD symptom group. At 48 hours after the session, ABC-H scores compared with baseline decreased by 56.4% in the high ADHD symptom group and by 66.3% in the low ADHD symptom group. This study provides initial evidence for the possible potential of the Empowered Brain system to reduce ADHD-related symptoms, such as hyperactivity, inattention, and impulsivity, in school-aged children, adolescents, and young adults with ASD. This digital smartglasses intervention can potentially be targeted at a broader array of mental health conditions that exhibit transdiagnostic attentional and social communication deficits, including schizophrenia and bipolar disorder. Further research is required to understand the clinical importance of these observed changes and to conduct longitudinal studies on this intervention with control groups and larger sample sizes.


[Mild and severe organic brain disorders in childhood].
An outline of the classification of minor brain dysfunction and severe brain syndrome is followed by a short historical sketch of the development of the concept of minor brain dysfunction and a description of the clinical and diagnostic relevance of this concept. After evaluation of specific therapeutic approaches, selected empirical studies are reported to elucidate the course of the disorder. The issue concerning the existence of a behavioral syndrome that characterizes severe brain disorders is discussed. The presentation of selected research findings on psychopathological and neuropsychological sequelae of severe brain traumas provides the argument for the final discussion. The necessity of qualifying a highly specific, narrowly defined psychopathological syndrome in childhood is emphasized.


Activation of the Interferon Pathway is Dependent Upon Autoantibodies in African-American SLE Patients, but Not in European-American SLE Patients.
In systemic lupus erythematosus (SLE), antibodies directed at RNA-binding proteins (anti-RBP) are associated with high serum type I interferon (IFN), which plays an important role in SLE pathogenesis. African-Americans (AA) are more likely to develop SLE, and SLE is also more severe in this population. We hypothesized that peripheral blood gene expression patterns would differ between AA and European-American (EA) SLE patients, and between those with anti-RBP antibodies and those who lack these antibodies. Whole blood RNA from 33 female SLE patients and 16 matched female controls from AA and EA ancestral backgrounds was analyzed on Affymetrix Gene 1.0 ST gene expression arrays. Ingenuity Pathway Analysis was used to compare the top differentially expressed canonical pathways amongst the sample groups. An independent cohort of 116 SLE patients was used to replicate findings using quantitative real-time PCR (qPCR). Both AA and EA patients with positive anti-RBP antibodies showed over-expression of similar IFN-related canonical pathways, such as IFN Signaling (P = 1.3 × 10(-7) and 6.3 × 10(-11) in AA vs. EA respectively), Antigen Presenting Pathway (P = 1.8 × 10(-5) and 2.5 × 10(-6)), and a number of pattern recognition receptor pathways. In anti-RBP negative (RBP-) patients, EA subjects demonstrated similar IFN-related pathway activation, whereas no IFN-related pathways were detected in RBP-AA patients. qPCR validation confirmed similar results. Our data show that IFN-induced gene expression is completely dependent on the presence of autoantibodies in AA SLE patients but not in EA patients. This molecular heterogeneity suggests differences in IFN-pathway activation between ancestral backgrounds in SLE. This heterogeneity may be clinically important, as therapeutics targeting this pathway are being developed.


Mobile social media use and life satisfaction among adolescents: a moderated mediation model.
Adolescence is a sensitive transitional period accompanied by great physical, mental, and behavioral changes. Therefore, maintaining physical and mental health is crucial to the growth and development of adolescents. As one of the important indicators of mental health, the influencing factors of life satisfaction have been widely concerned by scholars. In recent years, with the rapid development of Internet technology, mobile social media has penetrated into all aspects of adolescents' life, which has a subtle impact on their physical and mental health. Existing studies have indicated that mobile social media use can affect adolescents' life satisfaction. However, little is known about the mediating and moderating mechanisms linking this association. This study developed a moderated mediation model to examine the mediating role of meaning in life and the moderating role of childhood psychological maltreatment. A total of 1,198 adolescents across four provinces and municipalities of China completed questionnaires on mobile social media use, life satisfaction, meaning in life, and childhood psychological maltreatment. After controlling for gender and age, the results demonstrated that mobile social media use was positively associated with life satisfaction and meaning in life among adolescents. Moreover, meaning in life fully mediated the association between mobile social media use and life satisfaction. Finally, the association between mobile social media use and life satisfaction, as well as that between mobile social media use and meaning in life, was moderated by childhood psychological maltreatment. Specifically, these associations are stronger for adolescents with high levels of psychological maltreatment. These findings shed light on the important mechanism underlying mobile social media use's effects on adolescents' life satisfaction, which is helpful to formulate targeted measures for improving adolescents' life satisfaction.


Hemostatic defects in massive transfusion: rapid diagnosis and management.
Severe hemostatic defects that occur during massive transfusion are related to the volume of blood transfused, preexisting hemostatic abnormalities, concomitant pathologic changes, and therapeutic maneuvers. The relative role of each factor in the bleeding can be rapidly determined by using routine clinical laboratory tests. This determination requires an understanding of the properties of selected clotting factors, what coagulation screening tests measure, how these tests behave as the levels of factors change, and test profiles characteristic of different defects. Screening tests include platelet count, prothrombin time, partial thromboplastin time, thrombin time, and fibrinogen level. These tests are generally available on an emergent basis and can be completed within 25 minutes. The pattern associated with hemodilution is universal in massive transfusion. Patterns characteristic of the other pathologic processes that may be encountered are simply superimposed on the characteristics of hemodilution. Successful management of the contributing causes of bleeding depends on the administration of the appropriate blood components in the dose necessary to ensure that the levels of platelets and clotting factors are returned to and maintained at hemostatic levels.


Molecular chaperone Hsp90 associates with resistance protein N and its signaling proteins SGT1 and Rar1 to modulate an innate immune response in plants.
SGT1 and Rar1 are important signaling components of resistance (R) gene-mediated plant innate immune responses. Here we report that SGT1 and Rar1 associate with the molecular chaperone Hsp90. In addition, we show that Hsp90 associates with the resistance protein N that confers resistance to tobacco mosaic virus. This suggests that Hsp90-SGT1-Rar1 and R proteins might exist in one complex. Suppression of Hsp90 in Nicotiana benthamiana plants shows that it plays an important role in plant growth and development. In addition, Hsp90 suppression in NN plants compromises N-mediated resistance to tobacco mosaic virus. Our results reveal a new role for SGT1- and Rar1-associated chaperone machinery in R gene-mediated defense signaling.


Neurotensin receptor down-regulation induced by dexamethasone and forskolin in rat hypothalamic cultures is mediated by endogenous neurotensin.
Neurotensin (NT) has been shown to be involved in neuroendocrine regulation, and the presence of both the peptide and its receptors has been demonstrated in the hypothalamus. In the present study, we show that hypothalamic neurons in primary cultures express the neurotensin receptor (NTR) and we examined a possible regulation of this receptor by glucocorticoids and activators of adenylate cyclase. In the hypothalamic cultures, 125I-NT bound to a single class of binding sites, presenting a selectivity similar to that observed for the high-affinity NTR previously described in the adult rat brain. Radioautographic studies demonstrated that these 125I-NT binding sites were present on 3% of the neurons. A 48-h treatment with forskolin (fsk) decreased 125I-NT binding by 30%. No effect of dexamethasone (dex) alone was found on that parameter. However, a combined treatment with both agents led to a 40% decrease in 125I-NT binding, corresponding to a reduced number of binding sites, and to a 68% decrease in the amount of NTR mRNA. In parallel, the dex plus forsk treatment increased NT release in the incubation medium. Moreover, the decreases in 125I-NT binding and NTR mRNA induced by this treatment were abolished in the presence of an anti-NT antibody or SR 48692, a non-peptidic antagonist of NTR, suggesting that the down-regulation of NTR observed after dex plus fsk treatment was mediated by the release of endogenous NT. Agonist-induced down-regulation of the NTR in this system was confirmed by the application of an exogenous NT analogue, JMV 449. The present findings indicate that, in hypothalamic cultures, dex and fsk indirectly down-regulate NTR expression via the release of endogenous NT.


Descriptive survey of women's childbirth experiences in two state hospitals in KwaZulu-Natal.
Giving birth is one of the most important events in a woman's life and is a highly individualistic and unique experience. The study aimed to describe women's childbirth experiences in two state hospitals in KwaZulu-Natal. A non-experimental, quantitative, descriptive survey of low-risk mothers was conducted in two state hospitals by using the Childbirth Experience Questionnaire (CEQ). With a response rate of 96%, 201 questionnaires were completed and returned. The highest mean score of the four dimensions of the CEQ was for the dimension of Professional Support (3.1). The results of the individual dimension items scoring the highest positive response were: I felt that I handled the situation well (147; 74%) (Own Capacity); I felt very well cared for by my midwife (165; 82%) (Professional Support); 151 respondents (76%) scored the item My impression of the team's medical skill made me feel secure as the highest positive experience (Perceived Safety); and I felt I could have a say in the choice of pain relief (105; 52%) (Participation). The relationship between demographic variables (age, level of education, parity, antenatal clinic attendance, induction of labour, augmentation and duration of labour) and respondents' scores of the CEQ dimensions was calculated, and only the dimension of Perceived Safety and duration of labour (≥ 12 hours) were found to be significant (p = 0.026). From the women perspectives, the study results described childbirth experience as multi-dimensional experience and subjective. Both positive and negative experiences coexisted in all dimensions of the CEQ, with the dimension of Professional Support scoring the highest positive response. To maintain a positive birth experience, the study suggests that women should be involved and equipped with knowledge on the process of childbirth.


20 mA bidirectional laser triggering in planar devices based on vanadium dioxide thin films using CO(2) laser.
By utilizing a CO2 laser centered at ~10.6 μm as an optical stimulus, we demonstrated bidirectional laser triggering in a two-terminal planar device based on a highly resistive vanadium dioxide (VO2) thin film. The break-over voltage of the VO2-based device was measured as large as ~294.8 V, which resulted from the high resistivity of insulating VO2 grains comprising the thin film and the large electrode separation of the device. The bidirectional current switching of up to 20 mA was achieved by harnessing the dramatic resistance variation of the device photo-thermally induced by the laser illumination. The transient responses of laser-triggered currents were also analyzed when laser pulses excited the device at a variety of pulse widths and repetition rates. In the transient responses, a maximum switching contrast between off- and on-state currents was measured as ~7067 with an off-state current of ~2.83 μA, and rising and falling times were measured as ~30 and ~16 ms, respectively, for 100 ms laser pulses.


Vaccine efficacy against primary and metastatic cancer with in vitro-generated CD103+ conventional dendritic cells.
Type 1 conventional dendritic cells (cDC1s) possess efficient antigen presentation and cross-presentation activity, as well as potent T cell priming ability. Tissue-resident cDC1s (CD103+ cDC1s in mice, CD141+ cDC1s in humans) are linked with improved tumor control, yet the efficacy of immunotherapy using this population is understudied. We generated murine CD103+ cDC1s in vitro and examined their expression of cDC1-related factors, antigen cross-presentation activity, and accumulation in tumor-draining lymph nodes (TdLNs). The antitumor efficacy of the in vitro-generated CD103+ cDC1s was studied in murine melanoma and osteosarcoma models. We evaluated tumor responses on vaccination with CD103+ cDC1s, compared these to vaccination with monocyte-derived DCs (MoDCs), tested CD103+ cDC1 vaccination with checkpoint blockade, and examined the antimetastatic activity of CD103+ cDC1s. In vitro-generated CD103+ cDC1s produced cDC1-associated factors such as interleukin-12p70 and CXCL10, and demonstrated antigen cross-presentation activity on stimulation with the toll-like receptor 3 agonist polyinosinic:polycytidylic acid (poly I:C). In vitro-generated CD103+ cDC1s also migrated to TdLNs following poly I:C treatment and intratumoral delivery. Vaccination with poly I:C-activated and tumor antigen-loaded CD103+ cDC1s enhanced tumor infiltration of tumor antigen-specific and interferon-γ+ CD8+ T cells, and suppressed melanoma and osteosarcoma growth. CD103+ cDC1s showed superior antitumor efficacy compared with MoDC vaccination, and led to complete regression of 100% of osteosarcoma tumors in combination with CTLA-4 antibody-mediated checkpoint blockade. In vitro-generated CD103+ cDC1s effectively protected mice from pulmonary melanoma and osteosarcoma metastases. Our data indicate an in vitro-generated CD103+ cDC1 vaccine elicits systemic and long-lasting tumor-specific T cell-mediated cytotoxicity, which restrains primary and metastatic tumor growth. The CD103+ cDC1 vaccine was superior to MoDCs and enhanced response to immune checkpoint blockade. These results indicate the potential for new immunotherapies based on use of cDC1s alone or in combination with checkpoint blockade.


L-band tunable external cavity laser based on 1.58 μm superluminescent diode integrated with spot-size converter.
We report a 1.58 μm superluminescent diode (SLD) with a spot-size converter (SSC) designed and fabricated as a light source for a tunable external cavity laser (T-ECL). The active section of the SLD is fabricated by using a planar buried heterostructure (PBH) for low-threshold current and high-output power operation at a low injection current. The SSC structure of the SLD is designed to possess a buried deep-ridge waveguide (BD-RWG) and show a beam of less divergence. The full-width at half maximum (FWHM) of the horizontal and vertical far-field patterns (FFPs), due to the beam of the less divergence, are 14° and 13°, respectively. We also confirm that an L-band T-ECL employing the SSC SLD operates well enough to prove the characteristics of high performance.


Memory structures for encoding and retrieving a piece of music: an ERP investigation.
This study examined behavioral and neural correlates of expert musical memory, specifically the hypothesis that particular bars within a complex piece of music would serve as structural markers for encoding to and retrieval from memory. Six pianists were asked to learn and memorize a set prelude by J.S. Bach for performance, and to identify bars that they employed for structuring the prelude into component sections. Following performance from memory, the participants took part in a visual recognition memory task, in which single bars from the prelude had to be distinguished from matched new bars. During the recognition task, the electroencephalogram (EEG) was recorded, and event-related potentials (ERPs) from correctly identified prelude stimulus trials were averaged according to their hypothesized status into "structural" and "nonstructural" bars. The results showed that correct identification of structural bars was significantly faster (and tended to display higher accuracy) than recognition of non-structural ones. In addition, recognition of structural bars was associated with a significantly greater negative ERP peak of 300-400 ms latency and a right centro-parietal scalp distribution. This mid-latency negativity appears to index processing of stimuli that served as cues for encoding and retrieval of a complex semantic structure, and is qualitatively and conceptually different from other previously identified recognition memory ERPs (such as the "old/new" effect), as well as from the classic N400 ERP. The data support existing theories of expert memory and music cognition.


Impact of bacteria on the phenotype, functions, and therapeutic activities of invariant NKT cells in mice.
Invariant NKT (iNKT) cells are innate-like lymphocytes that recognize glycolipid antigens in the context of the MHC class I-like antigen-presenting molecule CD1d. In vivo activation of mouse iNKT cells with the glycolipid alpha-galactosylceramide (alpha-GalCer) results in the acquisition of a hyporesponsive (anergic) phenotype by these cells. Because iNKT cells can become activated in the context of infectious agents, here we evaluated whether iNKT cell activation by microorganisms can influence subsequent responses of these cells to glycolipid antigen stimulation. We found that mouse iNKT cells activated in vivo by multiple bacterial microorganisms, or by bacterial LPS or flagellin, became unresponsive to subsequent activation with alpha-GalCer. This hyporesponsive phenotype of iNKT cells required IL-12 expression and was associated with changes in the surface phenotype of these cells, reduced severity of concanavalin A-induced hepatitis, and alterations in the therapeutic activities of alpha-GalCer. These findings may have important implications for the development of iNKT cell-based therapies.


Simulation of the conformation of the murein fabric: the oligoglycan, penta-muropeptide, and cross-linked nona-muropeptide.
The structure and conformation of the sacculus of bacteria at a scale much larger than just the component disaccharide penta-muropeptide is not well known and is crucially important for the understanding of bacterial growth and cell wall function. By computer simulations, the minimal energy conformations and the energy needed for stretching the component parts were found. The oligosaccharide chain, modeled as (GlcNAc-MurNAc)8 when under no tension, can assumed a variety of nearly iso-energetic conformations. These included a variety of bends and kinks, with the chain forming an irregular random coil. In the most relaxed and minimal energy state, the D-lactyl groups of the MurNAc (N-acetyl muramic acid) residues protruded at about an angle of 90 degrees relative to the D-lactyl groups of their immediate MurNAc neighbors in the same chain. The cell wall penta-muropeptide precursor is identical for Escherichia coli and Bacillus subtilis; it also adopted many conformations, each of an energy almost equal to the global minimum. The cross-bridged structure of the tail-to-tail linkage of disaccharide nona-muropeptide has a second type of association, in addition to the covalent cross-bridge, which has not been considered before. This is the ionic interaction between the free D-Ala and the free amino group of the m-A2 pm. In vivo, when the cross-bridge is stretched (in the computer to simulate growth), this pairing dissociates. The possible biological significance of this is that it exposes the underlying 'tail-to-tail' peptide bond to autolysis and will expose both the ends of the m-A2 pm and the D-AlaD-Ala groups that may then be able to react with nascent penta-muropeptides to form trimers. This suggests a new model for growth of the bacterial cell wall that depends on changes in the chemical conformation of the cross-bridge structure as it comes to bear stress.


Integrated FRET Polymers Spatially Reveal Micro- to Nanostructure and Irregularities in Electrospun Microfibers.
A spatial view of macroscopic polymer material properties, in terms of nanostructure and irregularities, can help to better understand engineering processes such as when materials may fail. However, bridging the gap between the molecular-scale arrangement of polymer chains and the spatially resolved macroscopic properties of a material poses numerous difficulties. Herein, an integrated messenger material that can report on the material micro- to nanostructure and its processes is introduced. It is based on polymer chains labeled with fluorescent dyes that feature Förster resonance energy transfer (FRET) dependent on chain conformation and concentration within a host polymer material. These FRET materials are integrated within electrospun polystyrene microfibers, and the FRET is analyzed by confocal laser scanning microscopy (CLSM). Importantly, the use of CLSM allows a spatial view of material nanostructure and irregularities within the microfibers, where changes in FRET are significant when differences in fiber geometries and regularities exist. Furthermore, changes in FRET observed in damaged regions of the fibers indicate changes in polymer conformation and/or concentration as the material changes during compression. The system promises high utility for applications where nano-to-macro communication is needed for a better understanding of material processes.


Interaction of fluorescently-labeled contractile proteins with the cytoskeleton in cell models.
To determine if a living cell is necessary for the incorporation of actin, alpha-actinin, and tropomyosin into the cytoskeleton, we have exposed cell models to fluorescently labeled contractile proteins. In this in vitro system, lissamine rhodamine-labeled actin bound to attachment plaques, ruffles, cleavage furrows and stress fibers, and the binding could not be blocked by prior exposure to unlabeled actin. Fluorescently labeled alpha-actinin also bound to ruffles, attachment plaques, cleavage furrows, and stress fibers. The periodicity of fluorescent alpha-actinin along stress fibers was wider in gerbil fibroma cells than it was in PtK2 cells. The fluorescent alpha-actinin binding in cell models could not be blocked by the prior addition of unlabeled alpha-actinin suggesting that alpha-actinin was binding to itself. While there was only slight binding of fluorescent tropomyosin to the cytoskeleton of interphase cells, there was stronger binding in furrow regions of models of dividing cells. The binding of fluorescently labeled tropomyosin could be blocked by prior exposure of the cell models to unlabeled tropomyosin. If unlabeled actin was permitted to polymerize in the stress fibers in cell models, fluorescently labeled tropomyosin stained the fibers. In contrast to the labeled contractile proteins, fluorescently labeled ovalbumin and BSA did not stain any elements of the cytoskeleton. Our results are discussed in terms of the structure and assembly of stress fibers and cleavage furrows.


Oxypurinol attenuates ischemia-induced hippocampal damage in the gerbil.
Oxypurinol, an inhibitor of the enzyme xanthine oxidase, reduced ischemic hippocampal damage and the associated hypermotility in Mongolian gerbils. Cerebral ischemia was induced in unanesthetized gerbils by a bilateral 5-min occlusion of the carotid arteries. Oxypurinol (40 mg/kg, IP), administered 20 min prior to carotid occlusion, prevented the increase in locomotor activity observed in saline-injected ischemic animals and significantly reduced the damage to, and loss of, CA1 hippocampal neurons observed 5 days postischemia. These findings suggest that oxypurinol may be useful for the prevention of cerebral ischemic damage.


Implementation of Fragment Ion Protection (FIP) during Ultraviolet Photodissociation (UVPD) Mass Spectrometry.
Ultraviolet photodissociation (UVPD) is a nonselective activation method in which both precursor and fragment ions may absorb photons and dissociate. Photoactivation of fragment ions may result in secondary or multiple generations of dissociation, which decreases the signal-to-noise ratio (S/N) of larger fragment ions owing to the prevalent subdivision of the ion current into many smaller, often less informative, fragment ions. Here we report the use of dipolar excitation waveforms to displace fragment ions out of the laser beam path, thus alleviating the extent of secondary dissociation during 193 nm UVPD. This fragment ion protection (FIP) strategy increases S/N of larger fragment ions and improves the sequence coverage obtained for proteins via retaining information deeper into the midsection of protein sequences.


Mitogenic effect by lipopolysaccharide and pokeweed lectin on density-inhibited chick embryo fibroblasts.
The B lymphocyte mitogens, bacterial lipopolysaccharide (LPS), pokeweed lectin, and tuberculin, induced proliferation in density-inhibited monolayer cultures of chick embryo fibroblasts. The stimulation was seen both as an early increase in sugar uptake and cell volume and later as an increase in thymidine incorporation and cell number. The concentration of LPS maximally stimulating fibroblasts was remarkably low, about 0.1-1 ng/ml. LPS preparation with very different sugar chains gave quite similar results indicating that the architecture of the hydrophilic carbohydrate part is not critical for the mitogenic effect.


Myocardial force-frequency defect in mitral regurgitation heart failure is reversed by forskolin.
Postoperative ejection phase parameters and patient survival rates for mitral valve replacement surgery are considerably lower than for similar aortic valve surgery. While chordal transection probably is the major contributor to the lowered values, there is also evidence for decreased preoperative myocardial contractile reserve in mitral regurgitation patients. This study characterizes abnormalities in the force-frequency relation that may underlie impaired function of myocardium isolated from mitral regurgitation patients with New York Heart Association class II-III heart failure. Left ventricular epicardial myocardium was obtained by surgical biopsy during mitral valve replacement surgery in patients with mitral regurgitation heart failure (left ventricular ejection fraction, 0.64 +/- 0.05) and during coronary artery bypass surgery in patients with normal ventricular function. The steady-state twitch tension versus frequency relation was measured in myocardial strip preparations (37 degree C, 12 to 228 min-1) in the absence and presence of forskolin. Relative to normal function, peak isometric twitch tension in mitral regurgitation is depressed by 50% (P < .02) and 74% (P < .003) at contraction frequencies of 60 min-1 and 168 min-1, respectively. The slope of the tension-frequency curve is blunted and its peak is shifted to a lower frequency (mitral regurgitation: 134 min-1; normal function: 173 min-1; P < .02). The myosin heavy chain concentration did not differ between mitral regurgitation and normal function strips (53 +/- 4 versus 54 +/- 4 nmol/g blotted wt). Forskolin (0.5 mumol/L) completely reversed the tension depression, blunting, and the lowered peak frequency in the mitral regurgitation preparations. Preoperatively, myocardial tension generation in mitral regurgitation patients is severely depressed, and the force-frequency curve is blunted and has a negative slope in the exercise range of heart rates. The reversal of these defects by forskolin suggests that abnormal excitation-contraction coupling may underlie the decreased contractile reserve in mitral regurgitation patients.


Live Cell Imaging of Enzymatic Turnover of an Adenosine 5'-Tetraphosphate Analog.
The hydrolysis of nucleotides is of paramount importance as an energy source for cellular processes. In addition, the transfer of phosphates from nucleotides onto proteins is important as a post-translational protein modification. Monitoring the enzymatic turnover of nucleotides therefore offers great potential as a tool to follow enzymatic activity. While a number of fluorescence sensors are known, so far, there are no methods available for the real-time monitoring of ATP hydrolysis inside live cells. We present the synthesis and application of a novel fluorogenic adenosine 5'-tetraphosphate (Ap4) analog suited for this task. Upon enzymatic hydrolysis, the molecule displays an increase in fluorescence intensity, which provides a readout of its turnover. We demonstrate how this can be used for monitoring cellular processes involving Ap4 hydrolysis. To this end, we visualized the enzymatic activity in live cells using confocal fluorescence microscopy of the Ap4 analog. Our results demonstrate that the Ap4 analog is hydrolyzed in lysosomes. We show that this approach is suited to visualize the lysosome distribution profiles within the live cell and discuss how it can be employed to gather information regarding autophagic flux.


Management of anxiety and pain perception in women undergoing office hysteroscopy: a systematic review.
The aim of this review is to provide an overview of the literature about the perception and management of anxiety and pain in women undergoing an office hysteroscopic procedure. We performed a systematic literature search in Embase, PubMed/MEDLINE, Cochrane Library and Web of Science for original studies written in English (registered in PROSPERO 2019-CRD42019132341), using the terms 'hysteroscopy' AND 'pain' AND 'anxiety' published up to January 2019. Only original articles (randomized, observational and retrospective studies) about management of anxiety and pain related to the hysteroscopic procedure were considered eligible. Our literature search produced 84 records. After exclusions, 11 studies including 2222 patients showed the following results: (a) pain experienced during hysteroscopy is negatively affected by preprocedural anxiety; (b) pharmacological interventions seem to be help in reducing pain during hysteroscopy; (c) waiting time before the procedure is a significant factor affecting patients' anxiety; (d) music during the procedure may be helpful in reducing anxiety. The utilization of office hysteroscopy is hampered by varying levels of anxiety and pain perceived by women who are candidates for the procedure. For these reasons, it is essential to identify effective pharmacological and non-pharmacological strategies to alleviate these factors. We recommend further studies especially focusing on non-pharmacological interventions to facilitate the dissemination of good clinical practices among hysteroscopists.


The diverse CB1 and CB2 receptor pharmacology of three plant cannabinoids: delta9-tetrahydrocannabinol, cannabidiol and delta9-tetrahydrocannabivarin.
Cannabis sativa is the source of a unique set of compounds known collectively as plant cannabinoids or phytocannabinoids. This review focuses on the manner with which three of these compounds, (-)-trans-delta9-tetrahydrocannabinol (delta9-THC), (-)-cannabidiol (CBD) and (-)-trans-delta9-tetrahydrocannabivarin (delta9-THCV), interact with cannabinoid CB1 and CB2 receptors. Delta9-THC, the main psychotropic constituent of cannabis, is a CB1 and CB2 receptor partial agonist and in line with classical pharmacology, the responses it elicits appear to be strongly influenced both by the expression level and signalling efficiency of cannabinoid receptors and by ongoing endogenous cannabinoid release. CBD displays unexpectedly high potency as an antagonist of CB1/CB2 receptor agonists in CB1- and CB2-expressing cells or tissues, the manner with which it interacts with CB2 receptors providing a possible explanation for its ability to inhibit evoked immune cell migration. Delta9-THCV behaves as a potent CB2 receptor partial agonist in vitro. In contrast, it antagonizes cannabinoid receptor agonists in CB1-expressing tissues. This it does with relatively high potency and in a manner that is both tissue and ligand dependent. Delta9-THCV also interacts with CB1 receptors when administered in vivo, behaving either as a CB1 antagonist or, at higher doses, as a CB1 receptor agonist. Brief mention is also made in this review, first of the production by delta9-THC of pharmacodynamic tolerance, second of current knowledge about the extent to which delta9-THC, CBD and delta9-THCV interact with pharmacological targets other than CB1 or CB2 receptors, and third of actual and potential therapeutic applications for each of these cannabinoids.


Structure of the glycoprotein gene of sonchus yellow net virus, a plant rhabdovirus.
The nucleotide sequence of the glycoprotein (G) gene of sonchus yellow net virus (SYNV), a plant rhabdovirus, was determined from viral genomic and mRNA cDNA clones. The G cistron is 2045 nucleotides (nt) long and the G protein mRNA open reading frame (ORF), as determined from the cDNA sequence, contains 1896 nt and encodes a protein of 632 amino acids. Immunoblots with antibodies elecited against the purified glycoprotein from virus particles reacted with a fusion protein produced in Escherichia coli, indicating that the cloned ORF encodes the G protein. The 5' end of the G protein mRNA corresponds to nt 5111, relative to the 3' end of the viral (minus sense) genome, as determined by primer extension from mRNA isolated from infected plants, and extends to nt position 7155 on the genomic RNA. A 34-nt untranslated 5' leader sequence and a 115-nt untranslated 3' end flank the ORF on the mRNA. The gene junctions on either side of the G gene on the genomic RNA are identical to those previously described for other SYNV genes and are similar to sequences separating genes of animal rhabdoviruses. The predicted molecular weight of the G protein is 70,215 Da, a value less than the 77,000 Da estimated for the glycosylated G protein from virus particles. The deduced amino acid sequence of the SYNV G protein shares little direct relatedness with the G proteins of other rhabdoviruses, but appears to contain a similar signal sequence, a transmembrane anchor domain, and glycosylation signals. In addition, the SYNV G protein contains a putative nuclear targeting site near the carboxy terminus, which may be involved in transit to the nuclear membrane prior to morphogenesis.


The strength of association between psychological factors and clinical outcome in tendinopathy: A systematic review.
Tendinopathy is often a disabling, and persistent musculoskeletal disorder. Psychological factors appear to play a role in the perpetuation of symptoms and influence recovery in musculoskeletal pain. To date, the impact of psychological factors on clinical outcome in tendinopathy remains unclear. Therefore, the purpose of this systematic review was to investigate the strength of association between psychological factors and clinical outcome in tendinopathy. A systematic review of the literature and qualitative synthesis of published trials was conducted. Electronic searches of ovid MEDLINE, ovid EMBASE, PsychINFO, CINAHL and Cochrane Library was undertaken from their inception to June 2020. Eligibility criteria included RCT's and studies of observational design incorporating measurements of psychological factors and pain, disability and physical functional outcomes in people with tendinopathy. Risk of Bias was assessed by two authors using a modified version of the Newcastle Ottawa Scale. High or low certainty evidence was examined using the GRADE criteria. Ten studies of observational design (6-cross sectional and 4 prospective studies), involving a sample of 719 participants with tendinopathy were included. Risk of bias for the included studies ranged from 12/21 to 21/21. Cross-sectional studies of low to very low level of certainty evidence revealed significant weak to moderate strength of association (r = 0.24 to 0.53) between psychological factors and clinical outcomes. Prospective baseline data of very low certainty evidence showed weak strength of association between psychological factors and clinical outcome. However, prospective studies were inconsistent in showing a predictive relationship between baseline psychological factors on long-term outcome. Cross sectional studies report similar strengths of association between psychological factors and clinical outcomes in tendinopathy to those found in other musculoskeletal conditions. The overall body of the evidence after applying the GRADE criteria was low to very low certainty evidence, due to risk of bias, imprecision and indirectness found across included studies. Future, high quality longitudinal cohort studies are required to investigate the predictive value of baseline psychological factors on long-term clinical outcome.


[Hypertriglyceridemia-induced pancreatitis treated with insulin in a nondiabetic patient].
Heparin and/or insulin stimulate lipoprotein lipase and are known to decrease serum triglyceride level. However, their efficacy in hypertriglyceridemia-induced acute pancreatitis in nondiabetic patients is not well documented. We report a case of hypertriglyceridemia-induced pancreatitis in 43-year-old nondiabetic woman in whom treatment with insulin was accompanied by reduction in serum triglyceride level and the resolution of pancreatitis. She presented to the emergency department with abdominal pain and biochemical evidence of acute pancreatitis. Her medical history was unremarkable. There was no history of alcohol consumption, and biliary imaging was not remarkable. Subsequent laboratory investigation revealed marked hypertriglyceridemia (1,951 mg/dL), impaired fasting glucose, and normal HbAlc level. The Ransons score and APATCH II score were 1 and 4. Abdominal CT showed diffuse enlargement of pancreas, peripancreatic fat infiltration, and multiple fluid collections around the pancreas. We treated the patient with the infusion of 5% dextrose and 1.5 unit/hr regular insulin to reduce serum triglyceride level. The level of serum triglyceride was decreased to 305 mg/dL on day 5. During the remainder of hospitalization, her clinical symptoms and laboratory values gradually improved.