[Cloning and function analysis of high mobility group box 1 (HMGB1) protein of Schistosoma japonicum (Mainland strain)].
To clone and express a high mobility group box 1(HMGB1) protein of Schistosomajaponicum (Mainland strain) and analyze its function. The DNA fragment of open reading frame encoding Sj HMGB 1 protein was amplified by RT-PCR from the mRNA of S. japonicum worms, then it was subcloned into the expression vector pET28a(+) to form the recombinant expression plasmid SjHMGB1-pET28a. The recombinant expression plasmid was transformed into the component E. coli BL21(DE3), and the tranformant containing recombinant expression plasmid was induced with IPTG to express the recombinant protein SjHMGB1. The recombinant SjHMGB1 protein was purified by affinity chromatography with nickel chelating affinity chromatography agarose gel. The Gel retard experiment and animal immunization were performed to analyze the DNA binding capacity and the immunologic property of recombinant SjHMGB1. The expression levels of HMGB1 in different life cycle stages of S. japonicum were analyzed by Western bloting and RT-PCR. Female ICR mice were immunized with the recombinant SjHMGB1 protein and infected with 45 +/- 2 cercariae of S. japonicum after three immunizations. Forty-two days post-infection, the worms and eggs of S. japonicum were recovered from the portal vein and liver tissue, respectively. The worm and egg reduction rates were calculated respectively. A 530 bp of specific DNA fragment was amplified from mRNA of S. japonicum by RT-PCR, which was the open reading frame (ORF) encoding SjHMGBlprotein confirmed by DNA sequencing analysis. The recombinant expression plasmid SjHMGB1-pET28a was constructed by cloning the ORF of SjHMGB1 into a expression vector pET28a(+). The bacterium transformants containing the recombinant plasmid expressed a soluble recombinant protein about 28 kDa after induced by IPTG, and the recombinant SjHMGB1 protein was purified by nickel chelating affinity chromatography. The gel retard experiment showed that the recombinant SjHMGB1 protein could bind to both supercoiled DNA and linear DNA, and the recombinant protein immunized mice produced high titers of antiserum IgG. Western bloting indicated that the recombinant SjHMGB1 protein was recognized specifically by the S. japonicum-infected mice serum. Above results showed that the recombinant SjHMGB1 protein possessed both functional activity and immunogenicity as the natural protein. RT-PCR and Western blot results showed that SjHMGB1 was abundantly expressed in the adult and egg stages whereas barely detectable in the cercaria stage. The immune protection experiment showed that the recombinant SjHMGB1 induced mice to produce high titers of specific antibody IgG but failed to conduct an effective immune protection against S. japonicum. The gene encoding HMGB1 from S. japonicum and the soluble recombinant SjHMGB1 protein with natural functional activity are obtained, and the recombinant SjHMGB1 has a high immunogenicity but is not able to induce an effective immune protection against S. japonicum.


Antibody nanoparticle dispersions formed with mixtures of crowding molecules retain activity and in vivo bioavailability.
Monoclonal antibodies continue to command a large market for treatment of a variety of diseases. In many cases, the doses required for therapeutic efficacy are large, limiting options for antibody delivery and administration. We report a novel formulation strategy based on dispersions of antibody nanoclusters that allows for subcutaneous injection of highly concentrated antibody (≈ 190 mg/mL). A solution of monoclonal antibody 1B7 was rapidly frozen and lyophilized using a novel spiral-wound in-situ freezing technology to generate amorphous particles. Upon gentle stirring, a translucent dispersion of approximately 430 nm protein clusters with low apparent viscosity (≈ 24 cp) formed rapidly in buffer containing the pharmaceutically acceptable crowding agents such as trehalose, polyethylene glycol, and n-methyl-2-pyrrolidone. Upon in vitro dilution of the dispersion, the nanoclusters rapidly reverted to monomeric protein with full activity, as monitored by dynamic light scattering and antigen binding. When administered to mice as an intravenous solution, subcutaneous solution, or subcutaneous dispersion at similar (4.6-7.3 mg/kg) or ultra-high dosages (51.6 mg/kg), the distribution and elimination kinetics were within error and the protein retained full activity. Overall, this method of generating high-concentration, low-viscosity dispersions of antibody nanoclusters could lead to improved administration and patient compliance, providing new opportunities for the biotechnology industry.


Characterization of AKR murine leukemia virus sequences in AKR mouse substrains and structure of integrated recombinant genomes in tumor tissues.
A specific cDNA probe of AKR murine leukemia virus (AKR-MLV) was prepared to detect AKR-MLV sequences in normal and tumor tissues in a variety of AKR mouse substrains. AKR strains contained up to six endogenous AKR-MLV genomes. All substrains tested had one AKR-MLV locus in common, and closely related substrains had several proviruses integrated in an identical site. Virus-induced tumors in the AKR/FuRdA and AKR/JS strains showed a reintegration pattern of AKR-MLV sequences unique for the individual animal, suggesting a monoclonal origin for the outgrown tumors. An analysis of tumor DNAs from the AKR/FuRdA and AKR/JS substrains with restriction enzymes cleaving within the proviral genome revealed a new EcoRI restriction site and BamHI restriction site not present in normal tissues. The positions of these sites corresponded both with cleavage sites of EcoRI and BamHI in integrated Moloney recombinants and with the structure of isolated AKR mink cell focus-forming viruses. All tumors analyzed to data contain nearly identical integrated recombinant genomes, suggesting a causal relationship between the formation of recombinants and the leukemogenic process.


Cognitive load during speech perception in noise: the influence of age, hearing loss, and cognition on the pupil response.
The aim of the present study was to evaluate the influence of age, hearing loss, and cognitive ability on the cognitive processing load during listening to speech presented in noise. Cognitive load was assessed by means of pupillometry (i.e., examination of pupil dilation), supplemented with subjective ratings. Two groups of subjects participated: 38 middle-aged participants (mean age = 55 yrs) with normal hearing and 36 middle-aged participants (mean age = 61 yrs) with hearing loss. Using three Speech Reception Threshold (SRT) in stationary noise tests, we estimated the speech-to-noise ratios (SNRs) required for the correct repetition of 50%, 71%, or 84% of the sentences (SRT50%, SRT71%, and SRT84%, respectively). We examined the pupil response during listening: the peak amplitude, the peak latency, the mean dilation, and the pupil response duration. For each condition, participants rated the experienced listening effort and estimated their performance level. Participants also performed the Text Reception Threshold (TRT) test, a test of processing speed, and a word vocabulary test. Data were compared with previously published data from young participants with normal hearing. Hearing loss was related to relatively poor SRTs, and higher speech intelligibility was associated with lower effort and higher performance ratings. For listeners with normal hearing, increasing age was associated with poorer TRTs and slower processing speed but with larger word vocabulary. A multivariate repeated-measures analysis of variance indicated main effects of group and SNR and an interaction effect between these factors on the pupil response. The peak latency was relatively short and the mean dilation was relatively small at low intelligibility levels for the middle-aged groups, whereas the reverse was observed for high intelligibility levels. The decrease in the pupil response as a function of increasing SNR was relatively small for the listeners with hearing loss. Spearman correlation coefficients indicated that the cognitive load was larger in listeners with better TRT performances as reflected by a longer peak latency (normal-hearing participants, SRT50% condition) and a larger peak amplitude and longer response duration (hearing-impaired participants, SRT50% and SRT84% conditions). Also, a larger word vocabulary was related to longer response duration in the SRT84% condition for the participants with normal hearing. The pupil response systematically increased with decreasing speech intelligibility. Ageing and hearing loss were related to less release from effort when increasing the intelligibility of speech in noise. In difficult listening conditions, these factors may induce cognitive overload relatively early or they may be associated with relatively shallow speech processing. More research is needed to elucidate the underlying mechanisms explaining these results. Better TRTs and larger word vocabulary were related to higher mental processing load across speech intelligibility levels. This indicates that utilizing linguistic ability to improve speech perception is associated with increased listening load.


The shadow price of CO2 emissions in China's iron and steel industry.
As China becomes the world's largest energy consumer and CO2 emitter, there has been a rapidly emerging literature on estimating China's abatement cost for CO2 using a distance function approach. However, the existing studies have mostly focused on the cost estimates at macro levels (provinces or industries) with few examining firm-level abatement costs. No work has attempted to estimate the abatement cost of CO2 emissions in the iron and steel industry. Although some have argued that the directional distance function (DDF) is more appropriate in the presence of bad output under regulation, the choice of directions is largely arbitrary. This study provides the most up-to-date estimate of the shadow price of CO2 using a unique dataset of China's major iron and steel enterprises in 2014. The paper uses output quadratic DDF and investigates the impact of using different directional vectors representing different carbon mitigation strategies. The results show that the mean CO2 shadow price of China's iron and steel enterprises is very sensitive to the choice of direction vectors. The average shadow prices of CO2 are 407, 1226 and 6058Yuan/tonne respectively for the three different direction vectors. We also find substantial heterogeneity in the shadow prices of CO2 emissions among China's major iron and steel enterprises. Larger, listed enterprises are found to be associated lower CO2 shadow prices than smaller, unlisted enterprises.


Transparent and Flexible Mayan-Pyramid-based Pressure Sensor using Facile-Transferred Indium tin Oxide for Bimodal Sensor Applications.
Transparent and conducting flexible electrodes have been successfully developed over the last few decades due to their potential applications in optoelectronics. However, recent developments in smart electronics, such as a direct human-machine interface, health-monitoring devices, motion-tracking sensors, and artificially electronic skin also require materials with multifunctional properties such as transparency, flexibility and good portability. In such devices, there remains room to develop transparent and flexible devices such as pressure sensors or temperature sensors. Herein, we demonstrate a fully transparent and flexible bimodal sensor using indium tin oxide (ITO), which is embedded in a plastic substrate. For the proposed pressure sensor, the embedded ITO is detached from its Mayan-pyramid-structured silicon mold by an environmentally friendly method which utilizes water-soluble sacrificial layers. The Mayan-pyramid-based pressure sensor is capable of six different pressure sensations with excellent sensitivity in the range of 100 Pa-10 kPa, high endurance of 105 cycles, and good pulse detection and tactile sensing data processing capabilities through machine learning (ML) algorithms for different surface textures. A 5 × 5-pixel pressure-temperature-based bimodal sensor array with a zigzag-shaped ITO temperature sensor on top of it is also demonstrated without a noticeable interface effect. This work demonstrates the potential to develop transparent bimodal sensors that can be employed for electronic skin (E-skin) applications.


Longer-Term Omega-3 LCPUFA More Effective Adjunct Therapy for Tuberculosis Than Ibuprofen in a C3HeB/FeJ Tuberculosis Mouse Model.
Advancement in the understanding of inflammation regulation during tuberculosis (TB) treatment has led to novel therapeutic approaches being proposed. The use of immune mediators like anti-inflammatory and pro-resolving molecules for such, merits attention. Drug repurposing is a widely used strategy that seeks to identify new targets to treat or manage diseases. The widely explored nonsteroidal anti-inflammatory drug (NSAID) ibuprofen and a more recently explored pharmaconutrition therapy using omega-3 long-chain polyunsaturated fatty acids (n-3 LCPUFAs), have the potential to modulate the immune system and are thus considered potential repurposed drugs in this context. These approaches may be beneficial as supportive therapy to the already existing treatment regimen to improve clinical outcomes. Here, we applied adjunct ibuprofen and n-3 LCPUFA therapy, respectively, with standard anti-TB treatment, in a C3HeB/FeJ murine model of TB. Bacterial loads, lung pathology, lung cytokines/chemokines and lung lipid mediators were measured as outcomes. Lung bacterial load on day 14 post-treatment (PT) was lower in the n-3 LCPUFA, compared to the ibuprofen group (p = 0.039), but was higher in the ibuprofen group than the treated control group (p = 0.0315). Treated control and ibuprofen groups had more free alveolar space initially as compared to the n-3 LCPUFA group (4 days PT, p= 0.0114 and p= 0.002, respectively); however, significantly more alveolar space was present in the n-3 LCPUFA group as compared to the ibuprofen group by end of treatment (14 days PT, p = 0.035). Interleukin 6 (IL-6) was lower in the ibuprofen group as compared to the treated control, EPA/DHA and untreated control groups at 4 days PT (p = 0.019, p = 0.019 and p = 0.002, respectively). Importantly, pro-resolving EPA derived 9-HEPE, 11-HEPE, 12-HEPE and 18-HEPE lipid mediators (LMs) were significantly higher in the EPA/DHA group as compared to the ibuprofen and treated control groups. This suggests that n-3 LCPUFAs do improve pro-resolving and anti-inflammatory properties in TB, and it may be safe and effective to co-administer as adjunct therapy with standard TB treatment, particularly longer-term. Also, our results show host benefits upon short-term co-administration of ibuprofen, but not throughout the entire TB treatment course.


Treatment of infections by staphylococci and other gram-positive bacteria with teicoplanin: an open study.
Teicoplanin, 200-400 mg (3-6 mg/kg) daily iv or im, was used to treat 71 episodes of infection. The average duration of treatment was 22 days. The 64 evaluable episodes comprised 24 skin/soft tissue, 20 osteoarticular, ten urinary tract and one ventriculo-atrial shunt infections; one case of primary bacteraemia, three of endocarditis, two of pneumonia and three of pleural empyema. Fifty-five episodes were treated with teicoplanin monotherapy and nine with teicoplanin in association to other antibiotics. Overall 61% (39/64) of the evaluable infections were cured, 25% (16/64) improved and 14% (9/64) failed. Staphylococcus aureus was the most frequent pathogen, with 46 isolates. Infections by both methicillin-sensitive and resistant Staph. aureus strains showed favourable clinical and microbiological responses to teicoplanin. Side effects were observed in eight of the 64 episodes (12.5%). Bronchospasm was observed in two other cases at the beginning of therapy and the antibiotic administration was discontinued. Teicoplanin is an effective and well tolerated antibiotic for infections by Gram-positive bacteria, and it is effective against methicillin-resistant staphylococci.


A note on the accuracy of PAC-likelihood inference with microsatellite data.
Stephens and Donnelly have introduced a simple yet powerful importance sampling scheme for computing the likelihood in population genetic models. Fundamental to the method is an approximation to the conditional probability of the allelic type of an additional gene, given those currently in the sample. As noted by Li and Stephens, the product of these conditional probabilities for a sequence of draws that gives the frequency of allelic types in a sample is an approximation to the likelihood, and can be used directly in inference. The aim of this note is to demonstrate the high level of accuracy of "product of approximate conditionals" (PAC) likelihood when used with microsatellite data. Results obtained on simulated microsatellite data show that this strategy leads to a negligible bias over a wide range of the scaled mutation parameter theta. Furthermore, the sampling variance of likelihood estimates as well as the computation time are lower than that obtained with importance sampling on the whole range of theta. It follows that this approach represents an efficient substitute to IS algorithms in computer intensive (e.g. MCMC) inference methods in population genetics.


Recipient hemodynamics during non-heart-beating donor kidney transplantation are major predictors of primary nonfunction.
Non-heart-beating donor (NHBD) kidneys may substantially expand the donor pool, but many transplant centers are reluctant to use these kidneys because of the relatively high incidence of primary nonfunction (PNF). In heart-beating donor kidneys, intravascular fluid depletion during transplant surgery is associated with delayed graft function (DGF). Therefore, we studied the effect of the recipients' hemodynamic status on the outcome of 177 NHBD kidney transplantations. Independent statistically significant predictors of PNF were average central venous pressure (CVP) below 6 cmH(2)O (adjusted odds ratio (AOR) 3.1 (95% CI: 1.4-7.1), p=0.007), average systolic blood pressure below 110 mmHg (AOR 2.6 (95% CI: 1.1-5.9), p=0.03) and pre-operative diastolic blood pressure below 80 mmHg (AOR 2.4 (95% CI: 1.0-5.9), p=0.05). Donor characteristics were not independently associated with PNF (p>0.10). In a subgroup analysis of 56 paired kidneys, 29% of the recipients with the lower CVP of the pair experienced PNF compared with 11% of their counterparts with higher CVP (p=0.09). Our study indicates that recipient hemodynamics during transplant surgery are major predictors of PNF. Therefore, improving recipient hemodynamics by expansion of the intravascular volume is expected to enhance the results of NHBD kidney transplantations and may enlarge the donor pool by increasing the acceptance of NHBD kidneys.


The role of oral processing in dynamic sensory perception.
Food oral processing is not only important for the ingestion and digestion of food, but also plays an important role in the perception of texture and flavor. This overall sensory perception is dynamic and occurs during all stages of oral processing. However, the relationships between oral operations and sensory perception are not yet fully understood. This article reviews recent progress and research findings on oral food processing, with a focus on the dynamic character of sensory perception of solid foods. The reviewed studies are discussed in terms of both physiology and food properties, and cover first bite, mastication, and swallowing. Little is known about the dynamics of texture and flavor perception during mastication and the importance on overall perception. Novel approaches use time intensity and temporal dominance techniques, and these will be valuable tools for future research on the dynamics of texture and flavor perception.


Addressing medically underserved populations through maternal-fetal transport: a geographic analysis.
Objective: Despite regionalization of maternal care in the USA, there is little contemporary information on characteristics and utilization of maternal-fetal transport. We used geographic analysis to investigate referral and transportation patterns of the maternal-fetal transport service at our institution.Methods: This is a retrospective cohort study of all calls taken by our maternal-fetal transfer service. Call logs were abstracted, and diagnoses, gestational ages, referring hospital, and mode of transportation were analyzed. The United States Health Resources and Services Administration's Medically Underserved Area (MUA) designations were used to identify hospitals in these areas. Geographic information system software was used to analyze and map geographic variables, including frequency of transfers from each hospital, distance traveled, mode of transfer, and MUAs.Results: From November 2012 to March 2017, there were 835 telephone consults without transfer and 1682 patients transferred from 103 hospitals. Preterm labor was the most common diagnosis (n = 338, 20%), followed by hypertensive disorders (n = 231, 14%). There were 738 transfers (44%) from MUAs, and 20 (19%) of hospitals were critical access hospitals, accounting for 121 (7%) transfers. One-way trips from a referring hospital accounted for 659 patients, and the transport team was dispatched in 1023 cases. The median distance traveled was 24 mi. For hospitals within 50 mi, the mean difference in transport time between air and ground was <1 h, yet there were 73 air round trips for hospitals <50 mi from our hospital.Conclusion: The transfer system is robust and supports underserved hospitals; however, the service could be deployed more efficiently.


Apathy as a behavioural marker of cognitive impairment in Parkinson's disease: a longitudinal analysis.
Understanding the longitudinal course of non-motor symptoms, and finding markers to predict cognitive decline in Parkinson's disease (PD), are priorities. Previous work has demonstrated that apathy is one of the only behavioural symptoms that differentiates people with PD and intact cognition from those with mild cognitive impairment (MCI-PD). Other psychiatric symptoms emerge as dementia in PD develops. We explored statistical models of longitudinal change to detect apathy as a behavioural predictor of cognitive decline in PD. We followed 104 people with PD intermittently over 2 years, undertaking a variety of motor, behavioural and cognitive measures. We applied a linear mixed effects model to explore behavioural factors associated with cognitive change over time. Our approach goes beyond conventional modelling based on a random-intercept and slope approach, and can be used to examine the variability in measures within individuals over time. Global cognitive scores worsened during the two-year follow-up, whereas the longitudinal evolution of self-rated apathy scores and other behavioural measures was negligible. Level of apathy was negatively (- 0.598) correlated with level of cognitive impairment and participants with higher than average apathy scores at baseline also had poorer cognition. The model indicated that departure from the mean apathy score at any point in time was mirrored by a corresponding departure from average global cognitive score. High levels of apathy are predictive of negative cognitive and behavioural outcomes over time, suggesting that apathy may be a behavioural indicator of early cognitive decline. This has clinical and prognostic implications.


Nuclear Magnetic Resonance and Metadynamics Simulations Reveal the Atomistic Binding of l-Serine and O-Phospho-l-Serine at Disordered Calcium Phosphate Surfaces of Biocements.
Interactions between biomolecules and structurally disordered calcium phosphate (CaP) surfaces are crucial for the regulation of bone mineralization by noncollagenous proteins, the organization of complexes of casein and amorphous calcium phosphate (ACP) in milk, as well as for structure-function relationships of hybrid organic/inorganic interfaces in biomaterials. By a combination of advanced solid-state NMR experiments and metadynamics simulations, we examine the detailed binding of O-phospho-l-serine (Pser) and l-serine (Ser) with ACP in bone-adhesive CaP cements, whose capacity of gluing fractured bone together stems from the close integration of the organic molecules with ACP over a subnanometer scale. The proximity of each carboxy, aliphatic, and amino group of Pser/Ser to the Ca2+ and phosphate species of ACP observed from the metadynamics-derived models agreed well with results from heteronuclear solid-state NMR experiments that are sensitive to the 13C-31P and 15N-31P distances. The inorganic/organic contacts in Pser-doped cements are also contrasted with experimental and modeled data on the Pser binding at nanocrystalline HA particles grown from a Pser-bearing aqueous solution. The molecular adsorption is driven mainly by electrostatic interactions between the negatively charged carboxy/phosphate groups and Ca2+ cations of ACP, along with H bonds to either protonated or nonprotonated inorganic phosphate groups. The Pser and Ser molecules anchor at their phosphate/amino and carboxy/amino moieties, respectively, leading to an extended molecular conformation across the surface, as opposed to an "upright standing" molecule that would result from the binding of one sole functional group.


Coupled oscillator model of the dopaminergic neuron of the substantia nigra.
Calcium imaging using fura-2 and whole cell recording revealed the effective location of the oscillator mechanism on dopaminergic neurons of the substantia nigra, pars compacta, in slices from rats aged 15-20 days. As previously reported, dopaminergic neurons fired in a slow rhythmic single spiking pattern. The underlying membrane potential oscillation survived blockade of sodium currents with TTX and was enhanced by blockade of voltage-sensitive potassium currents with TEA. Calcium levels increased during the subthreshold depolarizing phase of the membrane potential oscillation and peaked at the onset of the hyperpolarizing phase as expected if the pacemaker potential were due to a low-threshold calcium current and the hyperpolarizing phase to calcium-dependent potassium current. Calcium oscillations were synchronous in the dendrites and soma and were greater in the dendrites than in the soma. Average calcium levels in the dendrites overshot steady-state levels and decayed over the course of seconds after the oscillation was resumed after having been halted by hyperpolarizing currents. Average calcium levels in the soma increased slowly, taking many cycles to achieve steady state. Voltage clamp with calcium imaging revealed the voltage dependence of the somatic calcium current without the artifacts of incomplete spatial voltage control. This showed that the calcium current had little or no inactivation and was half-maximal at -40 to -30 mV. The time constant of calcium removal was measured by the return of calcium to resting levels and depended on diameter. The calcium sensitivity of the calcium-dependent potassium current was estimated by plotting the slow tail current against calcium concentration during the decay of calcium to resting levels at -60 mV. A single compartment model of the dopaminergic neuron consisting of a noninactivating low-threshold calcium current, a calcium-dependent potassium current, and a small leak current reproduced most features of the membrane potential oscillations. The same currents much more accurately reproduced the calcium transients when distributed uniformly along a tapering cable in a multicompartment model. This model represented the dopaminergic neuron as a set of electrically coupled oscillators with different natural frequencies. Each frequency was determined by the surface area to volume ratio of the compartment. This model could account for additional features of the dopaminergic neurons seen in slices, such as slow adaptation of oscillation frequency and may produce irregular firing under different coupling conditions.


Directional and nondirectional spectral reflection from the human fovea.
A model of the directional and nondirectional reflection spectrum of the human fovea is developed, incorporating reflectors, absorbers, and a wavelength-dependent optical Stiles-Crawford effect (OSCE). Data from 102 healthy subjects between 18 and 75 years obtained with the fundus reflection analyzer (FRA), an imaging spectrograph that measures the directional reflection profile of the human fovea in the pupil plane from 400 to 950 nm, were analyzed. Subgroups of young (<40 years) and old (>50 years) observers were defined. Mean results of the young group defined a template for directionality versus wavelength. For the whole group, mean reflection at 550 nm from the cones was 2.12%, from the retinal pigment epithelium 0.56%, and from the choroid 7.92%. Lens density, cone disc reflection, and blood layer thickness showed significant trends versus age. The model for the first time simultaneously describes the spectra of the directional and nondirectional reflection of the human fovea. Rayleigh scatter losses of the media and in preretinal layers were assumed zero in the nondirectional pathway. Mean density of the macular pigment of a subgroup (53 subjects, 19 to 75 years) correlated significantly with independent data from reflectance and autoflourescence images obtained by scanning laser ophthalmoscope (SLO) and data from flicker photometry.


Phosphatidylcholine and cholesterol inhibit phosphatidate-mediated calcium traversal of liposomal bilayers.
Rates of phosphatidic acid- (PA-) mediated Ca2+-traversal are maximal in 'passive bilayers' void of lipid CO and OH groups: dietherphosphatidylcholine (diether-PC) or OH-blocked cholesterol liposomes. Phosphatidylcholine (PC) as bilayer matrix causes 99% inhibition, while 45 mol% cholesterol in passive bilayers inhibits by about 70%. Possibly, the absence of CO and OH groups causes a dehydration of the 'hydrogen belts', i.e., the membrane strata occupied by hydrogen bond acceptors (CO of phospholipids) and donors (OH of cholesterol, sphingosine) and thereby facilitates the formation of dehydrated Ca(PA)2, the ionophoric vehicle; or (our preferred explanation) PC engages in a (non-ionophoric) Ca(PA X PC) complex and thus reduces the concentration of the ionophore, while cholesterol competes with Ca2+ for the CO groups of phosphatidic acid by hydrogen-bonding. The Ca2+-traversal rates realized in bilayers with modified hydrogen belts lend support to the speculation that a Ca(PA)2 ferry may be of physiological importance, e.g., in membranes (such as myelin) containing much ether phospholipid (plasmalogen); and that Ca2+-membrane association and traversal may be controlled by the composition of the hydrogen belts.


Ursolic acid attenuates oxidative stress-mediated hepatocellular carcinoma induction by diethylnitrosamine in male Wistar rats.
Hepatocellular carcinoma is the most common primary cancer of the liver in Asian countries. For more than a decade natural dietary agents including fruits, vegetables and spices have drawn a great deal of attention in the prevention of diseases, preferably cancer. Ursolic acid is a natural triterpenoid widely found in food, medicinal herbs, apple peel and other products it has been extensively studied for its anticancer and antioxidant properties. The purpose of this study was to evaluate the effect of ursolic acid in diethylnitrosamine (DEN) induced and phenobarbital promoted hepatocarcinogenesis in male Wistar rats. Antioxidant status was assessed by alterations in level of lipid peroxides and protein carbonyls. Damage to plasma membranes was assessed by levels of membrane and tissue ATPases. Liver tissue was homogenized and utilized for estimation of lipid peroxides, protein carbonyls and glycoproteins. Anticoagulated blood was utilized for erythrocyte membrane isolation. Oral administration of UA 20 mg/kg bodyweight for 6 weeks decreased the levels of lipid peroxides and protein carbonyls at a significance of p< 0.05. Activities of membrane and tissue ATPases returned to normal after UA administration. Levels of glycoproteins were also restored after treatment. Histopathological observations were recorded. The findings from the above study suggest the effectiveness of UA in reducing the oxidative stress mediated changes in liver of rats. Since UA has been found to be a potent antioxidant, it can be suggested as an excellent chemopreventive agent in overcoming diseases like cancer which are mediated by free radicals.


Mechanisms Underlying the Selective Therapeutic Efficacy of Carbamazepine for Attenuation of Trigeminal Nerve Injury Pain.
Different peripheral nerve injuries cause neuropathic pain through distinct mechanisms. Even the site of injury may impact underlying mechanisms, as indicated by the clinical finding that the antiseizure drug carbamazepine (CBZ) relieves pain because of compression injuries of trigeminal but not somatic nerves. We leveraged this observation in the present study hypothesizing that because CBZ blocks voltage-gated sodium channels (VGSCs), its therapeutic selectivity reflects differences between trigeminal and somatic nerves with respect to injury-induced changes in VGSCs. CBZ diminished ongoing and evoked pain behavior in rats with chronic constriction injury (CCI) to the infraorbital nerve (ION) but had minimal effect in rats with sciatic nerve CCI. This difference in behavior was associated with a selective increase in the potency of CBZ-induced inhibition of compound action potentials in the ION, an effect mirrored in human trigeminal versus somatic nerves. The increase in potency was associated with a selective increase in the efficacy of the NaV1.1 channel blocker ICA-121431 and NaV1.1 protein in the ION, but no change in NaV1.1 mRNA in trigeminal ganglia. Importantly, local ICA-121431 administration reversed ION CCI-induced hypersensitivity. Our results suggest a novel therapeutic target for the treatment of trigeminal neuropathic pain.SIGNIFICANCE STATEMENT This study is based on evidence of differences in pain and its treatment depending on whether the pain is above (trigeminal) or below (somatic) the neck, as well as evidence that voltage-gated sodium channels (VGSCs) may contribute to these differences. The focus of the present study was on channels underlying action potential propagation in peripheral nerves. There were differences between somatic and trigeminal nerves in VGSC subtypes underlying action potential propagation both in the absence and presence of injury. Importantly, because the local block of NaV1.1 in the trigeminal nerve reverses nerve injury-induced mechanical hypersensitivity, the selective upregulation of NaV1.1 in trigeminal nerves suggests a novel therapeutic target for the treatment of pain associated with trigeminal nerve injury.


Impact of an online survivorship primer on clinician knowledge and intended practice changes.
The number of adult cancer survivors in the USA is expected to double by the year 2050. A call for increased survivorship care and provider training came from the Institute of Medicine (IOM) in the form of a landmark report in 2006. A shortage of physicians complicates the burden of survivorship care. The purpose of this effort was to design, develop, and evaluate a fully accredited, evidence-based continuing medical education (CME) and continuing education (CE) intervention to address the established knowledge gap for breast cancer, prostate cancer, colorectal cancer, and non-Hodgkin lymphoma survivorship education. Delivered through the Medscape Education (WebMD) platform, the course covered epidemiology, survivor issues, and currently available guidelines using illustrative patient cases. Knowledge gain was evaluated using a pretest-posttest design. Program evaluation was assessed by survey. Additional areas examined included post-intervention inquiry regarding expected changes to clinical practice. The results of this educational intervention demonstrated the effectiveness of internet-based CME/CE for cancer survivorship. Learning gain was significant (p < 0.0005). Effect size (d = 1.71) suggested extremely high practical significance, as the difference between the means was larger than 1 standard deviation. Significant knowledge gains were observed for each survivorship knowledge question across all clinical specialties studied. Nearly 100% of participants agreed that the course contributed to survivorship care and was organized effectively. Participants reported that the course was designed effectively (97.2%), and 68.1% responded in favor of adopting alternative communication strategies with patients and families upon completion of the course.


In vitro and ex vivo testing of tenofovir shows it is effective as an HIV-1 microbicide.
Tenofovir gel has entered into clinical trials for use as a topical microbicide to prevent HIV-1 infection but has no published data regarding pre-clinical testing using in vitro and ex vivo models. To validate our findings with on-going clinical trial results, we evaluated topical tenofovir gel for safety and efficacy. We also modeled systemic application of tenofovir for efficacy. Formulation assessment of tenofovir gel included osmolality, viscosity, in vitro release, and permeability testing. Safety was evaluated by measuring the effect on the viability of vaginal flora, PBMCs, epithelial cells, and ectocervical and colorectal explant tissues. For efficacy testing, PBMCs were cultured with tenofovir or vehicle control gels and HIV-1 representing subtypes A, B, and C. Additionally, polarized ectocervical and colorectal explant cultures were treated apically with either gel. Tenofovir was added basolaterally to simulate systemic application. All tissues were challenged with HIV-1 applied apically. Infection was assessed by measuring p24 by ELISA on collected supernatants and immunohistochemistry for ectocervical explants. Formulation testing showed the tenofovir and vehicle control gels were >10 times isosmolar. Permeability through ectocervical tissue was variable but in all cases the receptor compartment drug concentration reached levels that inhibit HIV-1 infection in vitro. The gels were non-toxic toward vaginal flora, PBMCs, or epithelial cells. A transient reduction in epithelial monolayer integrity and epithelial fracture for ectocervical and colorectal explants was noted and likely due to the hyperosmolar nature of the formulation. Tenofovir gel prevented HIV-1 infection of PBMCs regardless of HIV-1 subtype. Topical and systemic tenofovir were effective at preventing HIV-1 infection of explant cultures. These studies provide a mechanism for pre-clinical prediction of safety and efficacy of formulated microbicides. Tenofovir was effective against HIV-1 infection in our algorithm. These data support the use of tenofovir for pre-exposure prophylaxis.


Uncertainty and health-related quality of life 1 year after coronary angiography.
Little is known about illness-related uncertainty and decreased health-related quality of life in patients undergoing initial coronary angiography or about the long-term effects of uncertainty. To compare patients with and without high levels of uncertainty before angiography and to examine the influence of uncertainty on health-related quality of life 1 year after angiography. In a prospective, longitudinal study, measurements of perceived control, uncertainty, affective distress, and health-related quality of life were collected from 93 patients before angiography (baseline) and 1 year later. At baseline, patients were classified into high- and low-uncertainty groups by median split. At 1 year, analysis of variance was used to compare health-related quality of life and psychological outcomes in the 2 groups, and multiple linear regression with stepwise entry was used to identify independent determinants of health-related quality of life. Compared with patients with low baseline uncertainty, patients with high baseline uncertainty had higher levels of anxiety and depression and lower levels of perceived control and health-related quality of life 1 year after angiography. Baseline health-related quality of life, uncertainty, and life stress accounted for 54% of the variance in health-related quality of life, even when angiographic outcome was controlled for (P < .001). Baseline uncertainty was independently associated with health-related quality of life (beta = -0.25; 95% confidence interval, -9.40 to -0.05; P = .02). At initial angiography, high levels of uncertainty about illness portend negative health-related quality of life outcomes up to 1 year later.


Porphyrin-functionalized dendrimers: synthesis and application as recyclable photocatalysts in a nanofiltration membrane reactor.
The convergent synthesis of a series of porphyrin-functionalized pyrimidine dendrimers has been accomplished by a procedure involving the nucleophilic aromatic substitution (NAS) as a key reaction step. The resulting dendritic porphyrin catalysts show high activity in the light-induced generation of singlet oxygen ((1)O2) from ground-state oxygen. These materials are synthetically useful photosensitizers for the oxidation of various olefinic compounds to the corresponding allylic hydroperoxides. Catalytic activities and regio- and stereoselectivities of the dendritic photosensitizers are comparable to those observed for mononuclear porphyrin catalysts. Recycling of the dendrimer-enlarged homogeneous photocatalysts was possible by solvent-resistant nanofiltration (SRNF) by using an oxidatively stable membrane consisting of a polysiloxane polymer and ultrastable Y zeolite as inorganic filler. Moreover, this membrane technology provides a safe way to isolate the hydroperoxide products under very mild conditions. The membrane showed high retention for the macromolecular catalysts, even in chlorinated solvents, but some oxidative degradation of the porphyrin units of the dendrimer was observed over multiple catalytic runs.


OsGSTU34, a Bz2-like anthocyanin-related glutathione transferase transporter, is essential for rice (Oryza sativa L.) organs coloration.
Anthocyanins are a flavonoid compound known as one of the most important chromogenic substances. They play several functions, including health promotion and sustaining plants during adverse conditions. They are synthesized at the endoplasmic reticulum and sequestered in the vacuole. In this work, we generated knock-out lines of OsGSTU34, a glutathione transporter's tau gene family, with no transgene line and off-target through CRISPR/Cas9 mutagenesis and highlighted the loss of pigmentation in rice flowers, leaves, stems, shoots, and caryopsis. The anthocyanin quantification in the wild-type BLWT and mutant line BLG34-8 caryopsis showed that cyanidin-3-O-glucoside (C3G) and peonidin-3-O-glucoside (P3G) were almost undetectable in the mutant line. A tandem mass tag (TMT) labeling proteomic analysis was conducted to elucidate the proteomic changes in the BLWT and BLG34-8. The result revealed that 1175 proteins were altered, including 408 that were down-regulated and 767 that were upregulated. The accumulation of the OsGSTU34-related protein (Q8L576), along with several anthocyanin-related proteins, was down-regulated. The enrichment analysis showed that the down-regulated proteins were enriched in different pathways, among which the phenylpropanoid biosynthesis pathway, flavonoid biosynthesis metabolites, and anthocyanin biosynthesis pathway. Protein interaction network prediction revealed that glutathione-S-transferase (Q8L576) was connected to the proteins involved in the flavonoid and anthocyanin biosynthesis pathways, such as flavanone 3-dioxygenase 1 (Q7XM21), leucoanthocyanidin dioxygenase 1 (Q93VC3), 4-coumarate-CoA ligase 2 (Q42982), phenylalanine ammonia-lyase (P14717), chalcone synthase 1 (Q2R3A1), and 4-coumarate-CoA ligase 5 (Q6ZAC1). However, the expression of the most important anthocyanin biosynthesis gene was not altered, suggesting that only the transport mechanism was affected. Our findings highlight new insight into the anthocyanin pigmentation in black rice and provide new perspectives for future research.


Chronic work stress and decreased vagal tone impairs decision making and reaction time in jockeys.
The inverse relationship between acute stress and decision-making is well documented, but few studies have investigated the impact of chronic stress. Jockeys work exhaustive schedules and have extremely dangerous occupations, with safe performance requiring quick reaction time and accurate decision-making. We used the effort reward imbalance (ERI) occupational stress model to assess the relationship of work stress with indices of stress physiology and decision-making and reaction time. Jockeys (N=32) completed computerised cognitive tasks (Cogstate) on two occasions; September and November (naturally occurring lower and higher stress periods), either side of an acute stress test. Higher ERI was correlated with the cortisol awakening responses (high stress r=-0.37; low stress r=0.36), and with decrements in decision-making comparable to having a blood alcohol concentration of 0.08 in the high stress period (p<0.001) The LF/HF ratio of heart rate variability impacted the association of ERI with decision-making. Potentially, this may be attributed to a 'tipping point' whereby the higher ERI reported by jockeys in the high stress period decreases vagal tone, which may contribute to reduced decision-making abilities.


Isoform-specific O-glycosylation by murine UDP-GalNAc:polypeptide N-acetylgalactosaminyltransferase-T3, in vivo.
Multiple isoforms of UDP-GalNAc:polypeptide N-acetylgalactosaminyl- transferase (ppGaNTase) have been cloned and expressed from a variety of organisms. In general, these isoforms display different patterns of tissue-specific expression, but exhibit overlapping substrate specificities, in vitro . A peptide substrate, derived from the sequence of the V3 loop of the HIV gp120 protein (HIV peptide), has previously been shown to be glycosylated in vitro exclusively by the ppGaNTase-T3 (Bennett et al. , 1996). To determine if this isoform-specificity is maintained in vivo , we have examined the glycosylation of this substrate when it is expressed as a reporter peptide (rHIV) in a cell background (COS7 cells) which lacks detectable levels of the ppGaNTase-T3. Glycosylation of rHIV was greatly increased by coexpression of a recombinant ppGaNTase-T3. Overexpression of ppGaNTase-T1 yielded only partial glycosylation of the reporter. We have also determined that the introduction of a proline residue at the +3 position flanking the potential glycosylation site eliminated ppGaNTase-T3 selectivity toward rHIV observed both in vivo and in vitro .


Promoting proton coupled electron transfer in redox catalysts through molecular design.
Most bond-forming and -breaking redox reactions require the concomitant transfer of protons. Unassisted proton movement can result in kinetic and thermodynamic barriers that inhibit the rate of these reactions, leading to slow and/or inefficient catalysis. These barriers can be circumvented by effective proton management through molecular design. Different strategies for managing proton movement are discussed with examples from biological and synthetic systems. As proton management is particularly important in redox reactions for chemical fuel generation and utilization, the focus will be on catalysts for H-H and O-O bond formation and cleavage. However, we expect the approaches discussed herein will be general to most multi-electron, multi-proton reactions.


Anthropometric indices and measurements on primary school children of contrasting parental occupations.
The nutritional status in relation to parental occupation is a useful undertaking in predicting requirement of population groups. The investigation reported here was conducted to document the nutritional status of children 6-15 years of age of contrasting parental occupations living in Anambra State Community, Nigeria. The main weights of Anambra State boys at 6 years of age were 20.2 kg compared with teachers 19.5 kg and traders 15.7 kg. The weight achievement of the trader's at 15 years of age was 84% those of farmers' children. Similar conclusion may be drawn for height of both boys and girls for the contrasting occupation groups. The arm size of the farmers' children was initially low but hereafter it was similar to that of boys of other occupation groups, however the arm size of other civil servants (girls) was 26.3 cm compared with 21.1 cm for other businessmen's girls. On the other hand, the skinfold estimate of the teachers' boys (3.8 mm) and girls (3.8 mm) aged 10 years was statistically different (p less than 0.01) from the highest values (7.1 mm) for boys and (8.4 mm) girls at same age. Household diets were 90% of safe level of intake for civil servants and most farmers' children; that was accompanied by low dietary intake of riboflavin--81.3% for civil servants and 84% for farmers; thiamin 100% for civil servants and 79.3% for farmers. The Vitamin A intake of civil servants were also 85.7% safe level of adequacy. The information obtained was assessed from the point of view of its usefulness in the area surveyed for both the nutritionists and the clinicians and its long term benefit for Nigeria in National food policy and planning. These can be applied for other countries.


Amniotic membrane transplantation in ocular surface disorders.
Chronic ocular surface disorders, which can result in severe functional impairment, have been viewed for decades as untreatable diseases. In 1995, the reintroduction of amniotic membrane transplantation (AMT), either alone or associated with limbal stem cell transplantation, has offered new hope of using tissue and cell therapy strategies to repair ocular surface disorders. Amniotic membrane (AM) has been found to exert its effects by acting as a substrate for the growth of ocular surface epithelia, by suppressing inflammation and scarring and by serving as an anti-microbial barrier. Moreover, AM has recently been used as a substrate for ex vivo expansion of corneal epithelial cells for ocular surface reconstruction. Notwithstanding the substantial agreement among Authors regarding its clinical efficacy, there are still many uncertainties regarding the fate of grafted AM and consequently the mechanisms through which it exerts its long-term effects. Further studies including controlled clinical trials with numerous cases are required to understand which ocular surface conditions are certain to benefit from AM transplantation and how its mechanical properties interact with the mediators produced to favor ocular surface reconstruction.


Assessing and enhancing health care providers' response to domestic violence.
This study aimed to examine possible changes from 2008 to 2012 in the skills of health care staff in identifying and intervening in domestic violence (DV). A longitudinal descriptive study design with volunteer samples (baseline; n = 68, follow-up; n = 100) was used to acquire information regarding the present state and needs of the staff in practices related to DV. The results of the baseline survey were used as a basis for planning two interventions: staff training and drafting practical guidelines. Information was collected by questionnaires from nurses, physicians, and social workers and supplemented by responses from the interviews. The data were analysed using both quantitative and qualitative methods. A chi-square test was used to test the statistical significance of the data sets. In addition, participants' quotes are used to describe specific phenomena or issues. The comparison showed that overall a small positive change had taken place between the study periods. However, the participants were aware of their own shortcomings in identifying and intervening in DV. Changes happen slowly, and administrative support is needed to sustain such changes. Therefore, this paper offers recommendations to improve health care providers' response to DV. Moreover, there is a great need for evaluating the training programme used.


Effect of preoperative chemotherapy on the histopathological classification of gastric cancer.
In the era of individualized gastric cancer (GC) treatment, accurate determination of histological subtype becomes increasingly relevant. As yet, it is unclear whether preoperative chemotherapy may affect the histological subtype. The aim of this study was to assess concordance in histological subtype between pretreatment biopsies and surgical resection specimens before and after the introduction of perioperative treatment. Histological subtype was centrally determined in paired GC biopsies and surgical resection specimens of patients treated with either surgery alone (SA) in the Dutch D1/D2 study or with preoperative chemotherapy (CT) in the CRITICS trial. The histological subtype as determined in the resection specimen was considered the gold standard. Concordance rates and sensitivity and specificity of intestinal, diffuse, mixed, and "other" subtypes of GC were analyzed. In total, 105 and 515 pairs of GC biopsies and resection specimens of patients treated in the SA and CT cohorts, respectively, were included. Overall concordance in the histological subtype was 72% in the SA and 74% in the CT cohort and substantially higher in the diffuse subtype (83% and 86%) compared to the intestinal (70% and 74%), mixed (21% and 33%) and "other" subtypes (54% and 54%). In the SA cohort, sensitivities and specificities were 0.88 and 0.71 in the intestinal, 0.67 and 0.93 in the diffuse, 0.20 and 0.98 in the mixed, and 0.50 and 0.93 in the "other" subtypes, respectively. Our results suggest that accurate determination of histological subtype on gastric cancer biopsies is suboptimal but that the impact of preoperative chemotherapy on histological subtype is negligible.


[Interactions of inhibitor of free radical reactions and 2-deoxyribosyl macroradicals].
By steric computer modelling of five types of 2-deoxyribosyl radicals and their molecular products generated in DNA by radiation as a result of R + InH = RH + In reaction the displacement of DNA bases (maximum--for C'1 atoms and minimum for C'3 ones) has been determined. Literature data on the DNA decay in the irradiated cells, and in DNA frozen solutions as well as the data on radiolysis of compounds modelling separate DNA fragments allowed to offer a general scheme of the every 2-deoxyribosyl radical transformation and to calculate a balance between the intermediate and final molecular products (processes) of the 2-deoxyribosyl radiolysis. The DNA conformation change due to the 2-deoxyribosyl stereoisomers formation at C'3 and C'4 atoms is discussed as one of the suggested reasons for genetic radiation mutation.


Effect of dietary protein and minerals on calcium and zinc utilization.
The utilization of most minerals can be altered not only by varying the dietary levels of protein, carbohydrate, fiber, fat, and vitamins but also by varying the forms of macronutrients fed. Interactions among minerals are frequent and sometimes complex. Thus, all assessments of mineral requirements should include discussions of the effects of interactions on mineral bioavailability. Some of these interactions occur in the gut and affect the absorption of minerals. Other interactions affect the metabolism, transport and storage of minerals, and ultimately their excretion in urine or in endogenous gut secretions.


A mechanistic insight into benefits of aggregation induced emissive luminogens in cancer.
Exploration of advanced chemotheranostics that benefit from a combined in vivo strategy of cancer diagnosis and chemotherapy simultaneously is highly valued and will expose novel possibilities in modifying treatment and reduce side effects. In recent years, nanodrug delivery systems that incorporate aggregation-induced emissive luminogens (AIEgens) have been developed to track and monitor anticancer drug release, trace translocation processes and predict chemotherapeutic responses. There are several classes of AIEgen based chemotheranostics such us stimuli-responsive nanoprodrugs, pH-sensitive mesoporous silica nanocarriers, supramolecular polymer systems, drug encapsulated carriers, carrier-free nanodrugs, self-indicating drug delivery nanomachines and AIEgen-prodrug co-assembly. The present review conveys mechanistic insight into the benefits of AIEgens in the theranostic application by illustrating the recent breakthroughs in chemotheranostic nanomedicines that incorporate these unique fluorophores as signal reporters. The perspectives that can be further explored are also highlighted with the hope to instil more research interest in the advancement of AIE active cancer chemotheranostics for imaging and treatment in vivo.HIGHLIGHTSAggregation induced emissive materials (AIEgens) exhibit unique advantages over conventional luminogens for synergistic diagnosis and chemotherapy of cancer in vivo.The combination of AIE and nanotechnology offers an excellent platform to fabricate advanced chemotheranostics for cancer therapy.


Human alpha-fetoprotein and albumin: differences in free sulfhydryl groups.
The number of free sulfhydryl (SH) groups of both human alpha-fetoprotein (AFP) and albumin, isolated simultaneously from human cord serum, was determined by reacting the proteins with 5,5'-dithiobis (2-nitrobenzoic acid) (DTNB). A higher number of SH groups was consistently found with AFP than with albumin. When both proteins were pretreated with 100 mmol/l 2-mercaptoethanol, only approximately 0.5 SH groups were detected per molecule of human adult albumin, yet as many as 4 per molecule were found with AFP. Dithiothreitol was found to be more effective than 2-mercaptoethanol in SH group regeneration. The results suggested that some disulfide linkages of AFP could be reduced. Free SH groups regenerated by the treatment with SH reagent were found to disappear gradually during storage under aerobic conditions. The rate of disappearance differed between AFP and albumin.


SALL4 expression in germ cell and non-germ cell tumors: a systematic immunohistochemical study of 3215 cases.
The SALL4 transcription factor is associated with embryonic cell pluripotency and has been shown as a useful immunohistochemical marker for germ cell tumors. However, information of SALL4 distribution in normal human tissues and non-germ cell tumors is limited. In this study we examined normal human tissues and 3215 tumors for SALL4 expression using a monoclonal antibody 6E3 and automated immunohistochemistry. In a 10-week embryo, SALL4 was expressed in ovocytes, intestine, kidney, and some hepatocytes. In adult tissues, it was only detected in germ cells. SALL4 was consistently expressed in all germ cell tumors except some trophoblastic tumors and mature components of teratomas, in which it was selectively expressed in intestinal-like and some squamous epithelia. In non-germ cell carcinomas, SALL4 was detected in 20% of cases or more of serous carcinoma of the ovary, urothelial high-grade carcinoma, and gastric adenocarcinoma (especially the intestinal type). SALL4 was only rarely (≤ 5%) expressed in mammary, colorectal, prostatic, and squamous cell carcinomas. Many SALL4-positive carcinomas showed poorly differentiated patterns, and some showed positivity in most tumor cells mimicking the expression in germ cell tumors. SALL4 was commonly expressed in rhabdoid tumors of the kidney and extrarenal sites and in the Wilms tumor. Expression of SALL4 was rare in other mesenchymal and neuroendocrine tumors but was occasionally detected in melanoma, desmoplastic small round cell tumor, epithelioid sarcoma, and rhabdomyosarcoma. All hematopoietic tumors were negative. SALL4 is an excellent marker of nonteratomatous germ cell tumors, but it is also expressed in other tumors, sometimes extensively. Such expression may reflect stem cell-like differentiation and must be considered when using SALL4 as a marker for germ cell tumors. Observed lack of other pluripotency factors, OCT4 and NANOG, in SALL4-positive non-germ cell tumors can also be diagnostically helpful.


JAAD Grand Rounds quiz. Asymptomatic necrotic ulcer on leg.
At the conclusion of this learning activity, physician participants should be able to assess their own diagnostic and patient management skills and use the results of this exercise to help determine personal learning needs that can be addressed through subsequent CME involvement. Instructions for claiming CME credit appear in the front advertising section. See last page of Contents for page number. Instructions: In answering each question, refer to the specific directions provided. Because it is often necessary to provide information occurring later in a series that give away answers to earlier questions, please answer the questions in each series in sequence.


Long term exposure to environmental concentrations of diesel exhaust particles does not impact the phenotype of human bronchial epithelial cells.
Chronic exposure to diesel engine exhausts is associated with an increased risk of pulmonary diseases including lung cancer. Diesel engine exhausts contain large amounts of diesel exhaust particles (DEP) on which are adsorbed several carcinogenic compounds such as polycyclic aromatic hydrocarbons. Acute toxicity of high concentrations of DEP has been largely demonstrated in various in vitro cellular models. In contrast, the cellular and molecular impacts of low environmental concentrations of DEP on the phenotype of chronically exposed lung epithelial cells remain to be investigated. In the present study, we show that long term exposure (6 months) to 2 μg/ml (0.4 μg/cm2) DEP (standard reference material 1650b) increased cytochrome P4501A mRNA levels in the human bronchial epithelial BEAS-2B cell line. However, chronic exposure to DEP did not change cell morphology, trigger epithelial-mesenchymal transition or increase anchorage-independent cell growth. Moreover, DEP increase neither the levels of reactive oxygen species or those of γ-histone H2AX, nor the expression of interleukin-6 and interleukin-8. Our results thus demonstrate that the chronic exposure to low DEP concentrations could increase cytochrome P501A gene expression in BEAS-2B cells but did not induce molecular effects related to genotoxicity, oxidative stress or inflammation.


Monitoring mammalian mitochondrial translation with MitoRiboSeq.
Several essential components of the electron transport chain, the major producer of ATP in mammalian cells, are encoded in the mitochondrial genome. These 13 proteins are translated within mitochondria by 'mitoribosomes'. Defective mitochondrial translation underlies multiple inborn errors of metabolism and has been implicated in pathologies such as aging, metabolic syndrome and cancer. Here, we provide a detailed ribosome profiling protocol optimized to interrogate mitochondrial translation in mammalian cells (MitoRiboSeq), wherein mitoribosome footprints are generated with micrococcal nuclease and mitoribosomes are separated from cytosolic ribosomes and other RNAs by ultracentrifugation in a single straightforward step. We highlight critical steps during library preparation and provide a step-by-step guide to data analysis accompanied by open-source bioinformatic code. Our method outputs mitoribosome footprints at single-codon resolution. Codons with high footprint densities are sites of mitoribosome stalling. We recently applied this approach to demonstrate that defects in mitochondrial serine catabolism or in mitochondrial tRNA methylation cause stalling of mitoribosomes at specific codons. Our method can be applied to study basic mitochondrial biology or to characterize abnormalities in mitochondrial translation in patients with mitochondrial disorders.


Thyroid as a target of adjuvant autoimmunity/inflammatory syndrome due to mRNA-based SARS-CoV2 vaccination: from Graves' disease to silent thyroiditis.
As COVID-19 became a pandemic, the urgent need to find an effective treatment vaccine has been a major objective. Vaccines contain adjuvants which are not exempt from adverse effects and can trigger the autoimmune/inflammatory syndrome induced by adjuvants (ASIA). There is very little information about autoimmune endocrine disease and the ASIA after the use of mRNA-based SARS-CoV2 vaccination. We report three cases and also review the literature showing that the thyroid gland can be involved in the ASIA induced by the mRNA-based SARS-CoV2 vaccination. We present the first case to date of silent thyroiditis described in the context of SARS-CoV2 vaccination with Pfizer/BioNTech. Also, we discuss the first subacute thyroiditis in the context of SARS-CoV2 vaccination with the Moderna's vaccine. Finally, we provide another case to be added to existing evidence on Graves' disease occurring post-vaccination with the Pfizer/BioNTech vaccine. Adjuvants play an important role in vaccines. Their ability to increase the immunogenicity of the active ingredient is necessary to achieve the desired immune response. Both the Moderna and the Pfizer/BioNTech vaccines use mRNA coding for the SARS-CoV2 S protein enhanced by adjuvants. In addition, the cross-reactivity between SARS-CoV2 and thyroid antigens has been reported. This would explain, at least, some of the autoimmune/inflammatory reactions produced during and after SARS-CoV2 infection and vaccination. The autoimmune/inflammatory syndrome induced by adjuvants involving the thyroid could be an adverse effect of SARS-CoV2 vaccination and could be underdiagnosed.


Design and fabrication of a Gaussian fan-out optical interconnect.
A phase-retrieval algorithm is described and used to optimize the phase profile of a multilevel phase grating that achieves a two-dimensional Gaussian fan-out. Binary-optics technology is used to fabricate a surface-relief structure that implements this optimized phase profile. The results of the fabrication process are summarized, and the experimental measurements on the fan-out element are presented. Differences between theory and experiment arise from scalar-theory approximations used in the design algorithm. In order to illustrate the scalar-theory limitations, a rigorous formulation is used to analyze the diffraction efficiency and the reconstruction error of a representative binary surface-relief phase grating as a function of the period-to-wavelength ratio. The analysis serves to show qualitatively the effects of the period-to-wavelength ratio on the performance of free-space optical interconnects designed with a scalar-theory formalism.


Rapid local expression of interleukin-12, tumor necrosis factor alpha, and gamma interferon after cutaneous Francisella tularensis infection in tularemia-immune mice.
Francisella tularensis LVS is an effective live vaccine strain used for cutaneous vaccination against tularemia in man. In mice, injection of LVS causes invasive disease and subsequent development of immunity that is characterized by effective control of otherwise lethal doses of the organism. In the present investigation, it is shown that LVS-immune mice controlled an intradermal infection much more effectively than did naive mice; bacterial counts in skin samples were 1.5 to 2.0 log10 lower 24 h after injection and 6 log10 lower 72 h after injection in immune mice. Moreover, in contrast to naive mice, no bacteria were demonstrated in samples from livers and spleens of immune mice. By immunohistochemistry, skin samples from immune mice showed an intense staining for interleukin-12 (IL-12) and a moderate staining for tumor necrosis factor alpha (TNF-alpha) at 24 h postinoculation, after which staining for both cytokines faded. In naive mice, the staining for IL-12 was weak at all time points and no staining for TNF-alpha was observed. No staining for gamma interferon (IFN-gamma) was observed in any group before 72 h. At that time point, skin samples from immune mice showed moderate staining and skin samples from naive mice showed weak staining. Reverse transcriptase PCR showed an induction of mRNA of the three cytokines in the skin within the first day after injection. A quantitative analysis demonstrated higher IFN-gamma and TNF-alpha mRNA levels in immune mice at 24 h postinoculation. In conclusion, immunization with F. tularensis LVS conferred a capability to respond to cutaneous reinfection, with rapid local expression of IL-12, TNF-alpha, and IFN-gamma, and this expression was paralleled by containment and mitigation of the infection. The cytokine response may be part of a local barrier function of the skin, important to host protection against tularemia.


The use of bead beating to prepare suspensions of nuclei for flow cytometry from fresh leaves, herbarium leaves, petals and pollen.
"Bead beating" is commonly used to release DNA from cells for genomic studies but it was used here to prepare suspensions of plant nuclei for measurement of DNA amounts by flow cytometry. Plant material was placed in 2-ml screw-capped tubes containing beads of zirconia/silica (2.5 mm diameter) or glass (2.5 or 1.0 mm diameter) and 1 ml of lysis buffer. The tubes were mechanically shaken with an FP120 FastPrep Cell Disrupter to release intact nuclei from plant tissue by the impact of the beads. The nuclei were then stained with propidium iodide (PI) and analyzed by flow cytometry. The method was tested using fresh leaves, fresh petals and herbarium leaves of Rosa canina, leaves and pollen of R. rugosa, and fresh leaves of Petroselinum crispum, Nicotiana tabacum, and Allium cepa. Batches of 12 samples of fresh leaves were prepared, simultaneously, in 45 s by bead beating in the Cell Disrupter. In flow cytometry histograms, nuclei of fresh leaves gave G(1)/G(0) peaks with CVs of less than 3.0% and nuclei from fresh petals and herbarium leaves of R. canina, and pollen of the generative nuclei of R. rugosa gave peaks with coefficients of variation (CVs) of less than 4.0%. DNA amounts estimated from 24-month-old herbarium leaves, using P. crispum as an internal standard, were less than those of fresh leaves by a small but significant amount. Suspensions of nuclei can be prepared rapidly and conveniently from a diversity of tissues by bead beating. Exposure of laboratory workers to harmful substances in the lysis buffer is minimized.


Diffusivity anomaly in modified Stillinger-Weber liquids.
By modifying the tetrahedrality (the strength of the three body interactions) in the well-known Stillinger-Weber model for silicon, we study the diffusivity of a series of model liquids as a function of tetrahedrality and temperature at fixed pressure. Previous work has shown that at constant temperature, the diffusivity exhibits a maximum as a function of tetrahedrality, which we refer to as the diffusivity anomaly, in analogy with the well-known anomaly in water upon variation of pressure at constant temperature. We explore to what extent the structural and thermodynamic changes accompanying changes in the interaction potential can help rationalize the diffusivity anomaly, by employing the Rosenfeld relation between diffusivity and the excess entropy (over the ideal gas reference value), and the pair correlation entropy, which provides an approximation to the excess entropy in terms of the pair correlation function. We find that in the modified Stillinger-Weber liquids, the Rosenfeld relation works well above the melting temperatures but exhibits deviations below, with the deviations becoming smaller for smaller tetrahedrality. Further we find that both the excess entropy and the pair correlation entropy at constant temperature go through maxima as a function of the tetrahedrality, thus demonstrating the close relationship between structural, thermodynamic, and dynamical anomalies in the modified Stillinger-Weber liquids.


[201-thallium myocardial scintigraphy before and after percutaneous transluminal coronary angioplasty].
We describe our experience from the first 30 patients treated with percutaneous transluminal coronary angioplasty (PTCA) in the Cardiologic Department P, Gentofte Hospital. The main purpose was to evaluate the value of thallium scintigraphy performed before and three months after PTCA. After PTCA, 23 (77%) had no angina and 22 (73%) had normal exercise-ECG. On the basis of the scintigraphies before PTCA, the patients were divided into two groups: In group 1, five had normal scintigraphy and 14 reversible perfusion defects only. In group 2, all 11 patients had both persistent and reversible defects. In patients without persistent defects, significantly more were free from angina. No difference was observed between group 1 and 2 in percentage stenosis of the vessel or in reduction of the diameter. Significant reduction in the extent of reversible perfusion defects after PTCA was observed in both groups and also in the extent of persistent defects in group 2. In about 1/3 of the patients, the changes in percentage stenosis after PTCA and in the perfusion as judged by the scintigraphies differed. Therefore, as the two methods supplement each other, the main conclusions are: 1) Evaluation of the revascularization procedure is more reliable if a scintigraphy is obtained both before and after PTCA than if only coronary arteriography is performed. 2) Scintigraphy alone suffices for the three-month follow-up examination, partly because it gives better information about the myocardial perfusion than does the arteriography, partly because scintigraphy is a noninvasive procedure.


Ovarian dynamics and host use.
Oviposition behavior in herbivorous and frugivorous insects and parasitoids is dynamic at the level of the individual, responding to variation in host quality and availability. Patterns of variation in egg load in response to host presence and quality suggest that ovarian development also responds to variation in the host environment. Ovarian dynamics are mediated by feedback from oviposition, by host feeding, and by sensory input from the host. The last of these mechanisms, host sensory cuing, is known to occur in three major orders and provides strong evidence that ovarian dynamics are adaptive by design. Conditions favoring host effects on ovarian development include trade-offs between egg production and either survival or dispersal, uncertainty in the host environment, and a correlation in host conditions between the time that oogenesis is initiated and the time that eggs are laid. Some host defenses block ovarian development, suggesting that ovarian dynamics in host-specific insects should be viewed from a coevolutionary perspective.


Outbreak of Nipah-virus infection among abattoir workers in Singapore.
In March 1999, an outbreak of encephalitis and pneumonia occurred in workers at an abattoir in Singapore. We describe the clinical presentation and the results of investigations in these patients. Clinical and laboratory data were collected by systemic review of the case records. Serum and cerebrospinal fluid (CSF) samples were tested for IgM antibodies to Nipah virus with an IgM capture ELISA. Reverse-transcriptase PCR was done on the CSF and tissue samples from one patient who died. Eleven patients were confirmed to have acute Nipah-virus infection based on raised IgM in serum. Nipah virus was identified by reverse transcriptase PCR in the CSF and tissue of the patient who died. The patients were all men, with a median age of 44 years. The commonest presenting symptoms were fever, headache, and drowsiness. Eight patients presented with signs of encephalitis (decreased level of consciousness or focal neurological signs). Three patients presented with atypical pneumonia, but one later developed hallucinations and had evidence of encephalitis on CSF examination. Abnormal laboratory findings included a low lymphocyte count (nine patients), low platelet count, low serum sodium, and high aspartate aminostransferase concentration (each observed in five patients). The CSF protein was high in eight patients and white-blood-cell count was high in seven. Chest radiography showed mild interstitial shadowing in eight patients. Magnetic resonance imaging (MRI) showed focal areas of increased signal intensity in the cortical white marker in all eight patients who were scanned. The nine patients with encephalitis received empirical treatment with intravenous aciclovir and eight survived. Infection with Nipah virus caused an encephalitis illness with characteristic focal areas of increased intensity seen on MRI. Lung involvement was also common, and the disease may present as an atypical pneumonia.


Olfactory performance after crenotherapy in chronic rhinosinusitis in the elderly.
To evaluate the effectiveness of crenotherapy on the olfactory performance of elderly patients with chronic rhinosinusitis (CRS). A longitudinal case-control study of a cohort of elderly patients affected by CRS and olfactory dysfunction assessed with the Sniffin' Sticks (Burghart Medical Technology, Wedel, Germany) (SS) olfactory test. One hundred and thirty-seven elderly subjects with CRS were divided into two groups. The investigational arm (n = 69) underwent crenotherapy with hyperthermal water, rich in mineral salts, and the control group (n = 68) underwent NaCl 0.9% both for 12 days. At baseline and at 1 and 6 months after treatment, both groups underwent ear nose and throat assessment and SS. Self-report questionnaires were administered at baseline to evaluate the patients' own olfactory response, and after treatment to evaluate their degree tolerability. Olfactory performance was then evaluated in elderly subjects with hyposmia without CRS (n = 40) and in younger subjects with both hyposmia and CRS (n = 40). No adverse reactions were reported after crenotherapy. The SS total score showed that crenotherapy induced a statistically significant improvement in the olfactory function of both the elderly and the younger subjects with hyposmia and CRS. By contrast, no improvement was observed in the control arm and in the elderly with hyposmia without CRS. All subjects showed a good degree of tolerability. We demonstrated that crenotherapy effectively improves olfactory function in elderly patients with CRS. Finally, our study suggests that crenotherapy represents a safe therapeutic strategy for the treatment of CRS and olfactory dysfunction in the elderly.


Down-regulation of cdc2 in senescent human and hamster cells.
Senescent cells fail to respond to serum-induced signals for DNA synthesis. Because a central role for the p34cdc2 protein kinase is postulated in control of the cell cycle, we examined the status of this kinase in senescent cells and other growth-arrested cells. In growing human and Syrian hamster fibroblasts, three 35S-labeled proteins of 34-36 kDa were immunoprecipitated with p34cdc2 antiserum. Only the two slower migrating forms were phosphorylated as determined by 32P labelling. In senescent cells, which failed to incorporate [3H]thymidine, no p34cdc2 protein was synthesized and very little or no cdc2 mRNA was observed. When maintained for 48 h in 0.5% serum, young cells also retained only marginal cdc2 expression. After stimulation of low serum-arrested cells by addition of 10% serum, a time-dependent increase of cdc2 mRNA was observed, whereas serum stimulation of senescent cells did not increase cdc2 mRNA. In contrast to senescent and low serum-arrested cells, cdc2 mRNA was expressed at normal levels in cells partially growth arrested by isoleucine deficiency in G1, by aphidicolin at G1-S, by etoposide in G2, or by Colcemid in the M phase of the cell cycle, indicating that cdc2 down-regulation does not always occur upon growth arrest. Following transfection of a plasmid containing the human CDC2 gene into hamster cells, expression of human cdc2 failed to overcome the block to DNA synthesis in senescent cells. Although p34cdc2 was synthesized in the transfected cells, the multiple phosphorylated forms of the proteins were not observed. Taken together, these data support the concept that a chain of events leads to senescence. While p34cdc2 kinase may be one of the critical elements, other cell cycle controls are also involved.


Lumpy skin disease outbreak in cattle population of Chattogram, Bangladesh.
Lumpy skin disease (LSD) is an important viral disease causing significant economic losses in commercial livestock production. In mid-2019, an outbreak of LSD has been reported in cattle population from different parts of Bangladesh including Chattogram division. A cross-sectional surveillance study was undertaken from August 2019 to December 2019 to investigate the prevalence and associated risk factors of LSD in cattle in Chattogram district. A total of 3,327 cattle from 19 commercial farms were examined for the LSD specific skin lesions and associated risk factors. A total of 120 skin biopsies were collected from the suspected animal for the confirmation of the disease using molecular detection and histopathological examination. Partial genome sequencing and phylogenetic analyses were performed on selected viral isolates. The overall clinical prevalence of LSD in the study population was 10% (95% confidence interval [CI]: 9.4%-11%) where the highest farm level outbreak frequency was 63.33% (95% CI: 45.51%-78.13%) and the lowest 4.22% (95% CI: 3.39%-5.25%). Crossbred and female cattle showed a significantly higher prevalence of the disease compared to their counterparts. Introduction of new animals in farms was found to be one of the most significant risk factors in the transmission of the disease. All suspected skin biopsies were positive for LSD virus (LSDV) infection with granulomatous and pyogranulomatous dermatitis was revealed on histopathology. Phylogenetic analysis based on the inverted terminal repeat region of the LSDV gene suggested that the locally circulating strain was closely related to the strains isolated from the Middle East and North African countries. The data generated in this study would be beneficial to the field veterinarians and animal health decision makers in the country as well as it will aid in taking appropriate measures to prevent further relapse or outbreak of this disease in future.


Catalyst-site-controlled coordination polymerization of polar vinyl monomers to highly syndiotactic polymers.
This contribution reports a combined synthetic, kinetic, mechanistic, and theoretical/computational study of the recently discovered catalyst-site-controlled coordination polymerization of polar vinyl monomers [such as methyl methacrylate (MMA) and N,N-dimethylacrylamide (DMAA)] into highly syndiotactic polymers. Among the 12 C(s)-ligated ansa-cyclopentadienyl (Cp)-R(2)E(C,Si)-fluorenyl (Flu) group 4 metallocene catalyst systems examined-which varied in metal center, anion structure, bridging atom and substituents, and ligand substitution pattern-cationic ansa-metallocene ester enolate catalyst 6(+)[B(C(6)F(5))(4)](-), derived from the activation of the precatalyst [Ph(2)C(Cp)(2,7-(t)Bu(2)-Flu)]Zr[OC(O(i)Pr)=CMe(2)](2) with [Ph(3)C][B(C(6)F(5))(4)], stood out as the best catalyst in all aspects of the MMA polymerization at room temperature, including the highest activity (1554 h(-1) TOF), efficiency (98% I*), syndiotacticity (94% rr), and control (predicted number-average molecular weight and 1.14 molecular weight distribution). Kinetic and mechanistic results are consistent with a catalyst-site-controlled, monometallic coordination-addition mechanism, involving fast intramolecular addition within the catalyst-monomer complex leading to the resting eight-membered ester enolate chelate, followed by the rate-limiting ring-opening of the chelate to regenerate the active species. This work has also uncovered several unique features of this polymerization system that are in marked contrast to the propylene polymerization by analogous C(s)-ligated cationic alkyl catalysts: a constant syndiotacticity of PMMA produced over a wide polymerization temperature range (i.e., from 0 degrees C, 94% rr to 25 degrees C, 94% rr to 50 degrees C, 93% rr); insensitivity of its high activity, degree of control, and stereoselectivity to solvent polarity and structure of weakly coordinating anions; and deviation from a pure site-control mechanism at high [MMA]/[catalyst] ratios. Computational results provide theoretical support for the proposed monomer-assisted, catalyst-site epimerization, after an enantiofacial mistake, to a thermodynamically more stable resting state, which accounts for the observed higher than expected [mr] contents based on a pure site-controlled mechanism. DFT calculations rationalize why the Ph(2)C< bridged catalyst 6 exhibits higher stereoselectivity than other catalysts with the Me(2)C< or Me(2)Si< bridge: the bridge rigidity pushes the eta(3)-bound Flu ligand closer to the growing chain and the monomer, thereby increasing DeltaE(stereo) between the competing transition states for the addition of a monomer molecule to the opposite (correct and wrong) enantiofaces of the enolate growing chain. The relative polymerization activity of this catalyst series is shown to correlate with the relative energetics of the back-biting of the penultimate unit and ion-pair formation.


Degradation of Styrenic Plastics during Recycling: Accommodation of PP within ABS after WEEE Plastics Imperfect Sorting.
With the development of dark polymers for industrial sorting technologies, economically profitable recycling of plastics from Waste Electrical and Electronical Equipment (WEEE) can be envisaged even in the presence of residual impurities. In ABS extracted from WEEE, PP is expected to be the more detrimental because of its important lack of compatibility. Hence, PP was incorporated to ABS at different rates (2 to 8 wt%) with a twin-screw extruder. PP was shown to exhibit a nodular morphology with an average diameter around 1-2 µm. Tensile properties were importantly diminished beyond 4 wt% but impact resistance was decreased even at 2 wt%. Both properties were strongly reduced as function of the contamination rate. Various potential compatibilizers for the ABS + 4 wt% PP system were evaluated: PPH-g-MA, PPC-g-MA, ABS-g-MA, TPE-g-MA, SEBS and PP-g-SAN. SEBS was found the most promising, leading to diminution of nodule sizes and also acting as an impact modifier. Finally, a Design Of Experiments using the Response Surface Methodology (DOE-RSM) was applied to visualize the impacts and interactions of extrusion temperature and screw speed on impact resistance of compatibilized and uncompatibilized ABS + 4 wt% PP systems. Resilience improvements were obtained for the uncompatibilized system and interactions between extrusion parameters and compatibilizers were noticed.


Paternity after adolescent varicocele repair.
Varicocelectomy has long been a therapeutic modality used in the treatment of male infertility. In the past decade, adolescent varicocelectomy has become a frequent procedure to preserve testicular growth and to help prevent future infertility. Because our clinical population includes a large portion of orthodox Jews who traditionally marry early and are forbidden to use birth control by religious law, we thought that by studying our patients, we might be able to accelerate our follow-up regarding paternity. In addition, we wanted to learn whether adolescent varicocelectomy might have any negative impact. Questionnaires inquiring as to the marital and paternity status, postoperative course, and complications were sent to 50 patients who had undergone a unilateral or bilateral varicocele repair during adolescence and who were at least 21 years old at the time of this review. In addition, a careful chart review was performed to examine the perioperative and postoperative parameters of each respondent. Of the 43 responses (86% response rate), 18 of 18 patients who had attempted to father a child were successful. The remaining 25 were not married or had never attempted to father a child. In the paternity group, 10 of the fathers had undergone an Ivanissevich repair; the remaining 8 had a Palomo repair. Sixteen of the 18 had unilateral varicocelectomies, and 2 underwent bilateral repairs. Of those with a unilateral varicocele, the indication for surgery in 10 was a grade 2 to 3 varicocele associated with a >20% volume difference when compared with the right testicle. Three had 10% to 20% volume loss, whereas the remaining three had unusually large grade 3 varicoceles without concurrent volume difference. Varicocelectomy in the adolescent population has been proposed as a therapeutic intervention to preserve both fertility and testicular growth. Although not showing a cause-and-effect relationship, it is our contention that varicocelectomy in adolescence at worst does no harm and at best preserves fertility.


RNA polymerase II clustering through carboxy-terminal domain phase separation.
The carboxy-terminal domain (CTD) of RNA polymerase (Pol) II is an intrinsically disordered low-complexity region that is critical for pre-mRNA transcription and processing. The CTD consists of hepta-amino acid repeats varying in number from 52 in humans to 26 in yeast. Here we report that human and yeast CTDs undergo cooperative liquid phase separation, with the shorter yeast CTD forming less-stable droplets. In human cells, truncation of the CTD to the length of the yeast CTD decreases Pol II clustering and chromatin association, whereas CTD extension has the opposite effect. CTD droplets can incorporate intact Pol II and are dissolved by CTD phosphorylation with the transcription initiation factor IIH kinase CDK7. Together with published data, our results suggest that Pol II forms clusters or hubs at active genes through interactions between CTDs and with activators and that CTD phosphorylation liberates Pol II enzymes from hubs for promoter escape and transcription elongation.


Systemic listeriosis in caged canaries (Serinus canarius).
The occurrence of listeriosis in 12 caged canaries is described where 50% of the birds, including the female and all of the offspring, died within 2 weeks without clinical signs. At necropsy, multifocal necrotizing and partly granulomatous hepatitis, splenitis, myocarditis, interstitial nephritis, and exudative pericarditis with intra-lesional Listeria monocytogenes were the predominant findings as shown by histopathology and immunohistochemistry. Microbiology, serology and polymerase chain reaction revealed L. monocytogenes serotype 1/2a as the causative agent. Thus listeriosis has to be considered in the differential diagnosis for granulomas associated with mycobacteriosis, yersiniosis, coligranulomatosis or fungal infections.


Supporting community annotation and user collaboration in the integrated microbial genomes (IMG) system.
The exponential growth of genomic data from next generation technologies renders traditional manual expert curation effort unsustainable. Many genomic systems have included community annotation tools to address the problem. Most of these systems adopted a "Wiki-based" approach to take advantage of existing wiki technologies, but encountered obstacles in issues such as usability, authorship recognition, information reliability and incentive for community participation. Here, we present a different approach, relying on tightly integrated method rather than "Wiki-based" method, to support community annotation and user collaboration in the Integrated Microbial Genomes (IMG) system. The IMG approach allows users to use existing IMG data warehouse and analysis tools to add gene, pathway and biosynthetic cluster annotations, to analyze/reorganize contigs, genes and functions using workspace datasets, and to share private user annotations and workspace datasets with collaborators. We show that the annotation effort using IMG can be part of the research process to overcome the user incentive and authorship recognition problems thus fostering collaboration among domain experts. The usability and reliability issues are addressed by the integration of curated information and analysis tools in IMG, together with DOE Joint Genome Institute (JGI) expert review. By incorporating annotation operations into IMG, we provide an integrated environment for users to perform deeper and extended data analysis and annotation in a single system that can lead to publications and community knowledge sharing as shown in the case studies.


Long-term efficacy of upstream therapy with lipophilic or hydrophilic statins on antiarrhythmic drugs in patients with paroxysmal atrial fibrillation: comparison between atorvastatin and pravastatin.
There is little information available on the benefits of selection of statins as upstream therapy for the prevention of paroxysmal atrial fibrillation (AF). We compared the efficacy and safety of atorvastatin (A-group, n = 43) and pravastatin (P-group, n = 41) as upstream therapy in patients with paroxysmal AF and dyslipidemia. A total of 84 patients (45 men, mean age, 66 ± 9 years, mean follow-up, 49 ± 32 months) were retrospectively assigned to receive atorvastatin (n = 41;10 mg/day) or pravastatin (n = 43 ; 10 mg/day). Survival rates free from AF recurrence at 1, 6, 12, and 24 months were 93%, 74%, 60%, and 53% in A-group, and 88%, 49%, 37%, and 29%, respectively, in P-group (P = 0.029, A-group versus P-group). Survival rates free from conversion to permanent AF at 12, 36, 60, and 90 months were 100%, 100%, 98%, and 95% in A-group, and 100%, 95%, 88%, and 83%, respectively, in P-group (P = 0.063, A-group versus P-group). Using a logistic regression model, atorvastatin was found to be associated with a significantly reduced risk of AF recurrence in comparison to pravastatin (unadjusted odds ratio [OR] = 0.27, 95% confidence interval 0.11-0.68, P = 0.005). This association remained significant after adjustment for potentially confounding variables (OR = 0.26, 95% CI 0.08-0.86, P = 0.027). Using a logistic regression model, atorvastatin was not associated with a significantly reduced risk of converting to permanent AF in comparison to pravastatin (unadjusted OR = 0.29, 95% CI 0.05-1.50, P = 0.138), but this association did show a significant difference after adjustment for potentially confounding variables in a multivariate model (OR = 0.08, 95% CI 0.06-0.96, P = 0.046). Adverse effects requiring discontinuation of statins were observed in 1 case (2%, myalgia) in A-group, and 1 case (2%, elevation in CPK level ≥ 500 IU/L) in P-group, respectively (P = NS, A-group versus P-group). Atorvastatin, a lipophilic statin, was considered to be more effective in preventing recurrence of paroxysmal AF and conversion to permanent AF than pravastatin, a hydrophilic statin.


Genome sequences of Mx1, the first Myxococcus phage isolated, and Mx4, a generalized transducing myxophage.
Myxococcus xanthus is the best-studied member of the phylum Myxococcota, but the bacteriophages infecting it and their characterization remain limited. Here, we present complete genomes of Mx1, the first Myxococcus phage isolated, and of an Mx4 derivative widely used for generalized transduction, both unclassified Caudoviricetes with long, contractile tails.


Efficacy of a mobile hypertension prevention delivery platform with human coaching.
This pilot study evaluated the efficacy of a Hypertension Prevention Program (HPP) administered through a mobile application platform with human coaching (app) on reduction in blood pressure and weight in 50 adults with prehypertension or hypertension. Participants were recruited into a 24-week mobile application intervention to administer the HPP between January 2016 and July 2016. Dietary elements of the programme were based on the Dietary Approaches to Stop Hypertension. The programme included in-app human coaching with bi-weekly phone calls, meal logging, blood pressure tracking and educational material. Main outcome variables included change in systolic and diastolic blood pressure, hypertension category, and weight loss. Data were analysed between October 2016 and December 2016. The HPP yielded overall improvements in weight (-3.04±4.04 kg, P=<0.001), diastolic blood pressure (-5.06±11.89 mm Hg, P=0.004), and hypertension category (-0.48±0.74 mm Hg, P=<0.001). Sustained engagement of 80% resulted in significant reductions in systolic blood pressure (-7.75±12.56, P=<0.001) and weight (-3.73±4.01 kg, P<0.001) for programme completers, contributing to hypertension category change (-0.58±0.64 mm Hg, P<0.001). Mobile delivery of a lifestyle intervention for hypertension prevention showed short-term potential to reduce risk of hypertension, supporting the need for longer studies to investigate the use of mHealth lifestyle modification to reduce the risk of hypertension, a public health priority.


Macrophage-Immunomodulatory Actions of Bovine Whey Protein Isolate, Glycomacropeptide, and Their In Vitro and In Vivo Digests.
Whey protein isolate (WPI) consists of an array of proteins and peptides obtained as a byproduct of the cheesemaking process. Research suggests that WPI, along with its peptides such as glycomacropeptide (GMP), possesses immunomodulatory properties. These properties hold potential for alleviating the adverse effects of inflammatory conditions such as inflammatory bowel disease. Although promising, the immunoregulatory properties of the digested forms of WPI and GMP-those most likely to interact with the gut immune system-remain under-investigated. To address this knowledge gap, the current study examined the effects of in vitro-digested WPI and GMP, in vivo-digested WPI, and undigested WPI and GMP on the secretion of pro-inflammatory cytokines (TNF-α and IL-1β) in lipopolysaccharide-stimulated macrophage-like cells. Our results indicate that digested WPI and GMP reduced the expression of TNF-α and IL-1β, two pro-inflammatory cytokines. Whole WPI had no effect on TNF-α but reduced IL-1β levels. In contrast, in vivo-digested WPI reduced TNF-α but increased IL-1β. Undigested GMP, on the other hand, increased the secretion of both cytokines. These results demonstrate that digestion greatly modifies the effects of WPI and GMP on macrophages and suggest that digested WPI and GMP could help mitigate gastrointestinal inflammation. Further clinical studies are necessary to determine the biological relevance of WPI and GMP digestion products within the gut and their capacity to influence gut inflammation.