Radiotherapy of benign intracranial tumours.
We present the updated recommendations of the French Society for Radiation Oncology on benign intracranial tumours. Most of them are meningiomas, vestibular schwannomas, pituitary adenomas, craniopharyngiomas, and glomus tumours. Some grow very slowly, and can be observed without specific treatment, especially if they are asymptomatic. Symptomatic or growing tumours are treated by surgery, which is the reference treatment. When surgery is not possible, due to the location of the lesion, or general conditions, radiotherapy can be applied, as it is if there is a postoperative growing residual tumour, or a local relapse. Indications have to be discussed at a multidisciplinary panel, with precise evaluation of the benefit and risks of the treatments. The techniques to be used are the most modern ones, as multimodal imaging and image-guided radiation therapy. Stereotactic treatments, using fractionated or single doses depending on the size or the location of the tumours, are commonly realized, to avoid as much a possible the occurrence of late side effects.


The influence of protein concentration on key quality attributes of chickpea-based alternatives to cheese.
In response to consumer demands, plant protein ingredients are increasingly being used in the formulation of plant-based alternatives to cheese. The aim of this study was to determine the influence of protein concentration on key quality attributes of chickpea-based alternatives to cheese. Moreover, the age-induced changes in such attributes were assessed, with samples analysed after 1 month of storage. After characterisation of the ingredients, the chickpea-based formulations were prepared by blending chickpea flour and protein concentrate in different proportions to obtain four samples of increasing protein content (i.e., 8.68-21.5%). Formulations were developed at pH ∼4.5, and a moisture content of 50%, with shea butter used to obtain 15% fat content. The differential scanning calorimetry thermograms of the samples showed a main peak around 30 °C, corresponding to transition of the shea butter, and a smaller peak around 70 °C related to starch gelatinisation. Analysis of microstructure showed formation of a protein matrix with more extensive protein structure at high protein concentration. Furthermore, none of the chickpea-based samples melted under the testing conditions and all samples showed increasing values for adhesiveness, springiness and cohesiveness with increasing protein content. However, hardness was the highest for the sample with the lowest protein content, likely due to starch retrogradation. After storage, hardness increased further for all samples. This work improves our understanding of the role of chickpea protein in developing plant-based alternatives to cheese and the challenges therein.


Orchidaceae-Derived Anticancer Agents: A Review.
Species of orchids, which belong to the largest family of flowering plants, are commonly used in folk medicine for the treatment of infections and tumors. However, little is known about the actual chemical composition of these plants and their anticancer properties. In this paper, the most recent literature on orchid-derived bioactive substances with anticancer properties is reviewed. For the assessment, previous papers on the anticancer activity of Orchidaceae published since 2015 were considered. The papers were found by exploring electronic databases. According to the available data, many species of orchids contain potential antitumor chemicals. The bioactive substances in a relatively insignificant number of orchids are identified, and most studies are on Asian taxa. Broader research on American and African species and the correct identification of samples included in the experiments are essential for evaluating the usefulness of orchids as a plant family with vast anticancer potential.


Design and pharmacological activity of glycinamide and N-methoxy amide derivatives of analogs and constitutional isomers of valproic acid.
A series of glycinamide conjugates and N-methoxy amide derivatives of valproic acid (VPA) analogs and constitutional isomers were synthesized and evaluated for anticonvulsant activity. Of all compounds synthesized and tested, only N-methoxy-valnoctamide (N-methoxy-VCD) possessed better activity than VPA in the following anticonvulsant tests: maximal electroshock, subcutaneous metrazol, and 6-Hz (32-mA) seizure tests. In mice, the ED(50) values of N-methoxy-VCD were 142 mg/kg (maximal electroshock test), 70 mg/kg (subcutaneous metrazol test), and 35 mg/kg (6-Hz test), and its neurotoxicity TD(50) was 118 mg/kg. In rats, the ED(50) of N-methoxy-VCD in the subcutaneous metrazol test was 36 mg/kg and its protective index (PI=TD(50)/ED(50)) was >5.5. In the rat pilocarpine-induced status epilepticus model, N-methoxy-VCD demonstrated full protection at 200mg/kg, without any neurotoxicity. N-Methoxy-VCD was tested for its ability to induce teratogenicity in a mouse strain susceptible to VPA-induced teratogenicity and was found to be nonteratogenic, although it caused some resorptions. Nevertheless, a safety margin was still maintained between the ED(50) values of N-methoxy-VCD in the mouse subcutaneous metrazol test and the doses that caused the resorptions. On the basis of these results, N-methoxy-VCD is a good candidate for further evaluation as a new anticonvulsant and central nervous system drug.


Cynarin alleviates intervertebral disc degeneration via protecting nucleus pulposus cells from ferroptosis.
Intervertebral disc degeneration (IVDD) leads to a series of degenerative spine diseases. Clinical treatment of IVDD is mainly surgery, lacking effective drugs to alleviate intervertebral disc degeneration. In this study, we analysed the mRNA sequencing dataset of human degenerative intervertebral disc tissues and revealed the participation of ferroptosis in IVDD. Furthermore, we confirmed that TNF-α, an important cytokine in IVDD, induces ferroptosis in nucleus pulposus cells. Subsequently, a ferroptosis inhibitors screening strategy using multiple ferroptosis indicators was developed. Through the screen of various natural compounds, cynarin, a natural product enriched in Artichoke, was discovered to inhibit ferroptosis of nucleus pulposus cells. Cynarin can dose-dependently inhibit the catabolism of nucleus pulposus cells, increase the expression of key ferroptosis-inhibiting genes (GPX4 and NRF2), inhibit the increment of cellular Fe2+, lipid peroxides, and reactive oxygen species. It can also prevent mitochondria shrinkage, reduce mitochondria cristae density in ferroptosis, and prevent IVDD in the rat model. In conclusion, cynarin is a potential candidate for the drug development for IVDD.


Experience of surgical subspecialty residents on general surgery rotations.
Surgical subspecialty residents complete 5-6 years of training which includes general surgery rotations. A lack of data exists evaluating these rotations. This study aims to identify discrepancies in subspecialty training and improve the quality of surgical education. Case logs for surgical subspecialty residents and general surgery residents at our institution were analyzed and queried for cases performed on general surgery rotations. A survey was distributed to subspecialty residents regarding their perceptions of these rotations. 50 residents were included in the study and the majority were male (n = 27, 54%). Subspecialty residents perform fewer cases per month compared to general surgery residents (13 vs 21, p < 0.001). 75% of subspecialty residents were satisfied with their experience on general surgery rotations. Subspecialty residents perform fewer operations on general surgery rotations. Despite this, most are satisfied with off-service rotations and believe they are an important part of their education.


Identification of geographic clusters for temporal heterogeneity with application to dengue surveillance.
Identifying transmission of hot spots with temporal trends is important for reducing infectious disease propagation. Cluster analysis is a particularly useful tool to explore underlying stochastic processes between observations by grouping items into categories by their similarity. In a study of epidemic propagation, clustering geographic regions that have similar time series could help researchers track diffusion routes from a common source of an infectious disease. In this article, we propose a two-stage scan statistic to classify regions into various geographic clusters by their temporal heterogeneity. The proposed scan statistic is more flexible than traditional methods in that contiguous and nonproximate regions with similar temporal patterns can be identified simultaneously. A simulation study and data analysis for a dengue fever infection are also presented for illustration.


An integrative systems genetic analysis of mammalian lipid metabolism.
Dysregulation of lipid homeostasis is a precipitating event in the pathogenesis and progression of hepatosteatosis and metabolic syndrome. These conditions are highly prevalent in developed societies and currently have limited options for diagnostic and therapeutic intervention. Here, using a proteomic and lipidomic-wide systems genetic approach, we interrogated lipid regulatory networks in 107 genetically distinct mouse strains to reveal key insights into the control and network structure of mammalian lipid metabolism. These include the identification of plasma lipid signatures that predict pathological lipid abundance in the liver of mice and humans, defining subcellular localization and functionality of lipid-related proteins, and revealing functional protein and genetic variants that are predicted to modulate lipid abundance. Trans-omic analyses using these datasets facilitated the identification and validation of PSMD9 as a previously unknown lipid regulatory protein. Collectively, our study serves as a rich resource for probing mammalian lipid metabolism and provides opportunities for the discovery of therapeutic agents and biomarkers in the setting of hepatic lipotoxicity.


Identity and tissue localization of free and conjugated ecdysteroids in adults of Dirofilaria immitis and Ascaris suum.
Adult males and females of the dog heartworm, Dirofilaria immitis, and of the swine parasite, Ascaris suum, were extracted, the free and polar conjugated ecdysteroid fractions separated and the latter hydrolysed enzymically. The ecdysteroids released by hydrolysis of the conjugates and the free hormones were analysed by radioimmunoassay, high-performance liquid chromatography on reversed phase and adsorption columns monitoring fractions by radioimmunoassay, and by gas-liquid chromatography/mass spectrometry (selected ion monitoring). In both species, males and females contained free and polar conjugated ecdysteroids, with evidence for the presence primarily of ecdysone and 20-hydroxyecdysone together with smaller amounts of 20,26-dihydroxyecdysone. Males and females of both species were then dissected into body fluid, reproductive system, gut and remaining body wall compartments, the ecdysteroids extracted, fractionated and analysed by radioimmunoassay and high-performance liquid chromatography monitoring fractions by radioimmunoassay. The results for both sexes in the two species were similar and indicated that ecdysteroids were not detectable in body fluids and that free ecdysteroids occurred in the reproductive system and the body wall, whereas polar conjugated ecdysteroids were detected in the reproductive system and the gut; a minor portion of the free ecdysteroids in A. suum was also apparently present in the gut. Further localization of the ecdysteroids in the body wall of A. suum females suggested that negligible immunoreactivity was associated with the circumpharyngeal nerve ring. The possible significance of the results is discussed.


Antigenic variants of rabies virus.
Monoclonal antibodies directed either against the nucleocapsid or the glycoprotein of rabies virus were reacted by indirect immunofluorescence and mouse neutralization tests with vaccine strains and rabies field isolates of various origin. Major antigenic determinants allow the grouping of viruses and minor ones the differentiation of virus strains. A protection study showed the existence in nature of certain antigenic variants against which conventional vaccines do not fully protect. The isolation of two such variants from 'imported' bats in the Federal Republic of Germany is reported here for the first time.


BNT162b2 vaccination induces durable SARS-CoV-2-specific T cells with a stem cell memory phenotype.
Vaccination against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is effective in preventing hospitalization from severe COVID-19. However, multiple reports of breakthrough infections and of waning antibody titers have raised concerns on the durability of the vaccine, and current vaccination strategies now propose administration of a third dose. Here, we monitored T cell responses to the Spike protein of SARS-CoV-2 in 71 healthy donors vaccinated with two doses of the Pfizer-BioNTech mRNA vaccine (BNT162b2) for up to 6 months after vaccination. We found that vaccination induced the development of a sustained anti-viral CD4+ and CD8+ T cell response. These cells appeared before the development of high antibody titers, displayed markers of immunological maturity and stem cell memory, survived the physiological contraction of the immune response, and persisted for at least 6 months. Collectively, these data show that vaccination with BNT162b2 elicits an immunologically competent and long-lived SARS-CoV-2–specific T cell population.


Mechanical Thrombectomy in Acute Ischemic Stroke-Patients with Wake-Up Stroke and the Elderly May Benefit as Well.
We have assessed to what extent age, sex, preintervention infarct size, time to groin puncture, and stroke severity are associated with outcome of mechanical thrombectomy in patients treated for acute ischemic stroke. All 192 patients treated with mechanical thrombectomy for acute ischemic stroke from September 2005 to December 2011 were included in this observational study. Main outcomes were independence, defined as a modified Rankin Scale score of 0-2, an improvement in stroke severity score post-thrombectomy, and the occurrence of symptomatic hemorrhage. Exposure variables were sex, age, intravenous thrombolysis, prethrombectomy stroke severity grading, pretreatment infarct size, time from onset of symptoms to groin puncture, thrombectomy device used, and the angiographic result from the thrombectomy. For patients above 50 years with no neurological symptoms before stroke onset, the proportion gaining independence was unrelated to age group, ranging from 49% to 54%. For patients younger than 50 years of age, the proportion was significantly higher (81%). Patients with wake-up stroke did not differ from other patients with regard to outcome. Our results emphasize that when patients are selected on the basis of angiographic and perfusion imaging, both older patients and patients with wake-up stroke may experience a favorable outcome after mechanical thrombectomy.


Retraction: Open resection compared to mini-invasive in colorectal cancer and liver metastases: a meta-analysis.
[This retracts the article DOI: 10.3389/fsurg.2021.726217.].


The effect of corruption on carbon emissions in developed and developing countries: empirical investigation of a claim.
The impact of corruption on carbon emissions is one of the main objectives of empirical studies on environmental economics. Recently, a theoretical discussion was conducted on the significant impact of reducing the level of corruption on environmental quality in developing rather than in developed countries. In this study, an empirical investigation of this claim was conducted using panel data which included 61 countries, between 2003 and 2016. The effects of corruption on carbon emission were considered using a panel threshold model. The threshold variable included human development index (HDI) which divided countries into developing and developed ones endogenously. According to the results, the value of threshold (γ = 0.753) was consistent with the categorization of UNDP countries by HDI. In developing countries, for each unit of increase in the corruption index -which means a decrease in corruption levels- we observe a 0.08 unit decrease in carbon emission while carbon emission is no longer affected by the corruption levels in developed countries and decrease in corruption does not have a significant effect on carbon emission levels.


Global trends in semen quality of young men: a systematic review and regression analysis.
Many studies have reported declines in semen quality mainly focused on total sperm counts (TSC) and sperm concentration (SC), ignoring the importance of progressive motile sperm (PR), total motile sperm (TM), and normal morphological sperm (NM). Therefore, we performed a comprehensive meta-analysis to explore the trend in semen quality of young men. We searched 3 English databases and 4 Chinese databases from January 1980 to August 2022. Random-effect meta-analyses and weighted linear regression models were conducted to perform the trend in semen quality. Finally, 162 eligible studies including 264,665 men from 28 countries were got between 1978 and 2021. Significant decreases were observed in TSC (- 3.06 million/year, 95% CI - 3.28 to - 2.84), SC (- 0.47 million/ml/year, 95% CI - 0.51 to - 0.43), and PR (- 0.15%/year, 95% CI - 0.20 to - 0.09), and there was an upward trend in TM (0.28%/year, 95% CI 0.24 to 0.32). The results of meta-regression analyses indicated that age, continent, income, WHO criteria, and abstinence time significantly impacted on TSC, SC, PR, and TM. Positive regression coefficients were observed in some categories suggesting that outcomes might not be declining and even increasing in these subgroups. Downward trends in semen quality among global young men were observed in our study, including TSC, SC, and PR. But TM did not appear to be trending down or even to be leveling off. More studies are needed to focus on the causes of the declines.


Reduced relative fitness in hatchery-origin Pink Salmon in two streams in Prince William Sound, Alaska.
Previous studies generally report that hatchery-origin Pacific Salmon (Oncorhynchus spp.) have lower relative reproductive success (RRS) than their natural-origin counterparts. We estimated the RRS of Pink Salmon (O. gorbuscha) in Prince William Sound (PWS), Alaska using incomplete pedigrees. In contrast to other RRS studies, Pink Salmon have a short freshwater life history, freshwater habitats in PWS are largely unaltered by development, and sampling was conducted without the aid of dams or weirs resulting in incomplete sampling of spawning individuals. Pink Salmon released from large-scale hatchery programs in PWS have interacted with wild populations for more than 15 generations. Hatchery populations were established from PWS populations but have subsequently been managed as separate broodstocks. Gene flow is primarily directional, from hatchery strays to wild populations. We used genetic-based parentage analysis to estimate the RRS of a single generation of stray hatchery-origin Pink Salmon in two streams, and across the odd- and even-year lineages. Despite incomplete sampling, we assigned 1745 offspring to at least one parent. Reproductive success (RS), measured as sampled adult offspring that returned to their natal stream, was significantly lower for hatchery- vs. natural-origin parents in both lineages, with RRS ranging from 0.03 to 0.47 for females and 0.05 to 0.86 for males. Generalized linear modeling for the even-year lineage indicated that RRS was lower for hatchery-origin fish, ranging from 0.42 to 0.60, after accounting for sample date (run timing), sample location within the stream, and fish length. Our results strongly suggest that hatchery-origin strays have lower fitness in the wild. The consequences of reduced RRS on wild productivity depend on whether the mechanisms underlying reduced RRS are environmentally driven, and likely ephemeral, or genetically driven, and likely persistent across generations.


Reinforcement Learning Enables Resource Partitioning in Foraging Bats.
Every evening, from late spring to mid-summer, tens of thousands of hungry lactating female lesser long-nosed bats (Leptonycteris yerbabuenae) emerge from their roost and navigate over the Sonoran Desert, seeking for nectar and pollen [1, 2]. The bats roost in a huge maternal colony that is far from the foraging grounds but allows their pups to thermoregulate [3] while the mothers are foraging. Thus, the mothers have to fly tens of kilometers to the foraging sites-fields with thousands of Saguaro cacti [4, 5]. Once at the field, they must compete with many other bats over the same flowering cacti. Several solutions have been suggested for this classical foraging task of exploiting a resource composed of many renewable food sources whose locations are fixed. Some animals randomly visit the food sources [6], and some actively defend a restricted foraging territory [7-11] or use simple forms of learning, such as "win-stay lose-switch" strategy [12]. Many species have been suggested to follow a trapline, that is, to revisit the food sources in a repeating ordered manner [13-22]. We thus hypothesized that lesser long-nosed bats would visit cacti in a sequenced manner. Using miniature GPS devices, aerial imaging, and video recordings, we tracked the full movement of the bats and all of their visits to their natural food sources. Based on real data and evolutionary simulations, we argue that the bats use a reinforcement learning strategy that requires minimal memory to create small, non-overlapping cacti-cores and exploit nectar efficiently, without social communication.


Metabolic responses to pyruvate kinase deletion in lysine producing Corynebacterium glutamicum.
Pyruvate kinase is an important element in flux control of the intermediate metabolism. It catalyzes the irreversible conversion of phosphoenolpyruvate into pyruvate and is under allosteric control. In Corynebacterium glutamicum, this enzyme was regarded as promising target for improved production of lysine, one of the major amino acids in animal nutrition. In pyruvate kinase deficient strains the required equimolar ratio of the two lysine precursors oxaloacetate and pyruvate can be achieved through concerted action of the phosphotransferase system (PTS) and phosphoenolpyruvate carboxylase (PEPC), whereby a reduced amount of carbon may be lost as CO2 due to reduced flux into the tricarboxylic acid (TCA) cycle. In previous studies, deletion of pyruvate kinase in lysine-producing C. glutamicum, however, did not yield a clear picture and the exact metabolic consequences are not fully understood. In this work, deletion of the pyk gene, encoding pyruvate kinase, was carried out in the lysine-producing strain C. glutamicum lysCfbr, expressing a feedback resistant aspartokinase, to investigate the cellular response to deletion of this central glycolytic enzyme. Pyk deletion was achieved by allelic replacement, verified by PCR analysis and the lack of in vitro enzyme activity. The deletion mutant showed an overall growth behavior (specific growth rate, glucose uptake rate, biomass yield) which was very similar to that of the parent strain, but differed in slightly reduced lysine formation, increased formation of the overflow metabolites dihydroxyacetone and glycerol and in metabolic fluxes around the pyruvate node. The latter involved a flux shift from pyruvate carboxylase (PC) to PEPC, by which the cell maintained anaplerotic supply of the TCA cycle. This created a metabolic by-pass from PEP to pyruvate via malic enzyme demonstrating its contribution to metabolic flexibility of C. glutamicum on glucose. The metabolic flux analysis performed illustrates the high flexibility of the metabolic network of C. glutamicum to compensate for external perturbation. The organism could almost maintain its growth and production performance through a local redirection of the metabolic flux, thereby fulfilling all anabolic and catabolic needs. The formation of the undesired overflow metabolites dihydroxyacetone and glycerol, in the deletion mutant, however, indicates a limiting capacity of the metabolism down-stream of their common precursor glyceraldehyde 3-phosphate and opens possibilities for further strain engineering.


Nystatin induces secretion of interleukin (IL)-1beta, IL-8, and tumor necrosis factor alpha by a toll-like receptor-dependent mechanism.
Nystatin is an antifungal compound with potent proinflammatory properties. Herein, we demonstrate that nystatin induces interleukin (IL)-1beta, IL-8, and tumor necrosis factor alpha secretion through its activation of toll-like receptor 1 (TLR1) and TLR2. Hence, a TLR-dependent mechanism could serve as the molecular basis for the proinflammatory properties of nystatin.


A comparative study of the in vitro antitumor effect of mannose-doxorubicin conjugates with different linkers.
In this work, five Man-DOX conjugates with different linkers were developed for targeted DOX delivery. The five Man-DOX conjugates with different linkers were characterized by 1 H NMR, HRMS, HPLC, UV-vis, and fluorescence spectroscopy. Man-Suc-DOX, Man-TDG-DOX, and Man-DG-DOX can self-assemble into near-spherical nanoparticles with hydrodynamic diameters of 150-200 nm and negative zeta potentials in deionized water, whereas Man-SS-DOX and Man-SeSe-DOX are hardly dispersed in deionized water. The self-assembly behaviors of Man-Suc-DOX, Man-TDG-DOX, and Man-DG-DOX were studied by dissipative particle dynamics simulation and the results show that Man-Suc-DOX, Man-TDG-DOX, and Man-DG-DOX all self-assemble into spherical particles with Man and linkers on the surfaces and DOX in the interiors. The in vitro drug release study shows that Man-Suc-DOX, Man-TDG-DOX, and Man-DG-DOX exhibit limited drug release, while Man-SS-DOX and Man-SeSe-DOX exhibit glutathione-responsive drug release. The cellular uptake study shows that Man-DG-DOX exhibits the highest cellular uptake amount on HepG2 cells. Finally, Man-DG-DOX exhibits the best in vitro antitumor effect against HepG2 cells among the five Man-DOX conjugates with different linkers. Although the in vitro antitumor activity of Man-DG-DOX is still lower than free DOX, Man-DG-DOX shows significant selectivity toward HepG2 cells. Man-DG-DOX might achieve selective DOX delivery for mannose receptor overexpressed tumors.


Biomarkers of persistent renal vulnerability after acute kidney injury recovery.
Acute kidney injury (AKI) is a risk factor for new AKI episodes, chronic kidney disease, cardiovascular events and death, as renal repair may be deficient and maladaptive, and activate proinflammatory and profibrotic signals. AKI and AKI recovery definitions are based on changes in plasma creatinine, a parameter mostly associated to glomerular filtration, but largely uncoupled from renal tissue damage. The evolution of structural and functional repair has been incompletely described. We thus aimed at identifying subclinical sequelae persisting after recovery from cisplatin-induced AKI in rats. Compared to controls, after plasma creatinine recovery, post-AKI kidneys showed histological alterations and attendant susceptibility to new AKI episodes. Tubular function (assessed by the furosemide stress test, FST) also remained affected. Lingering parenchymal and functional subclinical alterations were paralleled by tapering, but abnormally high levels of urinary albumin, transferrin, insulin-like growth factor-binding protein 7 (IGFBP7), tissue inhibitor of metalloproteinases-2 (TIMP-2) and, especially, the [TIMP-2]*[IGFBP7] product. As subclinical surrogates of incomplete renal recovery, the FST and the urinary [TIMP-2]*[IGFBP7] product provide two potential diagnostic tools to monitor the sequelae and kidney vulnerability after the apparent recovery from AKI.


Shifts in the neurobiological mechanisms motivating cocaine use with the development of an addiction-like phenotype in male rats.
The development of addiction is accompanied by a shift in the mechanisms motivating cocaine use from nucleus accumbens (NAc) dopamine D1 receptor (D1R) signaling to glutamate AMPA-kainate receptor (AMPA-R) signaling. Here, we determined whether similar shifts occur for NAc-D2R signaling and following systemic manipulation of D1R, D2R, and AMPA-R signaling. Male rats were given short-access (20 infusions/day) or extended-access to cocaine (24 h/day, 96 infusions/day, 10 days). Motivation for cocaine was assessed following 14 days of abstinence using a progressive-ratio schedule. Once responding stabilized, the effects of NAc-D2R antagonism (eticlopride; 0-10.0 μg/side) and systemic D1R (SCH-23390; 0-1.0 mg/kg), D2R (eticlopride; 0-0.1 mg/kg), and AMPA-R (CNQX; 0-1.5 mg/kg) antagonism, and NAc-dopamine-R gene expression (Drd1/2/3) were examined. Motivation for cocaine was markedly higher in the extended- versus short-access group confirming the development of an addiction-like phenotype in the extended-access group. NAc-infused eticlopride decreased motivation for cocaine in both the short- and extended-access groups although low doses (0.1-0.3 μg) were more effective in the short-access group and high doses (3-10 μg/side) tended to be more effective in the extended-access group. Systemic administration of eticlopride (0.1 mg/kg) was more effective in the extended-access group, and systemic administration of CNQX was effective in the extended- but not short-access group. NAc-Drd2 expression was decreased in both the short- and extended-access groups. These findings indicate that in contrast to NAc-D1R, D2R remain critical for motivating cocaine use with the development of an addiction-like phenotype. These findings also indicate that shifts in the mechanisms motivating cocaine use impact the response to both site-specific and systemic pharmacological treatment.


Expression of the nonstructural proteins of Sindbis virus in insect cells by a baculovirus vector.
The genome of Sindbis virus encodes the polypeptides that are required for the replication and transcription of the virus RNA in infected cells. These polypeptides are translated as a polyprotein that is co- and post-translationally cleaved by an autoproteinase to give rise to four polypeptides designated nsP1, nsP2, nsP3 and nsP4. We have initiated a study of the functions of these proteins by expressing them in the Autographa californica baculovirus polyhedrin expression system. Spodoptera frugiperda cells infected with the recombinant baculovirus synthesized the four Sindbis polypeptides. We used a complementation assay which measures chloramphenicol acetyltransferase (CAT) activity to demonstrate that these proteins were biologically active. The infected cells were transfected with a Sindbis defective RNA that contains the CAT gene downstream of the promoter for the synthesis of the viral subgenomic RNA. CAT activity was found only in cells that had been infected with the recombinant baculovirus, not with wild type baculovirus, indicating that the required Sindbis nsP activities were present. Sindbis virions grew poorly in S. frugiperda cells and self-replicating Sindbis RNAs produced only very low levels of biological activity. Our results suggest that these cells are defective in their ability to replicate Sindbis RNAs and that the block is partially overcome when the Sindbis nsP mRNA is expressed under the control of the baculovirus DNA.


Encephalitis with convulsive status in an immunocompetent pediatric patient caused by Bartonella henselae.
Cat scratch's disease caused by Bartonella henselae, is known to be a self-limited benign process in immunocompetent children. The association with neurologic manifestations is very uncommon especially in patient with no immunologic defects and in cases without specific treatment. A 7 years old male patient, without any immunocompromised defect, presented an atypic presentation of the cat scratch disease. The patient came to the hospital in two opportunities in a status epilepticus, in both cases the diagnosis was encephalitis by Bartonella henselae and the evolution with treatment was monitored with PCR (polymerase chain reaction) in cerebrospinal fluid and blood, as well as IFI (IgM, IgG) serology (indirect immunofluorescence). The patient had a favorable clinical and laboratory evolution for 6 months showing no recurrence of the disease.


[Immune response enhanced by genes encoding IFN-alpha 8 and T alpha 1 co-inoculated with HBV DNA vaccine].
To evaluate if the humoral immune response of hepatitis B DNA vaccine pVAX1-S2S could be enhanced by Talpha1 and/or IFNa expression plasmid co-inoculated. The following mammalian expression recombinant plasmids were constructed: the plasmid pVAX1-S2S expressing hepatitis B surface antigen S2S, the plasmid pVAX1-T/I co-expressing thymosin a and IFNalpha, the plasmid pVAX1-I/S2S co-expressing IFNalpha and S2S. These plasmids were inoculated intramuscularly into several BALB/c mice groups in different combinations. In the co-immunization group 1 (pVAX1-I/S2S), each mouse was inoculated with 100 microg of pVAX1-I/S2S; in the co-immunization group 2 (pVAX1-S2S) each mouse was co-inoculated with pVAX1-S2S and 50 microg of pVAX1-TI; in the control group each mouse was inoculated with 100 microg of pVAX1-S2S. All the immunizations were boosted at 2 and 4 week intervals; then the serum samples were collected to detect the anti-HBs and anti-preS2 strengths. 3, 5 and 8 weeks after the first inoculation, the positive rates of anti-HBs were 12.5%, 12.5%, 62.5% respectively in the co-immunization group 1 and 25%, 50%, 50% in the co-immunization group 2, while those in the control group were 0, 25%, 37.5%. The titers of anti-preS2 in co-immunization group 2 was 5 times higher than those in the other two groups. The data shows that Talpha1 and/or IFNalpha expression plasmid co-inoculated with pVAX1-S2S might act as an adjuvant to enhance the humoral immune response induced by pVAX1-S2S.


Laser desorption/ionization-mass spectrometry for the analysis of interphases in lithium ion batteries.
Laser desorption/ionization-mass spectrometry (LDI-MS) is introduced as a complementary technique for the analysis of interphases formed at electrode|electrolyte interfaces in lithium ion batteries (LIBs). An understanding of these interphases is crucial for designing interphase-forming electrolyte formulations and increasing battery lifetime. Especially organic species are analyzed more effectively using LDI-MS than with established methodologies. The combination with trapped ion mobility spectrometry and tandem mass spectrometry yields additional structural information of interphase components. Furthermore, LDI-MS imaging reveals the lateral distribution of compounds on the electrode surface. Using the introduced methods, a deeper understanding of the mechanism of action of the established solid electrolyte interphase-forming electrolyte additive 3,4-dimethyloxazolidine-2,5-dione (Ala-N-CA) for silicon/graphite anodes is obtained, and active electrochemical transformation products are unambiguously identified. In the future, LDI-MS will help to provide a deeper understanding of interfacial processes in LIBs by using it in a multimodal approach with other surface analysis methods to obtain complementary information.


Place and the Structuring of Cross-Sector Partnerships: The Moral and Material Conflicts Over Healthcare and Homelessness.
Local places, such as communities, cities, and towns, host many cross-cross sector partnerships, many geared primarily toward alleviating local social and environmental issues. Yet, existing literatures focus predominantly on largescale systemic impact and global challenges such as climate change, paying scant attention to the role of local, geographically bounded dynamics in shaping these partnerships. In this article, I conceptualize places as geographic locations imbued with specific meaning systems and material resources to unpack how local embeddedness shape the structure of cross-sector partnerships. Specifically, I investigate how place-based conflict, arising from tensions between the moral and material aspects of a partnership, can shape formalized aspects of organizational structure. These include the scope of operations, partners' roles, and shared resources. I unpack these relationships using a case study of Occupy Medical, a local partnership between the civic society and the local government in Eugene, Oregon, tackling the problem of providing healthcare to the homeless and other marginalized and disenfranchised communities. The analysis covers the nine-year period of 2011-2020 and spans three major restructurings of the organization, the latest prompted by the onset of the COVID-19 pandemic in March 2020. I theorize two forms of structural arrangements for cross-sector partnerships, confined and leveraged, and further elaborate on the role of cross-sector partnerships in crises response on the local level.


Genotypes of Helicobacter pylori in children with upper abdominal pain.
Studies related to genotypes of Helicobacter pylori infection and upper abdominal pain (UAP) in children are scarce all over the world. We prospectively analyzed the association between H. pylori infection and UAP in our study group of children and evaluated the vacA genotypes associated with this disorder. We investigated 34 children with UAP (group I) and another 110 children as controls without UAP (group II) for H. pylori infection, using antral biopsies by culture, rapid urease tests, histopathology and ureA polymerase chain reaction (PCR). Genotyping was performed using specific cagA and vacA primers on 52 H. pylori strains (group I = 21; 18 boys, mean age 10.33 +/- 2.47 years; group II = 31; boys 21, mean age 9.63 +/- 2.51 years). A significant association between H. pylori infection and UAP was observed when compared to controls (21/34; 61.8%vs 31/110; 28.2%; P = 0.0004). cagA positive H. pylori strains were detected in 20 (95.2%) children with UAP and in 28 (90.3%) controls. Mixed infection was detected in 25% of children with no significant difference between the groups. On univariate regression analysis, s1a, m1 alleles and s1a/m1 genotype of vacA had significant associations with UAP (P = 0.018, 0.015 and 0.007, respectively), while s2 and m2 alleles and the s2/m2 genotype were significantly more frequent in controls (P = 0.034, 0.001 and 0.034, respectively). H. pylori infection is strongly associated with UAP in children and a significantly higher proportion have s1a, m1 alleles and s1a/m1 genotype. The negative associations of vacA s2, m2 alleles and s2/m2 genotype with UAP indicate that they are unlikely to have an important role in this disorder.


Exosomal long noncoding RNA lnc-GNAQ-6:1 may serve as a diagnostic marker for gastric cancer.
Gastric cancer (GC) is one of the most aggressive cancers, with limited early diagnostic measures. Tumor-originated exosomal molecules are regarded as suitable candidates for non-invasive diagnosis. This study aimed to investigate the capacity of exosomal long noncoding RNA lnc-GNAQ-6:1 as a biomarker for early diagnosis of GC. In this study, we collected sera from 43 patients with gastric cancer and 27 healthy subjects, then exosomes were isolated using commercial kits. Particle size analysis, Western bloting and protein-based exosomes quantification were conducted to identify the isolated exosomes and to evaluate its yield and purity. Expression levels of exosomal lnc-GNAQ-6:1 were detected by quantitative reverse transcription PCR (qRT-PCR). The serum concentrations of traditional biomarker (CA72-4, CEA, and CA19-9) were measured via a chemiluminescent detection system.The receiver operating characteristic curve (ROC) and area under curve (AUC) were used to estimate the diagnostic capacity. Furthermore, we analyzed the potential relationship between serum exosomal lnc-GNAQ-6:1 expression and clinicopathological parameters of gastric cancer. The exosomes extracted in this study exhibited the typical exosome characteristics and purity. Patients with gastric cancer had the higher exosome yield than healthy volunteer. The results of qRT-PCR showed that compared with the healthy control, the expression of lnc-GNAQ-6:1 was significantly lower in the gastric cancer group. The area under the ROC curve is 0.732, which was higher than the diagnostic accuracy of CEA, CA 19-9 and CA72-4. However, the expression level of lnc-GNAQ-6:1 was not correlated with gender, age, tumor metastasis, serum carcinoembryonic antigen (CEA), carbohydrate antigen 19-9 (CA19-9), and carbohydrate antigen 72-4(CA72-4). Our data demonstrated that serum exosomal lnc-GNAQ-6:1 is lowly expressed in patient with gastric cancer and might be evaluated in larger studies as a new diagnostic marker for gastric cancer.


Lateral diffusion in inhomogeneous membranes. Model membranes containing cholesterol.
The problem of lateral diffusion in inhomogeneous membranes is illustrated by a theoretical calculation of the lateral diffusion of a fluorescent lipid probe in binary mixtures of phosphatidylcholine and cholesterol under conditions of temperature and composition such that this lipid mixture consists of alternating parallel domains of fluid and solid lipid, having separations that are small compared with the distance scale employed in photobleaching experiments. The theoretical calculations clearly illustrate how inhomogeneities in membrane composition affecting the lateral motion of membrane components on a small (10-100 nm) distance scale can give complex diffusive responses in experiments such as fluorescence photobleaching that employ comparatively macroscopic distances (10-100 micrometers) for the measurement of diffusive recovery. The theoretical calculations exhibit the unusual dependence of the apparent lateral diffusion coefficient of a fluorescent lipid probe on lipid composition in binary mixtures of cholesterol and phosphatidylcholines as reported by Rubenstein et al. (1979, Proc. Natl. Acad. Sci. U.S.A., 76:15-18).


Rare ovarian tumours. Other treatments for ovarian cancer.
The description of rare malignant ovarian tumours and the most suitable treatments. Alternative therapies different from intravenous chemotherapy are also explained. Literature review and ongoing trial information have been used to elaborate this guide. Each ovarian cancer type must be identified and treated properly from diagnostic to surgery, adjuvant treatment and metastatic disease. Hormonotherapy can be useful as an alternative treatment, especially in low-grade ovarian cancer and endometrioid subtype. Tumour characterisation is appropriated for treatment selection when targeted therapy is indicated. MEK inhibitors, tyrosine-kinase inhibitors, EGFR inhibitors, therapies against integrins, antibody-drug conjugates and other strategies are described. Antiangiogenics, PARP inhibitors and immunotherapy are discussed in other parts of this publication. Different ovarian cancer types must receive the appropriated treatment. Alternative therapies may be evaluated beyond the standard therapy, frequently in a clinical trial, and an individualised molecular study may help to find the best treatment.


The human microbiome in disease and pathology.
This narrative review seeks to examine the relationships between bacterial microbiomes and infectious disease. This is achieved by detailing how different human host microbiomes develop and function, from the earliest infant acquisitions of maternal and environmental species through to the full development of microbiomes by adulthood. Communication between bacterial species or communities of species within and outside of the microbiome is a factor in both maintenance of homeostasis and management of threats from the external environment. Dysbiosis of this homeostasis is key to understanding the development of disease states. Several microbiomes and the microbiota within are used as prime examples of how changes in species composition, particularly at the phylum level, leads to such diverse conditions as inflammatory bowel disease (IBD), type 2 diabetes, psoriasis, Parkinson's disease, reflux oesophagitis and others. The review examines spatial relationships between microbiomes to understand how dysbiosis in the gut microbiome in particular can influence diseases in distant host sites via routes such as the gut-lung, gut-skin and gut-brain axes. Microbiome interaction with host processes such as adaptive immunity is increasingly identified as critical to developing the capacity of the immune system to react to pathogens. Dysbiosis of essential bacteria involved in modification of host substrates such as bile acid components can result in development of Crohn's disease, small intestine bacterial overgrowth, hepatic cancer and obesity. Interactions between microbiomes in distantly located sites are being increasingly being identified, resulting in a 'whole of body' effect by the combined host microbiome.


Functional anatomy of the human lateral pterygoid muscle.
The authors studied the lateral pterygoid muscle of 179 fresh cadavers by both anatomical dissection and magnetic resonance imaging. The aim was to define the general morphology and the architectural organisation of the lateral pterygoid muscle. Dissection plane by plane, anatomical sections in different planes of space and dissections via multiple approaches on harvested blocks allowed the demonstration that the lateral pterygoid shows an architectural arrangement different from that which is usually described but similar to that, from its typically penniform structure of the masseter and the medial pterygoid. The anatomical situation of this muscle gives rise to numerous differences in opinion much to its morphology as to its functional examination. This study has shown that one part of the muscle has a typical penniform structure made up of alternating musculo-aponeurotic layers and by the absence of individual muscle bellies. In addition the particular shape of this muscle makes it useless to insert intramuscular electrodes in its only accessible portion, which makes the results of electromyographic studies debatable.


Enhancing the communication of functional neurological disorder diagnosis: a multidisciplinary education session.
Functional neurological disorder (FND) is a common diagnosis within neurology. Effective communication of the diagnosis is known to be an important part of treatment and can result in reduction or cessation of symptoms, as well as decreased healthcare utilisation. A single group education session, facilitated by professionals commonly involved in the care of patients with FND, was developed to further enhance patients' and relatives' understanding and acceptance of diagnosis. Patients and relatives attending a single education session were asked to complete self-report ratings of understanding of diagnosis, acceptance of diagnosis, belief in treatability and hopefulness regarding recovery, at the beginning and end of the session. Satisfaction data were also collected. Data were obtained from 193 patients and 153 relatives. Patients had experienced a median duration of symptoms of 4 years, and more than 80% of patients reported more than one functional neurological symptom. There were significant increases in terms of understanding, acceptance, belief in treatability and hopefulness for patients and relatives. Effect sizes ranged from large for improved understanding of FND to small-to-medium for increased agreement with FND diagnosis. High levels of satisfaction were also reported. A multidisciplinary, single group education session is an effective and acceptable method of increasing understanding and acceptance of an FND diagnosis, even for patients with a long duration of symptoms and high symptom burden. It could help improve readiness for further treatment.


Barriers to and facilitators of a healthy lifestyle for patients with gynecological cancer: a systematic review of qualitative and quantitative research with healthcare providers and patients.
The prevalence of an unhealthy lifestyle among patients with gynecological cancer is high and associated with increased risk of all-cause mortality. Although lifestyle changes have the potential to improve outcomes, lifestyle counseling is not routinely integrated into standard care. This review explores research on the barriers to and facilitators of both the promotion of healthy lifestyles by healthcare providers (HCPs) and healthy lifestyle changes by patients with gynecological cancer. The Theoretical Domains Framework (TDF) was used to deductively code the identified factors for a comprehensive understanding of the barriers and facilitators. A search across five databases yielded a total of 12,687 unique studies, of which 43 were included in the review. Of these 43, 39 included gynecological cancer patients and only 6 included HCPs. Among the barriers identified for HCPs, most studies evaluated barriers regarding weight loss counseling. Limited knowledge, reluctance to address weight loss, skepticism about the benefits, and workload concerns were commonly reported barriers for HCPs. HCPs will benefit from education and training in lifestyle counseling, including effective communication skills like motivational interviewing. Gynecological cancer patients lacked tools, support, knowledge, and faced mental health issues, environmental constraints, and physical limitations. The review emphasizes the importance of addressing these barriers and utilizing identified facilitators, such as social support, to promote and support healthy lifestyle behaviors on the part of patients and their promotion by HCPs. Future research should focus not only on patients but also on supporting HCPs and implementing necessary changes in current practices.


Comparison of the psychospiritual needs of Chinese heart transplant recipients at pre- and postoperative stages.
This project sought to compare Chinese heart transplant (HT) recipients' psychospiritual needs during the preoperative (PS) and the postoperative predischarge stage (PDS). Thirty HT recipients from two leading medical centers in Taiwan were recruited for this research. Data collected by face-to-face interview were analyzed by content analysis. The patients' ages ranged from 23 to 45 years (mean [M] +/- standard deviation [SD] = 28.5 +/- 4.1); 90% were married. The subjects' cardiac disease history ranged from 4.2 to 12.3 years (M +/- SD = 8.2 +/- 2.3). The waiting time list for HT ranged from 1.2 to 6.4 years (M +/- SD = 3.1 +/- 2.6). The subjects had the following six psychospiritual concerns: death (PS), unfulfilled family responsibilities (PS and PDS), unaccomplished personal life goals (PS and PDS), poor body image (PDS), loss of significant others' support and love (PDS), and discrimination by others (PDS). The following seven needs of help were reported: comforting the fear of failure in heart transplantation (PS and PDS), meeting unfulfilled family responsibilities (PS and PDS), accomplishing personal life goal (PS and PDS), religious support (PS and PDS), establishing confidence in body image (PDS), establishing positive relationship with significant others (PS and PDS), and preparation for dying with dignity when necessary (PS and PDS). The in-depth investigation on Chinese HT recipients' psychospiritual needs was first compared in PDS and PS. More distress and needs were found in PDS than PS. These findings were attributed by the dilemma of pursuing prospective future versus taking the risk of loss of life, valuing families' and health professionals' support system, and expecting grief and dying with dignity. Health professionals are encouraged to understand and provide stage-specific support to help them meet psycho-spiritual needs.


Acute arsenic ingestion.
A 21-year-old man presented in shock after ingesting 2 g of arsenic trioxide. He died within 37 hours despite intensive treatment that included intramuscular dimercaprol and hemodialysis. Hemodynamic and laboratory data are presented illustrating the multisystem toxicities of inorganic arsenic. Hemodialysis, previously described as an effective therapeutic adjunct, was shown to be ineffective in this case.


Assessment of different mechanical treatments for improving the anaerobic biodegradability of residual raspberry extrudate.
Mechanical treatments can be simple and feasible methods for enhancing the anaerobic digestion of lignocellulosic substrates. This work aims to relate the direct effect of five different mechanical treatments, i.e., variation in the size and number of particles, with the variations in the chemical composition and, subsequently, the effect over the anaerobic digestion of residual raspberry extrudate, which was used as a model substrate. A high variation in the number of particles and the particle size distribution was achieved depending on the mechanical treatment applied, reaching the highest number of particles for the treatments with knife mills and mortar (around 8000 particles per gram). The higher number of particles was related to higher solubilisation, including phenolic compounds and sugars. The combination of knife mills and mortar pretreatment, which presented the highest number of particles, resulted in a 66% more of polyphenols in comparison to the raw substrate. However, the presence of anthocyanins was higher in mechanical treatments with less effect. The enhancement of the anaerobic digestion was clearly related to the increment in the number of particles of small size after the mechanical treatments. The highest methane yield coefficient (236 ± 11 mL CH4/g volatile solids) was achieved for the raspberry extrudate treated with knife mills.


Clinical Presentations of Breast Disorders in Pregnancy and Lactation.
The breast tissue undergoes significant physiological change during pregnancy and lactation. These changes can give rise to some unique disorders during pregnancy , puerperium and lactation or exaggerate pre-existing conditions. Clinical examination becomes less reliable due to textural change and density of breast tissue as a result of hormonal changes. The main symptoms during pregnancy and lactation are breast pain, mastitis, lactational abscess, breast lump, and blood- stained nipple discharge.Lactational mastitis/ abscess must be treated without delay. Open incision and drainage of lactational abscess is rarely required, any lactational abscess should be treated with appropriate antibiotics and ultrasound guided aspiration of the pus.Any breast lump during pregnancy and lactation should be investigated with triple assessment. Pregnancy associated breast cancer (PABC ) must be ruled out. The choice of investigations and treatment needs careful consideration. While ultrasound is the investigation of choice, mammography can be performed with abdominal shielding if malignancy is suspected. Core biopsy is necessary for evaluation of any breast pathology but it comes with risk of infection, bleeding, hematoma and even milk fistula.The treating clinical specialist must be aware of certain unusual unique clinical conditions in pregnancy and lactation including accessory axillary breast tissue, gigantomastia and Raynaud's phenomenon.


Novel integrated optical sensor based on a grating coupler triplet.
A novel approach for accomplishing robust integrated optical biosensors is presented. The principle is based on a symmetric grating coupler structure with the inherent feature of compensating disturbances originating from different kinds of mechanical instabilities. The complete sensor system has no moving parts and provides the high sensitivities typical for integrated optical sensors based on grating couplers. The feasibility of this approach is demonstrated by determining the refractive index of liquids as well as by measuring the specific binding of biomolecules (anti-chicken IgG) to immobilized chicken IgG on the sensor chip surface.


Surgical treatment of radiation injury to the rectosigmoid.
Radiation-induced injury to the rectosigmoid, the estimated incidence of which was 5.5%, was studied in 92 patients. The condition was surgically treated in 75 cases. Colostomy alone was undertaken in 40 patients, 37 of whom were greatly improved with no need for further surgery, while two had progression of rectal radiation damage and one died. Excisional surgery was performed in 35 cases--Hartmann procedure in 25 and resection with primary anastomosis in 10 cases. Five of these 35 patients died of operative complications. Of the total 16 colonic anastomoses, four leaked. Our impression is that a differentiated policy of proximal colostomy and intestinal resection is safe and effective, although a significant number of the patients will have a permanent colostomy.


Interaction between vasodilators and vasopressin in internal mammary artery and clinical significance.
Arginine vasopressin (AVP) has recently been demonstrated as an alternative in the treatment of severe refractory vasodilatation in coronary artery bypass grafting. However, AVP may be a spasmogen for graft spasm. We compared the in vitro antispastic effect among calcium-channel antagonists (nifedipine, diltiazem, and verapamil), nitroglycerin, and the highly selective AVP (V1) receptor antagonist [1-deaminopenicillamine, 4-valine, 8-D-arginine] vasopressin. Human internal mammary artery segments (n = 218) were studied in organ baths. The inhibitory effects of the above vasodilators on AVP-mediated contraction were studied in two ways: relaxation with AVP precontraction and depression of the AVP-induced contraction after pretreatment with vasodilators. All three calcium-channel antagonists caused limited relaxation (18.3%+/-5.4% for nifedipine, n = 11; 22.2%+/-3.8% for verapamil, n = 10; and 26.2%+/-7.5% for diltiazem, n = 9). The plasma concentration of calcium-channel antagonists had no significant depression effect on the AVP-induced contraction. In contrast, [1-deaminopenicillamine, 4-valine, 8-D-arginine] vasopressin caused full (100%, n = 11) and nitroglycerin caused nearly full (93%+/-3%, n = 10) relaxation. Pretreatment with [1-deaminopenicillamine, 4-valine, 8-D-arginine] vasopressin (10(-8), 10(-7), or 10(-6) mol/L, respectively) significantly increased the effective concentration for 50% of the AVP-induced contraction (10(-8.6)+/-10(0.1) mol/L, p = 0.009; 10(-7.8)+/-10(0.07) mol/L, p = 0.000; or 10(-6.9)+/-10(0.11) mol/L, p = 0.000 versus the control, 10(-9.24)+/-10(0.16) mol/L). However, nitroglycerin only slightly depressed the AVP-induced contraction. [1-Deaminopenicillamine, 4-valine, 8-D-arginine] vasopressin may provide specific antispastic effect in either prophylaxis or treatment of the AVP-related vasospasm in the internal mammary artery. Nitroglycerin may be effective in treatment but has little effect on prophylaxis. Use of calcium-channel antagonists may have little benefit in AVP-related vasospasm.


Cadmium triggers kidney cell apoptosis of purse red common carp (Cyprinus carpio) without caspase-8 activation.
Caspase-8, the essential initiator caspase, is believed to play a pivotal role in death receptor-mediated apoptotic pathway. It also participates in mitochondria-mediated apoptosis via cleavage of proapoptotic Bid in mammals. However, its role in fish remains elusive in Cadmium-induced apoptotic pathway. In this study, we isolated the caspase-8 gene from common carp, one of the most important industrial aquatic animals in China using rapid amplification of cDNA ends (RACE). The deduced amino acid sequence of caspase-8 comprised 475 amino acids, which showed approximately 64.1% identity and 79.8% similarity to zebrafish (Danio rerio) caspase-8, possessed two conserved death effector domains, a large subunit and a small subunit. Phylogenetic analysis demonstrated that caspase-8 formed a clade with zebrafish caspase-8. In kidney, cadmium (Cd) exposure triggered apoptosis and increased caspase-3 and -9 activities, whereas it did not affect caspase-8 activity. Real-time quantitative PCR analysis revealed that caspase-8 transcriptional level was not significantly increased in kidney after exposure to Cd. Using Western blot analysis, no caspase-8 cleaved fragment was detected and no significant alteration of procaspase-8 level was found with the same Cd-treated condition. Moreover, the immunopositive staining was predominantly limited to the cytoplasm of renal tubular epithelial cells and no remarkable changes of immunoreactivities were observed using immunohistochemical detection after Cd treatment. The results reveal that Cd can trigger apoptosis, while it cannot activate caspase-8 in purse red common carp.


A pharmacological effect of oestrone on human epidermis.
Oestrone has been found to depress the incorporation of tritiated thymidine, proline and histidine in human skin in vitro. The significance of this is briefly discussed.


Neonatal sympathoadrenal response to acute hypoxia: impairment after experimental intrauterine growth retardation.
Sympathoadrenal system function may be altered following intrauterine growth retardation (IUGR). We tested the hypothesis that the growth-retarded newborn rat pup has increased basal sympathoadrenal activity under normoxic conditions and a blunted sympathoadrenal response to acute hypoxia. IUGR was established by uterine artery ligation on d 18 of gestation in Sprague-Dawley rats. Growth-retarded pups were chosen as those whose birth wt was more than 2 x SD below the mean birth wt of control pups delivered to sham-operated dams. At 24 +/- 12 h of age cardiac sympathetic neuronal activity (CSNA) was determined by 3H-norepinephrine tracer and alpha-methyltyrosine techniques. Adrenal medullary catecholamine synthesis (CAT SYN) was measured by 14C-tyrosine precursor methods, and adrenal catecholamine release (CAT REL) was determined using alpha-methyl-tyrosine. IUGR and control pups were studied over a 120-min period of normoxia or hypoxia (FiO2 = 0.09). Under normoxic conditions, the IUGR pups had increased CSNA and increased adrenal CAT SYN and CAT REL compared to controls. Adrenal CAT REL in normoxic IUGR pups was selective for epinephrine. In response to acute hypoxia, control pups had increased CSNA and increased adrenal CAT SYN and CAT REL compared to normoxic controls, with the proportion of norepinephrine and epinephrine released mimicking the ratio of the two amines in the adrenal. In contrast, in hypoxic IUGR pups CSNA and adrenal CAT SYN did not increase, and norepinephrine alone was released from the adrenal medulla.(ABSTRACT TRUNCATED AT 250 WORDS)


Pulmonary hypertension in pregnancy and its effects on the fetus.
Pulmonary hypertension (PH) is an important and potentially lethal diagnosis for pregnant patients. Although pregnancy is usually contraindicated in this condition and there is no standard algorithm for the treatment of pregnant patients with PH, studies in recent years have shown improvement in maternal outcomes for those with PH. Many factors have likely contributed to the improved outcomes, including earlier treatment by multidisciplinary teams. Pregnant patients with PH require specialized management throughout pregnancy, especially in the early post-partum period. Echocardiography is an important diagnostic tool to follow cardiac function in these patients. PH and its treatment during pregnancy has significant implications on maternal as well as fetal outcomes. In this review, PH management during pregnancy and the fetal implications are summarized.


Reduced postprandial energy expenditure and increased exogenous fat oxidation in young woman after ingestion of test meals with a low protein content.
Macronutrient composition of diets can influence energy balance in humans. We tested the hypothesis whether low protein content in single meals may induce lower values of energy expenditure (EE) and fat oxidation (FO) as compared to adequate protein content. Indirect calorimetry was combined with a breath test using naturally 13C-enriched corn oil to differentiate between postprandial exogenous and endogenous FO. Young women ingested single meals containing either 3.9% (low protein, LP) or 11.4% (adequate protein, AP) of total energy (~3100 kJ) as protein. Postprandial EE was 160 kJ/6 h lower (p < 0.01) after LP meals and diet induced thermogenesis (DIT) increased less (p < 0.001) as compared to AP meals. Total postprandial FO was not significantly different between meals (~17 g/6 h). However, exogenous postprandial FO was significantly (p < 0.01) higher (4.28 +/- 1.57 g/6 h) after exposure to LP meals as compared to AP meals (1.87 +/- 1.00 g/6 h). Less than 10% of ingested fat (50 g) was oxidized in the postprandial phase. The overall postprandial fat balance was approximately + 33 g. Breath tests using naturally 13C-labeled corn oil mirror exogenous FO. Low protein meals resulted in reduced postprandial EE and increased exogenous FO as compared to adequate protein meals without differences in total FO.


Molecular diversity and evolution of defensins in the manila clam Ruditapes philippinarum.
Four types of defensins were identified in Manila clam and designated as Rpdef1, Rpdef2, Rpdef3 and Rpdef4, which encoded a polypeptide of 49, 46, 45 and 42 amino acids, respectively. Sequence alignments indicated that Rpdef1 shared 46.9% identity with Rpdef2, 40.8% with Rpdef3, and 34.7% with Rpdef4. Analysis of transcript polymorphism showed that Rpdef3 accounted for about 60% frequency of Rpdefs occurrence in clams from three geographic origins (Dalian, Qingdao and Hangzhou). By quantitative real-time RT-PCR (qRT-PCR) analysis, the transcripts of Rpdefs were mainly detected in hemocytes and they responded sensitively to bacterial challenge in hemocytes. Evolutionary analysis indicated that all Rpdefs were under positive selection with positively selected basic amino acid residues detected in the C-terminal regions, which perhaps have a functional relevance by modifying the charge distribution of Rpdefs. The results also showed some lineages with dN/dS > 1, suggesting positive selection pressures existed in some lineages of phylogeny tree constructed by mollusk defensins. Overall, our results suggest that Rpdefs perhaps played important roles in host defense and positive selection is the major driving force in generating high diversity of defensins in the Manila clam.


The energy-independent K(+)-H(+)-exchange was measured in intact mitochondria and submitochondrial particles at different K(+)-concentration of the incubation medium. Correlation was found between H+-displacement and specific activity of monoamine oxidase. It is concluded, that the energy-independent K(+)-H(+)-exchange is localized in the outer membrane of mitochondria.


Zearalenone (ZEN) disrupts the anti-inflammatory response of bovine oviductal epithelial cells to sperm in vitro.
Dietary contamination by Zearalenone (ZEN) has a detrimental effect on bovine fertility. Recently, we showed a novel anti-inflammatory response of bovine oviductal epithelial cells (BOEC) to active sperm cells in vitro. The aim of the present study was to investigate the effect of ZEN exposure of BOEC on the immune-related cytokine expression in response to bovine sperm. At concentrations of 100 and 1000ng/mL, ZEN induced the expression of TNF and IL1B (pro-inflammatory cytokines) as well as IL8 (chemokine) in BOEC in a dose-dependent manner. Furthermore, ZEN induced PTGES expression and PGE2 secretion in BOEC. Sperm co-culture induced an anti-inflammatory response in BOEC with upregulation of TGFB, secretion of PGE2 and downregulation of TNF. Most importantly, ZEN at 1-1000ng/mL eliminated the response of BOEC to sperm. Estradiol-17β (5ng/mL) treatment did not produce the same effects as ZEN, suggesting that the response of BOEC to ZEN is, at least in part, not mediated by estrogen receptors. Taken together, ZEN can produce inflammatory effects on BOEC by stimulating the expressions of pro-inflammatory cytokines and disrupt the normal interaction between sperm and BOEC at the level of cytokine expressions and PGE2 production. Thus, exposure of the bovine oviduct to ZEN may negatively affect sperm survival and reduce fertility.


Reversed-phase and hydrophobic-interaction high-performance liquid chromatography of elapid cardiotoxins.
The separation of proteins by hydrophobic-interaction HPLC and reversed-phase HPLC depends upon differences in the hydrophobicity of accessible surface groups. The elution order of a group of snake venom cardiotoxins was found to vary between these two HPLC methods. Circular dichroism spectroscopy showed that the eluant acetonitrile-trifluoroacetic acid used for reversed-phase HPLC altered the conformation of the toxins, whereas the salt-buffer eluting medium used for hydrophobic-interaction HPLC did not affect toxin conformation. The retention times of cardiotoxins on reversed-phase HPLC are therefore influenced by their conformational instability in the eluting medium which causes partial or complete unfolding. Hydrophobic interaction is clearly the preferred method with which to correlate the "surface hydrophobicity" of cardiotoxins and their biological effects.


Transcriptomic analysis provides insights into molecular mechanisms of thermal physiology.
Physiological trait variation underlies health, responses to global climate change, and ecological performance. Yet, most physiological traits are complex, and we have little understanding of the genes and genomic architectures that define their variation. To provide insight into the genetic architecture of physiological processes, we related physiological traits to heart and brain mRNA expression using a weighted gene co-expression network analysis. mRNA expression was used to explain variation in six physiological traits (whole animal metabolism (WAM), critical thermal maximum (CTmax), and four substrate specific cardiac metabolic rates (CaM)) under 12 °C and 28 °C acclimation conditions. Notably, the physiological trait variations among the three geographically close (within 15 km) and genetically similar F. heteroclitus populations are similar to those found among 77 aquatic species spanning 15-20° of latitude (~ 2,000 km). These large physiological trait variations among genetically similar individuals provide a powerful approach to determine the relationship between mRNA expression and heritable fitness related traits unconfounded by interspecific differences. Expression patterns explained up to 82% of metabolic trait variation and were enriched for multiple signaling pathways known to impact metabolic and thermal tolerance (e.g., AMPK, PPAR, mTOR, FoxO, and MAPK) but also contained several unexpected pathways (e.g., apoptosis, cellular senescence), suggesting that physiological trait variation is affected by many diverse genes.


Analysis of banding patterns and mosaic configurations in a case of ring chromosome 15.
Cytogenetic studies on lymphocytes from a 14-year-old mentally retarded girl with somatic anomalies suggestive of a chromosomal abnormality revealed a ring chromosome 15. The long arm of the defective chromosome is broken at band q24 or q25. The silver staining technique for nucleolus organizer regions showed that the ring had lost the achromatic stalk and the satellite. The chromosomal mosaicism resulting from the structural instability of the ring chromosome was analyzed and compared with 6 cases reported in the literature. It is proposed that the clinical manifestations in the different patients with ring chromosome 15 result from both the deficiency in the long arm and the mosaic configurations.


Direct cytolysis by partially-purified preparations of immune interferon.
Mouse IFN gamma preparations purified 30-fold were found to have direct cytolytic activity against a number of tumor and normal cells. Cell killing was determined using a sensitive, rapid and accurate assay which employed very low numbers of cells and very small quantities of interferon. The cytolytic activity of IFN gamma on 11 murine tumor cell lines was investigated. A 20-fold difference was found between the most-sensitive cell type, P-388 lymphoma, versus the most resistant cell type, C127v leukemia. A number of normal mouse cells was also found to have low to intermediate sensitivity to the cytolytic action of IFN gamma. Human IFN gamma was also shown to have cytolytic activity which, like mouse IFN gamma, was relatively species-specific. Direct cytolysis was not found to be a characteristic of IFN-alpha/beta. Different mechanisms of action for the antiviral and cytolytic activities of IFN gamma are indicated because the cytolytic titer of IFN gamma did not parallel its antiviral titer on most cell types and increasing the cell number produced a decrease in the cytolytic titer and an increase in the anti-viral titer. High concentrations of IFN gamma (i.e., 2,900 units/ml) resulted in complete lysis of cells within 24 h, while lower concentrations (i.e., 700 units/ml) resulted in a reversible inhibition of cell growth during this time period. Evidence that the cytolytic substance in the IFN gamma preparation was IFN gamma include the following: (1) both antiviral and anticellular activities copurified through a 30-fold purification; and both activities were (2) relatively species-specific; (3) sensitive to heat; (4) inactivated by low pH and (5) neutralized by antibodies to IFN gamma. Therefore, we propose the possibility that direct cytolysis is yet another of IFN gamma's distinctive antivities.


Relationship Between Decreased Cortisol and Development of Chronic Pain in Traumatically Injured.
The relationship between pain and stress is widely accepted, yet the underlying neuroendocrine mechanisms are poorly understood. Cortisol secretion during a stress response, may distract attention from a painful stimulus, inhibiting pain. However, when pain is the stressor, cortisol secretion may intensify the pain experience and condition a fear-based memory of pain. This study attempts to determine the relationship between acute pain, chronic pain, and cortisol in the traumatically injured population. Secondary analyses of a prospective observational study with participants from a Midwestern Adult Level I Trauma Center post traumatic injury, with interview and serum cortisol taken at hospitalization (baseline) and 6 mo after discharge, was completed using Ward's Method hierarchical cluster analysis, Pearson's correlations, and linear regressions. Two major clusters were identified. The Chronic Pain group were those who had severe pain at discharge and continued to have severe pain as defined by Numeric Pain Score. The Resolved Pain group were those who had moderate pain at discharge and their pain improved or resolved. Pain score at discharge significantly, negatively correlated with baseline cortisol levels (r = -0.142, P = 0.02). Minority status, single individuals, low cortisol at baseline, and greater psychological distress at baseline significantly increased the likelihood of developing chronic pain. Low cortisol and greater psychological stress, which are also associated with minority status and single individuals, contribute to chronic pain in the traumatically injured population. Trauma victims without an adequate cortisol response to acute injury and pain are at risk for development of chronic pain after injury.


Sewage sludge acidogenic fermentation for organic resource recovery towards carbon neutrality: An experimental survey testing the headspace influence.
Volatile fatty acids (VFAs) produced by acidogenic digestion of sewage sludge are very interesting bio-products which can contribute to carbon neutrality of wastewater treatment plants. Studies on the production of VFAs from sewage sludge from fermenters with membrane are limited. In view of above, VFAs from a fermenter pilot plant equipped with a membrane bioreactor and fed with real sewage sludge has been monitored. The effect of headspace volume (HdV) on VFA production was studied for the first time to elucidate the optimal operation conditions. Specifically, three fermenter HdV values (namely, 20, 40 and 60% of the total volume) have been investigated. Results revealed that the HdV of 20% ensured the highest sCOD production (900 mgCOD/L) and VFA/COD ratio (45.4%). High value of HdV (namely, 40 and 60%) strongly decreased the acidogenic fermentation performance in terms of VFA production.


Ipomoea pes-tigridis L. extract accelerates wound healing in Wistar albino rats in excision and incision models.
An annual herb, Ipomoea pes-tigridis L. (Convolvulaceae) is widely used for its anti-inflammatory and anti-spasmodic properties in traditional medicine. As well as treating wounds, fever, skin disorders, and other ailments, it is also used for other purposes. This study investigated polyphenolic content, antioxidant activity, RP-HPLC, wound healing, and antioxidant enzyme activity. In terms of I. pes-tigridis potential for healing wounds, there is no scientific data available. Hence this study is designed to use animal models to investigate the ethnopharmacological report. The crude extracts of stem and leaf were subjected to phytochemicals, TPC, TTC, TFC, and free radical scavenging assays (DPPH, ABTS, etc). Excision and incision models were used to assess wound healing using the screened extracts (IPLEA, IPLM, IPSEA, and IPSM). Various tissue parameters (hydroxyproline, hexosamine, hexuronic acid content), as well as antioxidant enzyme activity (SOD, Catalase, GPX, LPO), were also examined. The maximum amount of polyphenolic content was found in IPLM (TPC- 118.86 ± 5.94 mg GAE/g, TTC - 75.25 ± 2.64 mg TAE/g, and TFC-25.73 ± 0.99 mg GAE/g) with significant IC50 value of 1.65 ± 0.87 μg/mL among all the extracts. Coumaric acid was reported high (92.86 mg/g) in RP-HPLC analysis of crude extract in IPLEA. The in vivo excision wound healing model revealed that 1% IPLM had better healing property with the maximum wound healing area (0.098 ± 0.03 cm) and wound concentration (95.56 ± 1.95%) was reported with the significance level of ***P < 0.001, **P < 0.01, *P < 0.05. In the incision model, IPLM represented maximum tensile strength (27500 gf). A significant functional effect of the granulation tissue parameters and enzyme antioxidants on the wound-healed area of dry tissue was also observed. Finally, the histopathological analysis showed enhanced re-epithelialization, fibroblast proliferation, and collagen synthesis in wound-treated animal tissue in both models. According to the present study, antioxidant-rich I. pes-tigridis promotes healthy cell regeneration while reducing inflammation and oxidative stress for wound healing. Additionally, it also enhances circulation and promotes healing.


Biochemical properties and progress in cancers of tRNA-derived fragments.
tRNA-derived small RNAs (tRFs), a kind of noncoding RNAs, are generated from transfer RNAs. tRFs have some types according to their source and sizes. They play important roles in cell life and carcinogenesis. In this paper, we review the biogenesis and biological properties. We also focus on current progress of tRFs and some tsRNAs such as tRF-Leu-CAG, which have been studied or will be further investigated in tumorgenesis and diagnostic biomarkers in the clinic.


Structure, gene order, and nucleotide composition of mitochondrial genomes in parasitic lice from Amblycera.
Parasitic lice have unique mitochondrial (mt) genomes characterized by rearranged gene orders, variable genome structures, and less AT content compared to most other insects. However, relatively little is known about the mt genomes of Amblycera, the suborder sister to all other parasitic lice. Comparing among nine different genera (including representative of all seven families), we show that Amblycera have variable and highly rearranged mt genomes. Some genera have fragmented genomes that vary considerably in length, whereas others have a single mt chromosome. Notably, these genomes are more AT-biased than most other lice. We also recover genus-level phylogenetic relationships among Amblycera that are consistent with those reported from large nuclear datasets, indicating that mt sequences are reliable for reconstructing evolutionary relationships in Amblycera. However, gene order data cannot reliably recover these same relationships. Overall, our results suggest that the mt genomes of lice, already know to be distinctive, are even more variable than previously thought.


Nitrogen defects-rich porous graphitic carbon nitride for efficient photocatalytic hydrogen evolution.
Graphitic carbon nitride (CN) is considered as a promising photocatalyst for solar energy conversion. However, low specific surface area and fast electrons and holes recombination restrict the photocatalytic applications of CN material. Herein, a nitrogen defect-rich and highly porous CN nanostructure (CN-LT) was prepared by combining two strategies, i.e., LiOH treatment and heat etching. The as-prepared nitrogen defect-rich porous CN-LT not only has a larger specific surface area, as compared with pristine CN, but also the photogenerated electron-hole separation was boosted remarkably. Using Pt as a co-catalyst and lactic acid aqueous solutions as sacrificial reagent under visible light irradiation (λ > 400 nm), the hydrogen evolution reaction (HER) rate for CN-LT (1.54 mmol h-1 g-1) was 19.25 times higher than that for pristine CN (0.08 mmol h-1 g-1). While subjecting pristine CN to heat etching under the same experimental conditions, excepting the use of LiOH (CN-T), an increase in HER rate of 7.5 times was obtained. Our current study may shed more light on the enhancement of the photocatalytic activity of bulk CN materials by altering their microstructure.


Decreased number and reduced NKG2D expression of Vδ1 γδ T cells are involved in the impaired function of Vδ1 γδ T cells in the tissue of gastric cancer.
In cancer patients, impaired function of immune cells--such as CD8(+) T cells, NK cells, and dendritic cells--reportedly results in tumor progression. Although γδ T cells also play a critical role in tumor defense, their function remains unclear in cancer patients. The frequency and function of γδ T cells in peripheral blood, normal gastric mucosa, and cancer tissue were evaluated by multicolor flow cytometry. We also determined NKG2D expression on γδ T cells in gastric cancer patients. The frequency of Vδ1 γδ T cells in gastric cancer tissue is significantly lower than in normal gastric mucosa; however, differences in the frequencies of Vδ2 and Vγ9 γδ T cells between normal gastric mucosa and gastric cancer tissue were not statistically significant. The Vδ1 γδ T cells from gastric cancer tissue produce significantly less IFN-γ than those from normal gastric mucosa do. Expression of NKG2D on Vδ1 γδ T cells from gastric cancer tissue was significantly lower than in normal gastric mucosa. We also found a significant correlation between NKG2D expression and IFN-γ production of Vδ1 γδ T cells in gastric cancer tissue. Vδ1 γδ T cells show decreased frequency and impaired function in gastric cancer tissue, for which decreased NKG2D expression might be one of the mechanisms. Modalities specifically targeting NKG2D in Vδ1 γδ T cells may provide a breakthrough treatment for gastric cancer patients.


Biogenic amine formation and microbial spoilage in chilled garfish (Belone belone belone)--effect of modified atmosphere packaging and previous frozen storage.
To evaluate biogenic amine formation and microbial spoilage in fresh and thawed chilled garfish. Storage trials were carried out with fresh and thawed garfish fillets at 0 or 5 degrees C in air or in modified atmosphere packaging (MAP: 40% CO2 and 60% N2). During storage, sensory, chemical and microbial changes were recorded and histamine formation by isolates from the spoilage microflora was evaluated at 5 degrees C. Photobacterium phosphoreum was responsible for histamine formation (>1000 ppm) in chilled fresh garfish. The use of MAP did not reduce the histamine formation. Strongly histamine-producing P. phosphoreum isolates formed 2080-4490 ppm at 5 degrees C, whereas below 60 ppm was formed by other P. phosphoreum isolates. Frozen storage inactivated P. phosphoreum and consequently reduced histamine formation in thawed garfish at 5 degrees C markedly. Photobacterium phosphoreum can produce above 1000 ppm of histamine in chilled fresh garfish stored both in air and in MAP. Freezing inactivates P. phosphoreum, extends shelf life and markedly reduces histamine formation in thawed MAP garfish during chilled storage. At 5 degrees C, more than 1000 ppm of histamine was formed in garfish; thus even when it is chilled this product represents a histamine fish-poisoning risk.


An association between hostility and poor adherence to treatment in patients suffering from depression.
Direction of hostility, as measured by the Hostility and Direction of Hostility Questionnaire, was found to be associated with poor compliance during a treatment trial of a new tricyclic antidepressant. An association with other factors such as the type and severity of depression was not found. There are other studies in the literature which support these observations although some controversy surrounds the role played by personality variables in adherence to treatment. Hostility has been shown to be a possible factor in the aetiology of depression, an important factor in the classification of depression, and a symptom of depression which improves with treatment. Hostility may have a particularly large importance in compliance with treatment for depression or may have been overlooked in studies in compliance with treatment for non-psychiatric disorders.


Effect of partial dietary replacement of fishmeal by yellow mealworm (Tenebrio molitor) larvae meal on the innate immune response and intestinal antioxidant enzymes of rainbow trout (Oncorhynchus mykiss).
The present study investigated the effects of 25 and 50% dietary inclusion of yellow mealworm, Tenebrio molitor (TM), larvae on the intestinal antioxidant enzymes, lipid peroxidation and immune system of rainbow trout, Oncorhynchus mykiss. Fish were fed for 90 days with a control diet (TM0) or with 2 test diets (TM25 and TM50) where 35 and 67% of the fishmeal was replaced by TM larvae meal respectively. At the end of the growth trial, humoral immune parameters were assessed in the sera of the fish and the activities of antioxidant enzymes were measured in intestine homogenates. The present results show a strong increase of the activity of the antioxidant intestinal enzymes and reduction of lipid peroxidation. Increased trypsin inhibition and faster antibacterial activity of the serum were also detected. These evidences could be due to the physiological similarities between the exoskeleton of parasites and insects. The present findings reinforce the previous results which revealed a positive effect of the dietary TM on rainbow trout growth and further confirm its adequacy for fishmeal partial replacement in aquafeed.


S-nitrosation of Cys-800 of HIF-1alpha protein activates its interaction with p300 and stimulates its transcriptional activity.
Hypoxia inducible factor 1 (HIF-1) is a heterodimeric transcriptional complex that plays pivotal role in the regulation of cellular utilization of oxygen as well as glucose and is an essential regulator of angiogenesis in solid tumor and ischemic disorders. Recently HIF-1alpha, a subunit of HIF-1 complex, was characterized as a potential target for S-nitrosation, providing no information about the impact of this posttranslational modification on the protein transactivation. Cys-800 of HIF-1alpha protein has reactive SH-group, which is critical for the recruitment of p300 co-activator that is necessary for transcriptional activity of HIF-1 complex. Here we report that S-nitrosation of Cys-800 activates HIF-1alpha-p300 interaction and stimulates protein transactivation. We have found that S-nitrosation of the HIF-1alpha C-terminal domain by nitric oxide derived from donors and nitric oxide synthase increases protein transcriptional activity. The increase of HIF-1 transcriptional activity was not observed in the case of Cys-800 substitution to Ala, though other protein thiol groups were nitrosated. Experiments with GST pull-down assay suggest that S-nitrosation of Cys-800 stimulates the recruitment of p300 co-activator protein to the HIF-1alpha C-terminal domain.


Particle separation by a moving air-liquid interface in a microchannel.
Particle separation is an important topic in microfluidic field and has recently gained significant attention in sample preparations for biological and chemical studies. In this paper, a novel particle separation method was proposed. In this method, the particles were separated by the air-liquid interface in a microchannel. The motion of the air-liquid interface was controlled with a syringe pump. Depending on the air-liquid interface speed, the liquid film thickness and the viscous force on particles were changed and the particles were separated by sizes. We observed the separation of 1.01 μm particles from the larger particles when the air-liquid interface speed was less than 11 μm/s, and the separation of both 1.01 μm and 5.09 μm particles from the larger particles when the interface speed was between 11 μm/s and 120 μm/s. When the speed was higher than 120 μm/s, the drag force of the liquid flow generated by the advancing interface on particles was so strong that the flow removed all particles off from the bottom channel wall and there were no particles left behind the advancing interface.


The reimbursement for expensive medicines: stakeholder perspectives on the SMA medicine nusinersen and the Dutch Coverage Lock policy.
The reimbursement for expensive medicines poses a growing challenge to healthcare worldwide. In order to increase its control over the costs of medicines, the Dutch government introduced the Coverage Lock (CL) policy in 2015. The CL postpones decisions regarding reimbursement of expensive medicines until detailed advice on i.e., cost-effectiveness has been given. The CL has been in place for six years, has raised many questions and concerns, but currently, no evaluation is known to the authors. A better understanding of the effects of the CL on all stakeholders involved may contribute to reflections on the CL process and help find ways to improve it. An evaluation of Dutch policy will also be relevant for other countries that aim to optimize reimbursement procedures for expensive treatments. To perform this evaluation, we focused on the CL procedure for the medicine nusinersen. Nusinersen is the first treatment for spinal muscular atrophy (SMA). Following EMA approval in May 2017, it was placed in the CL. The analysis of cost-effectiveness and added therapeutic value resulted in an advice for reimbursement limited to children younger than 9.5 years at the start of treatment; this was implemented from August 2018 onwards. Qualitative stakeholder perspective analysis of the CL procedure focusing on nusinersen with 15 stakeholders. Stakeholders raised key issues of the CL based on their experience with nusinersen: emotional impact of the CL, duration of the CL procedure, appropriateness of the CL procedure for different types of medicines, transparency of the CL, a wish for patient-centred decision-making and the lack of uniformity of access to expensive treatments. Stakeholders supported measures to control healthcare expenses and to ensure reasonable pricing. They considered the delay in access to therapies and lack of procedural transparency to be the main challenges to the CL. Stakeholders also agreed that the interests of patients deserve more attention in the practical implementation of the reimbursement decision. Stakeholders suggested a number of adjustments to improve the CL, such as a faster start with conditional reimbursement programs to ensure access and intensify European collaboration to speed up the assessment of the medicine.


Cloning of circadian rhythmic pathway genes and perturbation of oscillation patterns in endocrine disrupting chemicals (EDCs)-exposed mangrove killifish Kryptolebias marmoratus.
To investigate the effect of endocrine disrupting chemicals (EDCs) on the circadian rhythm pathway, we cloned clock and circadian rhythmic pathway-associated genes (e.g. Per2, Cry1, Cry2, and BMAL1) in the self-fertilizing mangrove killifish Kryptolebias marmoratus. The promoter region of Km-clock had 1 aryl hydrocarbon receptor element (AhRE, GTGCGTGACA) and 8 estrogen receptor (ER) half-sites, indicating that the AhRE and ER half sites would likely be associated with regulation of clock protein activity during EDCs-induced cellular stress. The Km-clock protein domains (bHLH, PAS1, PAS2) were highly conserved in five additional fish species (zebrafish, Japanese medaka, Southern platyfish, Nile tilapia, and spotted green pufferfish), suggesting that the fish clock protein may play an important role in controlling endogenous circadian rhythms. The promoter regions of Km-BMAL1, -Cry1, -Cry2, and -Per2 were found to contain several xenobiotic response elements (XREs), indicating that EDCs may be able to alter the expression of these genes. To analyze the endogenous circadian rhythm in K. marmoratus, we measured expression of Km-clock and other circadian rhythmic genes (e.g. Per2, Cry1, Cry2, and BMAL1) in different tissues, and found ubiquitous expression, although there were different patterns of transcript amplification during different developmental stages. In an estrogen (E2)-exposed group, Km-clock expression was down-regulated, however, a hydroxytamoxifen (TMX, nonsteroid estrogen antagonist)-exposed group showed an upregulated pattern of Km-clock expression, suggesting that the expression of Km-clock is closely associated with exposure to EDCs. In response to the exposure of bisphenol A (BPA) and 4-tert-octyphenol (OP), Km-clock expression was down-regulated in the pituitary/brain, muscle, and skin in both gender types (hermaphrodite and secondary male). In juvenile K. marmoratus liver tissue, expression of Km-clock and other circadian rhythmic pathway-associated genes showed a regular oscillation pattern over a period of approximately 24h during a 12L:12D cycle. However, the circadian rhythm of BPA-exposed juvenile K. marmoratus liver tissue was perturbed over a 12L:12D period. This study will aid in our understanding of how EDCs perturb endogenous circadian rhythms, particularly in BPA-exposed fish liver tissue.


Functions of the Vasa gene in Schistosoma japonicum as assessed by RNA interference.
Vasa, an enzyme belonging to the helicase family, contributes to the regulation of reproductive system development in many species. Thus, we hypothesized that the Vasa3 gene may function in the reproductive system of the parasite Schistosoma japonicum (S. japonicum), which is a major causative agent of schistosomiasis. It is a severe disease globally affecting humans and animals. To test this hypothesis, we firstly conducted whole mount in situ hybridization analyses and found that the S. japonicum Vasa3 (SjVasa3) gene was expressed mainly in the reproductive organs. We then explored the reproductive functions of Vasa3 in S. japonicum using RNA interference (RNAi) techniques. Coupled schistosomes collected from mice 28days post infection (dpi) were transfected three times with SjVasa3-specific small interfering RNA (siRNA) and cultured in vitro for up to 10days. As measured by quantitative PCR (qPCR) and Western blot analysis, levels of SjVasa3 mRNA and protein in Vasa siRNA treated worms were significantly reduced compared with untreated and scrambled siRNA treated worms. Confocal laser scanning microscopy (CLSM) images showed markedly siRNA induced changes in the morphology of the reproductive organs, especially in the female ovary, vitellarium and the male testes. SjVasa3 gene silencing also significantly reduced egg production. These data demonstrate that SjVasa3 is essential in reproductive organ development and egg production in S. japonicum, and could be a potential target for developing novel compounds to treat schistosomiasis.


Senescence and steroid hormone receptor reactivities in accessory sex glands of elderly rats (Sprague-Dawley) following exogenous hormonal therapy.
The aim of this study was to characterize the stromal and epithelial distribution of AR, ERα and ERβ reactivities in the different accessory sex glands of elderly rats and during strong hormonal changes. Ten month old male rats were divided into six senile groups and submitted to treatment: Senile/Control group (SC); Senile/Testosterone group (ST): Senile/Estrogen group (SE); Castrated group (CA); Castrated/Testosterone group (CT); Castrated/Estrogen group (CE). After a 30-day treatment, the prostatic ventral lobe (VL), dorsal lobe (DL) and coagulating gland (CG) samples were processed for immunohistochemistry and Western Blotting. The results showed that AR immunoreactivity was characterized in the epithelium of VL and DL in senile/control rats and senile rats submitted to exogenous hormonal therapy. AR reactivity in the coagulating gland was verified predominantly in the stromal cells in the different experimental groups. ERα reactivity occurred predominantly in the stromal compartment in all accessory sex glands. In the DL and CG, ERα immunoreactivities were intense in the groups which received testosterone (ST) and estrogen (SE). ERβ immunoreactivity in the CG was verified in the stromal compartment in the different experimental groups, showing a positive response to both increased testosterone and estrogen levels. ERβ reactivity, in the DL, was intensified in the stroma of senile rats with higher serum testosterone levels, and in senile rats with increased serum estrogen levels, especially in the glandular epithelium. Thus, the results revealed different distribution pattern of steroid hormone receptors in each one of the prostatic lobes in senescence, especially in the prostate dorsal lobe and coagulating gland, which is a fundamental factor due to the fact that major prostatic diseases occur in a later period of life.