Impairment of the mitochondrial electron transport chain due to sleep deprivation in mice.
It has been demonstrated that sleep deprivation is associated with altered expression of genes related to metabolic processes, response to stress and inflammation, circadian sleep/wake cycles, regulation of cell proliferation and various signaling pathways. However, the molecular mechanisms underlying these changes remain poorly understood. Thus, the present study aims to characterize the function of the mitochondrial electron transport chain in the brain using an animal model of paradoxical sleep deprivation (PSD). The question of whether sleep recovery (rebound) can reverse changes found after PSD is also addressed. Adult male inbred C57BL/6J mice were randomly distributed into three groups: home-cage control, PSD and sleep rebound groups. The PSD and rebound groups were subjected to PSD for 72 h. After this sleep deprivation period, the rebound group was returned to its home cage and allowed to sleep in an undisturbed and spontaneous fashion for 24h. The mitochondrial complex I-III, complex II, succinate dehydrogenase and complex II-III activities were then measured by spectrophotometric methods in sub-mitochondrial particles extracted from the prefrontal cortex, hippocampus, striatum and hypothalamus. Our results showed a significant decrease in the activity of complex I-III in the PSD and rebound groups as compared to the control group. The complex II and II-III activity were particularly decreased in the hypothalamus of the sleep rebound group. These results are consistent with the involvement of sleep in energy metabolism and corroborate previous experiments demonstrating the importance of the hypothalamus in sleep regulation.


Role of soluble epoxide hydrolase in the abnormal activation of fibroblast-like synoviocytes from patients with rheumatoid arthritis.
Rheumatoid arthritis (RA) is an autoimmune disease characterized by enigmatic pathogenesis. Polyunsaturated fatty acids (PUFAs) are implicated in RA's development and progression, yet their exact mechanisms of influence are not fully understood. Soluble epoxide hydrolase (sEH) is an enzyme that metabolizes anti-inflammatory epoxy fatty acids (EpFAs), derivatives of PUFAs. In this study, we report elevated sEH expression in the joints of CIA (collagen-induced arthritis) rats, concomitant with diminished levels of two significant EpFAs. Additionally, increased sEH expression was detected in both the synovium of CIA rats and in the synovium and fibroblast-like synoviocytes (FLS) of RA patients. The sEH inhibitor TPPU attenuated the migration and invasion capabilities of FLS derived from RA patients and to reduce the secretion of inflammatory factors by these cells. Our findings indicate a pivotal role for sEH in RA pathogenesis and suggest that sEH inhibitors offer a promising new therapeutic strategy for managing RA.


Psycho-educational group therapy in acute psychiatric units: creating a psychosocial culture. An update of spread and effectiveness of a psychosocial intervention in Italian psychiatric wards.
The implementation of a Cognitive-Behavioural Group Intervention (CBGI) in Italian general hospital psychiatric units started in the years 2000-2001 in two Italian regions. Over the years it has became more and more popular also in other psychiatric units located in the rest of the country. Based on the "stress-vulnerability-coping" theory, the CBGI is a replicable and innovative psychosocial intervention that promotes the active involvement of inpatients in decisions concerning their individual objectives and care. In the present article, the authors briefly describe this intervention and the main findings regarding its implementation in several psychiatric units in different Regions of Italy. The authors emphasize that such a psychosocial approach to inpatient care is needed because it can produce improved clinical outcomes, reduction in untoward events and increased staff and inpatient satisfaction. However, its introduction and use still represent a major cultural and managerial challenge in our country.


Association between statin use on delirium and 30-day mortality in patients with chronic obstructive pulmonary disease in the intensive care unit.
Delirium occurs frequently in patients with chronic obstructive pulmonary disease in the intensive care unit. Effective prevention and treatment strategies for delirium remain limited. We aimed to assess delirium and 30-day mortality in patients with chronic obstructive pulmonary disease who were statin and non-statin users. In this retrospective study, patients with chronic obstructive pulmonary disease were identified from the Medical Information Mart for Intensive Care database (MIMIC-IV). The primary exposure variable was the use of statins 3 days after entering the intensive care unit and the primary outcome measure was the presence of delirium. The secondary outcome measure was 30-day mortality. Since the cohort study was retrospective, we used an inverse probability weighting derived from the propensity score matching to balance different variables. Among a cohort of 2725 patients, 1484 (54.5%) were statin users. Before propensity score matching, the prevalence of delirium was 16% and the 30-day mortality was 18% in patients with chronic obstructive pulmonary disease. Statin use was significantly negatively correlated with delirium, with an odds ratio of 0.69 (95% CI 0.56-0.85, p < 0.001) in the inverse probability weighted cohort and 30-day mortality of 0.7 (95% CI 0.57-0.85, p < 0.001). Statin use is associated with a lower incidence of delirium and 30-day mortality in patients with chronic obstructive pulmonary disease in the intensive care unit.


The metabolic effects of mirabegron are mediated primarily by β3 -adrenoceptors.
The β3 -adrenoceptor agonist mirabegron is approved for use for overactive bladder and has been purported to be useful in the treatment of obesity-related metabolic diseases in humans, including those involving disturbances of glucose homeostasis. We investigated the effect of mirabegron on glucose homeostasis with in vitro and in vivo models, focusing on its selectivity at β-adrenoceptors, ability to cause browning of white adipocytes, and the role of UCP1 in glucose homeostasis. In mouse brown, white, and brite adipocytes, mirabegron-mediated effects were examined on cyclic AMP, UCP1 mRNA, [3 H]-2-deoxyglucose uptake, cellular glycolysis, and O2 consumption. Mirabegron increased cyclic AMP levels, UCP1 mRNA content, glucose uptake, and cellular glycolysis in brown adipocytes, and these effects were either absent or reduced in white adipocytes. In brite adipocytes, mirabegron increased cyclic AMP levels and UCP1 mRNA content resulting in increased UCP1-mediated oxygen consumption, glucose uptake, and cellular glycolysis. The metabolic effects of mirabegron in both brown and brite adipocytes were primarily due to actions at β3 -adrenoceptors as they were largely absent in adipocytes derived from β3 -adrenoceptor knockout mice. In vivo, mirabegron increased whole body oxygen consumption, glucose uptake into brown and inguinal white adipose tissue, and improved glucose tolerance, all effects that required the presence of the β3 -adrenoceptor. Furthermore, in UCP1 knockout mice, the effects of mirabegron on glucose tolerance were attenuated. Thus, mirabegron had effects on cellular metabolism in adipocytes that improved glucose handling in vivo, and were primarily due to actions at the β3 -adrenoceptor.


Diagnostic Evaluation of Co-Occurrence of Acanthamoeba and Fungi in Keratitis: A Preliminary Report.
To test the hypothesis that the coexistence of Acanthamoeba with other forms of microbial keratitis, especially fungal keratitis (FK), is more prevalent than suspected. A prospective diagnostic study whereby patients presenting with stromal keratitis were additionally tested for Acanthamoeba, irrespective of the initial diagnosis. In addition to the routine workup with Gram stain, KOH mount, and cultures on blood agar and potato dextrose agar, nonnutrient agar was included. Confocal microscopy was performed where feasible. Samples for polymerase chain reaction studies were also obtained. We present the preliminary report of the first 100 culture-positive cases. The primary outcome measured was the number of coexistent Acanthamoeba and FK. The secondary outcomes were the total number of Acanthamoeba cases detected and the correlation between clinical diagnosis and microbiological observations. Of the first 100 cases, 22 were culture positive for Acanthamoeba, of which 9 were associated with concurrent FK, 5 with bacterial keratitis, and 8 in isolation. However, only 2 cases were diagnosed clinically as Acanthamoeba, whereas 5 were Acanthamoeba suspects. An additional 4 cases of fungal/Acanthamoeba coexistence in keratitis were revealed purely by confocal microscopy. Acanthamoeba can coexist with other forms of microbial keratitis. The frequency of infection coexistent or otherwise is higher than reported, and the possibility of coinfection must be considered especially in unresponsive cases. Including nonnutrient agar and confocal microscopy in all cases of keratitis would perhaps translate into better treatment strategies and outcomes.


Surface strategies for catalytic CO2 reduction: from two-dimensional materials to nanoclusters to single atoms.
Redox catalysis, including photocatalysis and (photo)electrocatalysis, may alleviate global warming and energy crises by removing excess CO2 from the atmosphere and converting it to value-added resources. Nano-to-atomic two-dimensional (2D) materials, clusters and single atoms are superior catalysts because of their engineerable ultrathin/small dimensions and large surface areas and have attracted worldwide research interest. Given the current gap between research and applications in CO2 reduction, our review systematically and constructively discusses nano-to-atomic surface strategies for catalysts reported to date. This work is expected to drive and benefit future research to rationally design surface strategies with multi-parameter synergistic impacts on the selectivity, activity and stability of next-generation CO2 reduction catalysts, thus opening new avenues for sustainable solutions to climate change, energy and environmental issues, and the potential industrial economy.


Profiling and quantification of pluripotency reprogramming reveal that WNT pathways and cell morphology have to be reprogramed extensively.
Pluripotent state can be established via reprogramming of somatic nuclei by factors within an oocyte or by ectopic expression of a few transgenes. Considered as being extensive and intensive, the full complement of genes to be reprogrammed, however, has never been defined, nor has the degree of reprogramming been determined quantitatively. Here, we propose a new concept of reprogramome, which is defined as the full complement of genes to be reprogrammed to the expression levels found in pluripotent stem cells (PSCs). This concept in combination with RNA-seq enables us to precisely profile reprogramome and sub-reprogramomes, and study the reprogramming process with the help of other available tools such as GO analyses. With reprogramming of human fibroblasts into PSCs as an example, we have defined the full complement of the human fibroblast-to-PSC reprogramome. Furthermore, our analyses of the reprogramome revealed that WNT pathways and genes with roles in cellular morphogenesis should be extensively and intensely reprogrammed for the establishment of pluripotency. We further developed a new mathematical model to quantitate the overall reprogramming, as well as reprogramming in a specific cellular feature such as WNT signaling pathways and genes regulating cellular morphogenesis. We anticipate that our concept and mathematical model may be applied to study and quantitate other reprogramming (pluripotency reprogramming from other somatic cells, and lineage reprogramming), as well as transcriptional and epigenetic differences between any two types of cells including cancer cells and their normal counterparts.


[The quantitative and qualitative characteristics of the reaction of committed bone marrow cell precursors in rats to burn trauma].
The article describes the dynamics of colony-forming units in diffusion chamber of tubular bone marrow compartments 1 and 7 days after a burn. It was found that under normal conditions the activity of the axial part of the bone marrow is twice that of the cortical part. After a burn the inductive capacity of the cortical part becomes equal to that of the axial part. A wider qualitative spectrum of colonies occurs due to the effect of short-distance humoral factors (eosinophils, mast cells).


Trimetallic PtAuNi alloy nanoparticles as an efficient electrocatalyst for the methanol electrooxidation reaction.
Platinum is an excellent electrocatalyst. However, the disadvantages of Pt as an electrocatalyst lie in its poor earth abundance, high cost, and poor stability due to surface poisoning. Thus it remains a challenge to find suitable alternative electrocatalysts and/or reduce the use of Pt. Herein, we report the solvothermal synthesis of bimetallic (PtAu and PtNi) and trimetallic (PtAuNi) alloy nanoparticles (NPs) with controlled percentages of individual metals. With an optimized Ni content, the trimetallic (Pt66Au11Ni23) alloy NPs show superior electrocatalytic activity (in terms of lower onset oxidation potential and higher mass activity) not only to bimetallic alloy NPs (PtAu and PtNi) but also to commercial Pt/C (20% Pt loading) for methanol electrooxidation (MEO). This enhanced electrocatalytic activity is due to the synergistic role of different metals in MEO catalysis. In particular, the catalytic activity is found to be controlled by the balance between the adsorption of methanol species on Pt and the removal of carbonaceous species from the catalyst surface by Au and Ni, as demonstrated here. The multimetallic alloy of optimal individual content thereby not only reduces the Pt content in the catalyst but also exhibits higher electrocatalytic activity than Pt/C for MEO that is desirable for fuel cell applications.


The role of proteolytic enzymes derived from crude bacterial collagenase in the liberation of hepatocytes from rat liver. Identification of two cell-liberating mechanisms.
Crude bacterial collagenase was chromatographed on DEAE-cellulose to yield three peaks with proteolytic activity: an arginine esterase (DEAE-1), gelatinase (DEAE-2) and a caseinolytic activity (DEAE-3). The arginine esterase and gelatinase activity fractions were slightly contaminated with each other but neither possessed caseinolytic activity; the caseinolytic fraction was devoid of arginine esterase and gelatinase activities. In addition, crude collagenase was fractionated by ZnII-affinity chromatography to produce a gelatinase peak (ZnII peak 1), which was free from arginine esterase and caseinolytic activities. The four fractions were compared to crude collagenase in their ability to liberate rat hepatocytes by using either liver slices or a standard perfusion technique. Compared to crude collagenase (0.05-0.1% w/v), which produced 70-80% liver digestion with approximately 80% cell viability, digestion with equivalent quantities of the isolated enzymic activities was relatively poor. Gelatinase activity (ZnII peak 1) was wholly ineffective and DEAE-1 and DEAE-2 each possessed only slight digestive properties. Hepatocyte liberation by the caseinolytic activity, DEAE-3, was partially successful (30-40% digestion, 25-30% viability) but only a portion of liver tissue was digested regardless of the quantity of DEAE-3 used. However, by mixing certain fractions before perfusion two gelatinase-dependent, cell-releasing mechanisms were identified: (a) DEAE-3 with ZnII peak 1 and (b) DEAE-1 mixed with either DEAE-2 or ZnII peak 1. Each system compared creditably with the digestive properties of an equivalent activity of crude collagenase. At present we are attempting to determine any differences between hepatocytes produced by the two enzymic mechanisms.


Silver nanoparticles exert concentration-dependent influences on biofilm development and architecture.
To investigate the impact of silver nanoparticles (AgNPs) on the biofilm growth and architecture. Silver nitrate was reduced by d-maltose to prepare AgNPs in the presence of ammonia and sodium hydroxide. The physicochemical properties of AgNPs were characterized by transmission electron microscopy, ultraviolet-visible spectroscopy and inductively coupled plasma mass spectrometry. The development of biofilm with and without AgNPs was explored by crystal violet stain. The structures of mature biofilm were visually studied by confocal laser scanning microscopy and scanning electron microscopy. Bacterial cell, polysaccharide and protein within biofilm were assessed quantitatively by colony-counting method, phenol-sulphuric acid method and Bradford assay, respectively. The spherical AgNPs (about 30 nm) were successfully synthesized. The effect of AgNPs on Pseudomonas aeruginosa biofilm development was concentration-dependent. Biofilm was more resistant to AgNPs than planktonic cells. Low doses of AgNPs exposure remarkably delayed the growth cycle of biofilm, whereas high concentration (18 μg/mL) of AgNPs fully prevented biofilm development. The analysis of biofilm architecture at the mature stage demonstrated that AgNPs exposure at all concentration led to significant decrease of cell viability within treated biofilms. However, sublethal doses of AgNPs increased the production of both polysaccharide and protein compared to control, which significantly changed the biofilm structure. AgNPs exert concentration-dependent influences on biofilm development and structure, which provides new insight into the role of concentration played in the interaction between antibacterial nanoparticles and biofilm, especially, an ignored sublethal concentration associated with potential unintended consequences.


Digital Health Intervention to Support Refugees in Switzerland.
Experiences of war and persecution often lead to mental health problems, resulting in post-traumatic stress disorders. In this work, we design a digital platform that aims at helping refugees coming to Switzerland by providing exercises for their mental health and information about daily life in Switzerland. In collaboration with the Swiss Red Cross (SRC), we collected requirements and developed a concept for information provision through in this platform. The architecture of a progressive web application (PWA) was identified as to best fulfill the given requirements. Based on the collected requirements mockups were created. In user interviews, we received feedback regarding the future system. We learned that the platform should include an avatar, which guides the user through the entire platform and asks questions. All texts should be accessible by a read-aloud function and exercises should be provided as videos. In summary, we learned that it is essential to involve the future user group in the development process since it is characterized by cultural diversity that has to be considered in the development and design. Enriched by this input, the next step is to realize the application in terms of a prototype.


[Menopause knowledge and experience for a group of women].
This ethnographic study is aimed at understanding a group of women's knowledge about menopause and how they experience it. The methods used for data collection were participant observation, recorded interviews and previously produced drawings. Data were analyzed based on a theoretical framework using both the ethnographic method and biographic interpretativism. From the narratives the authors obtained the cultural categories and themes. Thus it was possible to conclude that menopause is understood as a result of a unique construction integrated to a network of meanings built by the group that condition knowledge and experience within certain cultural patterns, among them the fact that menopause means leaving womanhood behind.


Practical application of the 10-mg warfarin initiation nomogram.
Initiation of warfarin therapy is a clinical challenge. A 10-mg warfarin initiation nomogram was recently validated in a randomized controlled trial. We sought to determine the efficacy and safety of this 10-mg warfarin initiation nomogram in 'real-life' daily practice. A retrospective cohort including all outpatients beginning concurrent treatment with warfarin and low-molecular-weight heparin over a 24-month period in our Thrombosis Unit was reviewed. Eight hundred and forty-one patients were included; of them, 640 (76.1%) were started on the nomogram. The nomogram was entirely followed in 324 patients (38.5%). The efficacy and safety profile was similar to that observed in the original clinical trial; 86% of patients managed according to the nomogram reached the international normalized ratio target of 2.0-3.0 within 5 days. Mean duration of low-molecular-weight heparin treatment was 6.0 +/- 1.9 days, and 3.7% of patients had an international normalized ratio of at least 5.0 in the first 4 weeks of treatment. The 10-mg nomogram effectively results in an early therapeutic international normalized ratio with a good safety profile in 'real-life' daily practice.


Cardiovascular regulation by endogenous nitric oxide is essential for survival after acute haemorrhage.
Our previous studies have indicated that endogenous nitric oxide serves as a physiologically important inhibitor of vascular tone during acute haemorrhage. This vasodilator action attenuates the concomitant reflex adrenergic constriction and thereby prevents critical reduction of tissue blood flow. The present study aimed to evaluate the overall importance of this nitric oxide regulation for survival after acute haemorrhage. This was done by comparative observations of survival time and circulatory, metabolic and histopathological changes after an acute standardized lethal blood loss (45%) in cats exposed to nitric oxide synthase (NOS) inhibition and in matched control animals with intact nitric oxide regulation. NOS inhibition was instituted by intravenously administered N omega-nitro-L-arginine methyl ester. The survival time averaged 2 h 49 min in the NOS-blocked animals and 10 h 14 min in the control animals (P < 0.001). NOS inhibition thus reduced the posthaemorrhagic survival time to < 30% of that in the control cats. Haemorrhage in the NOS-blocked animals led to rapidly developing arterial hypotension, increased anaerobic metabolism, metabolic lactacidosis, hyperkalaemia, and morphological tissue damage especially in heart and liver, in spite of maintained arterial normoxia, which signifies tissue hypoxia caused by seriously impaired nutritional blood supply. At the time of death of the NOS-blocked cats, the control animals still exhibited a virtually normal circulatory/metabolic state. A much later, and more slowly developing circulatory/metabolic deterioration was observed in the control animals. These differences between the two groups of animals indicate that nitric oxide release, by its vasodilator action, to a significant extent helps to maintain an adequate nutritional blood supply to the tissues in acute haemorrhage.


Physiotherapy for New Actors: Disputes and Innovations in Care for Chronically Ill Children.
Based on interviews with physiotherapists, we explored the arguments produced by them about their actions and the relationships of a change that has been taking place in the profile of illness in children, where an increased prevalence of complex chronic conditions has been observed. Structured in a qualitative approach, data were generated based on two provocative affirmations that generated arguments by the professionals. Data analysis revealed statements that can be referred to as results of power struggles over the performance in this new field of knowledge. Physiotherapists seek to affirm themselves through their knowledge and by approaching other professional categories, especially Medicine, in order to feel recognized and valued. However, we see that these professionals use a simpler discourse with family and caregivers, creating a bond of trust and identification with them, which facilitates their care and access to children. We point out the need for the hospital environment to be recognized as a place for the action of other categories, besides Medicine and Nursing, and the valuation in pediatric physiotherapy.


Functional knockout of the TRPV1 channel has no effect on the exercise pressor reflex in rats.
The role played by the transient receptor potential vanilloid 1 (TRPV1) channel on the thin fibre afferents evoking the exercise pressor reflex is controversial. To shed light on this controversy, we compared the exercise pressor reflex between newly developed TRPV1+/+ , TRPV1+/- and TRPV1-/- rats. Carotid arterial injection of capsaicin (0.5 μg), evoked significant pressor responses in TRPV1+/+ and TRPV1+/- rats, but not in TRPV1-/- rats. In acutely isolated dorsal root ganglion neurons innervating the gastrocnemius muscles, capsaicin evoked inward currents in neurons isolated from TRPV1+/+ and TRPV1+/- rats but not in neurons isolated from TRPV1-/- rats. The reflex was evoked by stimulating the tibial nerve in decerebrated rats whose femoral artery was either freely perfused or occluded. We found no difference between the reflex in the three groups of rats regardless of the patency of the femoral artery. For example, the peak pressor responses to contraction in TRPV1+/+ , TRPV1+/- and TRPV1-/- rats with patent femoral arteries averaged 17.1 ± 7.2, 18.9 ± 12.4 and 18.4 ± 8.6 mmHg, respectively. Stimulation of the tibial nerve after paralysis with pancuronium had no effect on arterial pressure, findings which indicated that the pressor responses to contraction were not caused by electrical stimulation of afferent tibial nerve axons. We also found that expression levels of acid-sensing ion channel 1 and endoperoxide 4 receptor in the L4 and 5 dorsal root ganglia were not upregulated in the TRPV1-/- rats. We conclude that TRPV1 is not needed to evoke the exercise pressor reflex in rats whose contracting muscles have either a patent or an occluded arterial blood supply. KEY POINTS: A reflex arising in contracting skeletal muscle contributes to the increases in arterial blood pressure, cardiac output and breathing evoked by exercise. The sensory arm of the reflex comprises both mechanoreceptors and metaboreceptors, of which the latter signals that blood flow to exercising muscle is not meeting its metabolic demand. The nature of the channel on the metaboreceptor sensing a mismatch between supply and demand is controversial; some believe that it is the transient receptor potential vanilloid 1 (TRPV1) channel. Using genetically engineered rats in which the TRPV1 channel is rendered non-functional, we have shown that it is not needed to evoke the metaboreflex.


Jupiter's Radiation Belts: Can Pioneer 10 Survive?
Model calculations of Jupiter's electron and proton radiation belts indicate that the Galilean satellites can reduce particle fluxes in certain regions of the inner magnetosphere by as much as six orders of magnitude. Average fluxes should be reduced by a factor of 100 or more along the Pioneer 10 trajectory through the heart of Jupiter's radiation belts in early December. This may be enough to prevent serious radiation damage to the spacecraft.


The Relevance of Fluid and Blood Management Using Microcirculatory Parameters in Children Undergoing Craniofacial Surgery.
Perioperative management of bleeding in children can be challenging. Microvascular imaging techniques have allowed evaluating the effect of blood transfusion on the microcirculation, but little is known about these effects in children. We aimed to investigate the effects of blood management using macro- and micro-hemodynamic parameters measurement in children undergoing craniofacial surgery. This is a prospective observational repeated measurement study including fourteen children. The indications for blood transfusion were changes of hemoglobin/hematocrit (Hct) levels, the presence of signs of altered tissue perfusion and impaired microcirculation images. Total and perfused vessel densities, proportion of perfused vessels, microvascular flow index, and systemic parameters (hemoglobin, Hct, lactate, mixed venous oxygen saturation, K+, heart rate, mean arterial blood pressure) were evaluated baseline (T1), at the end of the surgical bleeding (T2) and end of the operation (T3). Four patients did not need a blood transfusion. In the other 10 patients who received a blood transfusion, capillary perfusion was higher at T3 (13[9-16]) when compared with the values of at T2 (11[8-12]) (P < 0.05) but only 6 patients reached their baseline values. Although blood transfusions increased Hct values (17 ± 2.4 [T2]-19 ± 2.8 [T3]) (P < 0.05), there was no correlation between microvascular changes and systemic hemodynamic parameters (P > 0.05). The sublingual microcirculation could change by blood transfusion but there was not any correlation between microcirculation changes, hemodynamic, and tissue perfusion parameters even with Hct values. The indication, guidance, and timing of fluid and blood therapy may be assessed by bedside microvascular analysis in combination with standard hemodynamic and biochemical monitoring for intraoperative bleeding in children.


Surfactin isoforms isolated from a mushroom derived Bacillus halotolerans DMG-7-2.
A new iso-C14 [Val2, Val7] surfactin isoform (1) together with eight known ones (2-9), was isolated from the culture of a mushroom derived bacterium, Bacillus halotolerans DMG-7-2. The structures of them were mainly elucidated by NMR and MS data, and the NMR data of 5 also was reported for the first time. The absolute configuration of 1 was determined by Marfey's analysis (for amino acid residues) and the 13C NMR calculation of the two plausible epimers of 1 (for fatty acid). Compounds 1-9 showed moderate cytotoxicity against two human cancer cell lines (A549, MCF-7) and mice microglial BV2 cells, the IC50 values ranged from 8.91 to 33.00 µM, and the IC50 values of the positive control 5-FU were 99.94, 71.49 and 0.12 µM, respectively.[Formula: see text].


Urofacial syndrome: A subset of neurogenic bladder dysfunction syndromes?
The urofacial syndrome is probably a subset of neurogenic bladder dysfunction syndromes characterized by detrusor-sphincter discoordination along with a characteristic inversion of facial expression with laughing. This characteristic facial expression can facilitate early detection of this disorder, which leads to poor bladder emptying with high residual urine, hydro-nephrosis with vesico-ureteral reflux and potentially renal failure if left untreated. The etiology of the urofacial syndrome is unknown. In our case, a 12-year-old boy of Middle-Eastern origin presented to the Outpatient Department of our hospital with left pyelonephritis, hydronephrosis and bladder dilatation. Voiding cystourethrography performed 15 days later revealed left vesicoureteral reflux. Cystoscopy revealed bladder trabeculation however an anatomic urethral obstruction was not noticed. Both, neurological examination and radiography of the lumbosacral spine were normal. Urodynamic evaluation revealed the typical findings of detrusor-sphincter discoordination.


Effects of highly or relatively selective 5-HT1A receptor agonists on lordosis in female rats.
To investigate the role of serotonin (5-HT) receptor 1A or 7 in regulating lordosis behavior in female rats, ovariectomized rats were treated with 3 kinds of receptor agonists and lordosis behavior was observed. The injected agents were the selective 5-HT1A receptor agonist, buspirone (BUS), the highly selective 5-HT1A receptor agonist, 8-hydroxy-2-(di-n-propylamino)tetralin ((+/-)8-OH-DPAT), and the 5-HT1A and 5-HT7 receptor agonist, (R)-8-hydroxy-2-(di-n-propylamino)tetralin ((+)8-OH-DPAT). A behavioral test was performed after ovariectomy and subcutaneous implantation of a silicon tube containing estradiol. Female rats in which the lordosis quotient (LQ) was over 70 were intraperitoneally injected with several doses of these agents. As a result, in the BUS group, the dose of 3 mg/kg bw, but not 1 mg/kg was effective for suppressing lordosis. On the other hand, an inhibitory effect was observed from 0.25 mg/kg and 0.5 mg/kg in the (+)8-OH-DPAT and (+/-)8-OH-DPAT groups, respectively. In the time-course experiment, in all drug-treated groups, LQ decreased to lower than 20 after 15 min and low LQ continued for 1 hr at least. Measurement of locomotor activity using an infrared sensor system showed no relation between the decrease in lordosis by these agents and spontaneous locomotion. These results indicate that 5-HT1A is strongly involved in the lordosis-inhibiting circuit of the serotonin neurons.


Overexpression of S-adenosylmethionine decarboxylase impacts polyamine homeostasis during development of Dictyostelium discoideum.
The polyamines putrescine, spermidine and spermine are essential polycations involved in the regulation of cellular proliferation. They exert dynamic effects on nucleic acids and macromolecular synthesis in vitro but their specific functions in vivo are poorly understood. Here, we have modulated the spermidine levels either by overexpressing the S-adenosylmethionine decarboxylase (samdc) gene or treating the cells with methylglyoxal-bis (guanylhydrazone) (MGBG), an inhibitor of SAMDC. In Dictyostelium, overexpression of SAMDC slowed cell proliferation, delayed development and arrested cells in the S-phase of the cell cycle. Treatment with MGBG reduced cell proliferation and stimulated development, but in samdcOE cells, it increased cell proliferation suggesting critical levels of spermidine to be important. In samdcOE cells, spermidine levels remained high throughout development but only small changes in the spermine levels were observed. Initial putrescine levels did increase but reverted to wild-type levels after the mound stage. As tight regulation of polyamine homeostasis is required, we identified genes that could be involved in its maintenance. In conclusion, we characterised samdcOE cells and observed the maintenance of polyamine homeostasis during the development of Dictyostelium cells.


Recombinant DnaK Orally Administered Protects Axenic European Sea Bass Against Vibriosis.
Vibrio anguillarum causes high mortality in European sea bass (Dicentrarchus labrax) larviculture and is a hindering factor for successful sustainable aquaculture of this commercially valuable species. Priming of the innate immune system through administration of immunostimulants has become an important approach to control disease outbreaks in marine fish larviculture. This study was conducted to evaluate immunostimulation by Escherichia coli HSP70 (DnaK) in axenic European sea bass larvae in order to protect the larvae against vibriosis. DnaK stimulates the immune response in crustaceans and juvenile fish against bacterial infections. The use of axenic fish larvae allows to study immunostimulation in the absence of an interfering microbial community. At 7 days post-hatching, larvae received a single dose of alginate encapsulated recombinant DnaK. Two non-treated control groups in which animals either received empty alginate microparticles (C1) or no alginante microparticles (C2 and C3) were included in the study. Eighteen hours later, all larvae, except the ones from group C3 (non-infected control) were challenged with V. anguillarum (105 CFU, bath infection). Mortality was daily recorded until 120 h post infection and at 18, 24, and 36 h post infection, larvae were sampled for expression of immune related genes. Results showed that V. anguillarum induced an immune response in axenic sea bass larvae but that the innate immune response was incapable to protect the larvae against deadly septicaemic disease. In addition, we showed that administration of alginate encapsulated DnaK to axenic European sea bass larvae at DAH7 resulted in a significant, DnaK dose dependent, upreglation of immune sensor, regulatory and effector genes. Significant upregulation of cxcr4, cas1 and especially of hep and dic was correlated with significant higher survival rates in V. anguillarum infected larvae. In the future recombinant DnaK might perhaps be used as a novel immunostimulant in sea bass larviculture.


Regression of Anterior Disk-Osteophyte Complex Following Cervical Laminectomy and Fusion for Cervical Spondylotic Myelopathy.
A retrospective case-control study. To investigate whether posterior cervical laminectomy and fusion modifies the natural course of anterior disk-osteophyte complex in patients with multilevel cervical spondylotic myelopathy. Dorsal migration of the spinal cord is the main purported mechanism of spinal cord decompression following cervical laminectomy and fusion but other potential mechanisms have received scant attention in the literature. This study was conducted to investigate whether cervical laminectomy and fusion affects the size of anterior disk-osteophyte complex. The medical records and radiographic imaging of 44 patients who underwent cervical laminectomy and fusion for cervical spondylotic myelopathy between 2006 and 2013 were analyzed. The size of the anterior disk-osteophyte complex was measured preoperatively and postoperatively on MR images taken at an interval of >3 months apart. A control group consisted of 20 nonoperatively treated advanced cervical spondylosis patients. Patients in the control met the same inclusion and exclusion criteria and also had sequential magnetic resonance imaging (MRI) taken at an interval of >3 months apart. The nonoperative and operative groups were statistically similar in the pertinent patient demographics and characteristics including sex, age, time to second MRI, size of anterior disk-osteophyte complex on baseline MRI, mean number of levels affected, and percentage of patients with T2 signal change. As expected the mJOA scores were significantly lower in the operative versus nonoperative cohort (13.6 vs. 16.5, P<0.01). A significant decrease in the size of anterior disk osteophyte was observed in the operative group postoperatively (P<0.01). In comparison, there was no statistically significant change in the size of the anterior disk-osteophyte complex in the control group (P>0.05). The magnitude of the change in disk size between the 2 groups was statistically significant (P<0.01). The findings of this study suggest that regression of anterior disk-osteophyte complex occurs following cervical laminectomy and fusion, and likely provides another mechanism of spinal cord decompression.


Protocols for ancient DNA typing.
Molecular analysis of fossil and archaeological remains has been established as a powerful tool in providing new insight in phylogenetic investigations. The overlapping set of molecular modifications and degradation that forensic samples share with archaeological specimen suggests the application of similar technical approaches to the respective biological material. Polymerase chain reaction is the molecular technique of choice for the retrieval of specimen deoxyribonucleic acid (DNA) molecules. Because of intrinsic sensitivity, potential contaminations from exogenous DNA sources must be monitored through the entire process by the introduction of multiple blank controls. Cloning and sequencing of polymerase chain reaction products often is the only way to discriminate between contaminations and endogenous sequences as well as to identify variable positions from nucleotide modifications/DNA polymerase errors. Phylogenetic analysis and investigations of the pattern of substitutions are an additional and necessary step to validate the retrieved sequence. Comparison with available related samples (modern or extinct) is critical to correctly validate the results and to avoid artifactual data.


Annelida (Oligochaeta and Aphanoneura) from the Natural Reserve of Isla Martín García (upper Río de la Plata estuary, Argentina): biodiversity and response to environmental variables.
The Island of Martín García--located in the Upper Río de la Plata, to the south of the Uruguay River--is an outcropping of the crystalline basement. Fourteen sampling sites were selected, five along the littoral section of the island and nine in inland ponds. Four major environmental variables were measured: water temperature, dissolved oxygen, electrical conductivity, and pH. A total of 34 species of Oligochaeta and Aphanoneura were found, 30 belonging to Naididae plus one species each of the Narapidae, Lumbricidae, Enchytraeidae, and Aeolosomatidae. The thirteen most frequent species were: A. leydigi (30%), N. bonettoi (13%), L. hoffmeisteri (11%), N. variabilis (10%), S. trivandrana (6.5%), A. pigueti (5.6%), D. sawayai (4.5%), D. digitata (3.5%), C. diastrophus (2.7%), A. costatus (2.5%), P. longiseta (2.0%), Enchytraeidae (1.5%), and A. p. paraguayensis (1.4%). UPGMA clustering of species based on their occurrence in different ecological conditions revealed two main species groups. Canonic-correspondence analysis (CCA) was conducted with the 15 most frequent and abundant species in the 9 sampling sites and the 4 environmental variables. Results from the CCA revealed that the order of fluctuation of the environmental variables during the sampling period was, from the greatest to the least: dissolved oxygen, conductivity, pH, and water temperature. Approximately 97.6% of the correlations between species and environmental variables were expressed on axis 1 of the ordination diagram. Species richness correlated with the four environmental variables in the following order, from the weakest to the strongest: water temperature, pH, electrical conductivity, and dissolved oxygen.


Differential roles for STIM1 and STIM2 in store-operated calcium entry in rat neurons.
The interaction between Ca(2+) sensors STIM1 and STIM2 and Ca(2+) channel-forming protein ORAI1 is a crucial element of store-operated calcium entry (SOCE) in non-excitable cells. However, the molecular mechanism of SOCE in neurons remains unclear. We addressed this issue by establishing the presence and function of STIM proteins. Real-time polymerase chain reaction from cortical neurons showed that these cells contain significant amounts of Stim1 and Stim2 mRNA. Thapsigargin (TG) treatment increased the amount of both endogenous STIM proteins in neuronal membrane fractions. The number of YFP-STIM1/ORAI1 and YFP-STIM2/ORAI1 complexes was also enhanced by such treatment. The differences observed in the number of STIM1 and STIM2 complexes under SOCE conditions and the differential sensitivity to SOCE inhibitors suggest their distinct roles. Endoplasmic reticulum (ER) store depletion by TG enhanced intracellular Ca(2+) levels in loaded with Fura-2 neurons transfected with YFP-STIM1 and ORAI1, but not with YFP-STIM2 and ORAI1, which correlated well with the number of complexes formed. Moreover, the SOCE inhibitors ML-9 and 2-APB reduced Ca(2+) influx in neurons expressing YFP-STIM1/ORAI1 but produced no effect in cells transfected with YFP-STIM2/ORAI1. Moreover, in neurons transfected with YFP-STIM2/ORAI1, the increase in constitutive calcium entry was greater than with YFP-STIM1/ORAI1. Our data indicate that both STIM proteins are involved in calcium homeostasis in neurons. STIM1 mainly activates SOCE, whereas STIM2 regulates resting Ca(2+) levels in the ER and Ca(2+) leakage with the additional involvement of STIM1.


Metabolic and health complications of obesity.
Overnutrition manifested by obesity has emerged as a major health problem in affluent countries. In spite of increased interest in fitness, obesity is on the increase in the United States. This is particularly so among children and adolescents. Although obesity is associated with many risk factors for diseases, the mechanisms whereby it enhances disease risk are not fully understood. Such an understanding is needed to develop strategies for management of these conditions. In this report we suggest that overnutrition produces clinical diseases only in individuals who already possess a metabolic weakness or "defect" in a given system. In the absence of such underlying defects, overnutrition, or obesity, is well tolerated. One of the most common consequences of obesity is dyslipidemia, that is, elevations of very low-density lipoprotein (VLDL) triglycerides and low-density lipoprotein (LDL) cholesterol and low concentrations of high-density lipoprotein (HDL) cholesterol. The major effect of overnutrition on lipoprotein metabolism is to stimulate the production of VLDL. For patients who have an underlying defect in lypolysis of VLDL triglycerides, hypertriglyceridemia will develop in the obese state. For those who have defective clearance of LDL, obesity will accentuate hypercholesterolemia. Both of these effects can be explained by overproduction of VLDL, due to obesity, combined with a genetic defect in clearance of VLDL or LDL. The mechanism whereby obesity causes a lowering of HDL cholesterol is uncertain, although it could enhance removal of HDL by an excess of adipose tissue. Another disease associated with obesity is cholesterol gallstones. The presence of obesity more than doubles the risk for gallstones. Two underlying factors increase the danger for gallstones: a deficiency of hepatic secretion of bile acids and a tendency for formation of cholesterol crystals in bile. Overnutrition promotes the synthesis of whole-body cholesterol, and the only route for excretion of this excess cholesterol is through the biliary tree.(ABSTRACT TRUNCATED AT 400 WORDS)


Inhibition of SUMO-independent PML oligomerization by the human cytomegalovirus IE1 protein.
In human cytomegalovirus-infected cells, the immediate-early IE1 protein disrupts the subnuclear structures known as the PML oncogenic domains or PODs, via the induction of PML desumoylation. This activity correlates with the functions of IE1 in transcriptional regulation and in the stimulation of lytic infection. Here, the effects of IE1 in induction of desumoylation of PML were characterized. IE1 did not interfere with the formation of sumoylated forms of PML in vitro. In in vitro assays using the sumoylated proteins, a SUMO-specific protease SENP1 desumoylated both PML and IE1. However, the IE1 proteins generated from bacteria or insect cells were unable to desumoylate PML in the same conditions. Although both IE1 and SUMO proteases such as SENP1, Axam and SuPr-1 efficiently desumoylated PML in co-transfection assays, they exerted different effects on the localization of PML. In cells transfected with either SENP1 or SuPr-1, the number of PML foci was reduced significantly and these remnant PML foci were devoid of SUMO-1 signals. However, in cells co-transfected with both SUMO proteases and IE1, these SUMO-independent PML foci were also completely disrupted. Furthermore, IE1, but not SENP1, was shown to disrupt the PML foci generated via transfection of a sumoylation-deficient mutant of PML. These data suggest that IE1 exhibits neither an inhibitory effect on sumoylation of PML nor intrinsic SUMO protease activity against PML in vitro. The finding that IE1 is capable of disrupting SUMO-independent PML aggregates suggests that inhibition of PML oligomerization by IE1 may play an important role in inducing PML desumoylation in vivo.


[3H-1-glutamate binding with the synaptic membranes isolated from the cerebral cortex and hippocampus of Krushinskiĭ-Molodkina strain rats].
Specific binding of 3H-L-glutamate to synaptic membranes isolated from the cerebral cortex and hippocamp of Wistar and Krushinsky-Molodkina (KM) rats examined both in a quiet awake state and after audiogenic seizures was compared. The dissociation constant (KD) values and binding capacity (Bmax) for KM rats did not differ significantly from the corresponding parameters of binding determined for Wistar rats (KD--89.8 +/- 18.1 and 102.6 +/- 12.5 nm, Bmax--1.23 +/- +/- 0.08 and 1.30 +/- 0.15 pmol/mg for the cortex and hippocamp, respectively). After audiogenic seizures the binding capacity of the hippocamp of KM rats was reduced by 30%. It is suggested that hippocampal glutamate receptors of KM rats are involved in the mechanism of convulsive activity formation.


l-Arginine supplementation in severe asthma.
BACKGROUNDDysregulation of l-arginine metabolism has been proposed to occur in patients with severe asthma. The effects of l-arginine supplementation on l-arginine metabolite profiles in these patients are unknown. We hypothesized that individuals with severe asthma with low fractional exhaled nitric oxide (FeNO) would have fewer exacerbations with the addition of l-arginine to their standard asthma medications compared with placebo and would demonstrate the greatest changes in metabolite profiles.METHODSParticipants were enrolled in a single-center, crossover, double-blind l-arginine intervention trial at UCD. Subjects received placebo or l-arginine, dosed orally at 0.05 mg/kg (ideal body weight) twice daily. The primary end point was moderate asthma exacerbations. Longitudinal plasma metabolite levels were measured using mass spectrometry. A linear mixed-effect model with subject-specific intercepts was used for testing treatment effects.RESULTSA cohort of 50 subjects was included in the final analysis. l-Arginine did not significantly decrease asthma exacerbations in the overall cohort. Higher citrulline levels and a lower arginine availability index (AAI) were associated with higher FeNO (P = 0.005 and P = 2.51 × 10-9, respectively). Higher AAI was associated with lower exacerbation events. The eicosanoid prostaglandin H2 (PGH2) and Nα-acetyl-l-arginine were found to be good predictors for differentiating clinical responders and nonresponders.CONCLUSIONSThere was no statistically significant decrease in asthma exacerbations in the overall cohort with l-arginine intervention. PGH2, Nα-acetyl-l-arginine, and the AAI could serve as predictive biomarkers in future clinical trials that intervene in the arginine metabolome.TRIAL REGISTRATIONClinicalTrials.gov NCT01841281.FUNDINGThis study was supported by NIH grants R01HL105573, DK097154, UL1 TR001861, and K08HL114882. Metabolomics analysis was supported in part by a grant from the University of California Tobacco-Related Disease Research Program program (TRDRP).


Path lengths in protein-protein interaction networks and biological complexity.
We investigated the biological significance of path lengths in 12 protein-protein interaction (PPI) networks. We put forward three predictions, based on the idea that biological complexity influences path lengths. First, at the network level, path lengths are generally longer in PPIs than in random networks. Second, this pattern is more pronounced in more complex organisms. Third, within a PPI network, path lengths of individual proteins are biologically significant. We found that in 11 of the 12 species, average path lengths in PPI networks are significantly longer than those in randomly rewired networks. The PPI network of the malaria parasite Plasmodium falciparum, however, does not exhibit deviation from rewired networks. Furthermore, eukaryotic PPIs exhibit significantly greater deviation from randomly rewired networks than prokaryotic PPIs. Thus our study highlights the potentially meaningful variation in path lengths of PPI networks. Moreover, node eccentricity, defined as the longest path from a protein to others, is significantly correlated with the levels of gene expression and dispensability in the yeast PPI network. We conclude that biological complexity influences both global and local properties of path lengths in PPI networks. Investigating variation of path lengths may provide new tools to analyze the evolution of functional modules in biological systems.


[A Case of Laparoscopic Abdominoperineal Resection for Rectal Cancer with Left-Sided Inferior Vena Cava].
A 66-year-old-man was hospitalizedfor the treatment of rectal cancer. Preoperative contrast-enhancedcomputedtomography revealedrectal cancer, left-sided inferior vena cava, and distant metastasis. He was diagnosed with cStage Ⅳb rectal cancer. Laparoscopic abdominoperineal resection with standard lymph node dissection was performed. There have been a few reports on laparoscopic colorectal resection for patients with left-sided inferior vena cava. We report a case of laparoscopic abdominoperineal resection for a patient with left-sided inferior vena cava.


Alpha-1 antitrypsin deficiency, SZ phenotype: a rare type of a rare disease. Case report.
Alpha-1 antitrypsin deficiency is one of the genetic diseases with a clear impact on the structure and function of the lung, rarely diagnosed and treated. We present the case of a 51-year-old female patient, heavy smoker, known with chronic obstructive pulmonary disease (COPD) for 12 years, untreated, who was hospitalized for the first time in our clinic having symptoms of a severe COPD exacerbation. She has significant cardiac disease (rheumatic mitral disease, with previous episodes of pulmonary edema and cardiac arrest) and hepatitis B. The patient is hypoxic, with severe mixed ventilatory dysfunction. During the hospitalisation she received treatment of the exacerbation and after that she received recommendation of chronic inhaled bronchodilator and corticosteroid treatment. The test for alpha-1 antitrypsin deficiency has detected a plasma of 63 mg/dl, SZ phenotype. The patient returns for a second evaluation. Functional tests are significantly improved (despite inconsistent treatment) with the impressive improvement of FEV7 values and identification by plethysmography of a restrictive syndrome. Echocardiography identifies mitral valve changes likely rheumatic, severe pulmonary hypertension. Computer tomography was performed, highlighting discrete interstitial changes and denying the existence of emphysema. Marked increase in FEV1 values supported adding bronchial asthma to the list of diagnosis and recommendation to continue inhaled corticosteroid combination bronchodilator as treatment. The particularity of the case is the rare phenotype, association of asthma and COPD as the clinical manifestation and the presence of comorbidities, which complicates the diagnosis and prognosis.


Arbutamine stress perfusion imaging in dogs with critical coronary artery stenoses: (99m)Tc-sestamibi versus (201)Tl.
Having previously shown that dobutamine reduces (99m)Tc-methoxyisobutylisonitrile (sestamibi [MIBI]) uptake in normal myocardium by elevating intracellular calcium, we hypothesized that arbutamine, which has less inotropic effect than dobutamine, might cause less reduction in MIBI uptake, thereby improving defect contrast. In this study using a canine model, we compared the effects of arbutamine stress on myocardial blood flow, myocardial MIBI uptake, and systolic thickening in the presence of a coronary artery stenosis. Arbutamine was infused (0.5-250 ng/kg/min) in 8 open-chest dogs with critical coronary stenoses that abolished flow reserve. At the time of peak arbutamine effect, MIBI (296 MBq), (201)Tl (27.75 MBq), and microspheres were coinjected. The dogs were killed 5 min later, and myocardial tracer activities and flow were quantified by well counting. Ex vivo imaging of heart slices was also performed. Arbutamine increased mean heart rate, peak positive left ventricular pressure and its first time-derivative, and normal-zone myocardial thickening. Stenotic zone flow and thickening did not increase during arbutamine infusion. MIBI uptake versus flow was significantly lower than (201)Tl uptake at the same flow values. By imaging, defect magnitude (stenotic/normal) was greater for (201)Tl than MIBI (0.57 vs. 0.77; P < 0.001) [corrected]. In the presence of coronary stenoses that abolished regional flow reserve, myocardial uptake of MIBI, compared with (201)Tl, significantly underestimated the arbutamine-induced flow heterogeneity. The attenuation of MIBI uptake and diminished defect contrast during arbutamine stress were comparable with those previously reported for dobutamine stress.


Carbohydrate and lipid metabolism in chronic spinal cord injury.
Abnormalities of carbohydrate and lipid metabolism are more common in the spinal cord injury (SCI) population than in the able-bodied population. This is an important consideration in the long-term care of individuals with SCI. Literature review. When compared with the able-bodied population, people with SCI are more likely to have oral carbohydrate intolerance, insulin resistance, elevated low-density lipoprotein cholesterol, and reduced high-density lipoprotein cholesterol, associated with increased prevalences of diabetes mellitus and cardiovascular disease. Because of increased risk factors for diabetes mellitus and heart disease in individuals with SCI, modifiable risk factors should be addressed, eg, obesity, inactivity, dietary factors, and smoking. To reduce mortality and morbidity associated with these risk factors, periodic screening for carbohydrate and lipid abnormalities is recommended, with appropriate therapeutic interventions.


An enzyme that selectively S-nitrosylates proteins to regulate insulin signaling.
Acyl-coenzyme A (acyl-CoA) species are cofactors for numerous enzymes that acylate thousands of proteins. Here, we describe an enzyme that uses S-nitroso-CoA (SNO-CoA) as its cofactor to S-nitrosylate multiple proteins (SNO-CoA-assisted nitrosylase, SCAN). Separate domains in SCAN mediate SNO-CoA and substrate binding, allowing SCAN to selectively catalyze SNO transfer from SNO-CoA to SCAN to multiple protein targets, including the insulin receptor (INSR) and insulin receptor substrate 1 (IRS1). Insulin-stimulated S-nitrosylation of INSR/IRS1 by SCAN reduces insulin signaling physiologically, whereas increased SCAN activity in obesity causes INSR/IRS1 hypernitrosylation and insulin resistance. SCAN-deficient mice are thus protected from diabetes. In human skeletal muscle and adipose tissue, SCAN expression increases with body mass index and correlates with INSR S-nitrosylation. S-nitrosylation by SCAN/SNO-CoA thus defines a new enzyme class, a unique mode of receptor tyrosine kinase regulation, and a revised paradigm for NO function in physiology and disease.


Estrogen receptors alpha and beta in rat decidua cells: cell-specific expression and differential regulation by steroid hormones and prolactin.
Estradiol is known to play an important role in the growth and differentiation of rat uterine stromal cells into decidual cells. In particular, this hormone with progesterone is necessary for blastocyst implantation and subsequent decidualization in the rat. Although binding experiments have demonstrated the presence of estrogen-binding sites, no evidence exists as to whether the rat decidua expresses both isoforms of the estrogen receptor (ER), alpha and beta. In this investigation, we analyzed the expression of decidual ERalpha and ERbeta, studied their regulation by PRL and steroid hormones and examined the ability of decidual ERp to transduce the estradiol signal to the progesterone receptor. Immunocytochemistry, RT-PCR, and Northern blot analysis showed that both ER species are coexpressed in the decidua during pseudopregnancy. Interestingly, these genes were preferentially found in a cell population localized in the antimesometrial site of the uterus where blastocyst implantation takes place. Using decidual cells in primary culture obtained from pseudopregnant rats and a decidua-derived cell line (GG-AD), we show a differential regulation of ERalpha and ERbeta by PRL and ovarian steroid hormones. Whereas PRL, estradiol, and progesterone all increased ERbeta messenger RNA (mRNA) expression in a dose-dependent manner, only PRL up-regulated the mRNA levels of ERa. Estradiol had no effect on ERalpha expression, whereas progesterone markedly decreased its mRNA levels. Interestingly, progesterone, which up-regulates the ability of PRL to signal to a PRL-regulated gene in mammary-gland derived cells, prevented PRL stimulation of decidual ERalpha and had no synergistic effect on ERbeta expression. The use of GG-AD cells, which express only ERbeta, allowed us to demonstrate that this receptor subtype is functional and transduces estradiol signal to the progesterone receptor. In summary, the results of this investigation have revealed that ERbeta is expressed in addition to ERalpha in the rat decidua, and that the expression of both ERs are cell specific and differentially regulated by PRL and steroids. One salient finding of this investigation is that progesterone down-regulates ERalpha, but concomitantly increases the expression of a functional ERbeta that mediates estradiol up-regulation of the decidual progesterone receptor.


Effect of thin-split encephalomyosynangiosis on transient neurological events in revascularization surgery for pediatric patients with moyamoya disease.
Transient neurological events (TNEs) are among the most important events after revascularization surgery in pediatric patients with moyamoya disease (MMD). Although hemodynamic changes and crying are representative factors of TNEs, brain compression by encephalomyosynangiosis (EMS) is another important cause of TNEs. Therefore, the authors assumed that making the EMS as thin as possible reduces the frequency of TNEs. However, thin-split EMS can lead to insufficient development of collateral vessels. This study aimed to evaluate the effects of thin-split EMS in combined revascularization surgery on postoperative outcomes in pediatric patients with MMD. The authors retrospectively included 56 consecutive combined revascularization surgeries in the anterior cerebral circulation in pediatric patients with MMD. These surgeries were classified into the former group and thin-split EMS group. The temporal muscle was halved in the former EMS group and split as thinly as possible in the thin-split EMS group. The authors performed between-group comparisons of postoperative stroke events and TNEs during the acute period and the development of collateral flow and stroke events during the chronic period. Former and thin-split EMS procedures were performed in 37 and 19 patients, respectively. TNEs without crying or hemodynamic changes were observed significantly less frequently in the thin-split EMS group than in the former EMS group (0 [0.0%] in the thin-split EMS group vs 9 [24.3%] in the former EMS group, p = 0.021). There were no significant between-group differences in the development of indirect bypass during the chronic period (good: 34 [91.9%] and poor: 3 [8.1%] in the former EMS group vs good: 16 [84.2%] and poor: 3 [15.8%] in the thin-split EMS group; p = 0.397). Additionally, there were no significant between-group differences in the incidence of chronic clinical events, including death or stroke. Thin-split EMS can reduce TNEs that do not accompany crying or hemodynamic changes. Furthermore, the procedure has sufficient prevention effects against stroke events during the chronic postoperative period in pediatric patients with MMD.


Effect of climate change on surface ozone over North America, Europe, and East Asia.
The effect of future climate change on surface ozone over North America, Europe, and East Asia is evaluated using present-day (2000s) and future (2100s) hourly surface ozone simulated by four global models. Future climate follows RCP8.5, while methane and anthropogenic ozone precursors are fixed at year-2000 levels. Climate change shifts the seasonal surface ozone peak to earlier in the year and increases the amplitude of the annual cycle. Increases in mean summertime and high-percentile ozone are generally found in polluted environments, while decreases are found in clean environments. We propose climate change augments the efficiency of precursor emissions to generate surface ozone in polluted regions, thus reducing precursor export to neighboring downwind locations. Even with constant biogenic emissions, climate change causes the largest ozone increases at high percentiles. In most cases, air quality extreme episodes become larger and contain higher ozone levels relative to the rest of the distribution.


Role of gastric mucosal folds in formation of focal ulcers in the rat.
Ulcers induced in rats by a variety of techniques often have a characteristic elongated shape or are focal and follow a linear pattern. The reason for this is unknown but may be related to the presence of gastric mucosal folds. Studies utilizing a rat gastric chamber preparation demonstrate that the apex of the mucosal fold is peculiarly sensitive to injury by corrosive agents in normotensive or hypotensive animals and in the completely ischemic mucosa. Focal gastric mucosal potential difference (PD) measurements utilizing a "sucrose gap" technique demonstrate that the PD at the crest of a mucosal fold is significantly lower than in the trough at the base of the fold, suggesting a local perfusion or permeability defect. The stomachs of animals submitted to three different ulcerogenic treatment demonstrated significantly more fold-related lesions that nonfold lesions. It is concluded that although different mechanisms may be responsible for the grossly different lesions, the mucosal fold, for a variety of reasons, appears particularly sensitive to injury.


Novel isoenzyme of 2-oxoglutarate dehydrogenase is identified in brain, but not in heart.
2-Oxoglutarate dehydrogenase (OGDH) is the first and rate-limiting component of the multienzyme OGDH complex (OGDHC) whose malfunction is associated with neurodegeneration. The essential role of this complex in the degradation of glucose and glutamate, which have specific significance in brain, raises questions about the existence of brain-specific OGDHC isoenzyme(s). We purified OGDHC from extracts of brain or heart mitochondria using the same procedure of poly(ethylene glycol) fractionation, followed by size-exclusion chromatography. Chromatographic behavior and the insufficiency of mitochondrial disruption to solubilize OGDHC revealed functionally significant binding of the complex to membrane. Components of OGDHC from brain and heart were identified using nano-high performance liquid chromatography electrospray tandem mass spectrometry after trypsinolysis of the electrophoretically separated proteins. In contrast to the heart complex, where only the known OGDH was determined, the band corresponding to the brain OGDH component was found to also include the novel 2-oxoglutarate dehydrogenase-like (OGDHL) protein. The ratio of identified peptides characteristic of OGDH and OGDHL was preserved during purification and indicated comparable quantities of the two proteins in brain. Brain OGDHC also differed from the heart complex in the abundance of the components, lower apparent molecular mass and decreased stability upon size-exclusion chromatography. The functional competence of the novel brain isoenzyme and different regulation of OGDH and OGDHL by 2-oxoglutarate are inferred from the biphasic dependence of the overall reaction rate versus 2-oxoglutarate concentration. OGDHL may thus participate in brain-specific control of 2-oxoglutarate distribution between energy production and synthesis of the neurotransmitter glutamate.


Expert panel consensus recommendations for ambulatory blood pressure monitoring in Asia: The HOPE Asia Network.
Hypertension is an important public health issue because of its association with a number of significant diseases and adverse outcomes. However, there are important ethnic differences in the pathogenesis and cardio-/cerebrovascular consequences of hypertension. Given the large populations and rapidly aging demographic in Asian regions, optimal strategies to diagnose and manage hypertension are of high importance. Ambulatory blood pressure monitoring (ABPM) is an important out-of-office blood pressure (BP) measurement tool that should play a central role in hypertension detection and management. The use of ABPM is particularly important in Asia due to the specific features of hypertension in Asian patients, including a high prevalence of masked hypertension, disrupted BP variability with marked morning BP surge, and nocturnal hypertension. This HOPE Asia Network document summarizes region-specific literature on the relationship between ABPM parameters and cardiovascular risk and target organ damage, providing a rationale for consensus-based recommendations on the use of ABPM in Asia. The aim of these recommendations is to guide and improve clinical practice to facilitate optimal BP monitoring with the goal of optimizing patient management and expediting the efficient allocation of treatment and health care resources. This should contribute to the HOPE Asia Network mission of improving the management of hypertension and organ protection toward achieving "zero" cardiovascular events in Asia.


Enzyme-Mediated Temporal Control over the Conformational Disposition of a Condensed Protein in Macromolecular Crowded Media.
Temporal regulation between input and output signals is one of the hallmarks of complex biological processes. Herein, we report that the conformational disposition of a protein in macromolecularly crowded media can be controlled with time using enzymes. First, we demonstrate the pH dependence of bovine serum albumin (BSA) condensation and conformational alteration in the presence of poly(ethylene glycol) as a crowder. However, by exploiting the strength of pH-modulatory enzymatic reactions (glucose oxidase and urease), the conversion time between the condensed and free forms can be tuned. Additionally, we demonstrate that the trapping of intermediate states with respect to the overall system at a particular α-helix or β-sheet composition and rotational mobility can be possible simply by altering the substrate concentration. Finally, we show that the intrinsic catalytic ability of BSA toward the Kemp elimination (KE) reaction is inhibited in the aggregated form but regained in the free form. In fact, the rate of KE reaction can also be actuated enzymatically in a temporal fashion, therefore demonstrating the programmability of a cascade of biochemical events in crowded media.


Clinical and Molecular Attributes and Evaluation of Pancreatic Cystic Neoplasm.
Intraductal papillary mucinous neoplasms (IPMNs) and mucinous cystic neoplasms (MCNs) are all considered "Pancreatic cystic neoplasms (PCNs)" and show a varying risk of developing into pancreatic ductal adenocarcinoma (PDAC). These lesions display different molecular characteristics, mutations, and clinical manifestations. A lack of detailed understanding of PCN subtype characteristics and their molecular mechanisms limits the development of efficient diagnostic tools and therapeutic strategies for these lesions. Proper in vivo mouse models that mimic human PCNs are also needed to study the molecular mechanisms and for therapeutic testing. A comprehensive understanding of the current status of PCN biology, mechanisms, current diagnostic methods, and therapies will help in the early detection and proper management of patients with these lesions and PDAC. This review aims to describe all these aspects of PCNs, specifically IPMNs, by describing the future perspectives.


Tropical peat subsidence rates are related to decadal LULC changes: Insights from InSAR analysis.
Peatlands in Indonesia are subject to subsidence in recent years, resulting in significant soil organic carbon loss. Their degradation is responsible for several environmental issues; however, understanding the causes of peatland subsidence is of prime concern for implementing mitigation measures. Here, we employed time-series Small BAseline Subset (SBAS) Interferometric Synthetic Aperture Radar (InSAR) using ALOS PALSAR-2 images to assess the relationship between subsidence rates and land use/land cover (LULC) change (including drainage periods) derived from decadal Landsat data (1972-2019). Overall, the study area subsided with a mean rate of -2.646 ± 1.839 cm/year in 2018-2019. The subsidence rates slowed over time, with significant subsidence decreases in peatlands after being drained for 9 years. We found that the long-time persistence of vegetated areas leads to subsidence deceleration. The relatively lower subsidence rates are in areas that changed to rubber/mixed plantations. Further, the potential of subsidence prediction was assessed using Random Forest (RF) regression based on LULC change, distance from peat edge, and elevation. With an R2 of 0.532 (RMSE = 0.594 cm/year), this machine learning method potentially enlarges the spatial coverage of InSAR method for the higher frequency SAR data (such as Sentinel-1) that mainly have limited coverage due to decorrelation in vegetated areas. According to feature importance in the RF model, the contribution of LULC change (including drainage period) to the subsidence model is comparable with distance from peat edge and elevation. Other uncertainties are from unexplained factors related to drainage and peat condition, which need to be accounted for as well. This work shows the significance of decadal LULC change analysis to supplement InSAR measurement in tropical peatland subsidence monitoring programs.


Chemical composition and antibacterial activity of Origanum saccatum P.H. Davis essential oil obtained by solvent-free microwave extraction: comparison with hydrodistillation.
The components of the essential oils (EOs) obtained by solvent-free microwave extraction (SFME) and hydrodistillation (HD) from endemic Origanum saccatum P.H. Davis were identified by using GC/MS. The main constituents of both EOs obtained by SFME and HD, respectively, from O. saccatum were p-cymene (72.5 and 70.6%), thymol (9.32 and 8.11%), and carvacrol (7.18 and 6.36%). The EO obtained by SFME contained substantially higher amounts of oxygenated compounds and lower amounts of monoterpenes than did the EO obtained by HD. The antibacterial activities of the EOs obtained by SFME and HD were evaluated with the disc diffusion method by comparison with 10 different bacterial strains. The antibacterial activity of the EO extracted by SFME was found to be more effective than that of the EO extracted by HD against seven of the tested bacteria.


Structure of the R3/I5 chimeric relaxin peptide, a selective GPCR135 and GPCR142 agonist.
The human relaxin family comprises seven peptide hormones with various biological functions mediated through interactions with G-protein-coupled receptors. Interestingly, among the hitherto characterized receptors there is no absolute selectivity toward their primary ligand. The most striking example of this is the relaxin family ancestor, relaxin-3, which is an agonist for three of the four currently known relaxin receptors: GPCR135, GPCR142, and LGR7. Relaxin-3 and its endogenous receptor GPCR135 are both expressed predominantly in the brain and have been linked to regulation of stress and feeding. However, to fully understand the role of relaxin-3 in neurological signaling, the development of selective GPCR135 agonists and antagonists for in vivo studies is crucial. Recent reports have demonstrated that such selective ligands can be achieved by making chimeric peptides comprising the relaxin-3 B-chain combined with the INSL5 A-chain. To obtain structural insights into the consequences of combining A- and B-chains from different relaxins we have determined the NMR solution structure of a human relaxin-3/INSL5 chimeric peptide. The structure reveals that the INSL5 A-chain adopts a conformation similar to the relaxin-3 A-chain, and thus has the ability to structurally support a native-like conformation of the relaxin-3 B-chain. These findings suggest that the decrease in activity at the LGR7 receptor seen for this peptide is a result of the removal of a secondary LGR7 binding site present in the relaxin-3 A-chain, rather than conformational changes in the primary B-chain receptor binding site.


Parents' knowledge, beliefs, and acceptance of the HPV vaccination in relation to their socio-demographics and religious beliefs: A cross-sectional study in Thailand.
Thailand has one of the world's highest prevalence of cervical cancer, mainly caused by the human papillomavirus (HPV). HPV infections can successfully be prevented by vaccination, which is available at a cost but not yet implemented in the national vaccination program. Parents play a critical role in deciding whether to vaccinate their child against HPV. Thus, the aim was to examine the association between parents' knowledge, beliefs, and acceptance of the HPV vaccination for their daughters, considering their socio-demographics and religious beliefs. A cross-sectional design was used among three schools in Thailand: Nakorn Phatom province (suburban) and Bangkok (urban). Parents of 9-12-year-old daughters completed the questionnaires, guided by the Health Belief Model. In total, 359 parents completed the questionnaires; of those, 301 were included in the final analyses. The ordinary least squares (OLS) regression analysis showed that background knowledge of HPV and the HPV vaccine was positively related to knowledge of HPV and cervical cancer. For beliefs, knowledge was positively associated with susceptibility (i.e., parents' perceived risk of an HPV infection/ related disease), severity, and benefit. However, knowledge was not significantly related to barriers. For acceptance, higher susceptibility and benefit were related to higher acceptance, and greater knowledge was associated with higher acceptance. Thus, we found associations between parents' knowledge, beliefs, and acceptance of the HPV vaccination for their daughters, considering their socio-demographics and religious beliefs. Parents, who reported religion as important, as opposed to those who did not, were more favorable toward the HPV vaccination. Four out of ten mothers had never undergone a cervical cancer screening, but most had accepted previous childhood vaccinations for their daughters. The overall acceptance of the vaccine was high, and we believe our results are promising for future implementation of the HPV vaccination in the national childhood vaccination program in Thailand.


Hypovitaminosis D exacerbates the DNA damage load in human uterine fibroids, which is ameliorated by vitamin D3 treatment.
Uterine fibroids (UFs) are the most common benign neoplastic threat to women's health and associated with DNA damage and genomic instability. Hypovitaminosis D is a known risk factor for UFs, especially among African Americans. Vitamin D3 has been shown to effectively inhibit UF phenotype, but its mechanisms remain unclear. We hypothesize that Vitamin D3 ameliorates UFs by recovering the damaged DNA repair system, thus inhibits tumor progression. We compared the DNA damage status and Vitamin D receptor (VDR) expression between normal myometrial and UF primary cells. Unrepaired DNA double-strand breaks (DSBs) accumulated but VDR expression decreased in UFs. The RNA and protein levels of key DNA repair members belonging to DNA DSB sensors (MRE11, NBS1, RAD50), mediators and effectors (CHECK2, BRCA1, RAD51) were downregulated in UFs compared with myometrial cells. VDR KD induced DSB accumulation and DNA damage response (DDR) defects in myometrial cells. Using the DNA damage PCR array, the expression of many additional DNA repair genes was downregulated in VDR KD cells. Treatment of UF cells with Vitamin D3 (100 nM) significantly decreased DNA damage and restored DDR concomitant with VDR induction. Notably, the PCR array demonstrated that among 75 downregulated genes after VDR KD, 67 (89.3%) were upregulated after vitamin D3 treatment. These studies demonstrate a novel link between DNA damage and the vitamin D3/VDR axis in UFs. Vitamin D3 suppresses the UF phenotype through orchestrated targeting at multiple molecules in DNA repair pathways, thus offering novel mechanistic insights into the clinical effectiveness of vitamin D3 on UFs.


[Evaluation of work conditions for teachers of medical schools].
Complex sanitary hygienic and sociologic evaluation of high school factors enabled to assign work conditions of medical institute lecturers according to P 2.2.2006 - to 05 for jeopardy and to 2 degree 3 class for hardiness and intensity of work, to define psychosocial risks as main hazards. The authors specified and suggested prophylactic system aimed to increase work efficiency of medical institute lecturers.


Response in BRCA1 mutation carrier with metastatic pancreatic adenocarcinoma treated with FOLFIRINOX.
A woman in her 50s previously treated for early-stage breast cancer, parotid mucoepidermoid carcinoma and Caroli's disease was diagnosed with stage IV pancreatic ductal adenocarcinoma (PDAC) metastatic to the liver and was found to harbour a BRCA1 germline mutation. She had palliative chemotherapy, initially with 5-fluorouracil, leucovorin, irinotecan and oxaliplatin, and then FOLFIRI and capecitabine, achieving a sustained near-complete response for at least 86 months. Chemotherapy was eventually discontinued when she was diagnosed with a tongue squamous cell carcinoma. Despite withholding systemic therapy, she has maintained a durable response. This is the first report in the English literature showing a sustained duration of response in a patient with PDAC and BRCA1 germline mutation.


[Efficiency of shorter chemotherapy courses for intrathoracic tuberculosis in children].
To develop differential chemotherapy regimes for intrathoracic tuberculosis in children aged 3-12 years, 255 children with active tuberculosis underwent clinical and X-ray examination. Of them, 120 children received shorter chemotherapy in an early intensive phase by using three drugs (isoniazid, rifampicin, and pyrazinamide supplemented by streptomycin) in complicated tuberculosis. A control group (n = 135) had therapy without pyrazinamide. Shorter courses of therapy were shown not only to reduce total treatment duration on an average to 6.4 and 9.2 months in uncomplicated and complicated tuberculosis, respectively, but to contribute to more perfect healing processes and resolution of abnormal changes in 85.3 and 60.0% of children with uncomplicated and complicated forms, respectively. At the same time shorter treatment is well tolerated. It shows much fewer side effects than does longer treatment (20 and 36%). There are no increases in the incidence of recurrent tuberculosis late at follow-up (2.7 and 2.7%, respectively).


Mode of Ca2+ action on ciliary beat frequency in single ovine airway epithelial cells.
1. We analysed the kinetics of coupling between cytoplasmic calcium ([Ca2+]i) and ciliary beat frequency (CBF) using simultaneous single cilium recording and single cell [Ca2+]i measurements from cultured ovine tracheal epithelial cells. 2. CBF and [Ca2+]i (indicated by fura-2) were measured at rest and in response to activation of the G-protein coupled M3 muscarinic receptor by 10 microM acetylcholine (ACh). 3. Fourier transform analysis of 3 s data segments of light intensity from phase-contrast microscopy showed no significant delay between changes in [Ca2+]i and CBF during a 2 min exposure to ACh and subsequent washout. 4. CBF time resolution was improved by computing instantaneous beat frequency. This revealed that CBF lagged the rapid increase in [Ca2+]i in response to ACh with a delay of less than 1 beat cycle (143 ms at 7 Hz). When CBF was estimated by an improved Fourier method, this delay was observed to be 70 +/- 30 ms (mean +/- s.e.m.; n = 20 cilia). During the slower return to baseline, a lag of 8 +/- 3.2 s was observed, indicative of hysteresis. 5. While calmodulin inhibitors (calmidazolium and W-7; each n = 5) decreased baseline CBF by an average of 1.1 +/- 0.1 Hz, they did not alter the kinetic relationship between [Ca2+]i and CBF. Similarly, phosphatase inhibitors (okadaic acid and cyclosporin A; each n = 5), changed neither baseline CBF nor the kinetic coupling between [Ca2+]i and CBF. 6. These data suggest that the timing of Ca2+ action on CBF in ovine airway epithelial cells, is unlikely to be determined by phosphorylation reactions involving calmodulin or kinase/phosphatase reactions. 7. A simple model for Ca2+ stimulation of CBF is presented. Fits of the model to the data suggest four or more Ca2+ ions bind cooperatively to speed up CBF.


Identification of mouse mutant cells exhibiting the plastic mutant phenotype.
The initial processes involved in radiation carcinogenesis have not been clearly elucidated. We isolated mouse mutant cells exhibiting plasticity in their mutation phenotypes. These mutant cells were originally isolated from an irradiated cell population as 6-thioguanine resistant (6TGR) mutants that were deficient in hypoxanthine phosphoribosyl transferase (Hprt, E.C.18.104.22.168) activity at the frequency of approximately 6.2 x 10(-5). Approximately 10% of 6TGR cells showed plasticity in their mutant phenotypes and reverted to HAT-resistant (HATR), which is Hprt-proficient, wild type phenotype. Eventually we identified the plastic mutants in the un-irradiated wild type cell population as well and found that ionizing irradiation enhanced the frequency of the plastic mutation approximately 24 times. Treatment with 5-aza-cytidine did not affect the plasticity of mutant phenotypes identified in this study, suggesting that DNA methylation was not involved in the plastic changes of the mutant phenotypes. The plastic mutant phenotype identified in our study is a new type of genomic instability induced by ionizing irradiation, and it is likely to be involved in one of the primary changes that occur in the process of radiation carcinogenesis, and may explain one element of carcinogenesis, which is composed of multi-stages.


Inosine/pyruvate/phosphate medium but not adenosine/pyruvate/phosphate medium introduces millimolar amounts of 5-phosphoribosyl 1-pyrophosphate in human erythrocytes. A 31P-n.m.r. study.
Incubation of human erythrocytes in medium containing inosine (10 mM), pyruvate (10 mM), phosphate (50 mM) and NaCl (75 mM) at pH 6.6 leads to a more than 1000-fold increase in the concentration of 5-phosphoribosyl 1-pyrophosphate (PRPP), as identified and quantified by 31P-n.m.r. spectroscopy. The accumulation is highly pH-dependent, with a maximum at extracellular pH 6.60, and the maximum value of 1.3-1.6 mmol/l of erythrocytes is attained within 1 h at 37 degrees C. PRPP was accumulated despite high concentrations of 2,3-bisphosphoglycerate (2,3-BPG), an inhibitor of PRPP synthetase. The concentration of PRPP correlated with the intracellular concentration of inorganic phosphate (Pi). Substitution of either adenosine or adenosine plus inosine for inosine in the medium did not lead to 31P-n.m.r.-detectable accumulation of PRPP. These results show that neither 2,3-BPG nor PRPP itself inhibits the synthesis of PRPP in the human erythrocyte. Adenosine, however, prevents the inosine-stimulated accumulation of PRPP.


Ophthalmic Emergencies and Effects of Different Quarantine Models During the COVID-19 Pandemic.
BACKGROUND The aim of this study was to evaluate the epidemiological features of the patients admitted to our emergency department (ED) during the COVID-19 pandemic in March, April, and May 2020, compare them with the patients in March, April, and May 2019, and to investigate the effect of various quarantine models. MATERIAL AND METHODS The records of 1206 eligible patients were reviewed. Age groups were divided according to their quarantine status. We recorded the reasons for patient presentation to the ED, the average number of patients in age groups according to quarantine days, and the problems reported by patients who presented to the ED according to quarantine days. RESULTS We enrolled 3016 of 3123 patients. The mean age was 36.4±17.3 years in 2019 and 37.8±16.4 years during the pandemic period (P=0.031). While 73.1% of the ED presentations were due to ocular trauma in 2019, it decreased to 70.7% in 2020. The proportion of those who presented to the ED during the weekend was 30.6% in 2019, but decreased to 23.9% in 2020 (<0.001). While 84.8% of trauma patients were ages 19-64 years in 2019, this rate increased to 88.9% during the pandemic (P=0.067). Non-trauma emergencies were more common than trauma emergencies in both periods in those over the age of 65 years. During the pandemic period, admissions to the ED were decreased in the quarantined age groups (P=0.001). CONCLUSIONS The changes in the number and characteristic of admissions for ophthalmic emergencies during the pandemic period may help planning allocation of healthcare personnel and resources in outpatient and emergency clinics.


The forkhead transcription factor FoxO regulates transcription of p27Kip1 and Bim in response to IL-2.
The cytokine IL-2 plays a very important role in the proliferation and survival of activated T cells. These effects of IL-2 are dependent on signaling through the phosphatidylinositol 3-kinase (PI3K) pathway. We and others have shown that PI3K, through activation of protein kinase B/Akt, inhibits transcriptional activation by a number of forkhead transcription factors (FoxO1, FoxO3, and FoxO4). In this study we have investigated the role of these forkhead transcription factors in the IL-2-induced T cell proliferation and survival. We show that IL-2 regulates phosphorylation of FoxO3 in a PI3K-dependent fashion. Phosphorylation and inactivation of FoxO3 appears to play an important role in IL-2-mediated T cell survival, because mere activation of FoxO3 is sufficient to trigger apoptosis in T cells. Indeed, active FoxO3 can induce expression of IL-2-regulated genes, such as the cdk inhibitor p27(Kip1) and the proapoptotic Bcl-2 family member Bim. Furthermore, we show that IL-2 triggers a rapid, PI3K-dependent, phosphorylation of FoxO1a in primary T cells. Thus, we propose that inactivation of FoxO transcription factors by IL-2 plays a critical role in T cell proliferation and survival.


Prevalence of metabolic syndrome and its association with physical capacity, disability, and self-rated health in Lifestyle Interventions and Independence for Elders Study participants.
To evaluate the prevalence of metabolic syndrome (MetS) and its association with physical capacity, disability, and self-rated health in older adults at high risk of mobility disability, including those with and without diabetes mellitus. Cross-sectional analysis. Lifestyle Interventions and Independence for Elders (LIFE) Study. Community-dwelling sedentary adults aged 70 to 89 at high risk of mobility disability (Short Physical Performance Battery (SPPB) score ≤9; mean 7.4 ± 1.6) (N = 1,535). Metabolic syndrome was defined according to the 2009 multiagency harmonized criteria; outcomes were physical capacity (400-m walk time, grip strength, SPPB score), disability (composite 19-item score), and self-rated health (5-point scale ranging from excellent to poor). The prevalence of MetS was 49.8% in the overall sample (83.2% of those with diabetes mellitus, 38.1% of those without). MetS was associated with stronger grip strength (mean difference (Δ) = 1.2 kg, P = .01) in the overall sample and in participants without diabetes mellitus and with poorer self-rated health (Δ = 0.1 kg, P < .001) in the overall sample only. No significant differences were found in 400-m walk time, SPPB score, or disability score between participants with and without MetS, in the overall sample or diabetes mellitus subgroups. Metabolic dysfunction is highly prevalent in older adults at risk of mobility disability, yet consistent associations were not observed between MetS and walking speed, lower extremity function, or self-reported disability after adjusting for known and potential confounders. Longitudinal studies are needed to investigate whether MetS accelerates declines in functional status in high-risk older adults and to inform clinical and public health interventions aimed at preventing or delaying disability in this group.


Adenovirus E3/19K promotes evasion of NK cell recognition by intracellular sequestration of the NKG2D ligands major histocompatibility complex class I chain-related proteins A and B.
The adenovirus (Ad) early transcription unit 3 (E3) encodes multiple immunosubversive functions that are presumed to facilitate the establishment and persistence of infection. Indeed, the capacity of E3/19K to inhibit transport of HLA class I (HLA-I) to the cell surface, thereby preventing peptide presentation to CD8(+) T cells, has long been recognized as a paradigm for viral immune evasion. However, HLA-I downregulation has the potential to render Ad-infected cells vulnerable to natural killer (NK) cell recognition. Furthermore, expression of the immediate-early Ad gene E1A is associated with efficient induction of ligands for the key NK cell-activating receptor NKG2D. Here we show that while infection with wild-type Ad enhances synthesis of the NKG2D ligands, major histocompatibility complex class I chain-related proteins A and B (MICA and MICB), their expression on the cell surface is actively suppressed. Both MICA and MICB are retained within the endoplasmic reticulum as immature endoglycosidase H-sensitive forms. By analyzing a range of cell lines and viruses carrying mutated versions of the E3 gene region, E3/19K was identified as the gene responsible for this activity. The structural requirements within E3/19K necessary to sequester MICA/B and HLA-I are similar. In functional assays, deletion of E3/19K rendered Ad-infected cells more sensitive to NK cell recognition. We report the first NK evasion function in the Adenoviridae and describe a novel function for E3/19K. Thus, E3/19K has a dual function: inhibition of T-cell recognition and NK cell activation.


Effect of dietary fish oil on azoxymethane-induced colon carcinogenesis in male F344 rats.
The effect of dietary intake of different levels of Menhaden fish oil on azoxymethane-induced carcinogenesis was examined in male F344 rats fed the semipurified diets. Starting at 5 weeks of age, groups of animals were fed the 5% corn oil (low corn oil) diet. At 7 weeks of age, all animals except the vehicle-treated controls were given s.c. injections of azoxymethane (15 mg/kg body weight/week for 2 weeks). After 4 days, groups of animals were fed the diets containing 4% Menhaden oil + 1% corn oil (low fish oil), 22.5% Menhaden oil + 1% corn oil (high fish oil), 5% corn oil, and 23.5% corn oil (high corn oil). Thirty-four weeks after azoxymethane injections, all animals were necropsied. High fish oil diet had no tumor promoting effect in the large intestine when compared to the high corn oil diet. There was no difference in large intestinal tumor incidence among the other dietary groups. The results of this study indicate that fish oils rich in highly polyunsaturated n-3 fatty acids do not enhance large bowel carcinogenesis and that the fatty acid composition of the dietary fat is one of the determining factors in large bowel carcinogenesis.


Low level exposure to the flame retardant BDE-209 reduces thyroid hormone levels and disrupts thyroid signaling in fathead minnows.
Polybrominated diphenyl ether (PBDE) flame retardants have been shown to disrupt thyroid hormone regulation, neurodevelopment, and reproduction in some animals. However, effects of the most heavily used PBDE, decabromodiphenyl ether (BDE-209), on thyroid functioning remain unclear. This study examined low-dose effects of BDE-209 on thyroid hormone levels and signaling in fathead minnows. Adult males received dietary exposures of BDE-209 at a low dose (∼3 ng/g bw-day) and high dose (∼300 ng/g bw-day) for 28 days followed by a 14-day depuration to evaluate recovery. Compared to controls, fish exposed to the low dose for 28 days experienced a 53% and 46% decline in circulating total thyroxine (TT4) and 3,5,3'-triiodothyronine (TT3), respectively, while TT4 and TT3 deficits at the high dose were 59% and 62%. Brain deiodinase activity (T4-ORD) was reduced by ∼65% at both doses. BDE-209 elevated the relative mRNA expression of genes encoding deiodinases, nuclear thyroid receptors, and membrane transporters in the brain and liver in patterns that varied with time and dose, likely in compensation to hypothyroidism. Declines in the gonadal-somatic index (GSI) and increased mortality were also measured. Effects at the low dose were consistent with the high dose, suggesting nonlinear relationships between BDE-209 exposures and thyroid dysfunction.


Statin synergizes with LPS to induce IL-1beta release by THP-1 cells through activation of caspase-1.
Mevalonate kinase deficiency (MKD) is a hereditary syndrome characterized by recurring episodes of fever and inflammation. Peripheral blood mononuclear cells from MKD patients secrete high levels of interleukin (IL)-1beta when stimulated with lipopolysaccharide (LPS), which is thought to be a primary cause of the inflammation. However, the link between a deficient mevalonate kinase and excessive IL-1beta release remains unclear. To investigate this we made use of a model in which monocytic cells (THP-1) were treated with simvastatin. Statins are compounds that inhibit 3-hydroxy-3-methylglutaryl CoA (HMG-CoA) reductase and thereby artificially impair the isoprenoid biosynthesis pathway, mimicking mevalonate kinase deficiency. Our study revealed that LPS-stimulated THP-1 cells treated with simvastatin had an increased caspase-1 mediated processing of proIL-1beta. This increased processing was caused by enhanced autoprocessing of caspase-1, rather than enhanced transcription or translation of caspase-1 or proIL-1beta. Simvastatin-induced activation of caspase-1 was caused by an impairment of non-sterol isoprenoid biosynthesis, as the isoprenyl intermediate GGPP could block activation of caspase-1 and mIL-1beta release. In addition, inhibition of both farnesyl pyrophosphate synthase and geranylgeranyltransferase I also induce mIL-1beta release. Taken together, these results demonstrate that simvastatin augments LPS-induced IL-1beta release post-translationally, by inducing caspase-1 activity. These findings suggest that MKD patients may have overactive caspase-1, causing enhanced IL-1beta processing and subsequent inflammation in response to bacterial components.