Epithelial Ca2+ and Mg2+ channels in health and disease.
A near constancy of the extracellular Ca(2+) and Mg(2+) concentration is required for numerous physiologic functions at the organ, tissue, and cellular levels. This suggests that minor changes in the extracellular concentration of these divalents must be detected to allow the appropriate correction by the homeostatic systems. The maintenance of the Ca(2+) and Mg(2+) balance is controlled by the concerted action of intestinal absorption, renal excretion, and exchange with bone. After years of research, rapid progress was made recently in identification and characterization of the Ca(2+) and Mg(2+) transport proteins that contribute to the delicate balance of divalent cations. Expression-cloning approaches in combination with knockout mice models and genetic studies in families with a disturbed Mg(2+) balance revealed novel Ca(2+) and Mg(2+) gatekeeper proteins that belong to the super family of the transient receptor potential (TRP) channels. These epithelial Ca(2+) (TRPV5 and TRPV6) and Mg(2+) channels (TRPM6 and TRPM7) form prime targets for hormonal control of the active Ca(2+) and Mg(2+) flux from the urine space or intestinal lumen to the blood compartment. This review describes the characteristics of epithelial Ca(2+) and Mg(2+) transport in general and highlights in particular the distinctive features and the physiologic relevance of these new epithelial Ca(2+) and Mg(2+) channels in (patho)physiologic situations.


Comprehensive Tandem-Mass-Spectrometry Coverage of Complex Samples Enabled by Data-Set-Dependent Acquisition.
Tandem mass spectrometry (MS/MS) is an invaluable experimental tool for providing analytical data supporting the identification of small molecules and peptides in mass-spectrometry-based "omics" experiments. Data-dependent MS/MS (DDA) is a real-time MS/MS-acquisition strategy that is responsive to the signals detected in a given sample. However, in analysis of even moderately complex samples with state-of-the-art instrumentation, the speed of MS/MS acquisition is insufficient to offer comprehensive MS/MS coverage of all detected molecules. Data-independent approaches (DIA) offer greater MS/MS coverage, typically at the expense of selectivity or sensitivity. This report describes data-set-dependent MS/MS (DsDA), a novel integration of MS1-data processing and target prioritization to enable comprehensive MS/MS sampling during the initial MS-level experiment. This approach is guided by the premise that in omics experiments, individual injections are typically made as part of a larger set of samples, and feedback between data processing and data acquisition can allow approximately real-time optimization of MS/MS-acquisition parameters and nearly complete MS/MS-sampling coverage. Using a combination of R, Proteowizard, XCMS, and WRENS software, this concept was implemented on a liquid-chromatograph-coupled quadrupole time-of-flight mass spectrometer. The results illustrate comprehensive MS/MS coverage for a set of complex small-molecule samples and demonstrate a strong improvement on traditional DDA.


[Dietary phospholipids: lipid metabolism and risk factors for cardiovascular diseases].
Eggs are a major source of phospholipids (PL) in the Western diet. Dietary PL have emerged as a potential source of bioactive lipids that may have widespread effects on pathways related to inflammation, cholesterol metabolism, and high-density lipoprotein (HDL) function. Based on pre-clinical studies, egg phosphatidylcholine (PC) and sphingomyelin appeared to regulate cholesterol absorption and inflammation. In clinical studies, egg PL intake is associated with beneficial changes in biomarkers related to HDL reverse cholesterol transport. Recently, egg PC was shown to be a substrate for the generation of trimethylamine N-oxide (TMAO), a gut microbe-dependent metabolite associated with increased cardiovascular disease (CVD) risk. More researches are warranted to examine potential serum TMAO responses to chronic egg ingestion and in different populations, such as diabetics. In this review, the recent basic science, clinical, and epidemiological findings examining egg PL intake and risk of CVD are summarized.


Research Frontiers in Oral Toxicities of Cancer Therapies: Osteoradionecrosis of the Jaws.
The deleterious effects of head and neck radiation on bone, with osteoradionecrosis (ORN) as the major disabling side effect of head and neck cancer treatment, are difficult to prevent and hard to treat. This review focuses on the current state of the science regarding the pathobiology, clinical impact, and management of ORN. With regard to the pathobiology underlying ORN, it is not yet confirmed whether the current radiation schedules by 3-dimensional conformal radiotherapy and intensity modified radiotherapy result in an unchanged, decreased, or increased risk of developing ORN when compared with conventional radiation treatment, the main risk factor being the total radiation dose delivered on any clinically significant surface of the mandible. With regard to the prevention of ORN, a thorough, early pre-irradiation dental assessment is still considered the first step to reduce the hazard of developing ORN post-radiotherapy, and hyperbaric oxygen (HBO) treatment reduces the risk of developing ORN in case of dental surgery in an irradiated field. With regard to the treatment of ORN, the focus is bidirectional: elimination of the necrotic bone and improving the vascularity of the normal tissues that were included in the radiation portal. The cure rate of limited ORN by conservative therapy is approximately 50%, and the cure rate of surgical approaches when conservative therapy has failed is approximately 40%. Whether it is effective to support conservative or surgical treatment with HBO as an adjuvant is not set. HBO treatment is shown to increase the vascularity of hard and soft tissues and has been reported to be beneficial in selected cases. However, in randomized clinical trials comparing the preventive effect of HBO on developing ORN with, eg, antibiotic coverage in patients needing dental surgery, the preventive effect of HBO was not shown to surpass that of a more conservative approach. More recently, pharmacologic management was introduced in the treatment of ORN with success, but its efficacy has to be confirmed in randomized clinical trials. The major problem of performing well-designed randomized clinical trials in ORN is having access to large numbers of patients with well-defined, comparable cases of ORN. Because many institutions will not have large numbers of such ORN cases, national and international scientific societies must be approached to join multicenter trials. Fortunately, the interest of funding organizations and the number researchers with an interest in healthy aging is growing. Research aimed at prevention and reduction of the morbidity of cancer treatment fits well within these programs.


Who let the dogs out? Exploring the spatial ecology of free-roaming domestic dogs in western Kenya.
The spatial ecology of free-roaming dogs determines their role in the transmission of zoonoses. This study describes the geographic range of and identifies sites frequently visited by free-roaming domestic dogs in western Kenya. Eight sites in Busia county, western Kenya, were selected. At each site, ten dog-keeping households were recruited, a questionnaire was administered, and a GPS logger was fixed around the neck of one dog in each household. Loggers were programmed to capture the dog's position every minute, for five consecutive days. Individual summaries of GPS recordings were produced, and the daily distance traveled was calculated. 50% and 95% utilization distribution isopleths were produced, and the area within these isopleths was extracted to estimate the size of the core and extended Home Ranges (HRs), respectively. Linear regression analyses were performed to identify factors associated with the movement parameters. The centroid points of the 10, 50, and 90% isopleths were reproduced, and the corresponding sites identified on the ground. Seventy-three dogs were included in the final analyses. The median daily distance traveled was 13.5km, while the median core and extended HRs were 0.4 and 9.3 ha, respectively. Older dogs had a larger extended HR and traveled more daily, while the effect of sex on dog movement depended on their neutering status. Dogs spent most of their time at their household; other frequently visited sites included other household compounds, fields, and rubbish dumps. One of the centroids corresponded to a field located across the international Kenya-Uganda border, emphasizing the fluidity across the border in this ecosystem. Multiple dogs visited the same location, highlighting the heterogeneous contact networks between dogs, and between dogs and people. The field data presented are of value both in understanding domestic dog ecology and resource utilization, and in contextualizing infectious and parasitic disease transmission models.


Epizootic vesicular stomatitis in Colorado, 1982: infection in occupational risk groups.
In 1982-1983, an epizootic of vesicular stomatitis occurred in the western United States. Veterinarians, research workers, and regulatory personnel who were exposed to vesicular stomatitis virus were examined for patterns of human infection and prevalence of vesicular stomatitis New Jersey serotype neutralizing antibody. Insight into the mechanism of transmission was sought by comparing activities of antibody-positive and antibody-negative persons. A statistically significant risk factor was a history of infected animals sneezing in the face of serosurvey participants. Elevated odds ratios were also calculated for those who usually examined the oral cavity of affected animals, had open wounds on hands or arms, and had exposure to saliva through the eye or skin. Relatively intimate direct contact was required; a higher risk was associated with examining horses than cattle. Neutralizing antibody prevalence was significantly higher among exposed persons with illness (23%) than in exposed persons without a history of clinical illness (7%). Overall, however, infectivity of VSNJ for humans during the epizootic was low.


A bibliometric analysis of COVID-19 publications in neurology by using the visual mapping method.
The characteristic symptom of coronavirus disease 2019 (COVID-19) is respiratory distress, but neurological symptoms are the most frequent extra-pulmonary symptoms. This study aims to explore the current status and hot topics of neurology-related research on COVID-19 using bibliometric analysis. Publications regarding neurology and COVID-19 were retrieved from the Web of Science Core Collection (WoSCC) on March 28 2022. The Advanced search was conducted using "TS = ('COVID 19' or 'Novel Coronavirus 2019' or 'Coronavirus disease 2019' or '2019-nCOV' or 'SARS-CoV-2' or 'coronavirus-2') and TS = ('neurology'or 'neurological' or 'nervous system' or 'neurodegenerative disease' or 'brain' or 'cerebra' or 'nerve')". Microsoft Excel 2010 and VOSviewer were used to characterize the largest contributors, including the authors, journals, institutions, and countries. The hot topics and knowledge network were analyzed by CiteSpace and VOSviewer. A total of 5,329 publications between 2020 and 2022 were retrieved. The United States, Italy, and the United Kingdom were three key contributors to this field. Harvard Medical School, the Tehran University of Medical Sciences, and the UCL Queen Square Institute of Neurology were the major institutions with the largest publications. Josef Finsterer from the University of São Paulo (Austria) was the most prolific author. Tom Solomon from the University of Liverpool (UK) was the most cited author. Neurological Sciences and Frontiers in Neurology were the first two most productive journals, while Journal of Neurology held the first in terms of total citations and citations per publication. Cerebrovascular diseases, neurodegenerative diseases, encephalitis and encephalopathy, neuroimmune complications, neurological presentation in children, long COVID and mental health, and telemedicine were the central topics regarding the neurology-related research on COVID-19. Neurology-related research on COVID-19 has attracted considerable attention worldwide. Research topics shifted from "morality, autopsy, and telemedicine" in 2020 to various COVID-19-related neurological symptoms in 2021, such as "stroke," "Alzheimer's disease," "Parkinson's disease," "Guillain-Barre syndrome," "multiple sclerosis," "seizures in children," and "long COVID." "Applications of telemedicine in neurology during COVID-19 pandemic," "COVID-19-related neurological complications and mechanism," and "long COVID" require further study.


Intestinal parasites in children with neoplasms.
Stool specimens taken from 50 children with malignancy and from 92 healthy children were investigated for intestinal parasites, using the modified formol ethyl acetate concentration method, and native-lugol, trichrome and Kinyoun acid-fast stain methods. Thirty-eight (76.0%) of the 50 patients had lymphoma or leukemia and were considered immunosuppressed. Several different parasites were found in 21 (42.0%) of the 50 patients with malignancy and in 16 (47.3%) of the 38 patients with immune deficiency compared to in only 16 (17.3%) of the 92 healthy children. The incidence of parasites in patients with malignancy or immunosuppression was significantly higher than in the healthy control group (p<0.01, p<0.01).


Increasing the transplant dose and repeating faecal microbiota transplantation results in the responses of male patients with IBS reaching those of females.
Faecal microbiota transplantation (FMT) performed with a proper protocol is a safe treatment for IBS that has high efficacy and durable effects. Females have been reported to respond better than males to FMT. The present study aimed at determining whether increasing the transplant dose or repeating FMT improve the responses of males to FMT. This study included 186 IBS patients (131 females and 55 males) who were randomized at a 1:1:1 ratio to receive 90 g of donor faeces once into the large intestine, once into the small intestine or twice into the small intestine. Patients completed five questionnaires that assessed their symptoms and quality of life, and provided faecal samples at baseline and at 3, 6 and 12 months after FMT. The faecal bacterial profile and dysbiosis index were determined using 16S rRNA gene PCR DNA amplification covering variable genes V3-V9. The response rates to FMT at all observation times did not differ significantly between females and males regardless of the transplant administration route or whether it was repeated. Faecal Alistipes levels were higher in females than in males at baseline and increased in both females and males after FMT. In the repeated group, the Alistipes levels did not differ between females and males after FMT. Increasing the transplant dose and repeating FMT results in the responses of male IBS patients to FMT reaching those of females regardless of the administration route. Alistipes spp. levels appear to play a role in this improvement.www.clinicaltrials.gov (NCT04236843).


An improved recombinant IgG immunoblot for serodiagnosis of Lyme borreliosis.
We have previously described the use of the following recombinant antigens for serodiagnostic immunoblots: p83/100, p39, OspC and p41 (flagellin) internal fragment [Wilske et al. (1993) Med Microbiol Immunol 182:255-270; Rossler et al. (1997) J Clin Microbiol 35:2752-2758]. In our currently used immunoblot p83/100 is derived from strain PKo (Borrelia afzelii), p39 (BmpA) and OspC from strains PKa2 (B. hurgdorferi sensu stricto), PKo and PBi (B. garinii), respectively; the p41 (flagellin) internal fragments were cloned from strains PKo and PBi. In this study we describe the use of two additional recombinantly expressed highly immunogenic proteins Osp 7 (derived from PKo) and p58 (derived from PBi). A clinically well-defined panel of sera from 147 Lyme borreliosis patients and 139 controls previously tested by a standardized whole cell lysate immunoblot [Hauser et al. (1997) J Clin Microbiol 35:1433-1444] was investigated in the recombinant immunoblot without (old recombinant immunoblot) and with Ospl7 and p58 (new recombinant immunoblot) for IgG antibodies. The sensitivity of the recombinant IgG immunoblot for diagnosis of stage II and stage III could be significantly improved by addition of Osp17 and p58 without loss of specificity. With the exception of sera from patients with erythema migrans the diagnostic sensitivity is comparable to the whole cell lysate IgG immunoblot. The main advantage of the recombinant immunoblot is the easy identification of diagnostic bands, whereas the identification of bands in the whole cell lysate immunoblot is difficult. The recombinant immunoblot is especially suitable where large series of sera need to be investigated.


Identification of Trypanosoma brucei AdoMetDC Inhibitors Using a High-Throughput Mass Spectrometry-Based Assay.
Human African trypanosomiasis (HAT) is a fatal infectious disease caused by the eukaryotic pathogen Trypanosoma brucei (Tb). Available treatments are difficult to administer and have significant safety issues. S-Adenosylmethionine decarboxylase (AdoMetDC) is an essential enzyme in the parasite polyamine biosynthetic pathway. Previous attempts to develop TbAdoMetDC inhibitors into anti-HAT therapies failed due to poor brain exposure. Here, we describe a large screening campaign of two small-molecule libraries (∼400,000 compounds) employing a new high-throughput (∼7 s per sample) mass spectrometry-based assay for AdoMetDC activity. As a result of primary screening, followed by hit confirmation and validation, we identified 13 new classes of reversible TbAdoMetDC inhibitors with low-micromolar potency (IC50) against both TbAdoMetDC and T. brucei parasite cells. The majority of these compounds were >10-fold selective against the human enzyme. Importantly, compounds from four classes demonstrated high propensity to cross the blood-brain barrier in a cell monolayer assay. Biochemical analysis demonstrated that compounds from eight classes inhibited intracellular TbAdoMetDC in the parasite, although evidence for a secondary off-target component was also present. The discovery of several new TbAdoMetDC inhibitor chemotypes provides new hits for lead optimization programs aimed to deliver a novel treatment for HAT.


Lecithin-sphingomyelin ratios in amniotic fluid of pregnancies with an anencephalic fetus.
As many investigators have shown that surfactant production in the developing human lung is subject to multihormonal regulation, the present authors determined the lecithin-sphingomyelin (L/S) ratio in amniotic fluid of pregnancies with an anencephalic fetus, in which there was known to be aberrant production of fetal pituitary, adrenal, and consequently, placental hormones. The L/S ratio in amniotic fluid from seven of eight pregnancies with an anencephalic fetus was substantially lower than that in amniotic fluids of pregnancies with a normal fetus at the same stage of gestation. The L/S ratio in amniotic fluid of an anencephalic fetus of a twin pregnancy (monochorionic diamniotic) at 34 weeks' gestation was low; the L/S ratio of the amniotic fluid of the normal fetus was high. These data are supportive of the view that fetal lung maturation is dependent, in part, upon normal function of the fetal pituitary and adrenal.


Photorepair of Either CPD or 6-4PP DNA Lesions in Basal Keratinocytes Attenuates Ultraviolet-Induced Skin Effects in Nucleotide Excision Repair Deficient Mice.
Ultraviolet (UV) radiation is one of the most genotoxic, universal agents present in the environment. UVB (280-315 nm) radiation directly damages DNA, producing cyclobutane pyrimidine dimers (CPDs) and pyrimidine 6-4 pyrimidone photoproducts (6-4PPs). These photolesions interfere with essential cellular processes by blocking transcription and replication polymerases, and may induce skin inflammation, hyperplasia and cell death eventually contributing to skin aging, effects mediated mainly by keratinocytes. Additionally, these lesions may also induce mutations and thereby cause skin cancer. Photolesions are repaired by the Nucleotide Excision Repair (NER) pathway, responsible for repairing bulky DNA lesions. Both types of photolesions can also be repaired by distinct (CPD- or 6-4PP-) photolyases, enzymes that specifically repair their respective photolesion by directly splitting each dimer through a light-dependent process termed photoreactivation. However, as photolyases are absent in placental mammals, these organisms depend solely on NER for the repair of DNA UV lesions. However, the individual contribution of each UV dimer in the skin effects, as well as the role of keratinocytes has remained elusive. In this study, we show that in NER-deficient mice, the transgenic expression and photorepair of CPD-photolyase in basal keratinocytes completely inhibited UVB-induced epidermal thickness and cell proliferation. On the other hand, photorepair by 6-4PP-photolyase in keratinocytes reduced but did not abrogate these UV-induced effects. The photolyase mediated removal of either CPDs or 6-4PPs from basal keratinocytes in the skin also reduced UVB-induced apoptosis, ICAM-1 expression, and myeloperoxidase activation. These findings indicate that, in NER-deficient rodents, both types of photolesions have causal roles in UVB-induced epidermal cell proliferation, hyperplasia, cell death and inflammation. Furthermore, these findings also support the notion that basal keratinocytes, instead of other skin cells, are the major cellular mediators of these UVB-induced effects.


Emergence of coherent structures and large-scale flows in motile suspensions.
The emergence of coherent structures, large-scale flows and correlated dynamics in suspensions of motile particles such as swimming micro-organisms or artificial microswimmers is studied using direct particle simulations. A detailed model is proposed for a slender rod-like particle that propels itself in a viscous fluid by exerting a prescribed tangential stress on its surface, and a method is devised for the efficient calculation of hydrodynamic interactions in large-scale suspensions of such particles using slender-body theory and a smooth particle-mesh Ewald algorithm. Simulations are performed with periodic boundary conditions for various system sizes and suspension volume fractions, and demonstrate a transition to large-scale correlated motions in suspensions of rear-actuated swimmers, or Pushers, above a critical volume fraction or system size. This transition, which is not observed in suspensions of head-actuated swimmers, or Pullers, is seen most clearly in particle velocity and passive tracer statistics. These observations are consistent with predictions from our previous mean-field kinetic theory, one of which states that instabilities will arise in uniform isotropic suspensions of Pushers when the product of the linear system size with the suspension volume fraction exceeds a given threshold. We also find that the collective dynamics of Pushers result in giant number fluctuations, local alignment of swimmers and strongly mixing flows. Suspensions of Pullers, which evince no large-scale dynamics, nonetheless display interesting deviations from the random isotropic state.


Iron Isotope Fractionation during Fe(II) Oxidation Mediated by the Oxygen-Producing Marine Cyanobacterium Synechococcus PCC 7002.
In this study, we couple iron isotope analysis to microscopic and mineralogical investigation of iron speciation during circumneutral Fe(II) oxidation and Fe(III) precipitation with photosynthetically produced oxygen. In the presence of the cyanobacterium Synechococcus PCC 7002, aqueous Fe(II) (Fe(II)aq) is oxidized and precipitated as amorphous Fe(III) oxyhydroxide minerals (iron precipitates, Feppt), with distinct isotopic fractionation (ε56Fe) values determined from fitting the δ56Fe(II)aq (1.79‰ and 2.15‰) and the δ56Feppt (2.44‰ and 2.98‰) data trends from two replicate experiments. Additional Fe(II) and Fe(III) phases were detected using microscopy and chemical extractions and likely represent Fe(II) and Fe(III) sorbed to minerals and cells. The iron desorbed with sodium acetate (FeNaAc) yielded heavier δ56Fe compositions than Fe(II)aq. Modeling of the fractionation during Fe(III) sorption to cells and Fe(II) sorption to Feppt, combined with equilibration of sorbed iron and with Fe(II)aq using published fractionation factors, is consistent with our resulting δ56FeNaAc. The δ56Feppt data trend is inconsistent with complete equilibrium exchange with Fe(II)aq. Because of this and our detection of microbially excreted organics (e.g., exopolysaccharides) coating Feppt in our microscopic analysis, we suggest that electron and atom exchange is partially suppressed in this system by biologically produced organics. These results indicate that cyanobacteria influence the fate and composition of iron in sunlit environments via their role in Fe(II) oxidation through O2 production, the capacity of their cell surfaces to sorb iron, and the interaction of secreted organics with Fe(III) minerals.


Benefits and cultural adaptations of psychosocial interventions for parents and their children with intellectual disabilities in low-and middle-income countries: A systematic review.
Evidence and awareness of the importance of culturally adapting psychosocial interventions is growing. The aim of this paper is to systematically review studies on cultural adaptations of psychosocial interventions for parents and their children with intellectual disabilities, in low- and middle-income countries. Studies were identified through electronic databases and searching bibliographies. The quality and cultural adaptations of thirteen studies focusing on parental trainings were analysed using standardised tools and frameworks. Findings suggest interventions reduce the risk of depression and stress and increase coping strategies and positive perceptions of family functioning. Parenting skills training may improve parent-child interactions and child development. However, these benefits should be interpreted cautiously due to methodological shortcomings. Most studies described efforts to make appropriate cultural adaptations to the interventions, but these adaptations were not comprehensive. High-quality cultural adaptations are crucial to providing meaningful interventions in different parts of the world.


High-speed in vivo calcium imaging reveals neuronal network activity with near-millisecond precision.
Two-photon calcium imaging of neuronal populations enables optical recording of spiking activity in living animals, but standard laser scanners are too slow to accurately determine spike times. Here we report in vivo imaging in mouse neocortex with greatly improved temporal resolution using random-access scanning with acousto-optic deflectors. We obtained fluorescence measurements from 34-91 layer 2/3 neurons at a 180-490 Hz sampling rate. We detected single action potential-evoked calcium transients with signal-to-noise ratios of 2-5 and determined spike times with near-millisecond precision and 5-15 ms confidence intervals. An automated 'peeling' algorithm enabled reconstruction of complex spike trains from fluorescence traces up to 20-30 Hz frequency, uncovering spatiotemporal trial-to-trial variability of sensory responses in barrel cortex and visual cortex. By revealing spike sequences in neuronal populations on a fast time scale, high-speed calcium imaging will facilitate optical studies of information processing in brain microcircuits.


Assessment of Common Preoxygenation Strategies Outside of the Operating Room Environment.
Preoxygenation prior to intubation aims to increase the duration of safe apnea by causing denitrogenation of the functional residual capacity, replacing this volume with a reservoir of oxygen. In the operating room (OR) the criterion standard for preoxygenation is an anesthetic circuit and well-fitting face mask, which provide a high fractional inspired oxygen concentration (FiO2 ). Outside of the OR, various strategies exist to provide preoxygenation. The objective was to evaluate the effectiveness of commonly used preoxygenation strategies outside of the OR environment. This was a prospective randomized unblinded study of 30 healthy staff volunteers from a major trauma center emergency department (ED) in Sydney, Australia. The main outcome measure is fractional expired oxygen concentration (FeO2 ) measured after a 3-minute period of tidal volume breathing with seven different preoxygenation strategies. The mean FeO2 achieved with the anesthetic circuit was 81.0% (95% confidence interval [CI] = 78.3% to 83.6%), bag-valve-mask (BVM) 80.1% (95% CI = 76.5% to 83.6%), BVM with nasal cannula (NC) 74.8% (95% CI = 72.0% to 77.6%), BVM with positive end-expiratory pressure valve (PEEP) 78.9% (95% CI = 75.4% to 82.3%), BVM + NC + PEEP 75.5% (95% CI = 72.2% to 78.9%), nonrebreather mask (NRM) 51.6% (95% CI = 48.8% to 54.4%), and NRM + NC 57.1% (95% CI = 52.9% to 61.2%). Preoxygenation efficacy with BVM strategies was significantly greater than NRM strategies (p < 0.01) and noninferior to the anesthetic circuit. In healthy volunteers, the effectiveness of BVM preoxygenation was comparable to the anesthetic circuit (criterion standard) and superior to preoxygenation with NRM. The addition of NC oxygen, PEEP, or both did not improve the efficacy of the BVM device.


The Arabidopsis CCCH protein C3H14 contributes to basal defense against Botrytis cinerea mainly through the WRKY33-dependent pathway.
Necrotrophic pathogens such as Botrytis cinerea cause significant crop yield losses. Plant CCCH proteins play important roles in pathogen resistance responses. However, the CCCH-mediated defense mechanisms against necrotrophic pathogens are unclear. Here, we report that the Arabidopsis CCCH protein C3H14 positively regulates basal defense against B. cinerea mainly by WRKY33 signaling. Simultaneous mutation of C3H14 and its paralog C3H15 resulted in enhanced susceptibility to B. cinerea, while C3H14 or C3H15 overexpression lines exhibited reduced susceptibility. A large number of differentially expressed genes (DEGs) were present in the c3h14c3h15 double mutant and C3H14 overexpression plants compared with wild-type plants at 24 hr post infection. These DEGs covered over one third of B. cinerea-responsive WRKY33 targets, including genes involved in jasmonic acid (JA)/ethylene (ET) signaling, and camalexin biosynthesis. Genetic analysis indicated that C3H14 mainly depended on WRKY33 to modulate defense against B. cinerea. Moreover, C3H14 activated the WRKY33-ORA59 and -PAD3 cascades to correspondingly control JA/ET- and camalexin-mediated defense responses. However, C3H14 was essential for B. cinerea-induced production of 12-oxo-phytodienoic acid and it also directly mediated ORA59-dependent JA/ET signaling after infection. Therefore, C3H14 may act as a novel transcriptional regulator of the WRKY33-mediated defense pathway.


Association of anxiety, sleepiness, and sexual dysfunction with restless legs syndrome in hemodialysis patients.
Restless legs syndrome (RLS) is characterized by unpleasant sensations, pain in the legs along with irresistible urges to move the legs when at rest. It is often accompanied by sleep disturbance. The purpose of this study was to assess the association of anxiety and sleepiness with sexual function in hemodialysis patients with and without RLS. Sociodemographic parameters, laboratory data of hemodialysis patients from three dialysis centers were collected prospectively. Anxiety, sleepiness, sexual function, and presence of RLS symptoms were assessed with standardized questionnaires as the RLS Diagnosis and Scale, Hamilton Anxiety Rating Scale, Epworth Sleepiness Scale (ESS), Arizona Sex Experiences Scale (ASEX). Univariate, regression tree method were used for statistical analysis. RLS was observed in 45.9% (n = 113) of hemodialysis patients (n = 246). The mean age of patients and duration of hemodialysis were 59.7 ± 14.0 and 4.9 ± 4.2 years, respectively. The correlation between Arizona Sexual Experiences Scale (ASEX) and sociodemographic features was significant (P < 0.0001). Patients with RLS had higher scores for anxiety (9.4 ± 7.8 with RLS and 6.8 ± 6.0 without), higher ESS (ESS, 6.6 ± 5.2 with RLS and 4.6 ± 4.0 without), and higher ASEX (24.6 ± 5.7 with RLS and 22.5 ± 6.8 without) than did those without RLS. The presence of RLS symptoms in hemodialysis patients was associated with sleepiness, anxiety, and sexual dysfunction. A regression tree method, which is a different statistical method, can help physicians estimate patients ASEX, RLS, ESS, and anxiety scores.


Updated thresholds for serum alanine aminotransferase level in a large-scale population study composed of 34 346 subjects.
The sensitivity of current upper limit of normal (ULN) of serum alanine aminotransferase (ALT) levels for detecting chronic liver disease has been challenged recently. To identify modulating factors for serum ALT levels and to refine its ULN threshold. We enrolled 34 346 consecutive subjects who completed the health check-up at Taipei Veterans General Hospital from 2002 to 2009. ULN was set for healthy ALT level to the 95th percentile of the reference healthy population. A group of 21 282 subjects were used as a training set to define an ULN with the highest sensitivity; afterwards, this ULN was validated in another set of 13 064 subjects. A reference healthy population was selected from the training set after excluding subjects with any abnormalities in independent risk factors associated with elevated serum ALT level (>40 IU/L) by multivariate analysis like body mass index, waist circumference, glucose, cholesterol, high-density lipoprotein-cholesterol, triglyceride, hepatitis B virus surface antigen, anti-hepatitis C virus antibody and fatty liver. The new ULN of serum ALT level defined as the 95% percentile in the healthy population were 21 IU/L and 17 IU/L for men and women respectively. These cut-off values had the highest Youden's index and areas under the corresponding receiver operating curves among four widely applied thresholds in both the training and validation sets. The suggested threshold of upper limit of normal provides better discrimination between healthy and unhealthy status. Viral hepatitis, metabolic syndrome and fatty liver are the major risk factors of elevated serum alanine aminotransferase levels.


Evaluation of potent inhibitors of dihydrofolate reductase in a culture model for growth of Pneumocystis carinii.
Many antifolates are known to inhibit dihydrofolate reductase from murine Pneumocystis carinii, with 50% inhibitory concentrations (IC50s) ranging from 10(-4) to 10(-11) M. The relationship of the potency against isolated enzyme to the potency against intact murine P. carinii cells was explored with 17 compounds that had proven selectivity for or potency against P. carinii dihydrofolate reductase. Pyrimethamine and one analog were inhibitory to P. carinii in culture at concentrations two to seven times the IC50s for the enzyme, suggesting that the compounds may enter P. carinii cells in culture. Methotrexate was a potent inhibitor of P. carinii dihydrofolate reductase, but the concentrations effective in culture were more than 1,000-fold higher than IC50s for the enzyme, since P. carinii lacks an uptake system for methotrexate. Analogs of methotrexate in which chlorine, bromine, or iodine was added to the phenyl ring had improved potency against the isolated enzyme but were markedly less effective in culture; polyglutamation also lowered the activity in culture but improved activity against the enzyme. Substitution of a naphthyl group for the phenyl group of methotrexate produced a compound with improved activity against the enzyme (IC50, 0.00019 microM) and excellent activity in culture (IC50, 0.1 microM). One trimetrexate analog in which an aspartate or a chlorine replaced two of the methoxy groups of trimetrexate was much more potent and was much more selective toward P. carinii dihydrofolate reductase than trimetrexate; this analog was also as active as trimetrexate in culture. These studies suggest that modifications of antifolate structures can be made that facilitate activity against intact organisms while maintaining the high degrees of potency and the selectivities of the agents can be made.


[Mechanism of the cellular transformation of newborn hamsters inoculated with herpes simplex type 2 virus].
Combined virological, antigenic, electron microscopic, and molecular biologic study of the role of an oncovirus, herpes simplex type 2 virus (HSV-2), in the mechanisms of transformation of HSV-2-infected hamster cells was carried out. No expression of genetic information of the oncovirus could be detected. Cell transformation was shown to be associated with persistence and realization of HSV-2 genetic information in the transformed cells.


Noncentrosymmetric (NCS) solid solutions: elucidating the structure-nonlinear optical (NLO) property relationship and beyond.
A systematic approach toward the discovering of novel functional noncentrosymmetric (NCS) materials revealing technologically useful applications is an ongoing challenge. This Frontiers article investigates a series of NCS solid solutions with respect to their crystal structure and second-harmonic generation (SHG) response. The solid solutions include NCS polar aluminoborates, Al5-xGaxBO9 (0.0 ≤ x ≤ 0.5), rare earth element-doped bismuth tellurites, Bi2-xRExTeO5 (RE = Y, Ce, and Eu; 0.0 ≤ x ≤ 0.2), layered perovskites, Bi4-xLaxTi3O12 (0.0 ≤ x ≤ 0.75: Aurivillius phases) and CsBi1-xEuxNb2O7 (0.0 ≤ x ≤ 0.2: Dion-Jacobson phases), calcium bismuth oxides, Ca4Bi6-xLnxO13 (Ln = La and Eu; x = 0, 0.06 and 0.12), and sodium lanthanide iodates, NaLa1-xLnx(IO3)4 (Ln = Sm and Eu; 0 ≤ x ≤ 1). The origin of SHG for all the NCS solid solutions is discussed and the detailed structure-nonlinear optical (NLO) property relationships are elucidated. In addition, photoluminescence (PL) properties and subsequent energy transfer mechanisms for NCS solid solutions are provided.


Prenatal exposure to airborne polycyclic aromatic hydrocarbons and children's intelligence at 5 years of age in a prospective cohort study in Poland.
In this prospective cohort study of Caucasian mothers and children in Krakow, Poland, we evaluated the role of prenatal exposure to urban air pollutants in the pathogenesis of neurobehavioral disorders. The objective of this study was to investigate the relationship between prenatal polycyclic aromatic hydrocarbon (PAH) exposure and child intelligence at 5 years of age, controlling for potential confounders suspected to play a role in neurodevelopment. A cohort of pregnant, healthy, nonsmoking women was enrolled in Krakow, Poland, between 2001 and 2006. During pregnancy, participants were invited to complete a questionnaire and undergo 48-hr personal air monitoring to estimate their babies' exposure, and to provide a blood sample and/or a cord blood sample at the time of delivery. Two hundred fourteen children were followed through 5 years of age, when their nonverbal reasoning ability was assessed using the Raven Coloured Progressive Matrices (RCPM). We found that higher (above the median of 17.96 ng/m3) prenatal exposure to airborne PAHs (range, 1.8-272.2 ng/m3) was associated with decreased RCPM scores at 5 years of age, after adjusting for potential confounding variables (n = 214). Further adjusting for maternal intelligence, lead, or dietary PAHs did not alter this association. The reduction in RCPM score associated with high airborne PAH exposure corresponded to an estimated average decrease of 3.8 IQ points. These results suggest that prenatal exposure to airborne PAHs adversely affects children's cognitive development by 5 years of age, with potential implications for school performance. They are consistent with a recent finding in a parallel cohort in New York City.


Case report: Huge amoebic liver abscesses in both lobes.
We describe the case of a patient who returned to China from Africa and underwent emergency open surgical drainage with evacuation of 600 mL of anchovy sauce-like fluid from hepatic lesions. Computed tomography scans and surgical findings indicated abscesses in both hemilivers and communication between them. Bacteriological investigation of the fluid yielded negative results, but DNA assay of the pus detected 18S rRNA genes of Entamoeba histolytica. Serum anti-amoebic antibodies were detected using an indirect fluorescent-antibody test. Consequently, anti-amoebic drugs were administered and drainage was performed, leading to improvement in the patient's condition. As is evident from this case, an amoebic liver abscess in the left hepatic lobe is rare but treatable.


Inflammatory responses induced by substance P in rat paw.
Substance P (SP) injection in the plantar region of rat hind paw caused a dose related inflammation, which reached a peak within 10 min of injection and declined after 60 min. Low doses (0.25-0.063 mg/kg) of SP-antagonists like (D-Pro2, D-Trp7,9)-SP and (D-Pro2, D-Phe7, D-Trp9)-SP pretreatment significantly inhibited the SP induced paw oedema, while higher doses (0.5-1 mg/kg) showed agonistic effects. Pretreatment with diphenhydramine alone or along with low doses of SP-antagonists was highly significant in blocking this inflammation, the latter combination being more effective than the former. Pretreatment with acute capsaicin produced a synergestic effect on SP induced paw oedema, while pretreatment with chronic capsaicin significantly inhibited this SP induced paw oedema. The results indicate involvement of histamine and possible therapeutic importance of capsaicin in SP mediated inflammatory type of responses.


Rational and evolutionary engineering approaches uncover a small set of genetic changes efficient for rapid xylose fermentation in Saccharomyces cerevisiae.
Economic bioconversion of plant cell wall hydrolysates into fuels and chemicals has been hampered mainly due to the inability of microorganisms to efficiently co-ferment pentose and hexose sugars, especially glucose and xylose, which are the most abundant sugars in cellulosic hydrolysates. Saccharomyces cerevisiae cannot metabolize xylose due to a lack of xylose-metabolizing enzymes. We developed a rapid and efficient xylose-fermenting S. cerevisiae through rational and inverse metabolic engineering strategies, comprising the optimization of a heterologous xylose-assimilating pathway and evolutionary engineering. Strong and balanced expression levels of the XYL1, XYL2, and XYL3 genes constituting the xylose-assimilating pathway increased ethanol yields and the xylose consumption rates from a mixture of glucose and xylose with little xylitol accumulation. The engineered strain, however, still exhibited a long lag time when metabolizing xylose above 10 g/l as a sole carbon source, defined here as xylose toxicity. Through serial-subcultures on xylose, we isolated evolved strains which exhibited a shorter lag time and improved xylose-fermenting capabilities than the parental strain. Genome sequencing of the evolved strains revealed that mutations in PHO13 causing loss of the Pho13p function are associated with the improved phenotypes of the evolved strains. Crude extracts of a PHO13-overexpressing strain showed a higher phosphatase activity on xylulose-5-phosphate (X-5-P), suggesting that the dephosphorylation of X-5-P by Pho13p might generate a futile cycle with xylulokinase overexpression. While xylose consumption rates by the evolved strains improved substantially as compared to the parental strain, xylose metabolism was interrupted by accumulated acetate. Deletion of ALD6 coding for acetaldehyde dehydrogenase not only prevented acetate accumulation, but also enabled complete and efficient fermentation of xylose as well as a mixture of glucose and xylose by the evolved strain. These findings provide direct guidance for developing industrial strains to produce cellulosic fuels and chemicals.


Identifying Nuclear Matrix-Attached DNA Across the Genome.
Experimental approaches to define the relationship between gene expression and nuclear matrix attachment regions (MARs) have given contrasting and method-specific results. We have developed a next generation sequencing strategy to identify MARs across the human genome (MAR-Seq). The method is based on crosslinking chromatin to its nuclear matrix attachment sites to minimize changes during biochemical processing. We used this method to compare nuclear matrix organization in MCF-10A mammary epithelial-like cells and MDA-MB-231 breast cancer cells and evaluated the results in the context of global gene expression (array analysis) and positional enrichment of gene-regulatory histone modifications (ChIP-Seq). In the normal-like cells, nuclear matrix-attached DNA was enriched in expressed genes, while in the breast cancer cells, it was enriched in non-expressed genes. In both cell lines, the chromatin modifications that mark transcriptional activation or repression were appropriately associated with gene expression. Using this new MAR-Seq approach, we provide the first genome-wide characterization of nuclear matrix attachment in mammalian cells and reveal that the nuclear matrix-associated genome is highly cell-context dependent. J. Cell. Physiol. 232: 1295-1305, 2017. © 2016 Wiley Periodicals, Inc.


Escherichia coli BTC, a human cytochrome P450 competent tester strain with a high sensitivity towards alkylating agents: involvement of alkyltransferases in the repair of DNA damage induced by aromatic amines.
We report here on strain BTC, a new Escherichia coli mutagenicity tester strain for the expression of human cytochrome P450 (CYP) with an enhanced sensitivity for the detection of alkylating agents. This strain was developed first through knocking out of the genes ada and ogt in our previously developed strain BMX100, resulting in PD1000. Strain PD1000 demonstrated a significantly higher detection sensitivity towards several alkylating agents such as N-nitrosodiethylamine (NNdEA), N-nitrosodi-n-propylamine (NNdPA), 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) and N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). Unexpectedly, this strain also showed an enhanced sensitivity towards 2-aminoanthracene (2AA), 4-aminobiphenyl (4AbPh), 2-aminofluorene (2AF) and 2-nitroanthracene (2NA) mutagenicity. Subsequently, our previously developed bi-plasmid system for the co-expression of a specific human CYP form (CYP1A2, 2A6 or 2E1) with human NADPH-cytochrome P450 reductase (RED) was introduced in strain PD1000, resulting in strains BTC1A2, BTC2A6 and BTC2E1, respectively. The mutagenicity of NNdEA and NNK was successfully detected with strains BTC2A6 and BTC2E1 and with strains BTC1A2 and BTC2A6, respectively, in contrast to the corresponding MTC (ada+ ogt+) CYP strains. The (ada- ogt-) deficient strain BTC1A2 also showed an enhanced sensitivity towards the detection of 2AA mutagenicity, when compared with the proficient repair strain MTC1A2. This enhancement was much more pronounced with strain PD1000 using the rat liver S9 fraction than with strain BTC1A2.


A new species of Bachia Gray, 1845 (Squamata: Gymnophthalmidae) from the Eastern Brazilian Cerrado, and data on its ecology, physiology and behavior.
A new species of Bachia of the bresslaui group, Bachia geralista sp. nov., is described from Planalto dos Gerais, an old and partially dissected plateau extending along the Cerrados of Bahia, Minas Gerais and Tocantins states, Brazil. The new species is morphologically similar to B. bresslaui, with which it has been confused; however head scalation resembles other species from sandy spots within the Cerrado (B. psamophila and B. oxyrhina). Like in B. psamophila and B. oxyrhina, the shovel-shaped snout of the new species is highly prominent, a typical trait of psammophilous habits in other gymnophthalmids. The examination of specimens of B. bresslaui from several populations within the Cerrado revealed great variation among localities, leading to the reidentification of a specimen from Utiariti, Mato Grosso, previously referred to in the literature as the second record of B. bresslaui, as the recently described B. didactyla, suggesting that cryptic diversity might remain still undiscovered within this genus in the Cerrado. Despite occurring in a relatively open Cerrado, thermal physiology of Bachia geralista sp. nov. restricts its occurrence to shaded microhabitats within this habitat.


[Tissue engineering of the small joint].
We have developed a model of human phalanges and small joints that can be formed in a tissue engineering manner by suturing three different cell-polymer constructs. The polymer scaffolds supported growth of osteoblasts, chondrocytes, or tenocytes after the models were implanted in athymic nude mice. The constructs in vivo developed through vascularization, formed new bone, cartilage, and tendon with characteristic gene expression and protein synthesis, and maintained the shape of human phalanges with joints. As additional studies proceed, this approach will clinically become important.


Use of quantitative ultrasonography for noninvasive surveillance of the third metacarpal bone in racing and training Thoroughbreds.
OBJECTIVE-To use quantitative ultrasonography to evaluate the association between the speed of sound (SOS) at 9 sites in the third metacarpal bone (MCIII) of racing Thoroughbreds with workload accumulation and the effect that MCIII failure has on this association. ANIMALS-Sixty-two 2- and 3-year-old Thoroughbreds in racing condition. PROCEDURES-Cumulative work index (CWI) was used to calculate total workload (CWI(total)) and also 3 independent CWIs for the various gaits (ie, trot [CWI(trot)], gallop [CWI(gallop)], and race [CWI(race)]) used during training and racing. Speed of sound was monitored in horses during the 2007 racing season and compared with the CWIs via regression analysis. Sex, age, limb, and MCIII failure were included as covariates in the model. RESULTS-SOS was significantly associated with CWI(total) at 8 sites and with independent CWIs of the various gaits at all 9 sites. Progression of SOS in MCIIIs with workload differed significantly in horses with clinical signs of metacarpal bone failure, compared with results for horses with clinically normal MCIIIs, in 1 site by use of CWI(total) and in 5 sites by use of the independent CWIs for the various gaits. CONCLUSIONS AND CLINICAL RELEVANCE-These results indicated that SOS in the MCIII of racing Thoroughbreds followed a constant pattern of progression as workload accumulated. With the development of more precise quantitative ultrasonography devices, SOS corrected for amount of activity may be used to identify horses at risk of bone failure.


Chance of surgery in adult congenital heart disease.
Background Young patients with congenital heart disease reaching adulthood face mandatory transition to adult cardiology. Their new cardiologist needs to assess the chances of major future events such as surgery. Using a large national registry, we assessed if patient characteristics at the age of 18 years could predict the chance of congenital heart surgery in adulthood. Design and methods Of 10,300 patients from the CONCOR national registry, we used general patient characteristics at age 18 years, underlying congenital heart defect, history of complications, and interventions in childhood as potential predictors of congenital heart surgery occurring from age 18 years up to age 40 and 60 years. Cox regression was used to calculate hazard ratios with 95% confidence intervals. Analyses were performed separately for all congenital heart surgery and for valvular surgery alone. Results Altogether 2427 patients underwent congenital heart surgery after age 18 years, 1389 of whom underwent valvular surgery. Underlying heart defect, male sex, multiple defects, childhood endocarditis, supraventricular arrhythmia, aortic complications and paediatric cardiovascular surgery, independently predicted adult congenital heart surgery. The mean chance of congenital heart surgery was 22% up to age 40 and 43% up to age 60 years; individual chances spanned from 9-68% up to age 40 and from 19-93% up to age 60 years. Conclusion At the time of transition from paediatric to adult cardiology, an easily obtainable set of characteristics of patients with congenital heart disease can meaningfully inform cardiologists about the patient's individual chance of surgery in adulthood. Our findings warrant validation in other cohorts.


Kinetic Model of Incipient Hydride Formation in Zr Clad under Dynamic Oxide Growth Conditions.
The formation of elongated zirconium hydride platelets during corrosion of nuclear fuel clad is linked to its premature failure due to embrittlement and delayed hydride cracking. Despite their importance, however, most existing models of hydride nucleation and growth in Zr alloys are phenomenological and lack sufficient physical detail to become predictive under the variety of conditions found in nuclear reactors during operation. Moreover, most models ignore the dynamic nature of clad oxidation, which requires that hydrogen transport and precipitation be considered in a scenario where the oxide layer is continuously growing at the expense of the metal substrate. In this paper, we perform simulations of hydride formation in Zr clads with a moving oxide/metal boundary using a stochastic kinetic diffusion/reaction model parameterized with state-of-the-art defect and solute energetics. Our model uses the solutions of the hydrogen diffusion problem across an increasingly-coarse oxide layer to define boundary conditions for the kinetic simulations of hydrogen penetration, precipitation, and dissolution in the metal clad. Our method captures the spatial dependence of the problem by discretizing all spatial derivatives using a stochastic finite difference scheme. Our results include hydride number densities and size distributions along the radial coordinate of the clad for the first 1.6 h of evolution, providing a quantitative picture of hydride incipient nucleation and growth under clad service conditions.


A clustering performance measure based on fuzzy set decomposition.
Clustering is primarily used to uncover the true underlying structure of a given data set and, for this purpose, it is desirable to subject the same data to several different clustering algorithms. This paper attempts to put an order on the various partitions of a data set obtained from different clustering algorithms. The goodness of each partition is expressed by means of a performance measure based on a fuzzy set decomposition of the data set under consideration. Several experiments reported in here show that the proposed performance measure puts an order on different partitions of the same data which is consistent with the error rate of a classifier designed on the basis of the obtained cluster labelings.


Unforeseen decreases in dissolved oxygen levels affect tube formation kinetics in collagen gels.
The availability of oxygen (O(2)) is a critical parameter affecting vascular tube formation. In this study, we hypothesize that dissolved oxygen (DO) levels in collagen gels change during the three-dimensional (3D) culture of human umbilical vein endothelial cells (HUVECs) in atmospheric conditions and that such changes affect the kinetics of tube formation through the production of reactive oxygen species (ROS). We demonstrate a decrease in O(2) tension during 3D cultures of HUVECs. Noninvasive measurements of DO levels during culture under atmospheric conditions revealed a profound decrease that reached as low as 2% O(2) at the end of 24 h. After media replacement, DO levels rose rapidly and equilibrated at ∼15% O(2), creating a reoxygenated environment. To accurately estimate DO gradients in 3D collagen gels, we developed a 3D mathematical model and determined the Michaelis-Menten parameters, V(max) and K(m), of HUVECs in collagen gels. We detected an increase in ROS levels throughout the culture period. Using diphenyliodonium to inhibit ROS production resulted in the complete inhibition of tube formation. Interference RNA studies further showed that hypoxia-inducible factors (HIFs)-1α and -2α are not involved in the formation of 3D tubes in collagen gels. We conclude that ROS affect the tubulogenesis process through HIFα-independent pathways, where the levels of ROS are influenced by the uncontrolled variations in DO levels. This study is the first demonstration of the critical and unexpected role of O(2) during 3D in vitro culture models of tubulogenesis in atmospheric conditions.


In vivo measurement of apparent trabecular bone structure of the radius in women with low bone density discriminates patients with recent wrist fracture from those without fracture.
The purpose of this cross-sectional case-control study was to determine whether indices of bone structure at the 4% site of the radius discriminate women who have sustained a recent low-energy fracture of the contralateral distal radius (n = 21) from women with similar bone density and no history of fracture (n = 21). Images of the distal forearm were acquired using peripheral quantitative computed tomography (pQCT) and were analyzed using in-house-developed software to determine indices of trabecular bone structure (average hole size [H(A)], maximum hole size, and connectivity index). The same images were analyzed using commercial software to determine bone density, mass, geometry, and torsional strength. The fracture group had significantly larger H(A) (p = 0.03). No other bone variable differed between groups. Individual H(A) values were compared to the mean value for young adult women (n = 42). The odds ratio (95% confidence interval) associated with an H(A) >2 SDs above the young adult mean was 5.4 (1.2-24.3). Thus, estimation of in vivo trabecular bone porosity by measuring the average diameter of the pQCT-imaged marrow spaces in the ultradistal radius identifies individuals with low bone mass most at risk for osteoporotic fracture.


Bile Acids in Autoimmune Liver Disease: Unveiling the Nexus of Inflammation, Inflammatory Cells, and Treatment Strategies.
As bile acids not solely play an essential role in nutrition absorption, but also in regulating metabolic functions as well as immune response, bile acids and their signaling pathways are increasingly acknowledged as potential therapeutic targets in the context of chronic liver diseases. Bile acid receptors such as G protein bile acid-activated receptor 1 and farnesoid X receptor are expressed in different immune cells engaged in innate immunity. Recently, a series of studies have revealed distinct functions of bile acids and bile acid receptors within the adaptive immune system. In addition, a variety of molecules targeting bile acid receptors and transporters are currently in advanced stages of clinical development. Autoimmune liver diseases including conditions like primary biliary cholangitis, primary sclerosing cholangitis, and autoimmune hepatitis can lead to chronic inflammation, fibrosis, and even cirrhosis and liver failure. In this review, we focus on the role of bile acids in the inflammatory aspects of autoimmune liver diseases.


Oxidative Approach Enables Efficient Access to Cyclic Azobenzenes.
Azobenzenes are versatile photoswitches that have found widespread use in a variety of fields, ranging from photopharmacology to the material sciences. In addition to regular azobenzenes, the cyclic diazocines have recently emerged. Although diazocines have fascinating conformational and photophysical properties, their use has been limited by their synthetic accessibility. Herein, we present a general, high-yielding protocol that relies on the oxidative cyclization of dianilines. In combination with a modular substrate synthesis, it allows for rapid access to diversely functionalized diazocines on gram scales. Our work systematically explores substituent effects on the photoisomerization and thermal relaxation of diazocines. It will enable their incorporation into a wide variety of functional molecules, unlocking the full potential of these emerging photoswitches. The method can be applied to the synthesis of a new cyclic azobenzene with a nine-membered central ring and distinct properties.


Sponge-derived polybrominated diphenyl ethers and dibenzo-p-dioxins, irreversible inhibitors of the bacterial α-d-galactosidase.
An integrated in vitro and in silico approach was applied to evaluate the potency of hydroxylated polybrominated diphenyl ethers (OH-PBDEs) and spongiadioxins (OH-PBDDs) isolated from Dysidea sponges on the activity of the recombinant α-d-galactosidase of the GH36 family. It was revealed for the first time that all compounds rapidly and apparently irreversibly inhibited the bacterial α-d-galactosidase. The structure-activity relationship study in the series of OH-PBDEs showed that the presence of an additional hydroxyl group in 5 significantly enhanced the potency (IC50 4.26 μM); the increase of bromination in compounds from 1 to 3 increased their potency (IC50 41.8, 36.0, and 16.0 μM, respectively); the presence of a methoxy group decreased the potency (4, IC50 60.5 μM). Spongiadioxins 6, 7, and 8 (IC50 16.6, 33.1, and 28.6 μM, respectively) exhibited inhibitory action comparable to that of monohydroxylated diphenyl ethers 1-3. Docking analysis revealed that all compounds bind in a pocket close to the catalytic amino acid residues. Molecular docking detected significant compound-enzyme interactions in the binding sites of α-d-galactosidase. Superimposition of the enzyme-substrate and the enzyme-inhibitor complexes showed that their binding sites overlap.


Developing a translational discourse to communicate uncertainty in flood risk between science and the practitioner.
The language and tools of risk and uncertainty estimation in flood risk management (FRM) are rarely optimized for the extant communication challenge. This paper develops the rationale for a pragmatic semiotics of risk communication between scientists developing flood models and forecasts and those professional groups who are the receptors for flood risk estimates and warnings in the UK. The current barriers to effective communication and the constraints involved in the formation of a communication language are explored, focusing on the role of the professional's agenda or "mission" in creating or reducing those constraints. The tools available for the development of this discourse, for both flood warnings in real time and generalized FRM communications, are outlined. It is argued that the contested ownership of the articulation of uncertainties embedded in flood risk communications could be reduced by the development of a formally structured translational discourse between science and professionals in FRM, through which process "codes of practice" for uncertainty estimation in different application areas can be developed. Ways in which this might take place in an institutional context are considered.


Ischemic preconditioning of renal tissue: identification of early up-regulated genes.
Given the important effects of ischemic preconditioning (IPC) in minimizing tissue damage induced by sustained ischemia in several tissues, this study evaluated the effect of IPC in preserving renal function and identified up-regulated genes after 30 min of preconditioning. IPC induced by 2, 3 and 4 min of ischemia, intercalated by 5 min of reperfusion, induced a measurable protection of renal function and morphology. The improved functional and histological parameters occurred in parallel with up-regulation of 39 genes, as evaluated by subtractive hybridization; for 13 of them we could show, by RNAse protection assay, a significant increase in mRNA levels. These genes code for chaperones/chaperonins and cytoskeleton proteins that could be involved in preservation of protein folding and cellular structures after sustained ischemia; proteins related to oxidative metabolism that might be relevant for cellular use of alternate sources of energy or for faster recovery of ATP levels in this condition, and proteins that are putative scavengers of oxidant products. Summarizing, ischemic preconditioning induced up-regulation of genes that code proteins whose functional roles suggest their involvement in the tolerance of the preconditioned tissue to sustained ischemia.


Prostatic luminal cell differentiation and prostatic steroid-binding protein (PBP) gene expression are differentially affected by neonatal castration.
Although normal prostatic development is androgen-dependent, the prostate continues to grow in the neonate despite castration. However, the manner in which neonatal growth of the prostate occurs, in the absence of the testis, remains largely unknown. The purpose of this study was to examine the differentiation of prostatic epithelial cells after neonatal castration. Immunohistochemistry was utilized to detect the expression of differentiation products: basal-cell cytokeratin (CK 5), luminal-cell cytokeratin (CK 18), and prostatic steroid-binding protein (PBP), a ventral prostate-specific marker indicative of secretory function in luminal cells. The reverse transcription-polymerase chain reaction was used to detect transcription products of the three polypeptide subunits of PBP, designated C1, C2, and C3. Rats were castrated on day 5 after birth, and ventral prostates were collected on day 14. Dihydrotestosterone was injected (100 microg/animal every 2 days) in castrated animals to determine if PBP expression could be initiated by androgen. Although no major effects of castration were detected on the differentiation of stromal or basal cells (which differentiate prior to day 5), castration had a pronounced effect on luminal-cell differentiation. Castration inhibited PBP protein expression, but did not affect the expression of luminal-cell cytokeratin (CK 18) protein. Furthermore, castration reduced C1, C2, and C3 transcription. Androgen replacement to castrated animals allowed for the initiation of PBP expression, although its onset was delayed. These observations indicate that the testis is not necessary for prostatic luminal-cell differentiation, but is necessary for full expression of luminal-cell secretory phenotype. Furthermore, our study suggests that factors of testicular origin, in addition to androgen, are needed for proper timing of PBP expression. This investigation establishes that the cytological and the physiological differentiation of the rat prostate are differentially regulated.


The Injury Tolerance of the Tibia Under Off-Axis Impact Loading.
During a frontal collision, there are a range of lower extremity postures that the vehicle's occupant may assume, potentially changing the way load is transmitted to this region of the body. While most experimental studies on the tibia's injury tolerance assume that load is directed along the leg's long axis, the effects of off-axis loading due to non-standard postures have not been well quantified, and commonly-cited injury criteria such as the Tibia Index do not directly account for posture. Therefore, twelve cadaveric tibias (paired from six donors) were subjected to off-axis impact loading in a custom-built test apparatus. One specimen from each pair was held at an angle of 15° relative to the direction of loading, while the contralateral was held at an angle of 30°, with these angles representing ankle plantarflexion and corresponding knee extension in a vehicle occupant. Specimens held at 30° fractured at lower forces than the specimens held at 15° (mean force = 5.8 vs. 7.5 kN). This indicates that posture should be incorporated into injury criteria for the tibia in future safety assessments instead of using a single force value based on axial impacts.


[Leptin--a new weight-loss agent?].
The relatively constant level of body fat in the adult mammal is explained by the lipostat theory as follows: A factor is released from adipose tissue in amounts that reflect total body fat content. This factor is then registered by the hypothalamic centres that regulate appetite. The theory has gained particular support from parabiotic experiments in genetically fat mice, amongst whom the so-called ob/ob mice appear to produce too little of the factor, whilst signal transduction to the satiety centre appears to be defective in the so-called db/db mice. The two genetic defects have recently been clarified, and it has been shown that the normal ob gene nodes for a protein called leptin. Leptin is released from adipose tissue, and its plasma concentration reflects total body fat. The normal db-gene codes for a hypothalamic receptor for leptin. These crucial discoveries have greatly supported the correctness of the lipostat theory. At present, extensive investigations of the roles of leptin and the leptin receptor in human adiposity are in progress.


Membrane association of VP22, a herpes simplex virus type 1 tegument protein.
Tegument proteins of herpes simplex virus type 1 (HSV-1) are hypothesized to contain the functional information required for the budding or envelopment process proposed to occur at cytoplasmic compartments of the host cell. One of the most abundant tegument proteins of HSV-1 is the U(L)49 gene product, VP22, a 38-kDa protein of unknown function. To study its subcellular localization, a VP22-green fluorescent protein chimera was expressed in transfected human melanoma (A7) cells. In the absence of other HSV-1 proteins, VP22 localizes to acidic compartments of the cell that may include the trans-Golgi network (TGN), suggesting that this protein is membrane associated. Membrane pelleting and membrane flotation assays confirmed that VP22 partitions with the cellular membrane fraction. Through truncation mutagenesis, we determined that the membrane association of VP22 is a property attributed to amino acids 120 to 225 of this 301-amino-acid protein. The above results demonstrate that VP22 contains specific information required for targeting to membranes of acidic compartments of the cell which may be derived from the TGN, suggesting a potential role for VP22 during tegumentation and/or final envelopment.


Critical age windows in the impact of lifetime smoking exposure on respiratory symptoms and disease among ever smokers.
Despite extensive knowledge of smoking effects on respiratory disease, there is no study including all age windows of exposure among ever smokers. The objective of this study was to assess the effects from smoking exposure in utero, early childhood, adolescence and adulthood on respiratory health outcomes in adult male and female ever smokers. Respiratory health outcomes were assessed in 10,610 participants of the European Community Respiratory Health Survey (ECRHS) I who reported a history of ever smoking by questionnaire. The associations of maternal smoking in utero, maternal smoking during childhood, age of smoking debut and pack-years of smoking with respiratory symptoms, obstructive diseases and bronchial hyperreactivity were analysed using generalized linear regression, non-linearity between age of smoking debut and outcomes were assessed by Generalized additive mixed models. Respiratory symptoms and asthma were more frequent in adults if their mother smoked during pregnancy, and, in men, also if mother smoked in childhood. Wheeze and ≥3 respiratory symptoms declined with later smoking debut among women [≤10 years: OR = 3.51, 95% CI 1.26, 9.73; 11-12 years: 1.57[1.01-2.44]; 13-15 years: 1.11[0.94-1.32] and ≤10 years: 3.74[1.56-8.83]; 11-12 years: 1.76[1.19-2.56]; 13-15 years: 1.12[0.94-1.35], respectively]. Effects of increasing number of packyears were pronounced in women (Chronic Obstructive Pulmonary Disease (COPD): OR/10 packyears women: 1.33 [1.18, 1.50], men: 1.14 [1.04, 1.26] pinteraction = 0.01). Among ever smokers, smoking exposure in each stage of the lifespan show persistent harmful effects for adult respiratory health, while women appeared to be more vulnerable to an early age of smoking debut and amount of smoking in adulthood.


New Recommendation on Biological Materials Could Hamper Muscular Dystrophy Research.
The new 'Recommendation of the Committee of Ministers to member States on research on biological materials of human origin', adopted in Europe in May 2016 is confusing and lacks specificity on the research use of biomaterials taken from persons not able to consent. It is possible to interpret the relevant clauses in a restrictive manner and doing so would hamper biobank research, by requiring researchers or biobank curators to examine individual records in detail, to check they are adhering to the Recommendation. This would be particularly problematic for muscular dystrophy and other rare disease research, the progress of which relies increasingly on the sharing of biomaterials and data internationally, as it will add complexity to the logistics of biomaterials and data sharing and introduce barriers for researchers preparing biomaterials for sharing. Such barriers are contradictory to EC policies on promoting and funding rare disease research and removing barriers to better care and treatment. Such policies work in concert with international progress in rare disease research, in particular the NIH's Rare Diseases Clinical Research Network and Genetic and Rare Diseases Information Centre. The rare disease community has in recent years worked to create a common framework of harmonised approaches to enable the responsible, voluntary, and secure sharing of biomaterials and data. These efforts are supported by the European Commission in such moves as FP7 funding to advance rare disease research and the introduction of National Plans for rare disease; and are bolstered by similar efforts in the USA via the Clinical and Translational Science Awards Program and the NIH/NCATS Patient Registry developments. Introducing Recommendations from the Committee of Ministers, containing clauses which are incompatible to the efforts to advance rare disease research, seems counter-productive.


Leptin infusion and obesity in mouse cause alterations in the hypothalamic melanocortin system.
The objectives of this study were to identify potential alterations in gene expression of melanocortin-4 receptor (MC4-R), proopiomelanocortin (POMC), and Agouti-related protein (AgRP) in mouse hypothalamus under a chronic peripheral infusion of leptin or at early (8 weeks) and advanced (16 weeks) phases of diet-induced obesity. Control or diet-induced obesity mice (8 or 16 weeks of high-fat diet) were either treated or not treated with leptin. Metabolic features were analyzed and expression of the genes of interest was measured by quantitative reverse transcriptase-PCR (RT-qPCR) and western blot. We reported that in control mice, but not in obese mice, leptin infusion induced an increase in POMC mRNA level as well as in MC4-R mRNA level suggesting that leptin could act directly and/or through alpha-melanocyte-stimulating hormone (alpha-MSH). This hypothesis was reinforced after in vitro studies, using the mouse hypothalamic GT1-7 cell line, since both leptin and Norleucine(4), D-Phenylalanine(7)-alpha-MSH (NDP-alpha-MSH) treatments increased MC4-R expression. After 8 weeks of high-fat diet, nondiabetic obese mice became resistant to the central action of leptin and their hypothalamic content of POMC and AgRP mRNA were decreased without modification of MC4-R mRNA level. After 16 weeks of high-fat diet, mice exhibited more severe metabolic disorders with type 2 diabetes. Moreover, hypothalamic expression of MC4-R was highly increased. In conclusion, several alterations of the melanocortin system were found in obese mice that are probably consecutive to their central resistance to leptin. Moreover, when the metabolic status is highly degraded (with all characteristics of a type 2 diabetes), other regulatory mechanisms (independent of leptin) can also take place.


The Nordic Football Injury Audit: higher injury rates for professional football clubs with third-generation artificial turf at their home venue.
Previously, no difference in acute injury rate has been found when playing football on artificial turf (AT) compared with natural grass (NG). To compare acute injury rates in professional football played on AT and NG at the individual player level; and to compare, at club level, acute and overuse injury rates between clubs that have AT at their home venue (AT clubs) and clubs that have NG (NG clubs). 32 clubs (AT, n=11; NG, n=21) in the male Swedish and Norwegian premier leagues were followed prospectively during the 2010 and 2011 seasons. Injury rate was expressed as the number of time loss injuries/1000 h and compared with rate ratio (RR) and 99% CI. No statistically significant differences were found in acute injury rates on AT compared with NG during match play (RR 0.98, 99% CI 0.79 to 1.22) or training (RR 1.14, 99% CI 0.86 to 1.50) when analysing at the individual player level. When analysing at the club level, however, AT clubs had a significantly higher acute training injury rate (RR 1.31, 99% CI 1.04 to 1.63) and overuse injury rate (RR 1.38, 99% CI 1.14 to 1.65) compared with NG clubs. At the individual player level, no significant differences were found in acute injury rates when playing on AT compared with NG. However, clubs with AT at their home venue had higher rates of acute training injuries and overuse injuries compared with clubs that played home matches on NG.


[Labelling of nif-plasmid pEA9 from Enterobacter agglomerans 339].
The authors describe the in vivo labelling of the plasmid pEA9 in Enterobacter agglomerans 339 with a kanamycin resistance gene. For labelling purposes the donor plasmid pST5 was constructed. This plasmid contains the nif ENX region from pEA9,in which a kanamycin resistance gene is cloned.pST5 was transformed into E.a.339 and subsequently cured from the host. Curing was achieved with AP medium. Fourty strains that had lost pST5,but retained the kanamycin resistance, could be isolated. It showed that none of these clones contained co-integrates of pST5 and pEA9. This is evident that in all clones the kanamycin resistance gene was integrated into pEA9 by homologous recombination.


UIS2: A Unique Phosphatase Required for the Development of Plasmodium Liver Stages.
Plasmodium salivary sporozoites are the infectious form of the malaria parasite and are dormant inside salivary glands of Anopheles mosquitoes. During dormancy, protein translation is inhibited by the kinase UIS1 that phosphorylates serine 59 in the eukaryotic initiation factor 2α (eIF2α). De-phosphorylation of eIF2α-P is required for the transformation of sporozoites into the liver stage. In mammalian cells, the de-phosphorylation of eIF2α-P is mediated by the protein phosphatase 1 (PP1). Using a series of genetically knockout parasites we showed that in malaria sporozoites, contrary to mammalian cells, the eIF2α-P phosphatase is a member of the PP2C/PPM phosphatase family termed UIS2. We found that eIF2α was highly phosphorylated in uis2 conditional knockout sporozoites. These mutant sporozoites maintained the crescent shape after delivery into mammalian host and lost their infectivity. Both uis1 and uis2 were highly transcribed in the salivary gland sporozoites but uis2 expression was inhibited by the Pumilio protein Puf2. The repression of uis2 expression was alleviated when sporozoites developed into liver stage. While most eukaryotic phosphatases interact transiently with their substrates, UIS2 stably bound to phosphorylated eIF2α, raising the possibility that high-throughput searches may identify chemicals that disrupt this interaction and prevent malaria infection.


Partial deletion of p75NTR in large-diameter DRG neurons exerts no influence upon the survival of peripheral sensory neurons in vivo.
The p75 neurotrophin receptor (p75NTR ) is required for maintaining peripheral sensory neuron survival and function; however, the underlying cellular mechanism remains unclear. The general view is that expression of p75NTR by the neuron itself is required for maintaining sensory neuron survival and myelination in the peripheral nervous system (PNS). Adopting a neuronal-specific conditional knockout strategy, we demonstrate the partial depletion of p75NTR in neurons exerts little influence upon maintaining sensory neuron survival and peripheral nerve myelination in health and after demyelinating neuropathy. Our data show that the density and total number of dorsal root ganglion (DRG) neurons in 2-month-old mice is not affected following the deletion of p75NTR in large-diameter myelinating neurons, as assessed by stereology. Adopting experimental autoimmune neuritis induced in adult male mice, an animal model of demyelinating peripheral neuropathy, we identify that deleting p75NTR in myelinating neurons exerts no influence upon the disease progression, the total number of DRG neurons, and the extent of myelin damage in the sciatic nerve, indicating that the expression of neuronal p75NTR is not essential for maintaining peripheral neuron survival and myelination after a demyelinating insult in vivo. Together, results of this study suggest that the survival and myelination of peripheral sensory neurons is independent of p75NTR expressed by a subtype of neurons in vivo. Thus, our findings provide new insights into the mechanism underpinning p75NTR -mediated neuronal survival in the PNS.


Surgical revascularization for 'atherosclerotic' coronary artery aneurysms associated with multiple obstructive coronary artery disease.
Coronary artery aneurysms (CAA) were originally described in a series of post-mortem studies but are now more commonly observed during the widely used coronary angiography. This article reports on four adult patients with CAA associated with obstructive coronary atherosclerosis. Arterial and venous conduits were successfully performed in all four. The aetiology and treatment are discussed.


Mitogenome phylogeographic analysis of a planktonic crustacean.
Phylogeography places population genetics in an explicitly spatial context, and in doing so attempts to reconstruct the historical and contemporary evolutionary processes acting across a species range through space and time. Here we present the phylogeographical structure of Daphnia magna as determined for full mitochondrial genomes from samples of 60 populations throughout much of the species known range, including Europe, the Middle East, and Asia. Contrary to previous analyses, the present analysis of the mitochondrial genome reveals coarse-grained (continental scale) evidence for spatial structure, and in particular a deep split between Western Eurasia and East Asian D. magna lineages. In contrast to previous analyses with nuclear genetic markers, our mitogenomic analysis reveals much less structure within lineages. We quantify divergence between species using the full mitochondrial genome sequence of a closely related species, D. similis. The distribution of European and Middle Eastern genetic diversity is consistent with a rapid demographic expansion following the end of the most recent ice age about 10,000 years before present. By estimating species wide distributions of dN/dS in mtDNA, we provide evidence that the effectiveness of purifying selection on protein coding genes in the mitochondrial genome of coastal rock pool populations, which have pronounced extinction-colonization dynamics, is reduced compared to larger and more stable non-rock pool populations. The present study adds important insights into the evolutionary history of a widely used model organism in ecology, evolution and ecotoxicology, and highlights the utility of phylogeographic analysis of organellar genomes to understand evolutionary processes.


Gene expression inhibitors for the treatment of liver fibrosis: drugs under preclinical and early clinical investigation.
Liver fibrosis represents an unmet medical condition with growing incidence and only limited therapeutic options. Interfering with dysregulated gene expression was considered a specific treatment approach, and we are here reviewing the current options to modulate transcription and translation with small molecule inhibitors of involved enzymes, transcription factors or by using non-coding RNA molecules (RNA interference) or DNA antisense oligonucleotides. Despite promising results in preclinical models, only limited data are available from studies in humans. This expert opinion provides a general overview of how to interfere with gene expression (transcription and translation) and highlighting recent achievements in liver fibrosis. Many compounds that were explored to modulate gene expression in liver fibrosis (models) were developed as anti-cancer agents. Their use in humans with impaired liver function is often impaired by the lack of specificity to inhibit only fibrosis-related genes in the liver and by associated general toxicity and narrow therapeutic windows. RNAi approaches show a higher degree of specificity and potentially less systemic toxicity. Clinical development in liver fibrosis requires close interaction between pharmaceutical companies and regulatory authorities to address topics like relevant (surrogate) endpoints to achieve meaningful readouts faster.


[Review of Research Advances in Medical Service Robotics].
With the progress of science and technology and the increase of clinical demand, medical robots have developed rapidly and played a important role in promoting the medical cause. Service robot is a branch of medical robot, which is mainly oriented to medical service and assistance needs, and has been applied in many medical scenarios and achieved demonstration effects. This research first describes the development of medical service robots, and then summarizes the key technologies and clinical applications of robots. Finally, it points out the challenges and directions that medical service robots face at present, and puts forward prospects for their further development in the medical field.


Hydrosalpinx functional surgery or salpingectomy? The importance of hydrosalpinx fluid in assisted reproductive technologies.
The first IVF baby, Louise Brown, was born in a natural cycle IVF of a woman who had bilateral tubal block making IVF the only option for having a child. The last 3 decades has seen astounding progress in the field of ART. Today thanks to ART, tubal disease and tubal factor infertility is easily overcome. The accepted theory today is that the hydrosalpinx fluid plays a causative role in the reduced pregnancy rate with ART. It is well known that the success of ART for patients with tubal disease with hydrosalpinx is reduced by half compared with patients without hydrosalpinx. Ideal would be removal of a hydrosalpinx by laparoscopic salpingectomy to improve pregnancy rates. However in some cases this is not feasible due to dense pelvic adhesions making access difficult. In such cases it is recommended that even de-linking the tube from the uterus would help in improving the ART outcome. There is suggestion that sonographically visible hydrosalpinges and those affected bilaterally have a poorer prognosis than those seen incidentally at laparoscopy. While there is clinical evidence supporting the causative role of the fluid itself, there is a lack of knowledge as to how the fluid exerts its negative effects. It is generally believed that the fluid holds a key position in impairing implantation potential. The aim of this review is to highlight the importance of identifying hydrosalpinges and its association with reduced fertility outcome using assisted reproductive technologies. Here we have discussed the different options available for the same, and highlighted the current modes of treatment.


Changes in microtubules, microtubule-associated proteins, and intermediate filaments during the differentiation of HL-60 leukemia cells.
The cytoskeleton is composed mainly of microtubules (MT), microfilaments, and intermediate filaments (IF) that form a structural network which connects cellular membranes, cytoplasmic organelles, and the nucleus. Since the cytoskeleton may be involved in modulating signal transduction and in the morphological and structural changes that occur during cellular proliferation and differentiation, cytoskeletal changes were measured by immunofluorescence microscopy and fluorescence-activated cell sorter analysis during the differentiation of HL-60 leukemia cells induced by retinoic acid (RA). Differentiated HL-60 cells exhibited increased staining intensity and altered organization of MT and IF, as visualized by immunofluorescence microscopy with anti-tubulin monoclonal antibody and anti-vimentin antibody, respectively. A new procedure was developed and used to measure the content of the cytoskeletal components of HL-60 cells during the process of maturation. HL-60 cells were fixed with formaldehyde in an MT-stabilizing buffer, permeabilized using L-lysophosphatidylcholine, stained for immunofluorescent measurement with antibodies specific for particular cytoskeletal components, and analyzed by flow cytometry. Terminally differentiated cells produced by exposure to RA contained larger amounts of MT and the IF vimentin. During the course of the maturation process, a transient increase in the amounts of the microtubule-associated proteins, (MAPs) MAP2 and tau, occurred. An RA-supersensitive clone, designated HL-60/S4, and an RA-resistant clone, designated HL-60/R3, were developed by mutagenization and selection. Use of these clones supported the concept that the observed changes in MT, MAPs, and vimentin were associated with the differentiation process rather than being due to other effects produced by the retinoid. Thus, the findings suggest that changes in MT, MAPs, and IF are important to the terminal maturation of leukemia cells.


[A Molecular Genetic Approach to Analysis of Individual Differences in Professionally Important Qualities].
The authors present a theoretical analysis of a role of genetic polymorphisms in formation of individual differences influencing the quality of professional activity. The last 20 years have seen a rise in works dedicated to genetics of various cognitive and psychological functions. This review focuses on the psychological and cognitive traits affecting a subject's behavior during his work. Among them are differences in working memory performance, emotional reactions and some personality traits, alterations in prefrontal cortex functions, a number of emotional reactions and personality traits, chronotype and sleep deprivation sensitivity. It has been shown that their individual variability is to a large extent genetically determined. The studies have demonstrated the importance of allele variations of a number of genes in adrenergic, serotonergic and especially dopaminergic systems as well as molecular clock genes for emotional and cognitive spheres. This review is designed for specialists working in the fields of psychogenetics, cognitive and psychological aspects of human factors, occupational safety.


HER2-targeted therapies for salivary gland cancers.
Salivary gland cancers (SGCs) are a heterogeneous group of rare tumors including various histological subtypes with different molecular profiling. Human epidermal growth factor receptor 2 (HER2) is one of the most intriguing and studied molecular alterations with prognostic and predictive roles. Indeed, HER2 overexpression is commonly correlated with aggressive histological subtypes and poorer prognosis. However, HER2 may represent the target of personalized treatment. We performed a literature review of use of anti-HER2 targeted agents for treatment of recurrent or metastatic SGCs. The efficacy and safety of anti-HER2 were firstly evaluated in patients affected with other solid tumors, mostly breast and gastric cancers. For SGCs the literature is mainly comprised of case reports or case series and small clinical trials. The most common used drug is trastuzumab in combination with chemotherapy (i.e. taxanes, capecitabine, carboplatin, eribulin) or with another anti-HER2 targeted agent (i.e. pertuzumab). The use of anti-HER2 therapies induces improvement in clinical responses, which are mostly durable. Besides, new anti-HER2 drugs such as antibody-drug conjugates (ADC) (i.e. trastuzumab emtansine, trastuzumab deruxtecan) have been introduced in this setting inducing further therapeutic advances. Anti-HER2 treatment strategy is emerging as potentially effective in selected HER2 overexpressing SGCs. However, prospective and multicentric clinical trials are needed to evaluate the efficacy of these therapeutic regimens within larger cohorts and to assess the most appropriate treatment sequence strategy.


Uncoupling protein 2 regulates glucagon-like peptide-1 secretion in L-cells.
To investigate whether uncoupling protein 2 (UCP2) affects oleic acid-induced secretion of glucagon-like peptide-1 (GLP-1) in L-cells. mRNA and protein expression of UCP2 were analyzed in human NCI-H716 cells, which serve as a model for enteroendocrine L-cells, by quantitative reverse transcription-polymerase chain reaction and Western blotting before and after treatment with oleic acid. Localization of UCP2 and GLP-1 in NCI-H716 cells was assessed by immunofluorescence labeling. NCI-H716 cells were transiently transfected with a small interfering RNA (siRNA) that targets UCP2 (siUCP2) or with a non-specific siRNA using Lipofectamine 2000. The concentrations of bioactive GLP-1 in the medium were measured by enzyme linked immunosorbent assay. Both GLP-1 and UCP2 granules were expressed mainly in the cytoplasm of NCI-H716 cells. NCI-H716 cells that secreted GLP-1 also expressed UCP2. Time-course experiments revealed that release of GLP-1 from NCI-H716 cells into the medium reached a maximum at 120 min and remained stable until at least 180 min after treatment with oleic acid (the level of GLP-1 increased about 2.3-fold as compared with the level of GLP-1 in the control cells, P < 0.05). In an experiment to determine dose dependence, stimulation of NCI-H716 cells with ≤ 8 mmol oleic acid led to a concentration-dependent release of GLP-1 into the medium; 10 mmol oleic acid diminished the release of GLP-1. Furthermore, GLP-1 secretion induced by oleic acid from NCI-H716 cells that were transfected with siUCP2 decreased to 41.8%, as compared with NCI-H716 cells that were transfected with a non-specific siRNA (P < 0.01). UCP2 affected GLP-1 secretion induced by oleic acid. UCP2 plays an important role in L-cell secretion that is induced by free fatty acids.


Mechanisms and specific directionality of autotrophic nitrous oxide and nitric oxide generation during transient anoxia.
The overall goal of this study was to determine the molecular and metabolic responses of chemostat cultures of model nitrifying bacteria to imposition of and recovery from transient anoxic conditions. Based on the study, a specific directionality in nitrous oxide (N(2)O) and nitric oxide (NO) production was demonstrated. N(2)O production was only observed during recovery to aerobic conditions after a period of anoxia and correlated positively with the degree of ammonia accumulation during anoxia. NO, on the other hand, was emitted mainly under anoxia. The production of NO was linked to a major imbalance in the expression of the nitrite reductase gene, which was overexpressed during transient anoxia. In contrast, genes coding for ammonia and hydroxylamine oxidation and nitric oxide reduction were generally under-expressed during transient anoxia. These results are different from the observed parallel expression and activity of nitrite and nitric oxide reductase in heterotrophic bacteria subjected to transient oxygen cycling. Unlike NO, the production of N(2)O could not be solely correlated to gene expression patterns and likely involved responses at the enzyme activity or metabolic levels. Based on experimental data, the propensity of the nitrifying cultures for N(2)O production is related to a shift in their metabolism from a low specific activity (q < q(max)) toward the maximum specific activity (q(max)).


Association of E-Cigarettes With Erectile Dysfunction: The Population Assessment of Tobacco and Health Study.
Smoking is independently associated with erectile dysfunction and cardiovascular disease. Given existing similarities in the constituents of e-cigarettes or ENDS and cigarettes, this study examines the association between ENDS use and erectile dysfunction. Data from Wave 4 (2016-2018) of the Population Assessment of Tobacco and Health study were analyzed in 2020. Male participants aged ≥20 years who responded to the erectile dysfunction question were included. Multivariable logistic regression models examined the association of ENDS use with erectile dysfunction within the full sample and in a restricted sample (adults aged 20-65 years with no previous cardiovascular disease diagnosis) while adjusting for multiple risk factors. The proportion of erectile dysfunction varied from 20.7% (full sample) to 10.2% (restricted sample). The prevalence of current ENDS use within the full and restricted samples was 4.8% and 5.6%, respectively, with 2.1% and 2.5%, respectively, reporting daily use. Current daily ENDS users were more likely to report erectile dysfunction than never users in both the full (AOR=2.24, 95% CI=1.50, 3.34) and restricted (AOR=2.41, 95% CI=1.55, 3.74) samples. In the full sample, cardiovascular disease history (versus not present) and age ≥65 years (versus age 20-24 years) were associated with erectile dysfunction (AOR=1.39, 95% CI=1.10, 1.77; AOR= 17.4, 95% CI=12.15, 24.91), whereas physical activity was associated with lower odds of erectile dysfunction in both samples (AOR range=0.44-0.58). The use of ENDS seems to be associated with erectile dysfunction independent of age, cardiovascular disease, and other risk factors. While ENDS remain under evaluation for harm reduction and smoking-cessation potential, ENDS users should be informed about the possible association between ENDS use and erectile dysfunction.