Proliferation, Characterization and Differentiation Potency of Adipose Tissue-Derived Mesenchymal Stem Cells (AT-MSCs) Cultured in Fresh Frozen and non-Fresh Frozen Plasma.
Mesenchymal stem cells (MSCs) have unique properties, including high proliferation rates, self-renewal, and multilineage differentiation ability. Their characteristics are affected by increasing age and microenvironment. This research is aimed to determine the proliferation, characteristics and differentiation capacity of adipose tissue-derived (AT)-MSCs at many passages with different media. The cell proliferation capacity was assayed using trypan blue. MSCs characterization (CD90, CD44, CD105, CD73, CD11b, CD19, CD34, CD45, and HLA-DR) was performed by flow cytometry, and cell differentiation was determined by specific stainings. Population doubling time (PDT) of AT-MSCs treated with fresh frozen plasma (FFP) and non-FFP increased in the late passage (P) (P15 FFP was 22.67 ± 7.01 days and non-FFP was 19.65 ± 2.27 days). Cumulative cell number was significantly different between FFP and non-FFP at P5, 10, 15. AT-MSCs at P4-15 were positive for CD90, CD44, CD105, and CD73, and negative for CD11b, CD19, CD34, CD45, and HLA-DR surface markers. AT-MSCs at P5, 10, 15 had potential toward adipogenic, chondrogenic, and osteogenic differentiation. Therefore, PDT was affected by increased age but no difference was observed in morphology, surface markers and differentiation capacity among passages. Cumulative cell number in FFP was higher in comparison with non-FFP in P5, 10, 15. Our data suggest that FFP may replace FBS for culturing MSCs.


The Benefit of Neuromuscular Blockade in Patients with Postanoxic Myoclonus Otherwise Obscuring Continuous Electroencephalography (CEEG).
Introduction. Myoclonus status epilepticus is independently associated with poor outcome in coma patients after cardiac arrest. Determining if myoclonus is of cortical origin on continuous electroencephalography (CEEG) can be difficult secondary to the muscle artifact obscuring the underlying CEEG. The use of a neuromuscular blocker can be useful in these cases. Methods. Retrospective review of CEEG in patients with postanoxic myoclonus who received cisatracurium while being monitored. Results. Twelve patients (mean age: 53.3 years; 58.3% male) met inclusion criteria of clinical postanoxic myoclonus. The initial CEEG patterns immediately prior to neuromuscular blockade showed myoclonic artifact with continuous slowing (50%), burst suppression with myoclonic artifact (41.7%), and continuous myogenic artifact obscuring CEEG (8.3%). After intravenous administration of cisatracurium (0.1 mg-2 mg), reduction in artifact improved quality of CEEG recordings in 9/12 (75%), revealing previously unrecognized patterns: continuous EEG seizures (33.3%), lateralizing slowing (16.7%), burst suppression (16.7%), generalized periodic discharges (8.3%), and, in the patient who had an initially uninterpretable CEEG from myogenic artifact, continuous slowing. Conclusion. Short-acting neuromuscular blockade is useful in determining background cerebral activity on CEEG otherwise partially or completely obscured by muscle artifact in patients with postanoxic myoclonus. Fully understanding background cerebral activity is important in prognostication and treatment, particularly when there are underlying EEG seizures.


Inflammation in sleep apnea: an update.
Obstructive sleep apnea (OSA) is a common disorder associated with cardiovascular disease (CVD). One theory to explain this relationship proposes that OSA can induce systemic inflammation, thereby inducing CVD. This theory is based on the premise that obesity is a pro-inflammatory state, and that physiological derangements during sleep in subjects with OSA further aggravate inflammation. In support of this theory, some clinical studies have shown elevated inflammatory biomarkers in OSA subjects, or improvement in these markers following treatment of OSA. However, the data are inconsistent and often confounded by the effects of comorbid obesity. Animal models of OSA have been developed, which involve exposure of rodents or cells to intermittent hypoxia, a hallmark feature of OSA. Several of these experiments demonstrate that intermittent hypoxia can stimulate inflammatory pathways and lead to cardiovascular or metabolic pathology. In this review, we review relationships between OSA and inflammation, with particular attention to studies published within the last year.


Axillary nodal metastasis in ovarian cancer: a report of three cases and review of literature.
Ovarian cancer represents a major global health burden that is rarely associated with distant metastasis. Axillary lymph node metastasis from ovarian cancer is rare and is reported only in few case reports in literature. We report three cases of ovarian carcinoma associated with axillary lymph node metastasis as well as a brief literature review. The pathologic subtype in one case was malignant mixed Mullerian tumor, while the other two cases were high-grade serous ovarian carcinoma. Axillary nodal metastasis was reported as a synchronous event in one case, while it was reported as recurrence events in the other two cases. Physicians should be aware of this uncommon mode of metastasis in ovarian cancer cases. Multi-disciplinary discussion is crucial in the management of these cases to offer them the best suitable treatment.


Neuroprotection by progesterone after transient cerebral ischemia in stroke-prone spontaneously hypertensive rats.
We investigated the neuroprotective effects of progesterone (P4) treatment in stroke-prone spontaneously hypertensive rats (SHRSPs) given 60-min transient middle cerebral artery occlusion (tMCAO). The treatment groups were: (1) Wistar-Kyoto (normotensive sham), (2) SHRSP (hypertensive sham), (3) tMCAO SHRSPs (SHRSP+tMCAO), and (4) SHRSP+tMCAO+P4. P4 (8mg/kg) was administered 1h after occlusion and then daily for 14days. We measured cerebral infarction volume, blood pressure and body weight. Behavioral outcomes were analyzed at post-stroke days 3, 9, and 14. To assess morphological protection we measured activation of microglia and astrocytes, oxidative stress, apoptosis, expression of vascular endothelial growth factor (VEGF), an angiogenic marker, and IL-1β, a marker of inflammation, on day 14 post-stroke. There was no effect of P4 on body weight or systolic blood pressure compared to the SHRSP+tMCAO group. However, grip strength and sensory neglect measures in the P4 group were improved compared to SHRSP+tMCAO. In addition, significantly larger infarct volumes were seen in the SHRSP+tMCAO group compared to SHRSP+tMCAO+P4. Increased markers of the injury cascade such as macrophages, activated astrocytes, superoxide anion and apoptotic cells observed in the SHRSP+tMCAO group were significantly decreased by P4. We conclude that, despite hypertensive comorbidity, P4 improves functional outcomes and attenuates stroke infarct in hypertensive rats by reducing superoxide anion expression and by decreasing inflammation and neuronal apoptosis.


Regulation of alpha and luteinizing hormone beta subunit messenger ribonucleic acids during stimulatory and downregulatory phases of gonadotropin-releasing hormone action.
Decreased gonadotropin responsiveness (downregulation) to gonadotropin-releasing hormone (GnRH) following chronic in vivo and in vitro exposure to GnRH or its agonist (GnRH-A) has been previously reported. In the present studies, changes in LH subunit mRNAs in rat pituitary monolayer culture during stimulatory and down regulatory phases of GnRH action are described. Rat pituitary cells in culture, pretreated with medium alone or GnRH-A (10(-6) M) for 48 h were extensively washed and treated with graded concentrations of GnRH [10(-9) to 10(-7)] for 4 h. Medium was assayed for luteinizing hormone (LH) immunoreactivity, and total cytoplasmic RNAs were extracted by the hot phenol-guanidinium isothiocyanate method. Subunit-specific mRNAs were quantified by dot-hybridization assay using 32P-labeled subunit-specific cDNA probes. Cells pretreated with medium alone showed a dose-dependent increase in medium LH immunoreactivity, but the alpha and LH beta mRNAs showed no change over the 4-h period. Cells pretreated with GnRH-A showed no significant increase in medium LH with GnRH treatment, thus demonstrating that the cells had been desensitized by prior GnRH-A treatment. Alpha and LH beta subunit mRNAs of cells pretreated with GnRH-A did not show any significant change with further GnRH treatment. In subsequent experiments, cells were incubated with medium alone or 10(-7) M GnRH for 4, 8, or 24 h. GnRH failed to increase subunit mRNAs after 4 and 8 h incubation; after 24 h, alpha subunit mRNA showed a modest but significant increase and beta subunit mRNA showed a modest decrease compared to controls.(ABSTRACT TRUNCATED AT 250 WORDS)


Management of Extraoral Complications in a Patient Treated with Four Zygomatic Implants.
This article details the significant complications associated with treatment of a patient who underwent maxillary reconstruction based entirely on zygomatic implants. The use of zygomatic implants is becoming increasingly widespread, and treatment has been shown to be predictable and generally free of surgical complications. However, extending from the maxillary alveolus, alongside or through the maxillary antrum, toward the zygoma or orbital rim, these long implants have the potential to create a range of complications that may be more serious than those encountered with conventional dental implants. In the case presented, complications included implant failure, prominence of the implant apices, recurrent loosening of the implant prosthesis, and, most significantly, extraoral infection associated with the apex of one implant. The authors describe the complications and remedial surgical treatment for this patient.


Polymer Vesicles with Integrated Photothermal Responsiveness.
Functionalized polymer vesicles have been proven to be highly promising in biomedical applications due to their good biocompatibility, easy processability, and multifunctional responsive capacities. However, photothermal-responsive polymer vesicles triggered by near-infrared (NIR) light have not been widely reported until now. Herein, we propose a new strategy for designing NIR light-mediated photothermal polymer vesicles. A small molecule (PTA) with NIR-triggered photothermal features was synthesized by combining a D-D'-A-D'-D configuration framework with a molecular rotor function (TPE). The feasibility of the design strategy was demonstrated through density functional theory calculations. PTA moieties were introduced in the hydrophobic segment of a poly(ethylene glycol)-poly(trimethylene carbonate) block copolymer, of which the carbonate monomers were modified in the side chain with an active ester group. The amphiphilic block copolymers (PEG44-PTA2) were then used as building blocks for the self-assembly of photothermal-responsive polymer vesicles. The new class of functionalized polymer vesicles inherited the NIR-mediated high photothermal performance of the photothermal agent (PTA). After NIR laser irradiation for 10 min, the temperature of the PTA-Ps aqueous solution was raised to 56 °C. The photothermal properties and bilayer structure of PTA-Ps after laser irradiation were still intact, which demonstrated that they could be applied as a robust platform in photothermal therapy. Besides their photothermal performance, the loading capacity of PTA-Ps was investigated as well. Hydrophobic cargo (Cy7) and hydrophilic cargo (Sulfo-Cy5) were successfully encapsulated in the PTA-Ps. These properties make this new class of functionalized polymer vesicles an interesting platform for synergistic therapy in anticancer treatment.


The nucleotide and predicted amino acid sequence of the fusion protein of recent isolates of canine distemper virus in Japan.
Analysis of the molecular properties of fusion (F) proteins of field isolates of canine distemper virus (CDV) by immunoprecipitation analysis revealed an identical molecular mass of F protein of 3 field isolates as well as the Onderstepoort laboratory strain. Sequencing showed that the F gene of a field isolate (the Yanaka strain) shared 90.1% and 95.7% identities with the Onderstepoort strain at nucleotide and amino acid levels, respectively. All of the 13 cysteine residues and 4 potential asparagine-linked glycosylation sites were completely conserved amongst these strains. These results indicate that the F proteins is much less heterogeneous than that observed in the hemagglutinin proteins of CDV.


Palladium-Catalyzed Asymmetric Tandem Denitrogenative Heck/Tsuji-Trost of Benzotriazoles with 1,3-Dienes.
The asymmetric denitrogenative cycloaddition has emerged as a powerful tool to build chiral aza-heterocyles. However, only one example of asymmetric denitrogenative cycloaddition of benzotriazole with unsaturated hydrocarbons has been explored so far, because the ring-opening of benzotriazole to generate α-imino metal carbenoid species is a thermodynamically unfavorable process. We herein report an efficient asymmetric denitrogenative cycloaddition of benzotriazoles with cyclic and acyclic 1,3-dienes enabled by Pd and chiral sulfonamide phosphine ligand. A variety of substituted hexahydrocarbazoles and indolines were delivered in good yields with high ee values. Interestingly, a pair of enantiomers could be obtained with the use of Xu1 and PC2 with the same absolute configuration. The synthetic utilities of the optically active hexahydrocarbazoles were also showcased.


Drosophila myosin-XX functions as an actin-binding protein to facilitate the interaction between Zyx102 and actin.
The class XX myosin is a member of the diverse myosin superfamily and exists in insects and several lower invertebrates. DmMyo20, the class XX myosin in Drosophila, is encoded by dachs, which functions as a crucial downstream component of the Fat signaling pathway, influencing growth, affinity, and gene expression during development. Sequence analysis shows that DmMyo20 contains a unique N-terminal extension, the motor domain, followed by one IQ motif, and a C-terminal tail. To investigate the biochemical properties of DmMyo20, we expressed several DmMyo20 truncated constructs containing the motor domain in the baculovirus/Sf9 system. We found that the motor domain of DmMyo20 had neither ATPase activity nor the ability to bind to ATP, suggesting that DmMyo20 does not function as a molecular motor. We found that the motor domain of DmMyo20 could specifically bind to actin filaments in an ATP-independent manner and enhance the interaction between actin filaments and Zyx102, a downstream component of DmMyo20 in the Fat signaling pathway. These results suggest that DmMyo20 functions as a scaffold protein, but not as a molecular motor, in a signaling pathway controlling cell differentiation.


Enterococcal endocarditis: can we win the war?
Treatment of enterococcal infections has long been recognized as an important clinical challenge, particularly in the setting of infective endocarditis (IE). Furthermore, the increase prevalence of isolates exhibiting multidrug resistance (MDR) to traditional anti-enterococcal antibiotics such as ampicillin, vancomycin and aminoglycosides (high-level resistance) poses immense therapeutic dilemmas in hospitals around the world. Unlike IE caused by most isolates of Enterococcus faecalis, which still retain susceptibility to ampicillin and vancomycin, the emergence and dissemination of a hospital-associated genetic clade of multidrug resistant Enterococcus faecium, markedly limits the therapeutic options. The best treatment of IE MDR enterococcal endocarditis is unknown and the paucity of antibiotics with bactericidal activity against these organisms is a cause of serious concern. Although it appears that we are winning the war against E. faecalis, the battle rages on against isolates of multidrug-resistant E. faecium.


Individualizing insulin therapy in the management of type 2 diabetes.
It is recognized that reducing hyperglycemia early on in disease progression has long-term benefits for patients with diabetes. Insulin therapy has greater potential to reduce hyperglycemia than other therapies; however, there is often a significant delay in insulin initiation and intensification. Insulin replacement therapy in type 2 diabetes should no longer be viewed as the treatment of last resort. With the development of modern insulin analogs, the field has evolved. Large clinical trials have improved our understanding of the potential benefits and risks associated with intensive glycemic control in different patient populations and highlighted the need for individualization of glycemic targets and treatment strategies. Current treatment guidelines recognize the important role of insulin therapy both early on and throughout the progression of type 2 diabetes.


Evaluating Nutrient Intake of Career Firefighters Compared to Military Dietary Reference Intakes.
The primary goals of the Dietary Reference Intakes (DRI) are to plan and assess nutrient intakes to promote health, reduce chronic disease, and prevent toxicity. Firefighters have unique nutrient needs compared to the public due to their job demands. The military provides the only published guidance for tactical athletes' nutrient needs. The purpose of this study was to determine whether firefighters were meeting the Military Dietary Reference Intakes (MDRI). A cross-sectional study was conducted in a sample of career firefighters (n = 150, 37.4 ± 8.4 year-old males) employed in Southern California. Data were gathered during baseline assessments from a Federal Emergency Management Agency-funded Firefighter Wellness Initiative. Participants were asked to log their food and beverage consumption over a 72-h period. Descriptive statistics (means, standard deviations, 95% confidence intervals) were calculated for all participant characteristics and average three-day nutrient intakes. A 95% confidence interval compared their nutrient intake to MDRI to identify differences in nutrient intakes, significance accepted at p = 0.05. Compared to MDRI reference values, firefighters consumed an inadequate amount of total calories, linolenic and alpha-linolenic fatty acid, fiber, vitamins D, E, and K, potassium, magnesium, zinc, and carbohydrates. Vitamin D, magnesium, and potassium had the greatest shortcomings (95.3%, 94.0%, and 98.7%, respectively, under MRDA). Thus, firefighters are not meeting the established MDRI for several key nutrients required to promote health, improve performance, and reduce chronic disease. Dietitians and health care providers may use the results of this study to help design health promotion programs for this population. Future research should develop a customized reference intake for firefighters.


Warming and drought combine to increase pest insect fitness on urban trees.
Urban habitats are characterized by impervious surfaces, which increase temperatures and reduce water availability to plants. The effects of these conditions on herbivorous insects are not well understood, but may provide insight into future conditions. Three primary hypotheses have been proposed to explain why multiple herbivorous arthropods are more abundant and damaging in cities, and support has been found for each. First, less complex vegetation may reduce biological control of pests. Second, plant stress can increase plant quality for pests. And third, urban warming can directly increase pest fitness and abundance. These hypotheses are not mutually exclusive, and the effects of temperature and plant stress are particularly related. Thus, we test the hypothesis that urban warming and drought stress combine to increase the fitness and abundance of the scale insect, Melanaspis tenebricosa, an urban tree pest that is more abundant in urban than rural areas of the southeastern U.S. We did this by manipulating drought stress across an existing mosaic of urban warming. We found support for the additive effect of temperature and drought stress such that female embryo production and body size increased with temperature and was greater on drought-stressed than watered trees. This study provides further evidence that drivers of pest insect outbreaks act in concert, rather than independently, and calls for more research that manipulates multiple abiotic factors related to urbanization and climate change to predict their effects on ecological interactions. As cities expand and the climate changes, warmer temperatures and drought conditions may become more widespread in the native range of this pest. These changes have direct physiological benefits for M. tenebricosa, and potentially other pests, that may increase their fitness and abundance in urban and natural forests.


Cellulase activity in commercial lipase preparations.
Polymeric membranes are increasingly used as supports for the immobilization of enzymes in bioreactors. One of the more common reactor types employed in lipase-catalyzed hydrolysis of oils, contains modified cellulose as a membrane material. We found that this type of material is readily attacked by cellulase present in several commercially available lipase preparations. This leads to membrane damage, reactor instability, and leakage. We conclude that cellulose membranes are not suitable as supports in bioreactors for the immobilization of these lipases. The development of alternative membranes is currently in progress.


Conformation and side chains environments of recombinant human interleukin-1 receptor antagonist (rh-IL-1ra) probed by raman, raman optical activity, and UV-resonance Raman spectroscopy.
The conformation and local environments of the side chains cysteines and aromatics of recombinant human interleukin-1 receptor antagonist (rh-IL-1ra) have been studied by visible Raman, Raman optical activity (ROA) and UVRR spectroscopy. The results reveal that the secondary structure of rh-IL-1ra is predominantly beta-sheet, which is consistent with conclusions from multinuclear NMR in solutions and X-ray diffraction analysis of crystals. It confirms that all four cysteines are in reduced state. Three cysteines are not hydrogen bonded and exposed. One cysteine is moderately hydrogen bonded and buried. This explains the earlier observation that only three cysteines were detectable using DTNB titration. No characteristic Raman band of disulfide bond was observed in the Raman spectra of rh-IL-1ra in both solution and in crystals. It rules out the supposition that there is one disulfide bond in rh-IL-1ra crystals based on X-ray diffraction. Raman and UVRR spectra of rh-IL-1ra exhibit canonical marker bands of tryptophan. They do not support the proposal that there is cation-pi interaction involving tryptophans in solutions and crystals. These results demonstrate that Raman spectroscopy offers certain advantages over X-ray diffraction for studies of detailed local environment and intermolecular interactions of side chains of proteins in solution and in crystals.


[Double-blind comparison of cefotetan and cefmetazole in complex urinary tract infections].
For the purpose of carrying out an objective evaluation of the clinical efficacy and safety of cefotetan (CTT), a new cephamycin-type injectable antibiotic, in the treatment of complicated urinary tract infections, cefmetazole (CMZ) was employed as the control drug in a comparative study performed by the double-blind method. CTT was administered in a dose of 0.5 g or 1 g, while CMZ was used in a dose of 1 g; both of these drugs were administered by intravenous drip infusion twice a day for 5 days. The total number of treated cases was 393; 105 patients in the CTT 1 g group, 97 patients in the CTT 2 g group, and 96 patients in the CMZ 2 g group. The judgment of the clinical effects of these 3 therapeutic regimens was carried out in accordance with the criteria for evaluation of clinical efficacy of antimicrobial agents on UTI (2nd edition). The results are as follows. With regard to the overall efficacy of the drug treatments, the calculated efficacy rates were 54.3% in the CTT 1 g group, 63.9% in the CTT 2 g group and 52.1% in the CMZ 2 g group. The differences between these efficacy rates were not statistically significant. Regarding the efficacy of the treatments in terms of the bacteriuria, the bacterial cultures were seen to become negative in 40% of the cases in the CTT 1 g group, 54.6% of the CTT 2 g group and 42.7% of the CMZ 2 g group. Again, there were no statistically significant differences between the groups. Concerning the efficacy of the drugs against pyuria, by combining the cases found to be normalized and those seen to show improvement, the improvement rates were calculated to be 41.9% for the CTT 1 g group, 37.1% for the CTT 2 g group, and 37.5% for the CMZ 2 g group. These pyuria improvement rates show no statistically significant differences, respectively. The incidences of occurrence of side effects and laboratory test abnormalities thought to be caused by the drug therapy were as follows; 1.5% and 5.3% in the CTT 1 g group, 1.5% and 3.1% in the CTT 2 g group and 0% and 6.9% in the CMZ 2 g group. There were no significant differences between the groups. Based on the above clinical results, it can be concluded that, in the treatment of complicated urinary tract infections, we can expect CTT to provide the same therapeutic efficacy as CMZ when its dosage is the same or even half thereof. In addition, the safety of CTT is equal to that of CMZ. Therefore, CTT is concluded to be a useful drug.


PVT1 Signaling Is a Mediator of Cancer Progression.
There is increasing evidence that PVT1 has oncogenic properties and regulates proliferation and growth of many cancers. Themolecular mechanisms of action of PVT1 are mediated, in part, by microRNAs (miRNAs). However, some well-established transcription factors involved in cancer cell proliferation share a common thread of microRNA associations with PVT1. Furthermore, these microRNAs are also involved in mechanisms that lead to the development of drug resistance in cancer cells. While several microRNAs have been implicated directly in PVT1-mediated tumorigenesis, significant steps need to be taken to elucidate these important relationships. We synthesize the current knowledge of the miRNAs and associated genes by which PVT1 contributes to tumorigenesis. Overall, the trend suggests a negative correlation of microRNA expression with PVT1. It is clear that future studies involving PVT1 should be carried out in conjunction with microRNA analysis and should include large scale lncRNA-miRNA-mRNA network analysis. Likewise, the relationship between established transcription factors such as p53 and MYC, and processes like epithelial-mesenchymal transition may offer valuable insight into the yet unknown mechanisms of PVTI-mediated cancer progression via microRNA-dependent signaling networks.


Automated mapping of Portulacaria afra canopies for restoration monitoring with convolutional neural networks and heterogeneous unmanned aerial vehicle imagery.
Ecosystem restoration and reforestation often operate at large scales, whereas monitoring practices are usually limited to spatially restricted field measurements that are (i) time- and labour-intensive, and (ii) unable to accurately quantify restoration success over hundreds to thousands of hectares. Recent advances in remote sensing technologies paired with deep learning algorithms provide an unprecedented opportunity for monitoring changes in vegetation cover at spatial and temporal scales. Such data can feed directly into adaptive management practices and provide insights into restoration and regeneration dynamics. Here, we demonstrate that convolutional neural network (CNN) segmentation algorithms can accurately classify the canopy cover of Portulacaria afra Jacq. in imagery acquired using different models of unoccupied aerial vehicles (UAVs) and under variable light intensities. Portulacaria afra is the target species for the restoration of Albany Subtropical Thicket vegetation, endemic to South Africa, where canopy cover is challenging to measure due to the dense, tangled structure of this vegetation. The automated classification strategy presented here is widely transferable to restoration monitoring as its application does not require any knowledge of the CNN model or specialist training, and can be applied to imagery generated by a range of UAV models. This will reduce the sampling effort required to track restoration trajectories in space and time, contributing to more effective management of restoration sites, and promoting collaboration between scientists, practitioners and landowners.


A new role for Holliday junction resolvase Yen1 in processing DNA replication intermediates exposes Dna2 as an accessory replicative helicase.
DNA replication is mediated by a multi-protein complex known as the replisome. With the hexameric MCM (minichromosome maintenance) replicative helicase at its core, the replisome splits the parental DNA strands, forming replication forks (RFs), where it catalyses coupled leading and lagging strand DNA synthesis. While replication is a highly effective process, intrinsic and oncogene-induced replication stress impedes the progression of replisomes along chromosomes. As a consequence, RFs stall, arrest, and collapse, jeopardizing genome stability. In these instances, accessory fork progression and repair factors, orchestrated by the replication checkpoint, promote RF recovery, ensuring the chromosomes are fully replicated and can be safely segregated at cell division. Homologous recombination (HR) proteins play key roles in negotiating replication stress, binding at stalled RFs and shielding them from inappropriate processing. In addition, HR-mediated strand exchange reactions restart stalled or collapsed RFs and mediate error-free post-replicative repair. DNA transactions at stalled RFs further involve various DNA editing factors, notably helicases and nucleases. A study by Ölmezer et al. (2016) has recently identified a role for the structure-specific nuclease Yen1 (GEN1 in human) in the resolution of dead-end DNA replication intermediates after RF arrest. This new function of Yen1 is distinct from its previously known role as a Holliday junction resolvase, mediating the removal of branched HR intermediates, and it becomes essential for viable chromosome segregation in cells with a defective Dna2 helicase. These findings have revealed greater complexity in the tasks mediated by Yen1 and expose a replicative role for the elusive helicase activity of the conserved Dna2 nuclease-helicase.


Time to Rehospitalization in Patients With Schizophrenia Receiving Long-Acting Injectable Antipsychotics or Oral Antipsychotics.
This study aimed to investigate and compare time to rehospitalization in patients with schizophrenia receiving long-acting injectable antipsychotics (LAIs) after discharge with those receiving oral antipsychotics. Additionally, the trend of LAIs prescription rates was investigated. Patients with schizophrenia (n = 13 087), who were discharged from the study hospital from 2006 to 2017, were followed-up under naturalistic conditions in the year after discharge. The primary outcome was time to rehospitalization. Survival analysis was used in the comparisons between LAIs and oral antipsychotics and between FGA-LAIs and SGA-LAIs. Simple linear regression and Cochrane-Armitage trend test were used to test whether a time trend existed for LAIs prescription rates. In the 1 year following discharge, patients in the LAIs group had a significantly lower rehospitalization rate and a significantly lengthened time to rehospitalization than those in the oral antipsychotics group. Rehospitalization rate and time to rehospitalization were not significantly different in patients receiving FGA-LAIs or SGA-LAIs. A significantly higher percentage of patients treated with FGA-LAIs received anticholinergic agents than those treated with SGA-LAIs. The LAIs prescription rate grew significantly from 2006 to 2017 by an average of 0.5% per year. LAIs were significantly superior to oral antipsychotics in reducing rehospitalization risk, whereas SGA-LAIs were comparable with FGA-LAIs in reducing rehospitalization risk. However, use of concomitant anticholinergic agents was less frequent in the SGA-LAIs group than in the FGA-LAIs group. Increase in LAIs prescription rate may be due to growing experiences and success among clinicians in treating patients with LAIs.


Acceptorless Dehydrogenation of Hydrocarbons by Noble-Metal-Free Hybrid Catalyst System.
A hybrid catalysis that comprises an acridinium photoredox catalyst, a thiophosphate organocatalyst, and a nickel catalyst-enabled acceptorless dehydrogenation of hydrocarbons is reported. The cationic nickel complex played a critical role in the reactivity. This is the first example of acceptorless dehydrogenation of hydrocarbons by base metal catalysis under mild reaction conditions of visible light irradiation at room temperature.


Comparison of treatment outcomes between involved-field and elective nodal irradiation in limited-stage small cell lung cancer.
The present study was performed to assess the usefulness of involved-field irradiation and the impact of (18)F-fluorodeoxyglucose-positron emission tomography-based staging on treatment outcomes in limited-stage small cell lung cancer. Eighty patients who received definitive chemoradiotherapy for limited-stage small cell lung cancer were retrospectively analyzed. Fifty patients were treated with involved-field irradiation, which means that the radiotherapy portal includes only clinically identifiable tumors. The other 30 patients were irradiated with a comprehensive portal, including uninvolved mediastinal and/or supraclavicular lymph nodes, so-called elective nodal irradiation. No significant difference was seen in clinical factors between the two groups. At a median follow-up of 27 months (range, 5-75 months), no significant differences were observed in 3 year overall survival (44.6 vs. 54.1%, P= 0.220) and 3 year progression-free survival (24.4 vs. 42.8%, P= 0.133) between the involved-field irradiation group and the elective nodal irradiation group, respectively. For patients who did not undergo positron emission tomography scans, 3 year overall survival (29.3 vs. 56.3%, P= 0.022) and 3 year progression-free survival (11.0 vs. 50.0%, P= 0.040) were significantly longer in the elective nodal irradiation group. Crude incidences of isolated nodal failure were 6.0% in the involved-field irradiation group and 0% in the elective nodal irradiation group, respectively. All isolated nodal failures were developed in patients who had not undergone positron emission tomography scans in their initial work-ups. If patients did not undergo positron emission tomography-based staging, the omission of elective nodal irradiation resulted in impaired survival outcomes and raised the risk of isolated nodal failure. Therefore, involved-field irradiation for limited-stage small cell lung cancer might be reasonable only with positron emission tomography scan implementation.


A global online study of haematopoietic stem cell transplantation in multiple sclerosis and other neurodegenerative disorders.
The objective of this study was to understand the uptake of hemopoietic stem cell transplantation (HSCT) in neuroimmunological disorders like multiple sclerosis (MS). An independent University affiliated research organization conducted a global online survey of people having had HSCT, examining demographics, treatment protocol, and effectiveness. Of 271 participants, useful data were available in 223; women aged 35-54 accounted for 73.5%. Most had a household income greater than US$50,000, and the majority of participants were from Australia and the United States. Nearly 94.6% of people suffer from MS. Most had their treatment in Russia (38.7%) and 78.1% had nonmyeloablative transplants. Nearly half of the participants spent between US$50,000 to US$74,999. There were 54.5% of neurologists who did not support their patients having HSCT. Around 85.5% of participants believed HSCT helped them manage their disease from weeks to years after transplantation, and treatment was recommended by 9.5% of participants. The average reduction in Expanded Disability Status Score after transplantation was 1.2 (95% CI: 0.97-1.41; N = 197; p < 0.01; t: 10.7, df: 196). Participants were supportive of HSCT despite the costs and would recommend it to others. The data suggest some benefit in minimizing disability in MS and provides justification for large randomized controlled trials.


Target sequences for cis-acting regulation within the dual promoter of the human c-myc gene.
Recombinant plasmids of the human c-myc promoter-leader region and the bacterial chloramphenicol acetyltransferase (cat) gene were constructed. After transfection into different rodent and human cells, the 862-base-pair (bp) PvuII fragment carrying both c-myc promoters and 350 bp of the untranslated leader conferred 1/15 to 1/30 of the CAT activity mediated by the simian virus 40 promoter. The presence of additional sequences upstream of the PvuII fragment had an overall negative effect on c-myc promoter activity detectable by titration analysis with small amounts of transfected plasmid DNA. The analysis of numerous deletion constructs in the c-myc promoter-leader region as well as S1 mapping experiments demonstrated that the high CAT activity depended largely on the presence of the second promoter. By cotransfection of c-myc-cat constructs with plasmids carrying different parts of the c-myc promoter locus, targets for positively acting cellular factors were identified. Two positive regulatory elements were mapped within the 862-bp PvuII fragment. One was localized within the 248-bp PvuII-SmaI fragment -101 to -349 bp upstream of the first cap site and the other within the 142-pb XhoI-NaeI fragment of the first exon, comprising positions -95 to +47 relative to the second cap site. We conclude that the dual promotor of the human c-myc gene represents a strong eucaryotic promotor regulated by cooperation of positively and negatively acting cellular transcription factors.


Behavioral effects of wolf presence on moose habitat selection: testing the landscape of fear hypothesis in an anthropogenic landscape.
Landscape of fear refers to the spatial variation in prey perception of predation risk, that under certain conditions, may lead to changes in their behavior. Behavioral responses of prey in relation to large carnivore predation risk have mainly been conducted in areas with low anthropogenic impact. We used long-term data on the distribution of moose in different habitat types in a system characterized by intensive management of all three trophic levels (silviculture, harvest of wolves and moose) to study effects on moose habitat selection resulting from the return of an apex predator, the wolf. We assumed that coursing predators such as wolves will cause an increased risk for moose in some habitat types and tested the hypotheses that moose will avoid open or young forest habitats following wolf establishment. After wolf recolonization, moose reduced their use of one type of open habitat (bog) but there was neither change in the use of the other open habitat type (clear-cut), nor in their use of young forest. Wolf establishment did not influence the use of habitat close to dense habitat when being in open habitats. Thus, the effect of wolves varied among habitat types and there was no unidirectional support for a behavioral effect of wolves' establishment on moose habitat use. Human-driven habitat heterogeneity, concentration of moose forage to certain habitat types, and the effects of a multiple predator guild on moose may all contribute to the results found. We conclude that the landscape of fear is likely to have weak ecological effects on moose in this system.


Bereaved hospice caregivers' perceptions of the end-of-life care communication process and the involvement of health care professionals.
This study was conducted to gain an understanding of bereaved hospice family caregivers' perceptions of the communication process from initial discussion of the need for end-of-life care and hospice through the decision to choose hospice and the transition home. One hundred eight bereaved hospice caregivers that were 36 months postpatient death responded to a quantitative mailed survey. Aspects of the end-of-life care communication process, including involvement of health care professionals at each step, were assessed. Serious illness and the need for hospice care were first mentioned to patients and caregivers by physicians. Often the caregivers recalled only one meeting in which hospice was discussed. Other than the physicians, social workers were the professionals most often present in at least one of these meetings. Nurses and social workers were both perceived to be helpful in the transition to home hospice. Social workers were reported to be most comfortable with discussion of end-of-life care, most knowledgeable, and most available during the communication process that led to a decision about end-of-life care. Suggestions that these caregivers had to improve communication with health care professionals were categorized from responses to an open-ended survey question as: the need to discuss vital information earlier and what to expect for care at end stage; be more forthcoming about reality of death; and treat people as individuals. Although many health care professionals play a role in the communication and transition to hospice care, physicians and social workers were found to be pivotal participants in this process. A coordinated team approach to end-of-life communication and decision-making may facilitate the process and possibly lead to increased and earlier referral to hospice.


Prevalence of allergic sensitization to conifer pollen in a high cypress exposure area.
Sensitization to Pinales (Cupressaceae and Pinaceae) has increased dramatically in recent years. The prevalence of sensitization in different geographic areas is related to exposure to specific pollens. To investigate the prevalence of allergy to different conifer pollens, describe the characteristics of patients with such allergy, and identify the involved allergens. Patients were recruited at five hospitals near Madrid. Extracts from conifer pollen were prepared and used in skin-prick testing. Wheal sizes were recorded, and serum samples obtained from patients with positive reactions to Cupressus arizonica and/or Pinus pinea. The specific immunoglobulin E value to C. arizonica and Cup a 1 was determined. Individual immunoblots for each patient and with a pool of sera were performed. Allergenic proteins were sequenced by using liquid chromatography-tandem mass spectrometry. Of 499 individuals included in the study, 17 (14%) had positive skin-prick test results to some conifer pollen extracts. Sixty-four patients had positive results to C. arizonica (prevalence 12.8%) and 11 had positive results to P. pinea (2.2%). All the patients had respiratory symptoms (61.4% during the C. arizonica pollination period), and 62.9% had asthma. Approximately 86% of the patients had positive specific immunoglobulin E results to C. arizonica and 92.3% had positive results to Cup a 1. Fourteen different bands were recognized by immunoblot; the most frequent bands were those detected at 43, 18, 16, and 14 kDa. All sequenced proteins corresponded to Cup a 1. Allergy to conifer pollen could be considered a relevant cause of respiratory allergy in central Spain. Asthma was more frequent than in other studies. We only identified Cup a 1 as involved in sensitization.


Assessment and care of visuospatial disorientation in a mixed dementia patient: a case study using objective measurements.
We present a case of a mixed dementia patient with visuospatial disorientation and a developmental disability. The patient was a 69-year-old man who was hospitalized for resistance to care, wandering, and a sleep disorder. He was in the advanced stages of dementia and had severe visual impairment due to cataracts. We used an integrated circuit monitoring system and Actiwatch to measure the distance moved per day and sleep-rest cycle, respectively. We administered a coloured paper test to assess the colour most easily recognized by the patient, which was red. We found that displaying his name in large red letters by his door enabled the patient to read his name. We also moved him to a private room next to the day room. The distance moved per day decreased significantly after the implementation of the care plan. However, after the room change, large urine spills were often observed in the bathroom, indicating visuospatial problems. We placed red adhesive tape onto the toilet bowl to make it more visible, which solved the urine spill problem. This case study demonstrates the efficacy of using simple assessment techniques to identify the remaining abilities of a dementia patient and taking simple steps to address the behavioural and psychological symptoms of dementia.


Fasting gastric fluid and fecal polyamine concentrations in premature infants.
The role of milk polyamines in the development of the gastrointestinal tract of human infants is presently unknown. Polyamine concentrations are higher in human milk than in infant formulas. The aim of the present study was to gather data on luminal polyamines by measuring gastric fluid and fecal polyamine concentrations in premature infants during the postnatal period. We further compared gastric fluid polyamine concentrations with those reported for milk and looked for possible relationships between luminal polyamine concentrations, age, and growth rate. High-performance liquid chromatography was used for the measurement of polyamine concentrations in both fecal and gastric fluid samples. Ninetieth centiles for gastric polyamines during the first week were 62, 28, 82, and 14 microM for putrescine, spermidine, spermine, and cadaverine, respectively. These values are higher than those reported for human milk and infant formulas. Polyamine concentrations were unrelated to either age or growth rate. Ninetieth centiles for fecal polyamines during the first week were 7668, 5176, 53, and 75 microM for cadaverine, putrescine, spermidine, and spermine, respectively. Fasting gastric fluid polyamine concentrations in premature infants are higher than those reported for either human milk or infant formulas. The high fecal cadaverine and putrescine concentrations are probably of bacterial origin.


Meta-analysis of double-blind placebo-controlled trials of antidepressants and benzodiazepines for patients with panic disorders.
Meta-analysis of 19 double-blind placebo-controlled trials of antidepressants (N = 13) and benzodiazepines (N = 6) for patients with panic disorders showed that active treatment had 25% greater success rate than placebo over a mean duration of 14 weeks. There were no statistically significant differences observed between treatment sub-groups (antidepressants--mean duration 16 weeks; and benzodiazepines--mean duration 7 weeks). On this basis antidepressants and benzodiazepines prescribed in clinical settings are likely to be equally effective in the short-term treatment of people with panic disorders.


An overview of pain assessment and management.
Pain is a distressing, subjective and complex phenomenon that all nurses will encounter in their clinical practice. Effective pain management requires nurses to undertake a structured assessment to identify the probable causes of pain and guide management. Interventions used to reduce pain can be varied and multimodal. This article provides an overview of pain including its definition, classifications, assessment and management. It emphasises the importance of a person-centred approach to care which reflects Margo McCaffery's seminal quote that pain is 'whatever the experiencing person says it is'.


Avalon© bicaval dual-lumen cannula for venovenous extracorporeal membrane oxygenation: survey of cannula use in France.
We conducted an epidemiologic survey in France on the use of bicaval dual-lumen cannulas for extracorporeal membrane oxygenation (ECMO). Every service that used the Avalon cannula was contacted. Practitioners answered questions concerning its practical usage and complications that were attributable to its usage. We report data for 52 instances of cannula usage. The primary indication was acute respiratory distress syndrome (ARDS) in 77% of cases. Of all of the patients who required cannulas, 46% died. The maximum flow was 2,175 ± 556 ml/minute for 20-Fr.-diameter cannulas, 3,207 ± 653 ml/minute for 23 Fr., 3,963 ± 729 ml/minute for 27 Fr., and 5,490 ± 984 ml/minute for 31 Fr. Surgeons placed the cannulas in 52% of cases, intensivists placed the cannulas in 23% of cases, and multidisciplinary teams placed the cannulas in 25% of cases. The mean insertion time was 26 ± 13 minutes, and insertion was performed under transesophageal electrocardiography (TEE) (67%), transthoracic echocardiography (TTE) (25%), fluoroscopy (4%), or no guidance (4%). The main complication was migration into the right ventricle. Problems with hemolysis were described in 21% of cases. No case of cannula thrombosis was found. No case of infection was reported. Bleeding was noted in 17% of cases. The mean time of use was 8 ± 7 days. Modifications to the supportive care system were required in 15% of cases. Monitoring was performed by chest x-rays (90%), TTE (42%), and TEE (46%). Five extubations occurred during the support period. Nine patients were mobilized. The use of this cannula yielded satisfactory results. We suggest placing these cannulas using TTE or TEE and recommend the use of large-caliber cannulas in hypoxemic patients.


Eotaxin-1/CCL11 promotes cellular senescence in human-derived fibroblasts through pro-oxidant and pro-inflammatory pathways.
Eotaxin-1/CCL11 is a pivotal chemokine crucial for eosinophil homing to the lungs of asthmatic patients. Recent studies also suggest that CCL11 is involved in the aging process, as it is upregulated in elderly, and correlated with shorter telomere length in leukocytes from asthmatic children. Despite its potential pro-aging effects, the precise contribution of CCL11 and the underlying mechanisms involved in the promotion of cellular senescence remains unclear. Therefore, the primary goal of this study was to explore the role of CCL11 on senescence development and the signaling pathways activated by this chemokine in lung fibroblasts. To investigate the targets potentially modulated by CCL11, we performed an in silico analysis using PseudoCell. We validated in vitro the activation of these targets in the human lung fibroblast cell line MRC-5 following rhCCL11 exposure. Finally, we performed differential gene expression analysis in human airway epithelial cells of asthmatic patients to assess CCL11 signaling and activation of additional senescent markers. Our study revealed that eotaxin-1/CCL11 promote reactive oxygen secretion (ROS) production in lung fibroblasts, accompanied by increased activation of the DNA damage response (DDR) and p-TP53 and γH2AX. These modifications were accompanied by cellular senescence promotion and increased secretion of senescence-associated secretory phenotype inflammatory cytokines IL-6 and IL-8. Furthermore, our data show that airway epithelial lung cells from atopic asthmatic patients overexpress CCL11 along with aging markers such as CDKN2A (p16INK4a) and SERPINE1. These findings provide new insights into the mechanisms underlying the pro-aging effects of CCL11 in the lungs of asthmatic patients. Understanding the role of CCL11 on senescence development may have important implications for the treatment of age-related lung diseases, such as asthma.


Clinical significance of assay of thyroid-stimulating antibody in Graves' disease.
We correlated thyroid-stimulating antibody (direct thyroid stimulation method) with the clinical course of 187 patients with Graves' disease. Of 64 patients with newly diagnosed hyperthyroidism, 59 were positive; 36 of 38 patients tested early in therapy were positive. Twenty-eight patients received antithyroid drug therapy, and thyroid-stimulating antibody was measured at cessation of therapy; 13 patients, negative in the assay, remained in remission, and of 15 relapsed cases 12 were positive. Of 57 patients previously treated by various means for hyperthyroidism, 34 were positive, and they primarily had had relapse after initial treatment, then ablative therapy causing hypothyroidism. Six of 10 patients with euthyroid ophthalmopathy had a positive assay and an abnormal thyrotropin-releasing hormone (TRH) test or thyroid suppression test; the four negative patients had a normal TRH test. Thus measurement of thyroid-stimulating antibody appears effectively to reflect activity of the underlying disturbance in Graves' disease.


Sexual Murderers: Sex Offender, Murderer, or Both?
Sexual murderers perpetrate homicide and rape/sexual abuse, but it is unclear whether they should primarily be considered homicide offenders, sexual offenders, or both. Most studies have merged together different types of non-homicidal sex offenders (NHSOs), neglecting to consider the potential differences between the nonviolent and violent sex offenders. Here, we suggest it is important to isolate those violent sex offenders who inflict severe physical injuries that could potentially lead to a lethal outcome. Therefore, the aim of the current study is to compare different measures of the criminal career on three groups of sex offenders: NHSOs, violent NHSOs, and sexual homicide offenders (SHOs) using data from 616 incarcerated male sex offenders in a Federal penitentiary in Canada. Interestingly, the group of sex offenders with the worst criminal career profile was not the SHOs, but the violent NHSOs. Violent NHSOs had the greatest number of prior convictions and the most varied and versatile criminal career. Therefore, we suggest that based on their criminal career, SHOs should be considered more as murderers than sex offenders. However, to fully answer this question, future studies should include a group of non-sexual homicide offenders.


Acute respiratory distress syndrome definition: do we need a change?
Since the first description of the acute respiratory distress syndrome (ARDS) in 1967, no specific clinical sign or diagnostic test has yet been described that identifies ARDS. Its diagnosis is based on a combination of clinical, hemodynamic, and oxygenation criteria. The purpose of this review is to examine the current definition for ARDS and to discuss why this definition may not be the most appropriate definition for this syndrome. We will briefly review our current understanding of ARDS, discuss the problems with its current diagnosis, and present clinical, pathological, and biochemical evidences supporting a more appropriate definition for ARDS. In addition, we will discuss recent efforts to identify biological markers for lung injury in pulmonary edema fluid and blood collected from critically ill patients. On the basis of current evidence, it is time for a change in the ARDS definition. A newer classification system that recognizes different severities of pulmonary dysfunction is needed. Such a system should be able to identify patients that would be most responsive to supportive therapies and those unlikely to benefit because of the severity of their disease.


Mothers' and fathers' executive function both predict emergent executive function in toddlerhood.
There are multivariate influences on the development of children's executive function throughout the lifespan and substantial individual differences can be seen as early as when children are 1 and 2 years of age. These individual differences are moderately stable throughout early childhood, but more research is needed to better understand their origins. To some degree, individual differences in executive function are correlated between mother and child, but no research to date has examined these associations prior to when children are preschool age, nor have any studies considered the role of fathers' and mothers' executive function in tandem. Here, we use a sample of 484 families (Mothers 89.2% white; Fathers 92.5% white) in three countries (UK, USA, Netherlands) to investigate the role of each parents' executive function on the development of children's (49.7% female) executive function from 14 (M = 14.42, SD = 0.57) to 24 (M = 24.47, SD = 0.78) months, as well as parenting practices that underlie these associations. Results of structural equation models suggest stability in some-but not all-components of executive function and growing unity between components as children age. We replicate extant findings such that mothers' executive function predicts children's executive function over and above stability and extend these findings to include associations between father and child skills. We find an additive role of fathers' EF, similar in magnitude to the role of mothers' EF. Finally, for both mothers and fathers we find that sensitivity and autonomy supportive practices mediate the relations between parents' and children's executive function.


Nocardia brasiliensis Infection Complicating Cryptogenic Organizing Pneumonia.
Pulmonary nocardiosis is a severe and uncommon opportunistic infection caused by Nocardia species. We present a patient with cryptogenic organizing pneumonia who was receiving long-term immunosuppressive therapy, whose treatment course was complicated by cutaneous and pulmonary nocardiosis. Tissue cultures confirmed Nocardia brasiliensis. Nocardiosis should be a diagnostic consideration for patients treated with long-term immunosuppression who have worsening pulmonary symptoms and relapsing pustular skin lesions.


Use of human perivascular stem cells for bone regeneration.
Human perivascular stem cells (PSCs) can be isolated in sufficient numbers from multiple tissues for purposes of skeletal tissue engineering. PSCs are a FACS-sorted population of 'pericytes' (CD146+CD34-CD45-) and 'adventitial cells' (CD146-CD34+CD45-), each of which we have previously reported to have properties of mesenchymal stem cells. PSCs, like MSCs, are able to undergo osteogenic differentiation, as well as secrete pro-osteogenic cytokines. In the present protocol, we demonstrate the osteogenicity of PSCs in several animal models including a muscle pouch implantation in SCID (severe combined immunodeficient) mice, a SCID mouse calvarial defect and a femoral segmental defect (FSD) in athymic rats. The thigh muscle pouch model is used to assess ectopic bone formation. Calvarial defects are centered on the parietal bone and are standardly 4 mm in diameter (critically sized). FSDs are bicortical and are stabilized with a polyethylene bar and K-wires. The FSD described is also a critical size defect, which does not significantly heal on its own. In contrast, if stem cells or growth factors are added to the defect site, significant bone regeneration can be appreciated. The overall goal of PSC xenografting is to demonstrate the osteogenic capability of this cell type in both ectopic and orthotopic bone regeneration models.


Paracetamol (acetaminophen) pharmacodynamics: interpreting the plasma concentration.
Interpretation of analgesic and antipyretic responses documented after paracetamol administration is confused because response is not directly related to concentration in the blood, but rather to an effect compartment. The effect compartment does not have real measurable concentrations, but concentrations equate approximately to those observed in the cerebrospinal fluid. A time delay exists before drug reaches the effect compartment, and the equilibration half-time between the central and effect compartment is described by a single first-order parameter (Teq or T(1/2)keo), reported to be approximately 1 h for paracetamol. Paediatric analgesic studies are limited because they have only explored postoperative pain after tonsillectomy or day-stay surgery. Other pain types and pain confounders have not been investigated. Adult studies are also similarly limited. Studies investigating antipyresis have not explored the maximum response, limiting the precision of any EC(50) estimate. The influence of the cyclical nature of fever or initial temperature is seldom accounted for in antipyretic studies. Target effect compartment concentrations of 5 mg/l for fever and 10 mg/l for pain do not seem unreasonable on the basis of current literature. Speed of onset may be shortened by giving a larger initial dose or improving absorption characteristics. Consequent plasma concentration achieved, differences in effect compartment equilibration times, and the shape of the effect compartment concentration-response curve help to explain differences between common analgesic/antipyretic drugs.


Footprint-free gene mutation correction in induced pluripotent stem cell (iPSC) derived from recessive dystrophic epidermolysis bullosa (RDEB) using the CRISPR/Cas9 and piggyBac transposon system.
Recessive dystrophic epidermolysis bullosa (RDEB) is a monogenic skin blistering disorder caused by mutations in the type VII collagen gene. A combination of biological technologies, including induced pluripotent stem cells (iPSCs) and several gene-editing tools, allows us to develop gene and cell therapies for such inherited diseases. However, the methodologies for gene and cell therapies must be continuously innovated for safe clinical use. In this study, we used the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 technology to correct the pathogenic mutation in RDEB-specific iPSCs, and the piggyBac transposon system so that no residual gene fragments remained in the genome of iPSCs after correcting the mutation. For homologous recombination (HR)-based gene editing using CRISPR/Cas9, we designed guide RNA and template DNA including homologous sequences with drug-mediated selection cassette flanked by inverted repeat sequences of the transposon. HR reaction using CRISPR/Cas9 was induced in RDEB-specific iPSCs, and mutation-corrected iPSCs (MC-iPSCs) was obtained. Consequently, the selection cassette in the genome of MC-iPSCs was removed by transposase expression. After CRISPR/Cas9-induced gene editing, we confirmed that the pathogenic mutation in RDEB-specific iPSCs was properly corrected. In addition, MC-iPSCs had no genetic footprint after removing the selection cassette by transposon system, and maintained their "stemness". When differentiating MC-iPSCs into keratinocytes, the expression of type VII collagen was restored. Our study demonstrated one of the safer approaches to establish gene and cell therapies for skin hereditary disorders for future clinical use.


Citrate enhances the protective effect of orally administered bismuth subnitrate against the nephrotoxicity of cis-diamminedichloroplatinum.
Attenuation of the renal toxicity of cis-diamminedichloroplatinum (CDDP) is important in the use of this effective but cytotoxic anticancer agent. We have previously shown that the renal toxicity of CDDP can be efficiently reduced by the induction of metallothionein (MT) by preadministration of bismuth compounds in mice. Bismuth subnitrate (BSN) is used as an antigastric ulcer agent and as an antidiarrheic agent, and is suitable for inducing MT in the kidney in cancer patients. However, due to the low absorption rate of Bi from the gastrointestinal tract, the efficacy of BSN in inducing renal MT is low. In the present study, we examined the effects of citrate as a vehicle for oral administration of BSN on the tissue distribution of Bi and induction of MT in the kidneys and tumors in mice inoculated with Meth-A fibrosarcoma. Renal levels of MT and Bi were markedly increased in the mice given BSN dissolved in citrate solution compared with those given BSN suspended in saline. On the other hand, the use of citrate increased Bi accumulation in the tumor only slightly and did not increase tumor MT levels. Administration of BSN with citrate efficiently depressed the renal toxicity of CDDP, but did not affect its antitumor activity. Since both BSN and citrate are used clinically as pharmaceuticals, the combination regimen of BSN and citrate may be readily applicable as a countermeasure against the adverse side effects of CDDP without affecting its antitumor activity.


Spatially Controlled Fabrication of Surface-Enhanced Raman Scattering Hot Spots through Photoinduced Dewetting of Silver Thin Films.
A well-designed narrow gap between noble metal nanostructures plays a prominent role in surface-enhanced Raman scattering (SERS) to concentrate electromagnetic fields at the local point, called a "hot spot". However, SERS-active substrate fabrication remains a substantial hurdle due to the high process cost and the difficulty of engineering efficient plasmonic hot spots at the target area. In this study, we demonstrate a simple photolithographic method for generating ultrasensitive SERS hot spots at desired positions. The solid-state dewetting of a Ag thin film (thickness of ∼10 nm) using a continuous-wave laser (∼1 MW/cm2) generates a closely packed assembly of hemispherical Ag nanoislands. Some of these nanoislands provide substantial plasmonic-field enhancement that is sufficient for single-molecule detection and plasmon-catalyzed chemical reaction. Such hot spot structures can be patterned on the substrate with a spatial resolution of better than 1 μm. In integrated analytical devices, the patterned SERS hot spots can be used as position-specific chemical-sensing elements.


Topological Avenue for Efficient Decontamination of Large Volumes of Fluids via UVC Irradiation of Packed Metamaterials.
Nowadays, metamaterials application enjoys notoriety in fluid decontamination and pathogen annihilation, which are frequently present in polluted fluids (e.g., water, blood, blood plasma, air or other gases). The depollution effect is largely enhanced by UVC irradiation. The novelty of this contribution comes from the significant increase by packing of the total surface of metamaterials in contact with contaminated fluids. Packed metamaterial samples are subjected to UVC irradiation, with expected advantages for implant sterilization and long-term prevention of nosocomial infections over large clinical areas. The novel aspect of the investigation consists of a combination of big and small elements of the metamaterial to optimize the above effects connected with fluids and irradiation. The big elements allow the radiation to penetrate deep inside the fluid, and the small elements optimally disperse this radiation toward deeper regions of the metamaterial. A packing scheme of smaller, in-between large metamaterial spheres and fibres is proposed for promoting enhanced depollution against pathogen agents. It is demonstrated that the total surface of metamaterials in contact with contaminated fluids/surface is significantly increased as a result of packing. This opens, in our opinion, new auspicious perspectives in the construction of novel equipment with high sensibility in the detection and decontamination of microorganisms.


Human umbilical arteries and veins: generation of leukotrienes and response to exogenous leukotrienes.
When human umbilical arteries and veins were challenged with calcium ionophore A23187 they released a leukotriene (LT)-like material. On bioassay using guinea-pig ileal smooth muscle, the release was equivalent to 37.4 +/- 7.5 ng LTD4 (mean +/- SEM) per gram of arterial tissue, and 29.0 +/- 4.7 ng LTD4/g of venous tissue. The LT was further purified by high pressure liquid chromatography, and characterised as LTC4 by bioassay and radioimmunoassay of purified fractions. Longitudinal strips from human umbilical arteries in vitro did not contract to doses of synthetic LTD4 in the range 5 X 10(-11) to 10(-9) mol.


Effects of electroacupuncture and gonadotropin-releasing hormone treatments on hormonal changes in anoestrous sows.
The therapeutic effects of acupuncture in treating reproductive disturbances of man and animals have been proven in the past and is used clinically already. However, the mechanism of such therapy is not known yet. In this study, anoestrous sows were used to investigate the mechanism. Anoestrous sows with luteal ovaries were allocated to three groups. Four sows received electroacupuncture treatment at Pai-Hui and Wei-Ken (acupuncture treated group; group 1). Three sows received electroacupuncture treatment at Chiang-Feng and Chou-Shu (acupuncture control group; group 2). Four received 50 micrograms of gonadotropin-releasing hormone (GnRH) intravenously (drug control group; group 3). The concentrations of luteinizing hormone, progesterone, oestradiol and cortisol in serum were measured by radioimmunoassay. Oestrus return was monitored during 14 days after treatment. At the end of this observation period, the number of animals returned to oestrus were 3, 1 and 1 in groups 1, 2 and 3, respectively. It suggests that only treatment with acupuncture at Pai-Hui and Wei-Ken had therapeutic effects in inducing oestrus. This finding is further supported by the changes of serum sex hormone concentrations. Serum LH concentrations decreased for about 2 hours after electro-acupuncture treatment both in groups 1 and 2, whereas those in group 3 increased sharply at 10 minutes, reached to a peak at 20 minutes and returned gradually to basal level between 4 and 6 hours after GnRH injection. Serum progesterone concentrations rose between 4 to 6 hours after treatment in groups 1 and 3 but not in group 2. Five sows become oestrus, showed a decreased progesterone level 2 days after treatments whereas the other six anoestrous sows did not. Oestradiol levels did not have meaningful changes during the blood sampling period of 5 to 7 days in these 3 groups. Cortisol levels elevated in 15 minutes after the electroacupuncture in groups 1 and 2. However, the increment of cortisol induced by the electroacupuncture was less than that induced by the first bleeding, indicating that the adrenal stimulation may not be the main reason of the therapeutic action. The results indicated that the electroacupuncture treatment and GnRH injection could alter the release of LH from the pituitary in different ways but only electroacupuncture at Pai-Hui and Wei-Ken has a specific action on ovary and a significant therapeutic effect. Therapeutic effects of electroacupuncture on reproductive disturbance may involve a synergism of somatic-ovary and uterus reflex and central nervous-endocrine system (the hypothalamo-pituitary-ovary axis).


Molecular characterisation of organic material in air fine particles (PM10) using conventional and reactive pyrolysis-gas chromatography-mass spectrometry.
Pyrolysis-gas chromatography-mass spectrometry (Py-GC-MS) was applied to study the composition of organic constituents in air particulate matter (PM10) collected inside an industrial area. A few milligrams of sampling filters containing air particles were pyrolysed at 700 degrees C directly (conventional) or after the addition of a derivatising reagent (tetramethylammonium hydroxide, TMAH, for pyrolysis-methylation; hexamethyldisilazane, HMDS, for pyrolysis-silylation). Py-GC-MS was also applied to synthetic polymers (poly(styrene-co-isoprene), polylimonene and polypinene) and vegetation samples (coniferous pollen, bark and resin) to identify markers indicative of possible precursors. Pyrolysates of PM10 showed the same suite of compounds in all the four seasons, dominated by hydrocarbons like styrene, limonene and clusters of isomeric alkenes with 14, 15 and 16 carbon atoms. Pyrolysis products of natural origin, including furaldehyde, benzeneacetonitrile, dehydroabietin and other diterpenoids were found, while no specific markers of synthetic rubbers were detected. The principal products released from reactive pyrolysis of PM10 were methyl or trimethylsilyl (TMS) derivatives of 1,6-anhydroglucose (levoglucosan), fatty acids, dehydroabietic acid and other resin acids along with hydroxy (di)carboxylic acids. Possible sources of the detected products (e.g. pine forest, biomass combustion) are discussed.


Regulation of luteinizing hormone release by pulsatile and continuous administration of gonadotropin-releasing hormone to superfused rat and hamster pituitary cells.
Regulation of LH release by GnRH was studied in superfused anterior pituitary cells from 30-day-old female rats or hamsters. Dispersed cells were cultured 4-6 days on Cytodex beads, then loaded into water-jacketed columns, and perfused with medium (0.5 ml/min) at 37 C. Three-minute fractions of effluent were assayed for LH by RIA. LH release was dose related between 10(-10) and 10(-7) M GnRH. Rat and hamster cells released LH at peak rates of 11.3 and 12.5 ng/(min X 10(6) cells), respectively, when first exposed to 10(-8) M GnRH. Short pulses (6 min) of 10(-8) M GnRH given at 30-min intervals had little effect on the rate of LH release by rat pituitary cells; however, if the interpulse interval was reduced to 12 min, release declined 72% by the fifth pulse. In contrast, pulses of 10(-6) M GnRH at 30-min intervals desensitized rat cells. Hamster cells were desensitized by 10(-8) M GnRH after a single pulse regardless of whether a second pulse was given 30 min or 2.5 h later. Similar desensitization also occurred at other doses (10(-9) and 10(-6) M). After five pulses at 30-min intervals, the LH release rate in hamster cells was depressed 65%. Release was depressed 80% by pulses at 12-min intervals. Thus, in rats, desensitization is both frequency and dose dependent, whereas in hamsters, it is independent of frequency and dose. Stimulation with 10(-6) M GnRH pulses completely overcame desensitization in both species. Continuous exposure of anterior pituitary cells to 10(-8) M GnRH caused an initial rapid LH release, followed by a steady decline in the rate of release from peak rates to baseline levels by 2.5 h in both species. A 6-min, 10(-6) M GnRH pulse given immediately after a 3-h 10(-8) M GnRH exposure rapidly stimulated the cells to release LH at rates up to 192% of initial rates. When these pulses were continued at 30-min intervals, additional desensitization occurred. This overcoming of desensitization shows that desensitized anterior pituitary cells are not refractory to GnRH and suggests that the GnRH regulation of LH release may involve more than one GnRH receptor-mediated phenomenon.


Emergence of sentinel node biopsy in breast cancer as standard-of-care in academic comprehensive cancer centers.
Ongoing clinical trials are addressing the accuracy and safety of sentinel node biopsy (SNB) in the treatment of breast cancer; however, SNB is already increasingly being used in clinical practice. This study examined the extent and time trends of the use of SNB in stage I and II breast cancer patients. Clinical data were collected from stage I and II (tumor size < or =5.0 cm) breast cancer patients (n = 3003) who were treated at five comprehensive cancer centers between July 1, 1997, and December 31, 2000. Axillary surgery was classified as SNB alone, SNB + axillary node dissection (AND), AND alone, or none. Patterns of use of axillary surgery were summarized as the percentage of patients receiving each surgery type. The statistical significance of time trends for the use of SNB alone was analyzed by logistic regression models. All statistical tests were two-sided. Overall, SNB alone was used in 13% of patients, SNB + AND in 22%, AND alone in 59%, and no axillary surgery in 6%. Use of SNB alone was statistically significantly associated with breast-conserving surgery of both smaller (< or =2 cm) and larger tumors (2-5 cm) (P<.001 for both associations). For stage I cancer patients treated with breast-conserving surgery (n = 1763), use of SNB increased statistically significantly over the study period, from 8% in 1997 to 9%, 14%, 15%, 22%, 42%, and 58% for the next six consecutive 6-month time intervals, respectively. After controlling for center, age, and comorbidity, the odds ratio for the use of SNB alone was 2.30 (95% confidence interval = 1.88 to 2.82) for each 6-month interval (P<.001). Widespread use of SNB outside the clinical trial setting suggests that oncologists at cancer centers in our study have accepted SNB as standard-of-care for the treatment of breast cancer. This acceptance, if it occurs in other cancer centers and community practice, may affect accrual and generalizability of ongoing clinical trials of SNB.


Isolated pulmonary embolism - a specific clinical entity?
Pulmonary embolism in classical meaning is a complication of deep vein thrombosis (usually in the leg veins), developing after a part of the thrombus dislodged and got wedged in pulmonary arteries. However, in half of the patients with pulmonary embolism, deep vein thrombosis is not found. One potential explanation is a different, less common location of the thrombus or previous complete embolization of the whole thrombotic mass. Another possibility is pulmonary artery thrombosis in situ, which is a specific clinical entity associated with some typical risk factors. It develops in the place of vascular injury, as a consequence of hypoxia, inflammatory changes, endothelial dysfunction and injury. Pulmonary artery thrombosis in situ can be a complication after lung resection, radiation therapy, chest trauma, in the patients with Behçet´s disease, sickle cell anemia, chronic obstructive pulmonary disease, tuberculosis or covid pneumonia. Pulmonary artery thrombosis in situ may differ from classical pulmonary embolism in prognosis as well as in therapeutic approach.


Assessment of neuroforaminal decompression in degenerative spinal stenosis.
Surgical decompression of spinal stenosis is a balance between adequate removal of bone and soft tissue for an effective decompression of neural structures and a sufficient retention of bone to maintain mechanical stability of the spine. To develop an objective, reproducible technique for the assessment of neuroforaminal decompression in the adult lumbar spine, facet sparing laminectomies were performed from L1-S1 on the lumbar spines of 59 human cadavers. A series of semirigid probes in 0.5-mm increments were passed into each successive intervertebral foramen bilaterally, beginning at L2-L3. The minimum space available for the nerve root within the foramen of the lumbar spine is 3.5 mm at L2-L3, 3.9 mm at L3-L4, 4.3 mm at L4-L5 and 5.1 mm at L5-S1, as determined by three independent observers. There were no significant differences in interobserver or intraobserver measurements. There were no complications related to passage of the probes. This study provides a method for the intraoperative determination of the adequacy of neuroforaminal decompression.


A Method to Reproduce Symmetry in Midfacial Reconstruction: A Report of 19 Cases.
Reconstruction of facial skin defects remains a clinical challenge. With aging, ptosis of tissue over fixed structures creates an important facial feature known as the tear trough. This study aimed to evaluate the efficacy and aesthetic outcome of a novel surgical technique that reproduced this facial feature while avoiding ectropion during midfacial skin defect repair. Nineteen patients with midfacial skin defects received local flap reconstruction combined with an anchoring suture. The flap was designed in a unilateral pedicled V-Y pattern. When the flap was advanced to cover the defect, one or two sutures that connected the dermis of the flap with the infraorbital periosteum were made to reproduce the tear trough line. Midfacial defects were successfully repaired with the V-Y flap in all 19 patients. No lower eyelid ectropion or conspicuous scars were noted in any of the patients. Further, the tear trough was successfully reconstructed in each patient. Facial symmetry was maintained with static positioning and animation. The combination of local V-Y flap reconstruction with anchoring sutures to reproduce facial feature lines is an effective technique in midfacial skin defect repair.


Multicenter comparison of three different analytical systems for evaluation of DNA banding patterns from Cryptococcus neoformans.
The enormous improvement of molecular typing techniques for epidemiological and clinical studies has not always been matched by an equivalent effort in applying optimal criteria for the analysis of both phenotypic and molecular data. In spite of the availability of a large collection of statistical and phylogenetic methods, the vast majority of commercial packages are limited by using only the unweighted pair group method with arithmetic mean algorithm to construct trees and by considering electrophoretic pattern only as migration distances. The latter method has serious drawbacks when different runs (separate gels) of the same molecular analysis are to be compared. This work presents a multicenter comparison of three different systems of banding pattern analysis on random amplified polymorphic DNA, (GACA)(4), and contour-clamped homogeneous electric field patterns from strains of Cryptococcus neoformans var. neoformans isolated in different clinical and geographical situations and a standard Saccharomyces cerevisiae strain employed as an outgroup. The systems considered were evaluated for their actual ability to(i) recognize identities, (ii) define complete differences (i.e., the ability to place S. cerevisiae out of the C. neoformans cluster), and (iii) estimate the extent of similarity among different strains. The ability to cluster strains according to the patient from which they were isolated was also evaluated. The results indicate that different algorithms do indeed produce divergent trees, both in overall topology and in clustering of individual strains, thus suggesting that care must be taken by individual investigators to use the most appropriate procedure and by the scientific community in defining a consensus system.


Cerebral Venous Sinus Thrombosis and Posterior Reversible Encephalopathy Syndrome Coexisting in a Woman: A Rare Coincidence.
Cerebral venous sinus thrombosis (CVST) and posterior reversible encephalopathy syndrome (PRES) are two rare diseases which may present with similar symptoms and signs. We report a case with coexisting PRES and CVST in a 34 years old postpartum female presented with multiple episodes of generalized seizures and bilateral vision loss after delivery. MRI brain and venography revealed left transverse sinus, sigmoid sinus and internal cerebral vein thrombosis with vasogenic edema in bilateral parieto-occipital, right temporal and left frontal area, which was suggestive of posterior reversible encephalopathy syndrome (PRES). She was treated with antihypertensive, low molecular weight heparin (LMWH), oral anticoagulant and responded well to the treatment.


'If I am playing football, I forget that I have this virus': the challenges and coping strategies of adolescents with perinatally acquired HIV in KwaZulu-Natal, South Africa.
Adolescents who have acquired HIV perinatally (ALHIV) face unique challenges in taking lifelong antiretroviral therapy (ART), but little is known about what factors affect their adherence over the course of their lifelong treatment journey. We conducted a qualitative study with ALHIV who had periods of poor adherence to ART in KwaZulu-Natal, South Africa using Participant-generated Visual Methodologies (PVM). Participants used photography to represent their perspectives and experiences. 14 individuals participated in the research process. We developed a framework and identified four social domains which combined with the adolescent's own experiences and sense of self to either support or undermine adherence. Periods of non-adherence were reported by all participants. Participants described the importance of supportive relationships and households as well as the benefits of ART as supporting adherence. The fear of inadvertent disclosure of their HIV status and the side-effects of ART were barriers to adherence. Possible interventions to support adolescents in their treatment journey are identified. Current models of adherence support fail to address the challenges to lifelong therapy ALHIV face. Ongoing education and honest communication with health care providers, interventions that build resilience together with peer support, have the potential to improve adherence in ALHIV.


Sol-gel SiO2 film contained Au/SiO2/quantum dot core/shell/shell nanostructures with plasmonic enhanced photoluminescence.
A sol-gel method has been developed to fabricate functional silica film with Au/SiO2/quantum dot (QD) core/shell/shell nanostructures which exhibited plasmonic enhanced photoluminescence (PL). Au nanoparticles (NPs) were homogeneously coated with a SiO2 shell by an optimal Stöber synthesis. Hydrophobic CdSe/ZnS QDs was transferred into water phase via a ligand exchange by a thin functional SiO2 layer consisted of partially hydrolyzed 3-aminopropyltrimethoxysilane (APS) sol. The Au/SiO2/QD core/shell/shell nanostructure was created by assembling the functional SiO2-coated QDs to the SiO2-coated Au NPs while QDs transferred into water phase. Those partially hydrolyzed APS molecules play an important role for the connection between the QDs and SiO2-coated Au NPs. The Au/SiO2/QD core/shell/shell nanostructures were embedded in functional sol-gel SiO2 films fabricated via spinning and dipping coating, in which the film revealed strong surface plasmon scattering and enhanced PL. Because of the dual functionality, the film is utilizable for various applications including biological and medical sensors, optical devices, and solar cells. This technique can serve as a general route for encapsulating a variety of nanomaterials in sol-gel films.


Reduction of immunogenicity by covalent modification of murine and rabbit immunoglobulins with oxidized dextrans of low molecular weight.
This communication describes the covalent modification of monoclonal and polyclonal antibodies with oxidized dextrans of low molecular weight to generate conjugates having low immunogenicity in vivo. Conjugation conditions were optimized to generate monomeric, dextran-modified antibodies, free of both high-molecular-weight polymeric aggregates and unmodified antibodies. Conjugates could be prepared with varying levels of dextran substitution. These conjugates retained optimal immunoreactivity as well as in vivo pharmacokinetics and tumor-localization properties. In addition, multiple i.v. administrations of dextran-modified antibodies in animals did not elicit a measurable immune response to either the antibody or the dextran portion of these conjugates.


Post-cessation weight concerns among women calling a state tobacco quitline.
Obese and overweight women who smoke are more likely to be concerned about weight gain following cessation, impacting ability to quit and relapse. To determine differences in weight concerns for underweight, normal weight, overweight, and obese female smokers by race/ethnicity. From March to November 2008, female adult tobacco users calling the Oklahoma Tobacco Helpline were asked questions to determine the prevalence of obesity and concern for cessation-related weight gain. A score of 50 or greater, where 0=not at all concerned and 100=very concerned, on one of two weight concerns questions defined the outcome. BMI was calculated from self-reported height and weight. For the current analyses in 2013, race, ethnicity, age, education, marital status, and tobacco use history were examined as covariates. Multiple logistic regression was used to calculate ORs and 95% CIs. A significant interaction between race and BMI was observed; thus, separate models were created for white (n=3,579); black (n=330); American Indian (n=441); and Hispanic (n=125) women. BMI was independently associated with weight concerns among all racial/ethnic groups, but the strength of the association varied. For black and Hispanic women, there was a particularly strong association between BMI and weight concerns among obese women (OR=9.55, 95% CI=5.05, 18.07, and OR=8.46, 95% CI=2.57, 27.83, respectively), although sample sizes were small. State quitlines should consider tailoring promotional efforts and treatment protocols to include concerns about weight gain, especially for obese African American and Hispanic smokers.


The organization and evolution of transfer RNA genes in Mycoplasma capricolum.
The genes for presumably all the tRNA species in Mycoplasma capricolum, a derivative of Gram-positive eubacteria, have been cloned and sequenced. There are 30 genes encoding 29 tRNA species. This number is the smallest in all the known genetic systems except for mitochondria. The sequences of 9 tRNA genes of them have been previously reported (1-3). Twenty-two genes are organized in 5 clusters consisting of nine, five, four and two genes (2 sets), respectively. The other eight genes exist as a single transcription unit. All the tRNAs are encoded each by a single gene, except for the occurrence of two tRNA(Lys)(TTT) genes. The arrangement of tRNA genes in the 9-gene cluster, the 5-gene cluster, the 4-gene cluster and one of the 2-gene clusters reveals extensive similarity with a part of the 21-tRNA gene cluster and/or the 16-tRNA gene cluster in Bacillus subtilis, respectively. The results suggest that the present M. capricolum tRNA genes have evolved from large tRNA gene clusters in the ancestral Gram-positive bacterial genome common to M. capricolum and B. subtilis, by discarding genes for redundant as well as non-obligate tRNAs, so that all the codons may be translated by as small a number of tRNAs as possible.


Second-to-fourth digit ratio (2D:4D) is unrelated to measures of somatic reproductive effort among young men from Cebu, the Philippines.
A low second-to-fourth (2D:4D) digit ratio, a retrospective marker of high prenatal androgens, predicts increased investment in costly sexually dimorphic traits in men in some studies, although results are mixed. Here we test the hypothesis that the association of low 2D:4D ratios with increased muscularity and decreased adiposity depends on current testosterone (T) levels, such that digit ratio will be a particularly strong predictor of outcomes among men exhibiting a mating-effort-oriented endocrinological profile (high T). We also test the association between 2D:4D and somatic traits independently of T. We related 2D:4D digit ratios, and their interaction with T, to handgrip strength, lean mass, arm muscle area, and skinfold thickness in a sample of young, childess men (20-22 y) from Cebu, Philippines (N = 623). Digit ratio did not significantly predict men's T-dependent somatic traits. Interactions between 2D:4D and morning T, similarly, did not predict male muscularity or adiposity. Although two of the interactions were significant or marginally significant (p < .1), after adjusting for multiple testing the evidence in support of our hypothesis was weak. We found no evidence that 2D:4D predicted measures of somatic reproductive effort in this sample of young men from Cebu, who as a group could be considered mostly mating-oriented. These relationships were also not contingent upon, or stronger, when considering the moderating effect of concurrent T levels. In this sample, 2D:4D was therefore either a poor proxy of prenatal androgen exposure or prenatal androgens had limited influence on adult somatic outcomes.


Serological surveillance of plague in dogs and cats, California, 1979-1991.
Following natural infection both cats and dogs develop antibodies to plague which can be measured for several months after infection. Besides being helpful in the diagnosis of plague in domestic carnivores, the antibody titer has important implications in outbreak investigation and surveillance programs. We report the first serological survey for plague in domestic carnivores conducted in California between 1979 and 1991 in five different settings or programs. A total of 4115 dogs and 466 cats were tested for plague antibody by the passive hemagglutination test. 86 dogs (2.09%) and 15 cats (3.22%) had plague antibody titers > or = 1:16. The percentage of positive dogs and cats were respectively 3.96% and 0% on reservations, 3.27% and 1.39% on military bases, 0.74% and 1.25% in Los Angeles County and 0% and 4.61% in veterinary clinics, but 41.38% and 41.2% from outbreak investigations. Titers ranged from 1:16 to 1:4096 in dogs and cats, but were low in dogs and cats in the Los Angeles County survey and on the military bases. Serologic testing of pets during human case investigation or increased rodent mortality should be regularly implemented, as well as dog surveys on reservations. Surveys of pet dogs in veterinary clinics did not appear worthwhile, even if selected from plague endemic regions. Veterinarians should report suspect cases in cats to public health authorities, that will improve plague surveillance and reduce the risk of humans contracting the disease from their pets.


Angiogenin Attenuates Scar Formation in Burn Patients by Reducing Fibroblast Proliferation and Transforming Growth Factor β1 Secretion.
Deep burn wounds have a high tendency to form hypertrophic scars. Previously, we found that angiogenin promoted neovascularization during deep burn wound healing. However, the association between angiogenin and scar formation is unclear. We obtained human burn scar tissues from patients who underwent scar surgery and examined the role of angiogenin in scar tissues and determined its effects in scar fibroblasts and on transforming growth factor β1 (TGF-β1) secretion. Our results showed an inverse correlation between angiogenin expression and scar severity. Next, we examined the effects of angiogenin in scar fibroblasts. We found that angiogenin was persistently expressed in human scar fibroblasts and that angiogenin expression significantly increased with time in the culture medium of scar fibroblasts. Treatment of scar fibroblasts with recombinant angiogenin significantly decreased their proliferation and TGF-β1 secretion. Moreover, angiogenin inhibited TGF-β1-mediated Smad2 signaling pathway. Our data suggest a negative role of angiogenin in fibroblast proliferation via TGF-β1-mediated Smad2 signaling pathway.


Adoption research: building blocks for the next decade.
Basic national and state adoption indicators are now unavailable and must await future data collection efforts. The focus of future efforts on the adoption of children in foster care is appropriate. Case-level adoption data with unique child indicators must be developed to advance the field and further clarify the odds of adoption and disruption. Available psychosocial research clearly indicates that adoption generally results in successful outcomes. Yet, more comparative research on differences between long-term outcomes for children who are adopted and those remaining in foster care would better estimate the value of adoption to children and the public. Rigorous research on strategies to successfully place children for adoption is essential.


Anionic cycloaromatization of 1-aryl-3-hexen-1,5-diynes initiated by methoxide addition: synthesis of phenanthridinones, benzo[c]phenanthridinones, and biaryls.
Treatment of 2-((Z)-6-substituted-3-hexene-1,5-diynyl)benzonitriles with sodium methoxide in refluxing methanol in the presence of a polar aprotic solvent, such as DMSO, HMPA, THF, or 18-crown-6, gave phenanthridinones in 21-77% yields. In these cases, addition of 10% DMSO into the reaction mixture gave the highest yield. On the other hand, methanolysis of 2-(2-(2-alkynylphenyl)ethynyl)benzonitriles under the same reaction conditions gave benzo[c]phenanthridinones in 31-57% yields. Methanolysis of (Z)-1-aryl-3-hexen-1,5-diynes in the presence of 2 equiv of tetrabutylammonium iodide gave biaryls in 14-64% yields. It is found that the reactions with aryl groups bearing electron-withdrawing groups proceeded at greater rates and gave better yields.


[Changes in mouse submaxillary glands after injection of isoproterenol in intact and depancreatized animals].
The changes in the relative weight, cell area and ultrastructure of the submaxillary glands (SMG) of CBA/C57BL mice and those in the content of immunoreactive insulin-like protein (IRILP) occurring in the organ were studied. The changes were examined on day 6 after isoproterenol (ISP) injection (23 mg/100 g bw) to healthy animals (group I), on day 13 after pancreatectomy and on day 6 after injection of the indicated dose of ISP (group II), and on day 13 after pancreatectomy without ISP injection (group III). Intact animals served as control. In all the three experimental groups, the ratio between the SMG weight and the total weight of animals showed a tendency towards increase. The highest increase was recorded in group II, where the weight of the SMG was 46.1% higher than the control value. In groups II and III, the area of acinar cells increased by 49.1 and 12.5%, respectively. The area of salivary tube granular ducts decreased by 12.6% in group II and slightly increased (by 4.7% much greater than in group III. Electron microscopy demonstrated that secretory activity of the granular duct cells was enhanced in all the three groups and that secretory extrusion occurred via the apocrine (in group II animals via the holocrine as well) cells. The radioimmunoassay data suggest that IRILP content in the SMG of controls and animals entering groups I-III was 685 +/- 50, 1125 +/- 125, 914 +/- 120 and 302 +/- 66 mu units/g weight, respectively. It is concluded that the synthesis and accumulation of IRILP in the SMG after ISP injection are activated and that the formation and extrusion of IRILP after ISP injection to diabetic animals are overtly activated. Presumably ISP injection to diabetic mice facilitates the stimulation of the compensatory function of the SMG as an IRILP-producing organ.