Calcium-Dependent Protein Kinase CPK1 Controls Cell Death by In Vivo Phosphorylation of Senescence Master Regulator ORE1.
Calcium-regulated protein kinases are key components of intracellular signaling in plants that mediate rapid stress-induced responses to changes in the environment. To identify in vivo phosphorylation substrates of CALCIUM-DEPENDENT PROTEIN KINASE1 (CPK1), we analyzed the conditional expression of constitutively active CPK1 in conjunction with in vivo phosphoproteomics. We identified Arabidopsis (Arabidopsis thaliana) ORESARA1 (ORE1), the developmental master regulator of senescence, as a direct CPK1 phosphorylation substrate. CPK1 phosphorylates ORE1 at a hotspot within an intrinsically disordered region. This augments transcriptional activation by ORE1 of its downstream target gene BIFUNCTIONAL NUCLEASE1 (BFN1). Plants that overexpress ORE1, but not an ORE1 variant lacking the CPK1 phosphorylation hotspot, promote early senescence. Furthermore, ORE1 is required for enhanced cell death induced by CPK1 signaling. Our data validate the use of conditional expression of an active enzyme combined with phosphoproteomics to decipher specific kinase target proteins of low abundance, of transient phosphorylation, or in yet-undescribed biological contexts. Here, we have identified that senescence is not just under molecular surveillance manifested by stringent gene regulatory control over ORE1 In addition, the decision to die is superimposed by an additional layer of control toward ORE1 via its posttranslational modification linked to the calcium-regulatory network through CPK1.


Alterations in peptide structure of vesicular stomatitis virus mutant and its central nervous system isolate.
Gradient SDS-polyacrylamide gel electrophoresis (PAGE) and proteolytic digestions were utilized to examine the virion proteins of two isolates of wild-type vesicular stomatitis virus (WT-VSV), WTATCC from the American Type Culture Collection and WTGL and Glasgow, as well as temperature-sensitive (ts) mutant ts G31 and a central nervous system (CNS) isolate of ts G31 designated ts G31BP. The WTATCC M protein differed in electrophoretic mobility and in its tryptic or chymotryptic peptide maps from the 125I-labelled M proteins in WTGL, ts G31 or ts G31BP. The M protein in the latter three viruses appeared identical using either tryptic or chymotryptic digestion procedures; however, limited digestion with V8 protease revealed a difference between the M protein of ts G31 and both WTGL and ts G31BP M proteins. The L, NS and G proteins all had identical tryptic and chymotryptic peptide maps in WTGL, ts G31 and ts G31BP virions. The N protein, however, was demonstrated to be distinctly different in the WTGL virion when compared with the ts G31 (or ts G31BP) virion by its tryptic peptide map. In addition, limited proteolytic digestion of the 125I-labelled N proteins revealed a different peptide structure in ts G31BP compared to N proteins of ts G31 or WTGL. The altered N protein in the CNS isolate, ts G31BP, is discussed in terms of its altered in vivo phenotype of labile viral RNA, and its potential role in the unique CNS disease associated with this virus.


Assessing teachers' positive psychological functioning at work: Development and validation of the Teacher Subjective Wellbeing Questionnaire.
This study reports on the initial development and validation of the Teacher Subjective Wellbeing Questionnaire (TSWQ) with 2 samples of educators-a general sample of 185 elementary and middle school teachers, and a target sample of 21 elementary school teachers experiencing classroom management challenges. The TSWQ is an 8-item self-report instrument for assessing teachers' subjective wellbeing, which is operationalized via subscales measuring school connectedness and teaching efficacy. The conceptualization and development processes underlying the TSWQ are described, and results from a series of preliminary psychometric and exploratory analyses are reported to establish initial construct validity. Findings indicated that the TSWQ was characterized by 2 conceptually sound latent factors, that both subscales and the composite scale demonstrated strong internal consistency, and that all scales demonstrated convergent validity with self-reported school supports and divergent validity with self-reported stress and emotional burnout. Furthermore, results indicated that TSWQ scores did not differ according to teachers' school level (i.e., elementary vs. middle), but that they did differ according to unique school environment (e.g., 1 middle school vs. another middle school) and teacher stressors (i.e., general teachers vs. teachers experiencing classroom management challenges). Results also indicated that, for teachers experiencing classroom challenges, the TSWQ had strong short-term predictive validity for psychological distress, accounting for approximately half of the variance in teacher stress and emotional burnout. Implications for theory, research, and the practice of school psychology are discussed.


Efficacy and safety testing. The clinical perspective.
The number of workplace substances is uncountable, and technical progress enforces the adaptation of substances such as cutting fluids to the new demands. For that reason new substances can hardly be tested in in vivo studies. In vitro models are widely used to test the effects of barrier creams since they are simple, rapid and safe. Since no animal model could perfectly mimic human percutaneous absorption, these tests are particularly recommended as screening procedures for barrier cream candidates. A number of in vivo methods exist whereupon conventional, non-invasive bioengineering methods, along with clinical scoring, provide the most accurate, highly reproducible assessment of the inflammatory response to irritants and allergens. Nevertheless, no general accepted procedure for the evaluation of skin protection products exists. It is essential that all the products applied to the skin (protective creams inclusive) should be clinically tested in order to verify their propensity for causing cutaneous reactions. Safety testing is a stepwise approach, comprising various in vitro and in vivo test models. The nature of the plausible biological or even toxic reactions that might occur and the types of tests designed to determine the safety of the topical formulations in men are described in this chapter.


Genetic deletion of trkB.T1 increases neuromuscular function.
Neurotrophin-dependent activation of the tyrosine kinase receptor trkB.FL modulates neuromuscular synapse maintenance and function; however, it is unclear what role the alternative splice variant, truncated trkB (trkB.T1), may have in the peripheral neuromuscular axis. We examined this question in trkB.T1 null mice and demonstrate that in vivo neuromuscular performance and nerve-evoked muscle tension are significantly increased. In vitro assays indicated that the gain-in-function in trkB.T1(-/-) animals resulted specifically from an increased muscle contractility, and increased electrically evoked calcium release. In the trkB.T1 null muscle, we identified an increase in Akt activation in resting muscle as well as a significant increase in trkB.FL and Akt activation in response to contractile activity. On the basis of these findings, we conclude that the trkB signaling pathway might represent a novel target for intervention across diseases characterized by deficits in neuromuscular function.


[High molecular weight chitosan and sodium alginate effect on secretory acid proteinase of Candida albicans].
The effect of high molecular weight chitosan (HMWCh) and sodium alginate (NaAL) on acid proteinase secretion of Candida albicans (one of culture collection and five isolates) was evaluated. The secretion of acid proteinase was induced in the presence and the absence of these polymers in different concentrations and their enzymatic activity was determined. HMWCh and NaAL significantly diminished the enzymatic activity (>76% for the collection strains and > 89% for the isolates, p < 0.05). HMWCh did not modify protein concentrations, but NaAL did. It can be concluded that both polymers can inhibit the proteinase activity of Candida albicans.


Microfluidic preparation of drug-loaded PEGylated liposomes, and the impact of liposome size on tumour retention and penetration.
Understanding the effect of liposome size on tendency for accumulation in tumour tissue requires preparation of defined populations of different sized particles. However, controlling the size distributions without changing the lipid composition is difficult, and differences in compositions itself modify distribution behaviour. Here, a commercial microfluidic format as well as traditional methods was used to prepare doxorubicin-loaded liposomes of different size distributions but with the same lipid composition, and drug retention, biodistribution and localization in tumour tissues were evaluated. The small (∼50 nm diameter) liposomes prepared by microfluidics and large (∼75 nm diameter) liposomes displayed similar drug retention in in vitro release studies, and similar biodistribution patterns in tumour-bearing mice. However, the extent of extravasation was clearly dependent on size of the liposomes, with the small liposomes showing tissue distribution beyond the vascular area compared to the large liposomes. The use of microfluidics to prepare smaller size distribution liposomes compared to sonication methods is demonstrated, and allowed preparation of different size distribution drug carriers from the same lipid composition to enable new understanding of tissue distribution in compositionally consistent materials is demonstrated.


Benefits of ozonation before activated carbon adsorption for the removal of organic micropollutants from wastewater effluents.
Advanced processes for the removal of organic micropollutants (OMPs) from wastewater effluents include adsorption onto activated carbon, ozonation, or a combination of both processes. The removal of 28 OMPs present in a real wastewater effluent was studied by ozonation coupled to activated carbon adsorption and compared to a sole adsorption. The influence of the specific ozone dose (0.09-1.29 gO3/gDOC) and the influence of the powdered activated carbon (PAC) dose (2, 5 and 10 mg/L) were first studied separately. OMPs removal increased with both the specific ozone dose (up to 80% for a dose higher than 0.60 gO3/gDOC) and the PAC dose. Ozonation performances decreased in presence of suspended solids, which were converted to dissolved organic carbon. A correction of the specific ozone dose according to the suspended solids levels, in addition to nitrite, should be considered. The influence of ozonation (0.09, 0.22, 0.94 and 1.29 gO3/gDOC) on OMPs adsorption was then assessed. OMPs adsorption didn't change at low specific ozone doses but increased at higher specific ozone doses due to a decrease in DOM adsorption and competition with OMPs. At low ozone doses followed by adsorption (0.22 gO3/gDOC and 10 mg/L PAC), the two processes appeared complementary as OMPs with a low reactivity toward ozone were well absorbed onto PAC while most OMPs refractory to adsorption were well eliminated by ozone. Improved removals were obtained for all compounds with these selected doses, reaching more than 80% removal for most OMPs while limiting the formation of bromate ion.


Cytotoxic edema is independent of NMDA ion channel activation following middle cerebral artery occlusion (MCAO). An in vivo autoradiographic and MRI study.
Massive glutamate release is an important factor leading to ionic imbalance after occlusive stroke, which in turn contributes to cytotoxic edema formation. Currently, measurements of cytotoxic edema using 'diffusion weighted' MRI, is being used in human stroke studies, as a 'surrogate' end point for neuroprotective drug trials, including studies with glutamate antagonists. However, it is not fully understood to what extent glutamate-mediated N-methyl-D-aspartate (NMDA) receptor activation is related to 'cytotoxic' edema formation, and thus, to what degree apparent diffusion coefficient (ADC) changes, assessed by magnetic resonance imaging with 'ACD mapping', represent NMDA receptor activation. To study this relationship, four cats underwent permanent middle cerebral artery occlusion (MCAO). Edema formation was investigated using MRI with 'ACD mapping', while NMDA receptor activation was simultaneously detected in the same animals, using radio labeled 125IodoMK-801, which binds only in activated and open NMDA channels. At 5 h post-occlusion, a large area of edema could be found with significantly lower ADC values in the core and penumbral area of the ischemic lesion when compared to contralateral values. On corresponding sections of the feline brains, increased 125I-MK-801 binding was found in the infarct penumbra. However, there was no significant topographical correlation between ADC values and measured radioactivity. The results indicate that there is not a significant linkage between NMDA receptor activation and 'cytotoxic' edema following permanent MCAO. The detection of a large area of NMDA channel activation within regions of low ADC does however indicate an area of 'penumbral' ischemia susceptible to treatment with NMDA channel blockers.


Impact of donor type on outcome of bone marrow transplantation for Wiskott-Aldrich syndrome: collaborative study of the International Bone Marrow Transplant Registry and the National Marrow Donor Program.
Human leukocyte antigen (HLA)-identical sibling bone marrow transplantation is an effective treatment for Wiskott-Aldrich syndrome. However, most children with this disease lack such donors and many patients receive transplants from alternative donors. This study compared outcomes of HLA-identical sibling, other related donor, and unrelated donor transplantation for Wiskott-Aldrich syndrome. The outcome of 170 transplantations for Wiskott-Aldrich syndrome, from 1968 to 1996, reported to the International Bone Marrow Transplant Registry and/or National Marrow Donor Program were assessed. Fifty-five were from HLA-identical sibling donors, 48 from other relatives, and 67 from unrelated donors. Multivariate proportional hazards regression was used to compare outcome by donor type and identify other prognostic factors. Most transplant recipients were younger than 5 years (79%), had a pretransplantation performance score greater than or equal to 90% (63%), received pretransplantation preparative regimens without radiation (82%), and had non-T-cell-depleted grafts (77%). Eighty percent received their transplant after 1986. The 5-year probability of survival (95% confidence interval) for all subjects was 70% (63%-77%). Probabilities differed by donor type: 87% (74%-93%) with HLA-identical sibling donors, 52% (37%-65%) with other related donors, and 71% (58%-80%) with unrelated donors (P =.0006). Multivariate analysis indicated significantly lower survival using related donors other than HLA-identical siblings (P =.0004) or unrelated donors in boys older than 5 years (P =.0001), compared to HLA-identical sibling transplants. Boys receiving an unrelated donor transplant before age 5 had survivals similar to those receiving HLA-identical sibling transplants. The best transplantation outcomes in Wiskott-Aldrich syndrome are achieved with HLA-identical sibling donors. Equivalent survivals are possible with unrelated donors in young children.


Primary structure of the ribosomal DNA intergenic spacer from the mosquito, Aedes albopictus.
We have determined the primary structure of a 4.7-kb portion of the ribosomal DNA intergenic spacer from cultured cells of the mosquito, Aedes albopictus. Immediately upstream from the 18S rRNA gene was a 753-bp sequence containing two regions similar to known RNA polymerase I promoters, each preceded by potential transcription termination signals. Upstream from this putative promoter region was a 3.15-kb tandem array of 17 direct repeats with a consensus sequence length of 201 bp. The 201-bp repeats contained imperfect antisense duplications of 11-bp core domain regions in the putative RNA polymerase I promoters, and sequences of possible significance in recombination. Farthest upstream of the 18S rRNA gene was an 803-bp region containing two copies each of 34-, 48-, and 64-bp elements separated by apparently unique sequence. This first detailed structural analysis of a ribosomal DNA intergenic spacer from a member of the lower Diptera has revealed features similar to those described for the higher Diptera as well as conserved motifs presumably critical to rRNA transcription.


Dysfunctional hippocampal inhibition in the Ts65Dn mouse model of Down syndrome.
GABAergic dysfunction is implicated in hippocampal deficits of the Ts65Dn mouse model of Down syndrome (DS). Since Ts65Dn mice overexpress G-protein coupled inward-rectifying potassium (GIRK2) containing channels, we sought to evaluate whether increased GABAergic function disrupts the functioning of hippocampal circuitry. After confirming that GABA(B)/GIRK current density is significantly elevated in Ts65Dn CA1 pyramidal neurons, we compared monosynaptic inhibitory inputs in CA1 pyramidal neurons in response to proximal (stratum radiatum; SR) and distal (stratum lacunosum moleculare; SLM) stimulation of diploid and Ts65Dn acute hippocampal slices. Synaptic GABA(B) and GABA(A) mediated currents evoked by SR stimulation were generally unaffected in Ts65Dn CA1 neurons. However, the GABA(B)/GABA(A) ratios evoked by stimulation within the SLM of Ts65Dn hippocampus were significantly larger in magnitude, consistent with increased GABA(B)/GIRK currents after SLM stimulation. These results indicate that GIRK overexpression in Ts65Dn has functional consequences which affect the balance between GABA(B) and GABA(A) inhibition of CA1 pyramidal neurons, most likely in a pathway specific manner, and may contribute to cognitive deficits reported in these mice.


JI017 Attenuates Oxaliplatin-Induced Cold Allodynia via Spinal TRPV1 and Astrocytes Inhibition in Mice.
Oxaliplatin, a well-known chemotherapeutic agent, can induce severe neuropathic pain, which can seriously decrease the quality of life of patients. JI017 is an herb mixture composed of Aconitum carmichaelii, Angelica gigas, and Zingiber officinale. Its anti-tumor effect has been reported; however, the efficacy of JI017 against oxaliplatin-induced allodynia has never been explored. Single oxaliplatin injection [6 mg/kg, intraperitoneal, (i.p.)] induced both cold and mechanical allodynia, and oral administration of JI017 (500 mg/kg) alleviated cold but not mechanical allodynia in mice. Real-time polymerase chain reaction (PCR) analysis demonstrated that the upregulation of mRNA of spinal transient receptor potential vanilloid 1 (TRPV1) and astrocytes following oxaliplatin injection was downregulated after JI017 treatment. Moreover, TRPV1 expression and the activation of astrocytes were intensely increased in the superficial area of the spinal dorsal horn after oxaliplatin treatment, whereas JI017 suppressed both. The administration of TRPV1 antagonist [capsazepine, intrathecal (i.t.), 10 μg] attenuated the activation of astrocytes in the dorsal horn, demonstrating that the functions of spinal TRPV1 and astrocytes are closely related in oxaliplatin-induced neuropathic pain. Altogether, these results suggest that JI017 may be a potent candidate for the management of oxaliplatin-induced neuropathy as it decreases pain, spinal TRPV1, and astrocyte activation.


Scale-up of ART in Malawi has reduced case notification rates in HIV-positive and HIV-negative tuberculosis.
Setting: For 30 years, Malawi has experienced a dual epidemic of human immunodeficiency virus (HIV) infection and tuberculosis (TB) that has recently begun to be attenuated by the scale-up of antiretroviral therapy (ART). Objective: To report on the correlation between ART scale-up and annual national TB case notification rates (CNR) in Malawi, stratified by HIV-positive and HIV-negative status, from 2005 to 2015. Design: A retrospective descriptive ecological study using aggregate data from national reports. Results: From 2005 to 2015, ART was scaled up in Malawi from 28 470 to 618 488 total patients, with population coverage increasing from 2.4% to 52.2%. During this time, annual TB notifications declined by 35%, from 26 344 to 17 104, and the TB CNR per 100 000 population declined by 49%, from 206 to 105. HIV testing uptake increased from 51% to 92%. In known HIV-positive TB patients, the CNR decreased from a high of 1247/100 000 to 710/100 000, a 43% decrease. In known HIV-negative TB patients, the CNR also decreased, from a high of 66/100 000 to 49/100 000, a 26% decrease. Conclusion: TB case notifications have continued to decline in association with ART scale-up, with the decline seen more in HIV-positive than HIV-negative TB. These findings have programmatic implications for national TB control efforts.


Discovery of Anti-SARS-CoV-2 Nsp9 Binders from Natural Products by a Native Mass Spectrometry Approach.
The search for effective antiviral agents against SARS-CoV-2 remains a critical global endeavor. In this study, we focused on the viral nucleocapsid protein Nsp9, which is a key player in viral RNA replication and an attractive drug target. Employing a two-pronged approach, an in-house natural product library was screened using native mass spectrometry to identify compounds capable of binding to Nsp9. From the initial screening, apart from the previously reported hit oridonin (protein binding ratio of 0.56 in the initial screening, Kd = 7.2 ± 1.0 μM), we have identified a second Nsp9-interacting compound, the diterpenoid ryanodine, with a protein binding ratio of 0.3 and a Kd of 48.05 ± 5.03 μM. To gain deeper insights into the binding interactions and to explore potential structural requirements, the collision-induced affinity selection mass spectrometry (CIAS-MS) approach allowed us to identify six known oridonin analogues produced by the plant Rabdosia rubescens, each with varying affinities to Nsp9. Native MS validation of their individual binding activities to Nsp9 revealed that all analogues exhibited reduced affinity compared to oridonin. Structural-activity relationship analysis highlighted key functional groups, including 1-OH, 6-OH, 7-OH, and the enone moiety, which are crucial for Nsp9 binding. Combined data from our native mass spectrometry and CIAS-MS approaches provide valuable insights into the molecular interactions between Nsp9 and these compounds.


Genetic association between vitamin D receptor gene and Saudi patients confirmed with Familial Hypercholesterolemia.
Familial Hypercholesterolemia (FH) is a common condition caused by inherited genetic abnormalities. Inadequate clearance of the circulating low-density lipoproteins (LDL) is the primary cause of the excessive concentrations of LDL seen in FH patients. The relation with vitamin D deficiency and vitamin D receptor (VDR) gene is well documented in the Saudi Arabia. The aim of this study was to investigate the role of molecular analysis studied between FH patients and fours polymorphisms associated with VDR gene in Saudi Population. In this case-control study, 120 patients were selected, and 50 patients were confirmed as FH and 70 subjects were confirmed as healthy controls. Genotyping was performed with polymerase chain reaction followed by restriction fragment length polymorphism analysis using ApaI, BsmI, TaqI and FokI polymorphisms in the VDR gene. The current study results confirmed no association between clinical characteristics studied between FH cases and controls (p>0.05). Hardy Weinberg Equilibrium analysis was present in ApaI and FokI polymorphisms (p<0.05). Only ApaI (C vs A: OR-15.1 (95% CI:5.78-39.41); p<0.001; AC+CC vs AA: OR-6.59 (95% CI:2.42-17.95); p=0.0006) and BsmI (G vs A: OR-2.88 (95% CI:1.54-5.38); p=0.0006 and AG+GG vs AA: OR-3.79 (95% CI:1.72-8.35); p=0.0007) polymorphisms showed both allele and genotype association between FH patients and controls. ANOVA analysis confirmed that TG levels were associated (p=0.02) with combination of heterozygous and homozygous genotypes present in all four polymorphisms studied in this population. ApaI and BsmI polymorphisms in the VDR gene showed association with FH patients in the Saudi Population.


Cooling it gets hot.
Conflict is an inevitable part of work and relationships. How it is handled determines what can and cannot be accomplished personally, professionally, and organizationally. The pressures and complexities of health care interactions breeds conflicts that require constant, skillful negotiation. Increasingly, health care leaders are recognizing that proficiency at recognizing sources of friction before trouble flares up and at managing and resolving disputes that do break out is an essential part of their executive "toolbox." Resources are now widely available for acquiring fundamental knowledge of dispute prevention and negotiation techniques, and for locating and engaging reliable expertise when third-party intervention appears necessarily to reconcile apparently intractable differences.


MIG-seq is an effective method for high-throughput genotyping in wheat (Triticum spp.).
MIG-seq (Multiplexed inter-simple sequence repeats genotyping by sequencing) has been developed as a low cost genotyping technology, although the number of polymorphisms obtained is assumed to be minimal, resulting in the low application of this technique to analyses of agricultural plants. We applied MIG-seq to 12 plant species that include various crops and investigated the relationship between genome size and the number of bases that can be stably sequenced. The genome size and the number of loci, which can be sequenced by MIG-seq, are positively correlated. This is due to the linkage between genome size and the number of simple sequence repeats (SSRs) through the genome. The applicability of MIG-seq to population structure analysis, linkage mapping, and quantitative trait loci (QTL) analysis in wheat, which has a relatively large genome, was further evaluated. The results of population structure analysis for tetraploid wheat showed the differences among collection sites and subspecies, which agreed with previous findings. Additionally, in wheat biparental mapping populations, over 3,000 SNPs/indels with low deficiency were detected using MIG-seq, and the QTL analysis was able to detect recognized flowering-related genes. These results revealed the effectiveness of MIG-seq for genomic analysis of agricultural plants with large genomes, including wheat.


Active temperature and velocity correlations produced by a swimmer suspension.
The agitation produced in a fluid by a suspension of microswimmers in the low Reynolds number limit is studied. In this limit, swimmers are modeled as force dipoles all with equal strength. The agitation is characterized by the active temperature defined, as in kinetic theory, as the mean square velocity, and by the equal-time spatial correlations. Considering the phase in which the swimmers are homogeneously and isotropically distributed in the fluid, it is shown that the active temperature and velocity correlations depend on a single scalar correlation function of the dipole-dipole correlation function. By making a simple medium-range order model, in which the dipole-dipole correlation function is characterized by a single correlation length k(0)(-1) it is possible to make quantitative predictions. It is found that the active temperature depends on the system size, scaling as L(4-d) at large correlation lengths L<<k(0)(-1), while in the opposite limit it saturates in three dimensions and diverges logarithmically with the system size in two dimensions. In three dimensions the velocity correlations decay as 1/r for small correlation lengths, while at large correlation lengths the transverse correlation function becomes negative at maximum separation r~L/2, an effect that disappears as the system increases in size.


Evolving complexity of complement-related diseases: C3 glomerulopathy and atypical haemolytic uremic syndrome.
The current review will discuss recent advances in our understanding of the pathology of C3 glomerulopathy and atypical haemolytic uremic syndrome (aHUS). C3 glomerulopathy and aHUS are associated with abnormalities of control of the alternative pathway of complement. Recent articles have provided new insights into the classification of C3 glomerulopathy and its relationship to idiopathic immune complex-mediated glomerulonephritis. They suggest that there may be considerable overlap in pathogenesis between these entities and have indicated novel ways in which classification may be improved. There is increasing evidence that monoclonal gammopathy may cause C3 glomerulopathy or aHUS in older patients and emerging evidence that treatment of the underlying plasma cell clone may ameliorate the kidney disease. Recent work has provided new insights into the causes of C3 glomerulopathy and aHUS, and the mechanism by which complement is dysregulated. This is of particular importance with the advent of new therapeutic agents which can specifically target different parts of the complement cascade.


Mechanisms for induction of L-selectin loss from T lymphocytes by a cryptococcal polysaccharide, glucuronoxylomannan.
Disseminated cryptococcosis is accompanied by cryptococcal polysaccharides in the serum and the lack of cellular infiltrates in infected tissues. Cryptococcal polysaccharides given intravenously to mice inhibit the influx of T lymphocytes into the sites of cell-mediated immune response. The focus here was to determine whether cryptococcal polysaccharides modulate the expression of molecules, such as L-selectin, that are important in extravasation of T cells. Cryptococcal glucuronoxylomannan (GXM), but not galactoxylomannan or mannoprotein, was found to cause loss of L-selectin from freshly isolated human T cells of both CD4 and CD8 subsets and from Jurkat cells. With the signaling-pathway inhibitors staurosporine (which inhibits protein kinase C) and herbimycin A (which inhibits protein tyrosine kinases), we showed that GXM or the cryptococcal culture filtrate antigen CneF directly induces L-selectin loss from CD4(+) and CD8(+) T cells via a herbimycin A-sensitive pathway(s) presumably involving one or more protein tyrosine kinases but not via a pathway involving protein kinase C. Loss of L-selectin from the T cells before the T cells have a chance to bind to L-selectin ligands on endothelial cells would be expected to prevent T-cell migration into inflamed tissues and/or lymph organs.


Human anatomy in wax during Florentine enlightenment.
The high mortality rate of mothers and children during delivery was not taken into consideration by science before the end of the XVII Century; gynaecology became an official discipline in the Schools of Medicine at the beginning of the next century. The knowledge of anatomy was fundamental for doctors but human cadavers were still difficult to obtain; thus the need of models for the teaching. Anatomical models were obtained by injection of salts and alcohol on bodies or part of bodies: the procedure was difficult and the preparation did not last for a long time. Models in wax proved to be scientifically reliable, and everlasting. Schools of anatomical wax modelling were established in Italy: first in Bologna and later in Florence at the Museum "La Specola", a laboratory which soon became famous all through Europe. Description follows of the technique and the material used for the construction of the models; the scientists and the workers in the laboratory. Short notes on Felice Fontana, Paolo Mascagni, Giuseppe Galletti, Clemente Susini, Luigi Calamai, Egisto Tortori. A last chapter is devoted to the XVII Century Sicilian wax modeller Gaetano Giulio Zumbo, mainly renouned for the "Plague waxes" and some anatomical heads: most of his works are exhibited in the Museum La Specola in Florence.


Coumarin and isocoumarin as serine protease inhibitors.
Serine proteases are attractive targets for the design of enzyme inhibitors since they are involved in the etiology of several diseases. Within the class of serine proteases, HLE is one of the most destructive enzymes in the body. It is implicated in the promotion or exacerbation of a number of diseases including pancreatitis, acute respiratory syndrome, rheumatoid arthritis, atherosclerosis, pulmonary emphysema, and cystic fibrosis. Thrombin, a trypsin-like serine protease, plays a dual role in thrombogenesis, including fibrin formation and platelet activation. As a result, thrombin constitutes one of the most widely studied targets for antithrombotic strategy. Numerous inhibitors of serine proteases have been reported during the past three decades. Among them, coumarin-type molecules displayed a high inhibitory potency towards various serine proteases. At that time, halomethyl dihydrocoumarins have been shown to behave as the first general suicide inhibitors of serine protease. These molecules inhibit several proteases such as human leucocyte elastase, porcine pancreatic elastase, thrombin, urokinase and human plasmin. Isocoumarins are very effective as mechanism-based inhibitors of serine proteases. Pharmacomodulation on the 3-alkoxy-4-chloroisocoumarins and the 3-alkoxy-7-amino-4-chloroisocoumarins led to strong inhibitors of numerous serine proteases such as HLE, human factor XIa and XIIa, thrombin, urokinase and kallikrein. Recently, a series of coumarins characterised by an alkyl, aryl ester, amide, thioester or ketone in the position 3 and an electrophilic chloromethyl moiety in the position 6 have been developed. These compounds were found to be high inhibitors of alpha-chymotrypin, HLE and human thrombin.


A role for PP1/NIPP1 in steering migration of human cancer cells.
Electrical gradients are present in many developing and regenerating tissues and around tumours. Mimicking endogenous electric fields in vitro has profound effects on the behaviour of many cell types. Intriguingly, specific cell types migrate cathodally, others anodally and some polarise with their long axis perpendicular to the electric vector. These striking phenomena are likely to have in vivo relevance since one of the determining factors during cancer metastasis is the ability to switch between attractive and repulsive migration in response to extracellular guidance stimuli. We present evidence that the cervical cancer cell line HeLa migrates cathodally in a direct current electric field of physiological intensity, while the strongly metastatic prostate cancer cell line PC-3-M migrates anodally. Notably, genetic disruption of protein serine/threonine phosphatase-1 (PP1) and its regulator NIPP1 decrease directional migration in these cell lines. Conversely, the inducible expression of NIPP1 switched the directional response of HeLa cells from cathodal to slightly anodal in a PP1-dependent manner. Remarkably, induction of a hyperactive PP1/NIPP1 holoenzyme, further shifted directional migration towards the anode. We show that PP1 association with NIPP1 upregulates signalling by the GTPase Cdc42 and demonstrate that pharmacological inhibition of Cdc42 in cells overexpressing NIPP1 recovered cathodal migration. Taken together, we provide the first evidence for regulation of directional cell migration by NIPP1. In addition, we identify PP1/NIPP1 as a novel molecular compass that controls directed cell migration via upregulation of Cdc42 signalling and suggest a way by which PP1/NIPP1 may contribute to the migratory properties of cancer cells.


Melastatin expression and prognosis in cutaneous malignant melanoma.
Melastatin (MLSN-1), a novel melanocyte-specific gene recently identified using a genomic approach, is expressed in murine and human melanoma cells at levels inversely proportional to metastatic rates in vivo. We studied the relationship between expression of melastatin mRNA in the primary cutaneous tumor and prognosis in patients with localized malignant melanoma. Melastatin mRNA was evaluated by in situ hybridization in primary cutaneous melanoma from 150 patients with localized disease (American Joint Committee on Cancer [AJCC] stage I and II). Multivariate Cox proportional hazards regression analysis was performed to assess the prognostic utility of melastatin mRNA expression while adjusting for other prognostic indicators. Uniform melastatin mRNA expression in the primary tumor was correlated with prolonged disease-free survival (P < .0001). Multivariate analysis revealed that melastatin status, mitotic rate, and tumor thickness influence disease-free survival independently. The 8-year disease-free survival rate in AJCC stage I patients whose tumors diffusely expressed melastatin mRNA was 100%, whereas in stage I patients with melastatin loss, the disease-free survival rate was 77% +/- 15% (median +/- SE). In patients with stage II disease whose tumors diffusely expressed melastatin mRNA, the 8-year disease-free survival rate was 90% +/- 7%, whereas in patients with melastatin loss, the disease-free survival rate was 51% +/- 8%. Downregulation of melastatin mRNA in the primary cutaneous tumor is a prognostic marker for metastasis in patients with localized malignant melanoma and is independent of tumor thickness and other variables. Used in combination, melastatin status and tumor thickness allow for the identification of subgroups of patients at high and low risk of developing metastatic disease.


Development of mental health first-aid guidelines for suicide risk: a Delphi expert consensus study in Argentina and Chile.
Suicide continues to pose a significant global public health challenge and ranks as one of the leading causes of death worldwide. Given the prevalence of suicide risk in the community, there is a significant likelihood of encountering individuals who may be experiencing suicidal thoughts or plans, creating an opening for non-health professionals to offer support. This study aims to culturally adapt the original Australian Mental Health First Aid Guidelines for suicide risk to the Chilean and Argentine context. A two-round Delphi expert consensus study was conducted involving two panels, one comprising individuals with personal experience in suicide thoughts/attempts or caregiving for those with such experiences (n = 18), and the other consisting of professionals specialized in suicide assessment and support for individuals at risk (n = 25). They rated a total of 179 items mainly derived from guidelines developed by Australian experts and translated into Spanish (168), and new items included by the research team (11). The panel members were requested to assess each item utilizing a five-point Likert scale. During the second round, items that received moderate approval in the initial round were re-evaluated, and new items suggested by the local experts in the first round were also subjected to evaluation in the next round. Inclusion in the final guidelines required an 80% endorsement as "essential" or "important" from both panels. Consensus of approval was reached for 189 statements. Among these, 139 statements were derived from the English-language guidelines, while 50 locally generated statements were accepted during the second round. A significant difference from the original guideline was identified concerning the local experts' reluctance to discuss actions collaboratively with adolescents. Furthermore, the local experts proposed the inclusion of an entirely new section addressing suicide risk in older individuals, particularly focusing on suicide methods and warning signs. A Delphi expert consensus study was conducted to culturally adapt mental health first aid guidelines for assessing suicide risk in Chile and Argentina. This study involved professionals and individuals with lived experience. While many items were endorsed, some related to inquiring about suicide risk and autonomy, particularly for adolescents, were not. An additional section for older individuals was introduced. Future research should explore the implementation and impact of these adapted guidelines in training courses. This is vital for enhancing mental health support and implementing effective suicide prevention strategies in Chile and Argentina.


Interactions between PIAS proteins and SOX9 result in an increase in the cellular concentrations of SOX9.
We have identified PIAS1 (protein inhibitor of activated STAT-1), -3, -xalpha, and -xbeta as SOX9-associated polypeptides using the Gal4-based yeast two-hybrid system and a cDNA library derived from a chondrocytic cell line. These PIAS proteins were shown to interact directly with SOX9 in two-hybrid, co-immunoprecipitation, and electrophoretic mobility shift assays. SOX9 was sumoylated in cotransfection experiments with COS-7 cells using PIAS and SUMO-1 (small ubiquitin-like modifier-1) expression vectors. SOX9 was also sumoylated in vitro by PIAS proteins in the presence of SUMO-1, the SUMO-activating enzyme, and the SUMO-conjugating enzyme. In COS-7 cells, PIAS proteins stimulated the SOX9-dependent transcriptional activity of a Col2a1 promoter-enhancer reporter. This increase in reporter activity was paralleled by an increase in the cellular levels of SOX9. Cotransfection with a SUMO-expressing vector further enhanced the transcriptional activity of this SOX9-dependent Col2a1 reporter in COS-7 cells, and this additional activation was inhibited in the presence of either SUMO-1 mutants or PIAS RING domain mutants or by coexpression of a desumoylation enzyme. Immunofluorescence microscopy of SOX9-transfected COS-7 cells showed that the subnuclear distribution of SOX9 became more diffuse in the presence of PIAS1 and SUMO-1. Our results suggest that, by controlling the cellular concentrations of SOX9, PIAS proteins and sumoylation may be part of a major regulatory system of SOX9 functions.


Use of phosphorus-32 in microassay for nucleic acid synthesis in Escherichia coli.
A method was developed to facilitate studies of ribonucleic acid and deoxyribonucleic acid synthesis in growing bacterial cultures of small volume. The nucleic acid fractions were separated by means of a modified Schmidt-Thannhauser procedure applied to micro quantities of P(32)-labeled cells collected on membrane filters. The validity and usefulness of the method are discussed.


Diet composition and the risk of type 2 diabetes: epidemiological and clinical evidence.
In the last 10 years nutritional research on diabetes has improved dramatically in terms of both number of studies produced and quality of methodologies employed. Therefore, it is now possible to attempt to provide the evidence on which nutritional recommendations for the prevention of type 2 diabetes could be based. We therefore performed a literature search and, among the papers published in indexed journals, we selected relevant epidemiological (mostly prospective) and controlled intervention studies. Lifestyle factors that have, so far, been consistently associated with increased risk of type 2 diabetes are overweight and physical inactivity. However, recent evidence from epidemiological studies has shown that the risk of type 2 diabetes is also associated with diet composition, particularly with: (1) low fibre intake; (2) a high trans fatty acid intake and a low unsaturated:saturated fat intake ratio; (3) absence of or excess alcohol consumption. All these factors are extremely common in Western populations and therefore the potential impact of any intervention on them is large: indeed, >90 % of the general population has one or more of these risk factors. The ability to correct these behaviours in the population is estimated to reduce the incidence of diabetes by as much as 87 %. Recent intervention studies have shown that type 2 diabetes can be prevented by lifestyle changes aimed at body-weight reduction, increased physical activity and multiple changes in the composition of the diet. Within this context, the average amount of weight loss needed is not large, about 5 % initial weight, which is much less than the weight loss traditionally considered to be clinically significant for prevention of type 2 diabetes. In conclusion, new emphasis on prevention by multiple lifestyle modifications, including moderate changes in the composition of the habitual diet, might limit the dramatic increase in incidence of type 2 diabetes envisaged worldwide.


Comparison of five major trichome regulatory genes in Brassica villosa with orthologues within the Brassicaceae.
Coding sequences for major trichome regulatory genes, including the positive regulators GLABRA 1(GL1), GLABRA 2 (GL2), ENHANCER OF GLABRA 3 (EGL3), and TRANSPARENT TESTA GLABRA 1 (TTG1) and the negative regulator TRIPTYCHON (TRY), were cloned from wild Brassica villosa, which is characterized by dense trichome coverage over most of the plant. Transcript (FPKM) levels from RNA sequencing indicated much higher expression of the GL2 and TTG1 regulatory genes in B. villosa leaves compared with expression levels of GL1 and EGL3 genes in either B. villosa or the reference genome species, glabrous B. oleracea; however, cotyledon TTG1 expression was high in both species. RNA sequencing and Q-PCR also revealed an unusual expression pattern for the negative regulators TRY and CPC, which were much more highly expressed in trichome-rich B. villosa leaves than in glabrous B. oleracea leaves and in glabrous cotyledons from both species. The B. villosa TRY expression pattern also contrasted with TRY expression patterns in two diploid Brassica species, and with the Arabidopsis model for expression of negative regulators of trichome development. Further unique sequence polymorphisms, protein characteristics, and gene evolution studies highlighted specific amino acids in GL1 and GL2 coding sequences that distinguished glabrous species from hairy species and several variants that were specific for each B. villosa gene. Positive selection was observed for GL1 between hairy and non-hairy plants, and as expected the origin of the four expressed positive trichome regulatory genes in B. villosa was predicted to be from B. oleracea. In particular the unpredicted expression patterns for TRY and CPC in B. villosa suggest additional characterization is needed to determine the function of the expanded families of trichome regulatory genes in more complex polyploid species within the Brassicaceae.


Fitness level and risk of weight gain in middle-age women: a prospective cohort study.
This study was conducted to determine if cardiorespiratory fitness at baseline, and changes in fitness, influence risk of weight gain (>or= 3 kg) over 20 months. Another aim was to ascertain if potential confounding factors, including age, education, strength training, energy intake, and weight, influence risk of weight gain. In a prospective study of 257 women, fitness (VO(2max)) was assessed using a graded, maximal treadmill test at baseline and follow-up. Energy intake was measured using 7-day, weighed food records. Subjects were divided into quartiles based on fitness. Risk ratios were used to show the risk of weight gain among those who were fit at baseline compared with their counterparts. Most women gained weight and 23% gained >or= 3 kg. Mean VO(2max) was 35.7 +/- 7.2 mL * kg(-1) * min(-1). Women with low-fitness at baseline had 3.18 times (95% CI: 1.46 to 6.93) greater risk, and moderately fit women had 2.24 times (95% CI: 1.04 to 4.82) greater risk of weight gain than women in the high-fitness quartile. Adjusting for potential confounders had little effect on results. High levels of fitness seem to help protect middle-aged women against weight gain, whereas low and moderate fitness increase risk of weight gain over time.


Teasdale's differential activation hypothesis: implications for mechanisms of depressive relapse and suicidal behaviour.
Teasdale's differential activation hypothesis (DAH) has been proposed as one account of cognitive vulnerability to depression. This view holds that important factors determining whether one's initial depression becomes more severe or persistent are the degree of activation, and content, of negative thinking patterns that become accessible in the depressed state. This phenomenon has been referred to as cognitive reactivity. Empirical support for the predictions of this model derives from a combination of cross-sectional and prospective studies. In this article, we evaluate this evidence with the goal of determining whether mood-induced cognitive reactivity can be considered a risk factor for depressive relapse/recurrence. Our review demonstrates sufficient evidence to consider cognitive reactivity as a potential causal risk factor for depressive relapse/recurrence. Furthermore, we extend the application of this model to the problem of suicidal relapse/recurrence including a review of preliminary support for this approach.


Sleep disturbance in relatives of palliative patients cared for at home.
The aim of the present pilot study was to investigate insomnia, sleep quality, and daytime sleepiness in relatives of dying patients cared for at home. The study has a descriptive, comparative, and cross-sectional design. The sample consisted of relatives of patients cared for through palliative home care in Uppsala County on 3 specific days. Relatives completed a questionnaire consisting of demographic questions, and items from the Insomnia Severity Index (ISI), the Epworth Sleepiness Scale (ESS), and the Richard Campell Sleep Questionnaire (RCSQ). Seventy-five relatives answered the questionnaire. The average total ISI score was 9.6, with 23% reporting moderate or severe clinical insomnia. The mean sleep duration was 6.5 hours, the mean assessed need of sleep was 8 hours, and the mean discrepancy was 1 hour. The total mean ESS score was 5.6 and only 15% of respondents reported excessive daytime sleepiness. Four percent scored very poor sleep quality, whereas 39% scored very good sleep quality (RCSQ). Two general age- and gender-related patterns were observed. Negative correlations were found between age and sleep problems, with younger relatives reporting more insomnia problems and more daytime sleepiness than did older relatives. The other general pattern was that womens' sleep quality was significantly inferior to that of men. A significant positive correlation was found between ISI and ESS, but not between RCSQ and ESS. The picture of the relatives' sleep condition is fairly complex. A minority reported clinical insomnia problems or excessive daytime sleepiness, and 73% reported getting less sleep than they wanted.


[Risk injuries at operating and delivery rooms. EPINETAC Project 1996-2000].
Surgical areas have long been considered risky with regard to occupational exposures to blood-borne pathogens. The objective of study was to describe and evaluate the risk of occupational exposure to blood-borne pathogens at operating and delivery rooms, from reports of injuries in health care workers. Transversal study of percutaneous injuries occurring in operating and delivery rooms which were registered in the Spanish surveillance system EPINETAC (Exposure Prevention Information Network Accidents) between 1996 and 2000. We recorded data from the exposed health care worker, from the accident itself and from the exposure source. The risk of exposition was calculated by logistic regression. The dependent variable was the exposition in operating/delivery rooms. We calculated the rate of exposure, total and by occupational categories, per 10,000 surgical procedures in 3 surgical specialties. There were 3,625 percutaneous injuries reported. The exposure risk was higher in midwives [OR 36.6 (CI 95% 19.61-68.52)] than in staff [OR 12.6 (CI 95% 10.21-15.71)] or training doctors [OR 12.8 (CI 95% 10.34-15.98)]. The highest risk turned up during use of material [OR 1.37 (CI 95% 1.05-1.79)] and during preparation of material for reuse [OR 1.81 (CI 95% 1.27-2.59)]. The exposure rate, in gynecologic procedures, was 34.36 injuries per 10,000, in digestive surgery it was 24.61 per 10,000, and in trauma surgery it was 18.92 per 10,000 surgical procedures. The risk of occupational exposure to blood-borne pathogens in staff and training doctors was higher in operating and delivery rooms than in others areas. Obstetric and gynecologic procedures exhibited the highest risk of exposure.


Biological markers in inflammatory bowel disease: practical consideration for clinicians.
The biomarkers are important in the Inflammatory Bowel Disease (IBD) to gain an objective measurement of disease activity and severity, as well as prognostic indicator and outcome of therapy. And they can be helpful to avoid invasive procedures. The ideal biomarker does not exist for IBD and it is likely that more than one biomarker will be needed. Biological markers potentially useful in IBD include acute-phase proteins, fecal markers, several antibodies and novel genetic determinants. The C-reactive protein (CRP) is the most studied and has been shown to be an objective marker of inflammation. CRP is a good marker of measuring disease activity in Crohn's disease (CD) and its levels can be used to guide therapy. The fecal markers (calprotectin and lactoferrin) may be helpful in differentiating patients with IBD from those with functional disorders and to predict clinical relapse. The panel of serologic markers (anti-Saccharomyces cerevisiae antibody, perinuclear anti-neutrophil cytoplasmic antibody, anti-OmpC and anti-I2 and antiglycan antibodies) for IBD can be used to stratify IBD patients into more homogeneous subgroups with respect to disease progression. Correlating serologic markers with genotypes and clinical phenotypes should enhance our understanding of the pathophysiology of IBD. The development of biomarkers in IBD will be very important in the future with the increasing utilization of novel methodological approaches like genomics and proteomics.


Synthesis, characterization, and biological studies of multidentate gold(i) and gold(iii) NHC complexes.
The synthesis and characterization of a novel macrocyclic Au(iii) N-heterocyclic carbene (NHC) imidazolyl complex, a novel macrocyclic tetra-NHC benzimidazole ligand, and the corresponding Ag(i) and Au(i) complexes are presented. Single-crystal X-ray diffraction analysis of the Au(i) benzimidazolyl complex 3 reveals an unusual structure, differing from the respective Au(i) imidazolyl complex 4. Both complexes have a Au4L2 composition; however, 3 has two C-Au(i)-C units acting as a connection between the two ligands with two Au(i) atoms being linearly coordinated inside the cavity of the macrocyclic ligand. In the case of complex 4, the structure shows a box-type coordination with all four Au(i) atoms being located between the two ligands. Stability studies in cell culture medium are performed for subsequent MTT assays and they show an unprecedented proton-to-deuterium exchange of the methylene bridge of the Au(iii) imidazolyl complex. In MTT assays, the tetranuclear acyclic Au(i) complex 5 displays the lowest IC50 values in MCF-7, PC3, and A2780cisR cells with a selective cytotoxicity for MCF-7 and A2780cisR cells.


A conditional likelihood approach for regression analysis using biomarkers measured with batch-specific error.
Measurement error is common in epidemiological and biomedical studies. When biomarkers are measured in batches or groups, measurement error is potentially correlated within each batch or group. In regression analysis, most existing methods are not applicable in the presence of batch-specific measurement error in predictors. We propose a robust conditional likelihood approach to account for batch-specific error in predictors when batch effect is additive and the predominant source of error, which requires no assumptions on the distribution of measurement error. Although a regression model with batch as a categorical covariable yields the same parameter estimates as the proposed conditional likelihood approach for linear regression, this result does not hold in general for all generalized linear models, in particular, logistic regression. Our simulation studies show that the conditional likelihood approach achieves better finite sample performance than the regression calibration approach or a naive approach without adjustment for measurement error. In the case of logistic regression, our proposed approach is shown to also outperform the regression approach with batch as a categorical covariate. In addition, we also examine a 'hybrid' approach combining the conditional likelihood method and the regression calibration method, which is shown in simulations to achieve good performance in the presence of both batch-specific and measurement-specific errors. We illustrate our method by using data from a colorectal adenoma study.


Better prediction of the location of alpha-turns in proteins with support vector machine.
We have developed a novel method named AlphaTurn to predict alpha-turns in proteins based on the support vector machine (SVM). The prediction was done on a data set of 469 nonhomologous proteins containing 967 alpha-turns. A great improvement in prediction performance was achieved by using multiple sequence alignment generated by PSI-BLAST as input instead of the single amino acid sequence. The introduction of secondary structure information predicted by PSIPRED also improved the prediction performance. Moreover, we handled the very uneven data set by combining the cost factor j with the "state-shifting" rule. This further promoted the prediction quality of our method. The final SVM model yielded a Matthews correlation coefficient (MCC) of 0.25 by a 10-fold cross-validation. To our knowledge, this MCC value is the highest obtained so far for predicting alpha-turns. An online Web server based on this method has been developed and can be freely accessed at http://bmc.hust.edu.cn/bioinformatics/ or http://184.108.40.206/.


Uncircumcision: a historical review of preputial restoration.
Throughout history, demands for restoration of the prepuce after circumcision were most commonly related to the political or religious persecution of the Jewish people. The first evidence for such a procedure is mentioned in the Bible: Under the reign of Antiochus IV (168 BC) Hellenistic ideals, such as public nakedness at athletic games or in public baths, emerged in Judea and forced Jews to stretch their shortened foreskins with a special weight, the Pondus Judaeus, to cover the glans (I. Maccabees 1). Similar efforts are reported in the Talmud during the time of Hadrian (132 AD). Celsius (25 BC-50 AD) was the first to give a detailed description of two surgical techniques for uncircumcision in his De medicina libri octo. Subsequent works, for example by Galen (131-200 AD) and Paulus Aeginata in the seventh century, only contained a repetition of these methods without presenting any new aspects. Ambroise Paré gave a new impetus in the sixteenth century, suggesting the insertion of a catheter into the distal urethra to guarantee free passage of urine during postoperative healing. In this past century, Johann Friedrich Dieffenbach was the first to dedicate a whole chapter to the problem of "posthioplastice" in a modern textbook of plastic surgery. Almost no written documents exist of uncircumcision during the Nazi era; nevertheless, surgical treatment seemed to be widespread as every circumcised man was in danger of being denounced as a Jew. Personal reports of patients and doctors performing surgical restoration of the prepuce are presented. Nowadays, reports on surgical foreskin restoration are still rare and alternative methods of nonsurgical skin-expansion have become more common. Several organizations were founded in America against routine infant circumcision and give advice to foreskin restoration seekers.


SSTR2A expression in medullary thyroid carcinoma is correlated with longer survival.
Medullary thyroid carcinoma (MTC) derives from the parafollicular C-cells of the thyroid gland. Somatostatin receptors (SSTRs) are expressed in various neuroendocrine tumours including MTC. The aim of this study was to evaluate SSTR2A as a prognostic factor for MTC, to study distribution of SSTR2A expression within tumours and to compare expression of SSTR2A between primary tumours and corresponding lymph node metastases. Patients who underwent surgery between 1988 and 2014 for MTC from five tertiary referral centres in The Netherlands were included. In total, primary tumours of 114 patients and lymph node metastases of 34 patients were analysed for expression of SSTR2A using a tissue microarray, and correlated with clinicopathological variables and survival. The mean age of patients was 45.5 years (SD 16.2), 55 patients were male (49.5%). Primary tumours of 58 patients (50.9%) showed SSTR2A expression. In multivariate Cox-regression analysis, SSTR2A positivity correlated independently with better overall survival (OS) (HR 0.3; 95% CI 0.1-1.0). In stage IV MTC patients, 10-year survival rates for SSTR2A-negative and positive patients were 43% and 96%, respectively. In 53.9% of patients with lymph node metastases, expression in primary tumour and lymph node metastases differed. SSTR2A expression is correlated with longer OS in MTC, especially for stage IV patients, suggesting that SSTR2A expression might be a useful prognostic factor in MTC. The SSTR2A status of the primary MTC does not predict expression in lymph node metastases.


The ocular aspects of bypass surgery of the carotid artery.
A new neurosurgical procedure, anastomosis of the superficial temporal artery to the middle cerebral artery, is being performed with increasing frequency. Over a 7-year period (1971 through 1977), 121 patients have undergone this operation in our institution. Ophthalmodynamometry may be of great help in identifying occlusion of an internal carotid artery. Progressing venous stasis retinopathy and ischemic orbital pain are believed to be indications for bypass surgery of the carotid artery.


Integrated thermal stabilization of a microring modulator.
A defect-enhanced silicon photodiode and heater are integrated with and used to thermally stabilize a microring modulator. These optoelectronic components are interfaced with external control circuitry to create a closed-looped feedback system for thermally stabilizing the microring modulator. The thermal stabilization system enables the microring modulator to provide error-free 5-Gb/s modulation while being subjected to thermal fluctuations that would normally render it inoperable.


Airway environment drives the selection of quorum sensing mutants and promote Staphylococcus aureus chronic lifestyle.
Staphylococcus aureus is a predominant cause of chronic lung infections. While the airway environment is rich in highly sialylated mucins, the interaction of S. aureus with sialic acid is poorly characterized. Using S. aureus USA300 as well as clinical isolates, we demonstrate that quorum-sensing dysfunction, a hallmark of S. aureus adaptation, correlates with a greater ability to consume free sialic acid, providing a growth advantage in an air-liquid interface model and in vivo. Furthermore, RNA-seq experiment reveals that free sialic acid triggers transcriptional reprogramming promoting S. aureus chronic lifestyle. To support the clinical relevance of our results, we show the co-occurrence of S. aureus, sialidase-producing microbiota and free sialic acid in the airway of patients with cystic fibrosis. Our findings suggest a dual role for sialic acid in S. aureus airway infection, triggering virulence reprogramming and driving S. aureus adaptive strategies through the selection of quorum-sensing dysfunctional strains.


Effect of graded Nrf2 activation on phase-I and -II drug metabolizing enzymes and transporters in mouse liver.
Nuclear factor erythroid 2-related factor 2 (Nrf2) is a transcription factor that induces a battery of cytoprotective genes in response to oxidative/electrophilic stress. Kelch-like ECH associating protein 1 (Keap1) sequesters Nrf2 in the cytosol. The purpose of this study was to investigate the role of Nrf2 in regulating the mRNA of genes encoding drug metabolizing enzymes and xenobiotic transporters. Microarray analysis was performed in livers of Nrf2-null, wild-type, Keap1-knockdown mice with increased Nrf2 activation, and Keap1-hepatocyte knockout mice with maximum Nrf2 activation. In general, Nrf2 did not have a marked effect on uptake transporters, but the mRNAs of organic anion transporting polypeptide 1a1, sodium taurocholate cotransporting polypeptide, and organic anion transporter 2 were decreased with Nrf2 activation. The effect of Nrf2 on cytochrome P450 (Cyp) genes was minimal, with only Cyp2a5, Cyp2c50, Cyp2c54, and Cyp2g1 increased, and Cyp2u1 decreased with enhanced Nrf2 activation. However, Nrf2 increased mRNA of many other phase-I enzymes, such as aldo-keto reductases, carbonyl reductases, and aldehyde dehydrogenase 1. Many genes involved in phase-II drug metabolism were induced by Nrf2, including glutathione S-transferases, UDP- glucuronosyltransferases, and UDP-glucuronic acid synthesis enzymes. Efflux transporters, such as multidrug resistance-associated proteins, breast cancer resistant protein, as well as ATP-binding cassette g5 and g8 were induced by Nrf2. In conclusion, Nrf2 markedly alters hepatic mRNA of a large number of drug metabolizing enzymes and xenobiotic transporters, and thus Nrf2 plays a central role in xenobiotic metabolism and detoxification.


[Morphological manifestations of acute stress reactions in the adrenal cortex of hypokinetic rats].
Adrenal changes in response to an acute stress-effect (5 h-immobilization stress) were investigated in female rats exposed to hypokinesia for 3 months. The rate of delipoidization in the adrenal cortex increased in the rats exposed to an acute stress after short-term (1-2 weeks) hypokinesia. The process of delipoidization did not advance in the rats exposed to an acute stress in the course of prolonged (2-3 months) hypokinesia. This does not yet prove the lack of the stress reaction but gives evidence that during prolonged hypokinesia the adrenals develop the capacity to react to an additional effect without the entire complex of morphological signs typical of an acute stress reaction. The immobilization test used to assess the state of the adrenal cortex has shown that it does not deteriorate even during 3-month hypokinesia (in this study).


Examination of the relationship between post-traumatic growth and social support and life satisfaction in wounded veterans in Turkey.
This study aims to examine the effects of social support and life satisfaction on post-traumatic growth in wounded veterans in Turkey. 1175 wounded veterans participated in the cross-sectional study. The sociodemographic questionnaire, the Post-Traumatic Growth Inventory, the Multidimensional Scale of Perceived Social Support, and the Satisfaction with Life Scale were used to collect data. Data were analyzed via SPSS. The results of the study show that the mean age is 43.04 ± 7.36. The participants are low in education and socioeconomic level. The mean of the Post-Traumatic Growth Inventory score is 43.44 ± 21.18, the mean of the Multidimensional Scale of Perceived Social Support score is 50.01 ± 15.23, and the mean of the Satisfaction With Life Scale score is 9.86 ± 4.04. The results of the study show that there is a positive relationship between social support and life satisfaction and post-traumatic growth. It can be suggested to develop psychosocial intervention strategies and service delivery systems to promote the life satisfaction, social support mechanisms of wounded veterans.


Theoretical insights into surface-phase transition and ion competition during alkali ion intercalation on the Cu4Se4 nanosheet.
The development of stable and efficient electrode materials is imperative and also indispensable for further commercialization of sodium/potassium-ion batteries (SIBs/PIBs) and new detrimental issues such as proton intercalation arise when utilizing aqueous electrolytes. Herein the electrochemical performance of the Cu4Se4 nanosheet was determined for both organic and aqueous SIBs and PIBs. By means of density functional theory calculation, Na+, K+ and H+ intercalations onto both sides of the Cu4Se4 nanosheet were revealed. The Cu4Se4 nanosheet well maintains its metallic electronic conductivity and the changes in lateral lattice parameters are within 4.66% during the whole Na+/K+ intercalation process for both SIBS and PIBs. The theoretical maximum Na/K storage capacity of 188.07 mA h g-1 can be achieved by stabilized second-layer adsorption of Na+/K+. The migration barriers of Na and K atoms on the Cu4Se4 nanosheet are 0.270 and 0.173 eV, respectively. It was discovered that Na/K- intercalation in the first layer is accompanied by a first-order surface phase transition, resulting in an intercalation voltage plateau of 0.659/0.756 V, respectively. The region of the two-surface phase coexistence for PIBs, is shifted toward a lower coverage when compared with that for SIBs. The partially protonated Cu4Se4 nanosheet (HxCu4Se4, x ≤ 10/9) was revealed to be structurally and thermodynamically stable. While the partially protonated Cu4Se4 nanosheet is favorable in acidic electrolytes (pH = 0) when protons and Na/K ions compete, we showed that Na+/K+ intercalated products may be preferred over H+ at low coverages in alkali electrolyte (pH = 14). However, the proton intercalation substantially decreases the battery capacity in aqueous SIBs and PIBs. Our work not only identifies the promising performance of Cu4Se4 nanosheets as an electrode material of SIBs and PIBs, but also provides a computational method for aqueous metal-ion batteries.


The management of radioactive wastes resulting from emergency situations on land and sea.
The term 'emergency' applies to unplanned events that have generated, or had the potential to generate, radioactive wastes that could not be handled locally. Reports of past emergencies form the basis for discussion of future trends in emergency situations, the factors that influence waste management, the technology of waste management and personnel requirements. The number of past emergencies has been small and these have not increased the volume of radioactive wastes significantly. The majority of future emergencies are likely to be associated with reactors as they have been in the past. Little change in the nature of the waste is expected, although the greater use of separated plutonium may create some new problems. The factors that influence emergency waste management fall into two categories, those that are concerned with the location of the emergency event, and those concerned with the characteristics of the wastes. As in the past, it can be expected that in most emergencies proven waste management techniques will be adapted to suit the local circumstances but there is a need for simpler and more economic methods. Trained personnel and comprehensive planning are cited as key factors in successful waste management operations.


Cell cycle-related kinase mediates viral-host signalling to promote hepatitis B virus-associated hepatocarcinogenesis.
Androgen receptor (AR) signalling contributes to male predominance in hepatocellular carcinoma (HCC), which is more pronounced in HBV-endemic areas. Cell cycle-related kinase (CCRK) is essential for AR-induced hepatocarcinogenesis but its molecular function in HBV-associated HCC remains obscure. To determine the molecular function of CCRK in HBV-associated HCC. Transcriptional regulation was assessed by chromatin immunoprecipitation, promoter mutation and luciferase reporter assays. Hepatocellular proliferation and tumourigenesis were examined by colony formation, soft agar assays and using HBV X protein (HBx) transgenic mice with low-dose exposure to diethylnitrosamine. Protein expressions were examined in clinical samples and correlated with patient survival by log-rank Mantel-Cox test. Overexpression of CCRK, but not its kinase-defective mutant, activated β-catenin/T cell factor signalling through phosphorylation of glycogen synthase kinase-3β (GSK-3β) at Ser9, led to upregulation of AR transcriptional activity and, subsequently, expression of HBx. The viral transactivator in turn induced CCRK expression through enhanced AR signalling, thus forming a positive regulatory loop. RNA interference silencing of CCRK, which suppressed the CCRK/GSK-3β/β-catenin/AR regulatory loop, significantly suppressed HBx-induced hepatocellular proliferation (p=0.001) and transformation (p<0.001) and remarkably reduced >80% diethylnitrosamine-mediated hepatocarcinogenesis in HBx transgenic mice. Finally, patients with HBV-associated HCC with concordant overexpression of CCRK, GSK-3β phosphorylation at Ser9, active dephosphorylated β-catenin and AR phosphorylation at Ser81 had poorer overall (HR=31.26, p<0.0001) and disease-free (HR=3.60, p<0.01) survival rates. Our findings highlight the critical role of CCRK in a self-reinforcing circuitry that regulates HBV-associated hepatocarcinogenesis. Further characterisation of this intricate viral-host signalling may provide new prognostic biomarkers and therapeutic targets for HCC treatment.


Pharmacological PPARγ modulation regulates sebogenesis and inflammation in SZ95 human sebocytes.
The nuclear receptor peroxisome proliferator-activated receptor gamma (PPARγ) controls the expression of genes involved in the regulation of lipid and glucose metabolism, cell proliferation/differentiation as well as inflammatory pathways. Pivotal studies in human sebocytes and isolated sebaceous glands have raised the interesting possibility that compounds acting on PPARγ can modulate sebaceous lipids and inflammation and, as such, may be useful in the treatment of acne. To investigate the role of this receptor in the regulation of lipid synthesis, proliferation and inflammation, we used the SZ95 sebaceous gland cell line stimulated with insulin. In sebocytes, insulin signaling activated the phosphatidylinositide 3-kinase-Akt (PI3K/Akt) and mammalian target of rapamycin (mTOR) pathways, which, in turn, induced high protein/lipid synthesis, increased cell growth and proliferation as well as inflammation. As regards lipogenesis, insulin initially stimulated the formation of unsaturated lipids and then the neosynthesis of lipids. The results showed, that the modulation of PPARγ, counteracted the insulin-induced altered lipogenesis, evident through a decrease in gene expression of key enzymes responsible for the synthesis of fatty acids, and through a reduction of lipid species synthesis analyzed by Oil/Nile Red staining and GC-MS. PPARγ modulation also regulated the insulin-induced proliferation, inhibiting the cell cycle progression and p21WAF1/CIP1 (p21) protein reduction. Moreover, the expression of inflammatory cytokines, induced by insulin or lipopolysaccharide (LPS), was down-modulated. In PPARγ-deficient cells or in the presence of GW9662 antagonist, all these observed effects were abolished, indicating that PPARγ activation plays a role in regulating alteration of lipogenesis, cell proliferation and inflammatory signaling. We demonstrated that selective modulation of PPARγ activity is likely to represent a therapeutic strategy for the treatment of acne.


Influence of diets containing casein, soy isolate, and soy concentrate on serum cholesterol and lipoproteins in middle-aged volunteers.
Fifty-seven healthy volunteers (mean age 46 yr) were fed for 45 days on diets containing 16% of energy as protein, 35% as fat (polyunsaturated/saturated fat ratio = 0.5) and about 375 mg cholesterol/day. Of the protein in the diets 60% was provided as caseinate, as soy protein isolate, or as soy protein concentrate. After a control period of 17 days during which all the subjects received the casein diet, 17 subjects continued on this diet for the next 28 days (test period), 20 subjects switched to the soy isolate diet, and the remaining 20 subjects switched to the soy concentrate diet. Serum cholesterol levels at the end of the control period were 207 +/- 36, 205 +/- 40, and 199 +/- 35 mg/dl (mean +/- SD) for the casein, isolate, and concentrate groups, respectively. Mean changes over the test period were -2 +/- 10, -8 +/- 12, and +1 +/- 10 mg/dl, respectively. Compared with the casein diet, the isolate diet caused a small, nonsignificant decline in both serum total cholesterol and low-density lipoprotein cholesterol (-6.5 mg/dl) and an increase in high-density lipoprotein cholesterol (+5.8 mg/dl) (p less than 0.05). These effects may have been more obvious if there had been no differences between groups in weight loss. No correlation was found between the response and the initial cholesterol level. No differences in lipoprotein composition were found between the casein and soy concentrate groups. Our data suggest that soy protein preparations do not have dramatic effects on the serum total cholesterol concentration in healthy subjects. However, pure soy protein might have some beneficial effect on the distribution of cholesterol over the lipoproteins. The lack of effect of the less refined soy protein concentrate suggests that the dietary fiber and other nonprotein components of soy concentrate do not have, at least in the short-term, a favorable effect on serum cholesterol and lipoproteins in healthy adults.


Allergy, atopy, and cancer: a prospective study of the 1981 Busselton cohort.
The associations among certain allergic disorders, atopy upon skin-prick testing, and specific cancers were evaluated in a prospective study. Information regarding history of asthma and hay fever was collected by questionnaire from 3,308 cancer-free participants in the 1981 Busselton Health Survey. A subset of 1,005 participants also underwent skin-prick testing. The cohort was followed for a new diagnosis of cancer or death until the end of 1999. Cox proportional hazards regression analysis was used to estimate adjusted hazard ratios (relative risks) for breast, prostate, colorectal, lung, and hematologic cancers and melanoma. Having a skin reaction to house dust mites nearly tripled the risk of prostate cancer (relative risk = 2.90, 95% confidence interval: 1.26, 6.68). History of asthma and hay fever were associated with a trend toward a reduced risk of colorectal cancer and increased risk of leukemia, but these results were not statistically significant. Hay fever was associated with melanoma risk in men but not in women. No association was found between breast and lung cancers and allergic disorders or atopy.


The effects of treatment on the direct costs of diabetes.
Treatment of diabetic complications consumes health care resources. Intensive therapy was shown by the Diabetes Control and Complications Trial (DCCT) to avert complications. Economic analyses and models have been used to evaluate the cost-effectiveness of intensive therapy for people with type 1 and type 2 diabetes. An economic analysis of the DCCT estimated the cost of intensive therapy to be two to three times greater than that of conventional therapy. In contrast, an economic model predicts that intensive therapy, as compared with conventional therapy, could reduce blindness from 34 to 20% or by 41%, end-stage renal disease from 24 to 7% or by 71%, and lower-extremity amputations from 7 to 4% or by 43%. Although intensive therapy is more expensive, when the costs of complications are factored in, it becomes cost-effective for treatment of type 1 diabetes. Similarly, a model to evaluate the cost-effectiveness of intensive therapy for people with type 2 diabetes found that the lifetime costs of general and diabetes-related medical care would be approximately two times greater. However, the reduction in lifetime costs of complications, which would produce substantial reductions in costs of treatment, largely offsets the difference. Intensive therapy for type 1 and type 2 diabetes may be more expensive than conventional therapy, but from an economic perspective, it is comparable in cost to pharmacological therapies for people with hypertension and hypercholesterolemia. From a health system viewpoint, intensive therapy represents a fruitful long-term financial investment.


Myopathic dropped head syndrome: a syndrome of mixed aetiology.
We report two patients with myopathic dropped head syndrome, a rare and interesting neuromuscular syndrome characterised by a predominant weakness of the neck extensor muscles. The first patient, a middle aged Chinese man, presented with progressive weakness of neck extension but his clinical course later stabilised despite a lack of response to corticosteroids. Muscle biopsy revealed a necrotising myopathy with no evidence of inflammation. This patient supports the existence of an idiopathic restricted non-inflammatory myopathy, a so called isolated neck extensor myopathy syndrome which is recognised to pursue a less progressive, more benign course. Our second patient had histopathological evidence for polymyositis; there was a favourable response to steroids. Our cases underscore the fact that there may be a spectrum of pathological processes associated with the myopathic dropped head syndrome ranging from non-inflammatory muscle necrosis to a full blown inflammatory myositis.


3-Methylmethcathinone Intoxication: Discrepancies Between Blood Concentrations and Clinical Outcomes-A Short Communication.
The absence of a correlation between the blood concentration of 3-methylmethcathinone (3-MMC) and clinical outcomes in intoxication cases has been attributed to stability issues. Indeed, a loss of more than 50%, 70%, and even 95% of 3-MMC in whole blood after 2 weeks of storage at 20°C, 4°C, and room temperature, respectively, has been reported in the past. Here, the authors report the case of a 43-year-old man who was hospitalized with generalized convulsive status epilepticus related to 3-MMC use with a plasma concentration of 9600 ng/mL (delay between sampling and analysis <72 hours). The stability of 3-MMC was evaluated in several biological specimens. Three quality control samples (human plasma, whole blood, and postmortem blood) spiked with 3-MMC were stored at -20°C and 4°C for 14 days. The initial analysis was performed on day 1 to establish the initial concentration of 3-MMC in each specimen type, and the samples were divided into 2 aliquots for storage under both conditions. Further analyses were performed on days 7 and 14 for each specimen, and the results were compared with those obtained on day 1. 3-MMC appeared relatively stable in whole and postmortem blood when stored at -20°C for 1 week, with losses of <3% in both matrices (0% and 2.5%, respectively). At +4°C, 3-MMC losses ranged from 25% to 53%. These results differ from others reported in the literature. Hence, it may be hypothesized that other elements should be considered to explain the discrepancy between the concentration and toxicity pointed out by the Toxicology community, especially the development of tolerance.


Wavelet packets-based blind watermarking for medical image management.
The last decade has witnessed an explosive use of medical images and Electronics Patient Record (EPR) in the healthcare sector for facilitating the sharing of patient information and exchange between networked hospitals and healthcare centers. To guarantee the security, authenticity and management of medical images and information through storage and distribution, the watermarking techniques are growing to protect the medical healthcare information. This paper presents a technique for embedding the EPR information in the medical image to save storage space and transmission overheads and to guarantee security of the shared data. In this paper a new method for protecting the patient information in which the information is embedded as a watermark in the discrete wavelet packet transform (DWPT) of the medical image using the hospital logo as a reference image. The patient information is coded by an error correcting code (ECC), BCH code, to enhance the robustness of the proposed method. The scheme is blind so that the EPR can be extracted from the medical image without the need of the original image. Therefore, this proposed technique is useful in telemedicine applications. Performance of the proposed method was tested using four modalities of medical images; MRA, MRI, Radiological, and CT. Experimental results showed no visible difference between the watermarked and the original image. Moreover, the proposed watermarking method is robust against a wide range of attacks such as JPEG coding, Gaussian noise addition, histogram equalization, gamma correction, contrast adjustment, and sharpen filter and rotation.


Technology Use Among Youth While Operating Farm Equipment.
Distracted driving by using modern technologies has been linked with increased risk of injuries, however, the current research has focused primarily on the use of technology while operating roadway vehicles. The purpose of this study was to explore the use of technology while operating farm equipment among members of an agricultural youth organization (Pennsylvania FFA Association members). An online survey was used to examine the use of technology while operating a tractor, ATV/UTV, skid steer, and truck/car. Survey responses with 10 types of technology use were analyzed in relation to a wide variety of factors including frequency of use, safety beliefs, safety concern, and seeing others using technology. One hundred forty-three youth completed the questionnaire in 2017. Eighty-eight percent of youth surveyed used technology to some extent while operating farm equipment. Self-reported technology use was most influenced by safety beliefs and seeing others using technology while operating farm equipment regardless of the farm equipment type. Findings from this study provide insights into factors behind technology use while engaged in farm work-related driving by youth.


Late Imaging with [1-(11)C]Acetate Improves Detection of Tumor Fatty Acid Synthesis with PET.
Tumors are often characterized by high levels of de novo fatty acid synthesis. The kinetics of acetate incorporation into tricarboxylic acid cycle intermediates and into lipids suggest that detection of tumors with [1-(11)C]acetate PET could be improved by imaging at later time points. The uptake and metabolism of [1-(11)C], [1-(13)C], and [1-(14)C]acetate were measured in mouse prostate and lung cancer models to investigate the time course of (11)C label incorporation into tumor metabolites. Radioactivity in the lipid fraction, as compared with the aqueous fraction, in extracts of C4-2B human prostate xenografts peaked at 90 min after [1-(14)C]acetate injection, which coincided with peak (13)C label incorporation into the fatty acids palmitate and stearate. Contrast between the tumor and tissues, such as blood and muscle, increased in PET images acquired over a period of 120 min after [1-(11)C]acetate injection, and Patlak plots were linear from 17.5 min after injection. Similar results were obtained in a genetically engineered K-ras(G12D); p53(null) lung cancer model, in which the mean tumor-to-lung ratio at 90 min after [1-(14)C]acetate injection was 4.4-fold higher than at 15 min. These findings suggest that when imaging de novo fatty acid synthesis with [1-(11)C]acetate it is preferable to measure uptake at later time points, when the effects of perfusion and (11)C incorporation into tricarboxylic acid cycle intermediates and bicarbonate are declining. The data presented here suggest that future clinical PET scans of tumors should be acquired later than 30 min, when tracer accumulation due to de novo fatty acid synthesis prevails.


Stereoretentive Ring-Opening Metathesis Polymerization to Access All-cis Poly(p-phenylenevinylene)s with Living Characteristics.
Poly(p-phenylenevinylene)s (PPVs), a staple of the conductive polymer family, consist of alternating alkene and phenyl groups in conjugation. The physical properties of this organic material are intimately linked to the cis/trans configuration of the alkene groups. While many synthetic methods afford PPVs with all-trans stereochemistry, very few deliver the all-cis congeners. We report herein a synthesis of all-cis PPVs with living characteristics via stereoretentive ring-opening metathesis polymerization (ROMP). Exquisite catalyst control allows for the preparation of homopolymers or diblock copolymers with perfect stereoselectivity, narrow dispersities, and predictable average molar masses. All-cis PPVs can then serve as light-responsive polymers through clean photoisomerization of the stilbenoid units.


Limited activation of the intrinsic apoptotic pathway plays a main role in amyloid-β-induced apoptosis without eliciting the activation of the extrinsic apoptotic pathway.
Amyloid-β (Aβ), a main pathogenic factor of Alzheimer's disease (AD), induces apoptosis accompanied by caspase activation. However, limited caspase activation and the suppression of the intrinsic apoptotic pathway (IAPW) are frequently observed upon Aβ treatment. In this study, we investigated whether these suppressive effects of Aβ can be overcome; we also examined the death-related pathways. Single treatments of cells with Aβ42 for up to 48 h barely induced caspase activation. In cells treated with Aβ42 twice for 2 h followed by 22 h (2+22 h) or for longer durations, the apoptotic protease activating factor-1 (Apaf-1) apoptosome was formed and caspases-3 and -9 were activated to a certain extent, suggesting the activation of the IAPW. However, the Aβ42-induced activation of the IAPW differed from that induced by treatment with other agents, such as staurosporine (STS) in that lower amounts of cytochrome c were released from the mitochondria, the majority of procaspase-9 in the Apaf-1 complex was not processed and caspase-3 was activated to a lesser extent in the peptide-treated cells. Thus, it seemed that the IAPW was not fully activated by Aβ42. The 30- and 41/43-kDa fragments derived from procaspase-8 were detected, which appear to be produced through the IAPW without death-inducing signaling-complex (DISC) formation, a key feature of the extrinsic apoptotic pathway (EAPW). Bid cleavage was observed only after caspase-3 activity reached its maximal levels, suggesting that the cleavage may contribute in a limited capacity to the amplification process of the IAPW in the Aβ-treated cells. Taken together, our data suggest that the IAPW, albeit functional only to a limited extent, plays a major role in Aβ42-induced apoptosis without the EAPW.


Improvements in Nanoparticle Tracking Analysis To Measure Particle Aggregation and Mass Distribution: A Case Study on Engineered Nanomaterial Stability in Incineration Landfill Leachates.
Numerous nanometrology techniques have been developed in recent years to determine the size, concentration, and a number of other characteristics of engineered nanomaterials (ENM) in environmental matrices. Among the many available techniques, nanoparticle tracking analysis (NTA) can measure individual particles to create a size distribution and measure the particle number. Therefore, we explore the possibility to use these data to calculate the particle mass distribution. Additionally, we further developed the NTA methodology to explore its suitability for analysis of ENM in complex matrices by measuring ENM agglomeration and sedimentation in municipal solid waste incineration landfill leachates over time. 100 nm Au ENM were spiked into DI H2O and synthetic and natural leachates. We present the possibility of measuring ENM in the presence of natural particles based on differences in particle refractivity indices, delineate the necessity of creating a calibration curve to adjust the given NTA particle number concentration, and determine the instruments linear range under different conditions. By measuring the particle size and the particle number distribution, we were able to calculate the ENM mass remaining in suspension. By combining these metrics together with transmission electron microscopy (TEM) analyses, we could assess the extent of both homo- and heteroagglomeration as well as particle sedimentation. Reporting both size and mass based metrics is common in atmospheric particle measurements, but now, the NTA can give us the possibility of applying the same approach also to aqueous samples.


Body piercing and tattoo: awareness of health related risks among 4,277 Italian secondary school adolescents.
The awareness of health risks associated with body art among secondary school pupils has never previously been studied in depth. A large sample of secondary school adolescents from the Veneto Region (North East Italy) were investigated in order to inform health education programs. 6 public secondary schools from each of the 7 Provinces of the Veneto Region were selected. All students attending the 1st, 3rd, and 5th school years were surveyed by an anonymous self administered questionnaire on their perception of health risks related to body art and other explanatory variables. Logistic regression analysis was employed, reporting adjusted Odds Ratios (OR) with 95% Confidence Interval (CI). Among 4,277 available students (aged 14-22 years), boys were consistently: less knowledgeable of infectious diseases related to body art (OR = 0.78; CI: 0.66, 0.94), less likely to be aware of the hygienic norms to be observed in a body art parlour (OR = 0.54; 0.44, 0.65), less likely to refer to a certified body art parlour (OR = 0.56; 0.48, 0.66), less likely to refer to a professional health care provider for complications related to body art (OR = 0.71; 0.59, 0.86). Students attending the first school year (baseline) had a lesser knowledge of body art related infectious diseases, were less likely to refer to a certified body art parlour, and to know the mandatory hygienic rules to be observed when performing body modifications. Interviewees from the provinces of Rovigo and Vicenza were less likely to be conscious of the health risks associated with body modifications, and those with tattoos were less knowledgeable about the infection risk (OR = 0.60; 0.42, 0.86) and less likely to refer to a professional health care provider in case of medical complication (OR = 0.68;0.48, 0.95). Students with piercings were less likely to refer to a certified practitioner for receiving body art (OR = 0.62; 0.50, 0.77) or therapy for medical complications (OR = 0.37; 0.29, 0.46). Health education programs should focus on males, pupils attending lower school years, living in specific Provinces of the Region, and with a positive attitude towards piercing or tattoo.


Clinical and Laboratory Findings of Middle East Respiratory Syndrome Coronavirus Infection.
There is a paucity of data regarding the differentiating characteristics of patients with laboratory-confirmed and those negative for Middle East respiratory syndrome coronavirus (MERS-CoV) in South Korea. This hospital-based retrospective study compared MERS-CoV-positive and MERS-CoV-negative patients. A total of seven positive patients and 55 negative patients with a median age of 43 years (P = 0.845) were included. No statistical differences were observed with respect to their sex and the presence of comorbidities. At the time of admission, headache (28.6% vs. 3.6%; odds ratio [OR], 10.60; 95% confidence interval [CI], 1.22-92.27), myalgia (57.1% vs. 9.1%; OR, 13.33; 95% CI, 2.30-77.24), and diarrhea (57.1% vs. 14.5%; OR, 7.83; 95% CI, 1.47-41.79) were common among MERS-CoV-positive patients. MERS-CoV-positive patients were more likely to have a low platelet count (164 ± 76.57 vs. 240 ± 79.87) and eosinophil (0.27 ± 0.43 vs. 2.13 ± 2.01; P = 0.003). Chest radiography with diffuse bronchopneumonia was more frequent in MERS-CoV-positive patients than in negative patients (100% vs. 62.5%; P = 0.491). The symptoms of headache, myalgia, and diarrhea, as well as laboratory characteristics, including low platelet counts and eosinophil, and chest X-ray showing diffuse bronchopneumonia might enhance the ability to detect patients in South Korea infected with MERS-CoV.


Reducing ammonia emissions and volatile fatty acids in poultry litter with liquid aluminum chloride.
This study was a pen trial in which the effects of adding different rates of liquid aluminum chloride (AlCl(3)) on litter pH, total volatile fatty acids (VFAs), and ammonia (NH(3)) fluxes was evaluated. Liquid AlCl(3) treatments used in this study were sprayed on the rice hull surface at rates of 100 g, 200 g, and 300 g liquid AlCl(3)/kg rice hulls; untreated rice hulls served as controls. Litter pH, total VFAs, and NH(3) fluxes were all lowered (P< 0.05) by all of the liquid AlCl(3) treatments compared with controls during certain times of the 5 week study. However, there were no significant differences among treatments on litter pH at the end of the study (from 3 to 5 weeks) or NH(3) fluxes at beginning of the study (0 to 3 weeks). Total VFAs were reduced 16 %, 29 %, and 53 % by 100 g liquid AlCl(3)/kg rice hulls, 200 g liquid AlCl(3)/kg rice hulls, and 300 g liquid AlCl(3)/kg rice hulls, respectively. Liquid AlCl(3)additions reduced NH(3) fluxes by 35 %, 57 % and 67 %, respectively, at the low, medium and high rates. In summary, these results indicate that adding liquid aluminum chloride to rice hulls would be a useful tool in reducing the negative environmental impact of poultry litter. It should be noted that the decreased VFA production and NH(3) volatilization was chiefly associated with reduction in litter pH.


Spatially resolved scattering correlation spectroscopy using a total internal reflection configuration.
In the paper, we present a novel single particle method, named spatially resolved scattering correlation spectroscopy (SRSCS), based on a total internal reflection (TIR) configuration and strong resonance light scattering (RLS) of silver nanoparticles (AgNPs). The principle of SRSCS is similar to fluorescence correlation spectroscopy (FCS), and it is based on measuring the RLS fluctuations in a small volume due to Brownian motion of single nanoparticles. We first established a highly sensitive SRSCS system. In the SRSCS system, a millimeter-scale hole is employed to efficiently separate nanoparticle scattering light from the background reflected beam, and an electron multiplying charge-coupled device (EMCCD) is used as an array detector. The SRSCS system was successfully used for detection and imaging of single AgNPs in solution. Furthermore, we developed the model of SRSCS according to the FCS method and systematically investigated the effects of certain factors such as particle concentration, viscosity of the solution, hardware and software binning and accumulation time on SRSCS measurements using AgNPs as a model sample. A series of calibration experiments were conducted, and the experimental data obtained were in good agreement with the SRSCS model. This new method is multiplexing, spatially resolved, and free of photobleaching and may become a useful method for study on heterogeneous systems, such as the motion of proteins on the cell membrane.