[Bioacoustic of the advertisement call of Ceratophrys cranwelli (Anura: Ceratophryidae)].
The advertisement call plays an important role in the life history of anuran amphibians, mainly during the breeding season. Call features represent an important character to discriminate species, and sound emissions are very effective to assure or reinforce genetic incompatibility, especially in the case of sibling species. Since frogs are ectotherms, acoustic properties of their calls will vary with temperature. In this study, we described the advertisement call of C. cranwelli, quantifying the temperature effect on its components. The acoustic emissions were recorded during 2007 using a DAT record Sony TCD-100 with stereo microphone ECM-MS907 Sony and tape TDK DAT-RGX 60. As males emit their calls floating in temporary ponds, water temperatures were registered after recording the advertisement calls with a digital thermometer TES 1300+/-0.1 degreeC. Altogether, 54 calls from 18 males were analyzed. The temporal variables of each advertisement call were measured using oscillograms and sonograms and the analyses of dominant frequency were performed using a spectrogram. Multiple correlation analysis was used to identify the temperature-dependent acoustic variables and the temperature effect on these variables was quantified using linear regression models. The advertisement call of C. cranwelli consists of a single pulse group. Call duration, Pulse duration and Pulse interval decreased with the temperature, whereas the Pulse rate increased with temperature. The temperature-dependent variables were standardized at 25 degreeC according to the linear regression model obtained. The acoustic variables that were correlated with the temperature are the variables which emissions depend on laryngeal muscles and the temperature constraints the contractile properties of muscles. Our results indicated that temperature explains an important fraction of the variability in some acoustic variables (79% in the Pulse rate), and demonstrated the importance of considering the effect of temperature in acoustic components. The results suggest that acoustic variables show geographic variation to compare data with previous works.


Annexin A2-S100A10 heterotetramer, a novel substrate of thioredoxin.
The binding of plasminogen activators and plasminogen to the cell surface results in the rapid generation of the serine protease plasmin. Plasmin is further degraded by an autoproteolytic reaction, resulting in the release of an angiostatin, A61 (Lys78-Lys468). Previously, we demonstrated that the annexin A2-S100A10 heterotetramer (AIIt) stimulates the release of A61 from plasmin by promoting the autoproteolytic cleavage of the Lys468-Gly469 bond and reduction of the plasmin Cys462-Cys541 disulfide (Kwon, M., Caplan, J. F., Filipenko, N. R., Choi, K. S., Fitzpatrick, S. L., Zhang, L., and Waisman, D. M. (2002) J. Biol. Chem. 277, 10903-10911). Mechanistically, it was unclear if AIIt promoted a conformational change in plasmin, resulting in contortion of the plasmin disulfide, or directly reduced the plasmin disulfide. In the present study, we show that AIIt thiols are oxidized during the reduction of plasmin disulfides, establishing that AIIt directly participates in the reduction reaction. Incubation of HT1080 cells with plasminogen resulted in the rapid loss of thiol-specific labeling of AIIt by 3-(N-maleimidopropionyl)biocytin. The plasminogen-dependent oxidation of AIIt could be attenuated by thioredoxin. Thioredoxin reductase catalyzed the transfer of electrons from NADPH to the oxidized thioredoxin, thus completing the flow of electrons from NADPH to AIIt. Therefore, we identify AIIt as a substrate of the thioredoxin system and propose a new model for the role of AIIt in the redox-dependent processing of plasminogen and generation of an angiostatin at the cell surface.


The trigeminal depressor response: a novel vasodepressor response originating from the trigeminal system.
Electrical stimulation within discrete sites of the spinal trigeminal complex in anesthetized or decerebrated rabbits results in arterial hypotension, often over 50 mm Hg, bradycardia of up to 60 beats/min, apnea, and gastric hypermotility, collectively termed the trigeminal depressor response (TDR). The threshold for the TDR is less than or equal to 10 muA and is graded up to 3-6 times threshold. It can only be elicited by trains of stimuli of low frequency (0.5-20 Hz); at 50 Hz the response disappears or becomes pressor. The bradycardia is only abolished by bilateral vagotomy combined with beta-adrenergic blockade, and thus results from combined excitation of cardio-vagal and inhibition of cardiac sympathetic nerves. The hypotension is unassociated with changes in cardiac output, does not change after blockade of the bradycardia, but disappears after alpha-adrenergic blockade and hence is entirely attributable to inhibition of ongoing sympathetic vasoconstrictor nerve activity. Below threshold stimulation the TDR can only be elicited from the root entry zone of the Vth nerve, from dorsal portions of the spinal tract of the Vth nerve, and to portions of the nucleus of the spinal tract, notably the nucleus caudalis. A TDR of reduced magnitude can also be elicited by low frequency stimulation of numerous branches of the Vth nerve arising from all three divisions and including the supra- and infra-orbital, the inferior alveolar, and lingual nerves. Bilateral electrolytic lesions of the nucleus tractus solitarii at the obex, with complete abolition of baroreceptor reflexes from carotid sinus and aortic depressor nerves, fail to alter the TDR elicited from the brain or from branches of the Vth nerve, or the vasodepressor responses elicited by electrical stimulation of the central ends of the IXth and Xth cranial nerves transescted distal to the branches of baro-receptor nerves. In contrast, caudal lesions of the trigeminal complex abolish the TDR elicited from brain and Vth nerve and substantially reduces the vasodepressor responses from the IXth and Xth nerves, without altering baroreceptor reflexes. We conclude that the TDR represents a heretofore recognized vasodepressor response dependent upon the spinal trigeminal complex which is at least in part anatomically distinct from pathways subserving arterial baroreceptor and somatic vasodepressor reflexes. The TDR can be reflexly elicited from widely distributed but yet unidentified receptors innervated by branches of the Vth and of the IXth and Xth cranial nerves other than those innervating arterial baroreceptors. It is of unknown function, but may be related to pain mechanisms.


Bovine papillomavirus with a mutation in the E2 serine 301 phosphorylation site replicates at a high copy number.
The E2 open reading frame of bovine papillomavirus type 1 (BPV-1) encodes at least three proteins with transcriptional regulatory properties. The full-length E2 open reading frame encodes a transcriptional transactivator, and the 3' region encodes two smaller polypeptides that repress E2-mediated transactivation. The full-length gene product is also required for viral DNA replication. We have demonstrated that the BPV-1 E2 polypeptides are phosphorylated primarily on two serine residues at a site adjacent to the carboxy-terminal DNA binding domain, which is common to all three E2 proteins (A. A. McBride, J. B. Bolen, and P. M. Howley, J. Virol. 63:5076-5085, 1989). These serine residues, at amino acid positions 298 and 301, were substituted with alanine residues in the context of the entire BPV-1 genome. The mutated BPV-1 genomes were introduced into rodent cell lines and assayed for focus formation, viral gene expression, and extrachromosomal viral DNA replication. Viral DNAs containing the E2 serine-to-alanine substitution mutants transformed C127 cells with efficiencies comparable to that of wild-type BPV-1. However, the viral genome containing the serine-to-alanine substitution at position 301 of the E2 polypeptide replicated to a copy number 20-fold higher than that of wild-type DNA.


Retrogenic ICOS Expression Increases Differentiation of KLRG-1hiCD127loCD8+ T Cells during Listeria Infection and Diminishes Recall Responses.
Following T cell encounter with Ag, multiple signals are integrated to collectively induce distinct differentiation programs within Ag-specific CD8(+) T cell populations. Several factors contribute to these cell fate decisions, including the amount and duration of Ag, exposure to inflammatory cytokines, and degree of ligation of cosignaling molecules. The ICOS is not expressed on resting T cells but is rapidly upregulated upon encounter with Ag. However, the impact of ICOS signaling on programmed differentiation is not well understood. In this study, we therefore sought to determine the role of ICOS signaling on CD8(+) T cell programmed differentiation. Through the creation of novel ICOS retrogenic Ag-specific TCR-transgenic CD8(+) T cells, we interrogated the phenotype, functionality, and recall potential of CD8(+) T cells that receive early and sustained ICOS signaling during Ag exposure. Our results reveal that these ICOS signals critically impacted cell fate decisions of Ag-specific CD8(+) T cells, resulting in increased frequencies of KLRG-1(hi)CD127(lo) cells, altered BLIMP-1, T-bet, and eomesodermin expression, and increased cytolytic capacity as compared with empty vector controls. Interestingly, however, ICOS retrogenic CD8(+) T cells also preferentially homed to nonlymphoid organs and exhibited reduced multicytokine functionality and reduced ability to mount secondary recall responses upon challenge in vivo. In sum, our results suggest that an altered differentiation program is induced following early and sustained ICOS expression, resulting in the generation of more cytolyticly potent, terminally differentiated effectors that possess limited capacity for recall response.


A new tomato pregnane glycoside from the overripe fruits.
A new pregnane glycoside has been isolated from the overripe fruits of Cherry tomato (Mini tomato), Lycopersicon esculentum var. cerasiforme (DUNAL) ALEF. The structure was determined to be 3-O-beta-lycotetraosyl 3beta-hydroxy-5alpha-pregn-16-en-20-one on the basis of spectroscopic analysis. The seasonal variation of the tomato saponin is discussed.


Method for estimating the cooperativity length in polymers.
The problem of estimating the size of the cooperatively rearranging regions (CRRs) in supercooled polymeric melts from an analysis of the α-process in ordinary relaxation experiments is addressed. The mechanism whereby a CRR changes its configuration is viewed as consisting of two distinct steps: a reduced number of monomers reaches initially an activated state, allowing for some local rearrangement; then, the subsequent regression of the energy fluctuation may take place through the configurational degrees of freedom, thus allowing for further rearrangements on larger length scales. The latter are indeed those to which the well-known Donth's scheme refers. Local readjustments are described in the framework of a canonical formalism on a stationary ensemble of small-scale regions, distributed over all possible energy thresholds for rearrangement. Large-scale configurational changes, instead, are described as spontaneous processes. Two main regimes are envisaged, depending on whether the role played by the configurational degrees of freedom in the regression of the energy fluctuation is significant or not. It is argued that the latter case is related to the occurrence of an Arrhenian dependence of the central relaxation rate. Consistency with Donth's scheme is demonstrated, and data from the literature confirm the agreement of the two methods of analysis when configurational degrees of freedom are relevant for the fluctuation regression. Poly(n-butyl methacrylate) is chosen in order to show how CRR size and temperature fluctuations at rearrangement can be estimated from stress relaxation experiments carried out by means of an atomic force microscopy setup. Cases in which the configurational pathway for regression is significantly hindered are considered. Relaxation in poly(dimethyl siloxane) confined in nanopores is taken as an example to suggest how a more complete view of the effects of configurational constraints would be possible if direct measurements of temperature fluctuations were combined with the proposed analysis.


Uncertainty Visualization by Representative Sampling from Prediction Ensembles.
Data ensembles are often used to infer statistics to be used for a summary display of an uncertain prediction. In a spatial context, these summary displays have the drawback that when uncertainty is encoded via a spatial spread, display glyph area increases in size with prediction uncertainty. This increase can be easily confounded with an increase in the size, strength or other attribute of the phenomenon being presented. We argue that by directly displaying a carefully chosen subset of a prediction ensemble, so that uncertainty is conveyed implicitly, such misinterpretations can be avoided. Since such a display does not require uncertainty annotation, an information channel remains available for encoding additional information about the prediction. We demonstrate these points in the context of hurricane prediction visualizations, showing how we avoid occlusion of selected ensemble elements while preserving the spatial statistics of the original ensemble, and how an explicit encoding of uncertainty can also be constructed from such a selection. We conclude with the results of a cognitive experiment demonstrating that the approach can be used to construct storm prediction displays that significantly reduce the confounding of uncertainty with storm size, and thus improve viewers' ability to estimate potential for storm damage.


Corneal ulceration at an urban African hospital.
During a one-year survey 283 corneal ulcers from 274 patients were seen at St John's Eye Unit of Baragwanath Hospital. Central bacterial ulcers constituted the largest problem, and the commonest isolate in this group was Streptococcus pneumoniae. Mycotic and dendritic keratitis were relatively uncommon, while marginal catarrhal ulceration secondary to chronic staphylococcal lid disease was frequently seen. The microbiology of the various ulcers is described, and the placing of organisms into classes is stressed in determining significance of isolates. Many of the patients were male Africans who were either manual labourers or unemployed. Half the patients had used topical antibiotics before presentation.


Molecular Characterization of Newcastle Disease Viruses Isolated from Chickens in Tanzania and Ghana.
Newcastle disease (ND) is one of the most challenging infectious diseases affecting poultry production in Africa, causing major economic losses. To date, Newcastle disease virus isolates from several African countries have been grouped into class II NDV genotypes I, IV, V, VI, VII, XI, XIII, XIV, XVII, XVIII and XXI. Although ND is endemic in many African countries, information on circulating genotypes is still scarce. In Tanzania, outbreaks with genotypes V and XIII have been reported. In West and Central Africa, genotypes XIV, XVII, and XVIII are the most predominant. To investigate other genotypes circulating in Tanzania and Ghana, we performed molecular genotyping on isolates from Tanzania and Ghana using the MinION, a third-generation portable sequencing device from Oxford Nanopore Technologies. Using the MinION, we successfully sequenced the NDV F gene hypervariable region of 24 isolates from Tanzania and four samples from Ghana. In Tanzania, genotypes V, VII and XIII were detected. All isolates from Ghana belonged to genotype XVIII. The data obtained in this study reflect the genetic diversity of NDV in Africa and highlight the importance of surveillance for monitoring the distribution of NDV genotypes and viral evolution.


The role of encoding strategies in contextual memory deficits in patients with bipolar disorder.
Contextual memory is important for the encoding and retrieval of episodic memory, which is often impaired in euthymic patients with bipolar disorder (BD). The objective was to investigate the effect of low and high cognitive support on encoding in an incidental contextual memory task in euthymic patients with BD. Twenty-three patients with a BD type I diagnosis (aged 23-63 years, 17 women and 6 men) and 29 healthy controls completed a recognition memory task for context (location of a recognised object). Participants were assigned to one of two incidental encoding conditions: (1) with a binding cue to encourage the association of the object to its location (judging the degree of appropriateness of an object in relation to its location) or (2) without a binding cue (judging daily use of objects). Patients showed a deficit in incidental contextual memory in the absence of a binding cue at encoding. Under incidental encoding with the binding cue, no differences were observed between the groups for contextual memory. Contextual memory deficits in BD patients were reduced by providing cognitive support at encoding. The role of this strategy should be investigated in larger samples to evaluate its use for cognitive remediation in BD patients.


Modification of the antigenic structure of human interferon alpha-2 by PH 2 treatment: a further support for the antigenic relationship between alpha and beta interferons.
We have recently reported that a unique antigenic structure, designated as Common epitope 1, was found to be shared by human recombinant IFN alpha-2 and the human fibroblast IFN beta. The Common epitope 1 was identified with the aid of a synthetic IFN alpha-2 fragment SH 132-137. Based on this observation, we proposed the hypothesis that an antigenic relationship should exist also between natural human leukocyte IFN alpha and natural human fibroblast IFN beta. However, we were not able to detect any Common epitope 1 in preparations of conventional human leukocyte IFN alpha. In the present study, we were looking for a possible explanation of absence of the Common epitope 1 in conventional leukocyte IFN alpha. First, we demonstrated its acid labile nature in the recombinant IFN alpha-2 molecule and second, we proposed that the pH 2 lability of this unique epitope might be responsible for the lack of antigenicity also in pH 2-treated (conventional) leukocyte IFN alpha preparations. Actually, when pH 2 non-treated leukocyte IFN alpha was examined, we succeeded in demonstration of the Common epitope 1 in IFN-preparation. Moreover, anti-serum against pH 2 non-treated IFN alpha was capable of neutralizing both the conventional i.e. pH 2-treated leukocyte IFN alpha and fibroblast IFN beta. It is concluded that the nomenclature distinguishing two classes (i.e. alpha and beta as class I and gamma as class II) of IFNs is more appropriate than the current official nomenclature distinguishing three antigenic classes of IFNs.


Bees explain floral variation in a recent radiation of Linaria.
The role of pollinators in floral divergence has long attracted the attention of evolutionary biologists. Although abundant studies have reported the effect of pollinators on flower-shape variation and plant speciation, the influence of pollinators on plant species differentiation during rapid radiations and the specific consequences of shifts among similar pollinators are not well understood. Here, we evaluate the association between pollinators and floral morphology in a closely related and recently diversifying clade of Linaria species (sect. Supinae subsect. Supinae). Our approach combined pollinator observations, functional floral morphometric measures and phylogenetic comparative analyses. The fauna visiting Linaria species was determined by extensive surveys and categorized by a modularity algorithm, and the size and shape of flowers were analysed by means of standard and geometric morphometric measures. Standard measures failed to find relationships between the sizes of representative pollinators and flowers. However, discriminant function analyses of geometric morphometric data revealed that pollination niches are finer predictors of flower morphologies in Linaria if compared with phylogenetic relationships. Species with the most restrictive flowers displayed the most slender spurs and were pollinated by bees with larger proboscides. These restrictive flower shapes likely appeared more than once during the evolutionary history of the study group. We show that floral variation can be driven by shifts between pollinators that have been traditionally included in a single functional group, and discuss the consequences of such transitions for plant species differentiation during rapid radiations.


A review study to evaluate holmium:YAG laser lithotripsy with flexible ureteroscopy in patients on ongoing oral anticoagulant therapy.
The objective of this study was to evaluate the safety and efficacy of flexible ureteroscopy (FURS) and holmium:YAG laser lithotripsy for the treatment of upper urinary tract stones in patients on active oral anticoagulants. The records of 1081 patients who underwent flexible ureteroscopic holmium:YAG (Ho:YAG) laser lithotripsy for upper ureteral and renal calculi from 1999 to 2015 were retrospectively reviewed. A total of 84 patients on continuous oral anticoagulation or antiplatelet therapy (warfarin, aspirin, or clopidogrel) were identified. Of these patients, 40 were on warfarin, 25 on aspirin, 11 on clopidogrel, and 8 on both aspirin and clopidogrel. The drugs were not discontinued. The baseline characteristics, indications for anticoagulation therapy, perioperative data, stone-free rate, and complications were documented. Evaluation of outcomes was assessed at 1-, 3-, and 6-month follow-up postoperatively. Mean stone size was 19.7 ± 9.4 (range 8 to 31 mm). Twenty patients had upper ureteral and 64 patients had intrarenal calculi. Two patients had bilateral renal calculi. Mean operation time was 78.2 ± 23.8 min (range 17 to 144 min). Two procedures (2.3%) in warfarin group were terminated due to persistent bleeding causing visual impairment. No transfusions were required. The mean serum hemoglobin levels did not change significantly (12.9 ± 3.7 to 12.2 ± 3.3 g/dL). No thromboembolic or cardiac adverse events were observed perioperatively. The double-j (DJ) ureteral catheterization time was 29.6 ± 9.3 days (range 14 to 68 days) and the hospital stay was 1.6 ± 0.6 days (range 1 to 4). The stone-free rate was 95.2% (80 patients) at 6 months. Flexible ureteroscopic Ho:YAG laser lithotripsy in patients requiring long-term anticoagulation therapy seems to be a safe and effective procedure and should be considered as a first-line treatment option in such patients for the surgical management of upper urinary tract stones.


Community incidence of hepatitis B and C among reincarcerated women.
The incarceration rate has increased 239% in the United States over the past 2 decades. This increase in incarceration has been fueled by the movement towards a criminal, rather than medical, response to the problem of drug dependence. For women in particular, incarceration and drug use are interdependent epidemics. Given that incarceration is common among drug-dependent persons, infectious diseases--including hepatitis B virus (HBV) and hepatitis C virus (HCV) infection--are prevalent among incarcerated persons. We sought to determine the incidence of HBV and HCV infection among recidivist women prisoners. From 1996 through 1997, excess from serum samples collected during HIV testing of female admittees to a state Department of Corrections facility were tested for HBV and HCV. Multiple samples obtained from women incarcerated multiple times during the study period were compared for incidence. Baseline prevalences of markers of HBV and HCV were 36% and 34%, respectively. Incidence rates for HBV and HCV infection among reincarcerated women were 12.2 and 18.2 per 100 person-years, respectively. The majority of the time spent between serial intakes was not spent in the correctional facility; thus, incident infections likely occurred in the community. Incidences of HBV and HCV infection among reincarcerated women were high. Prisons and jails can be efficient locations for the diagnosis, treatment, and prevention of hepatitis B and C through programs such as testing, counseling, education, vaccination, and linkage to medical and drug treatment services.


The Role of Ubiquitination and SUMOylation in Telomere Biology.
Telomeres are a unique structure of DNA repeats covered by proteins at the ends of the chromosomes that protect the coding regions of the genome and function as a biological clock. They require a tight regulation of the factors covering and protecting their structure, as they are shortened with each cell division to limit the ability of cells to replicate uncontrollably. Additionally, they protect the chromosome ends from DNA damage responses and thereby, prevent genomic instability. Telomere dysfunction can lead to chromosomal abnormalities and cancer. Therefore, dysregulation of any of the factors that regulate the integrity of the telomeres will have implications to chromosomal stability, replicative lifespan and may lead to cell transformation. This review will cover the main factors participating in the normal function of the telomeres and how these are regulated by the ubiquitin and SUMO systems. Accumulating evidence indicate that the ubiquitin and SUMO pathways are significant regulators of the shelterin complex and other chromatin modifiers, which are important for telomere structure integrity. Furthermore, the crosstalk between these two pathways has been reported in telomeric DNA repair. A better understanding of the factors contributing to telomere biology, and how they are regulated, is important for the design of new strategies for cancer therapies and regenerative medicine.


Constrained instruments and their application to Mendelian randomization with pleiotropy.
In Mendelian randomization (MR), inference about causal relationship between a phenotype of interest and a response or disease outcome can be obtained by constructing instrumental variables from genetic variants. However, MR inference requires three assumptions, one of which is that the genetic variants only influence the outcome through phenotype of interest. Pleiotropy, that is, the situation in which some genetic variants affect more than one phenotype, can invalidate these genetic variants for use as instrumental variables; thus a naive analysis will give biased estimates of the causal relation. Here, we present new methods (constrained instrumental variable [CIV] methods) to construct valid instrumental variables and perform adjusted causal effect estimation when pleiotropy exists and when the pleiotropic phenotypes are available. We demonstrate that a smoothed version of CIV performs approximate selection of genetic variants that are valid instruments, and provides unbiased estimates of the causal effects. We provide details on a number of existing methods, together with a comparison of their performance in a large series of simulations. CIV performs robustly across different pleiotropic violations of the MR assumptions. We also analyzed the data from the Alzheimer's disease (AD) neuroimaging initiative (ADNI; Mueller et al., 2005. Alzheimer's Dementia, 11(1), 55-66) to disentangle causal relationships of several biomarkers with AD progression.


[Influence of various physical forms of the feed on the energy metabolism of ruminants. 3. Investigation of the energetic utilisation of rations with roughage treated with various mechanical methods].
In a total of 170 experiments with the complete metabolism of fullgrown sheep the energy utilisation of rations with roughage treated with different mechanical methods was investigated. The roughage feedstuffs dried green fodder of vetch/grass, lucerne and rye grass and straw were given to 4 parallel animals in 9 rations with differing quotas of concentrates (0 to 60%). The roughage underwent the following 5 methods of treatment: 1) chopping; 2) chopping and pelleting; 3) grinding; 4) grinding and pelleting; 5) grinding and pelleting with concentrates. The results show that the bases given in the GDR system of feedstuff evaluation concerning the energetic evaluation of feedstuffs are to be applied to mechanically treated roughage too. In order to avoid negative effects on the energy metabolism one has to take care that the use of mechanically treated roughage does not cause dysfunctions of the rumen.


The application of ultrasound contrast agents in the characterization of renal tumors.
The aim of this article is to describe the current ultrasound (US) contrast agents employed in the characterization of renal tumors and to report our experience in the use of a contrast-specific ultrasound technique pulse inversion harmonic imaging (PIHI).A total of 23 renal masses were prospectively evaluated by conventional US, CDUS, PIHI and finally by helical-CT (HCT). The study was performed using a wideband convex array 2-5 MHz transducer and a US digital apparatus . PIHI scanning of each renal mass was performed before and after the injection of Levovist. To reach a definitive diagnosis HCT and/or histological findings were considered as the reference procedures. Eleven solid renal cell carcinomas (RCCs), one embryonal metanephric adenoma (EMA), seven angiomyolipomas (AMLs), and four cystic RCCs were analyzed. Solid RCCs revealed a higher contrast enhancement than AMLs with a typical pattern on conventional US. The EMA and AML with an atypical pattern revealed intense contrast enhancement during the arterial phase, progressively decreasing during the delayed phase. Cystic RCCs showed intense contrast enhancement on the peripheral thick wall during the arterial phase decreasing during the delayed phase. PIHI with Levovist may differentiate solid RCCs from AMLs with no hypervascular pattern while solid RCCs show a higher intensity of contrast enhancement than hypervascular AML.


The role of proteolysis in R gene mediated defence in plants.
SUMMARY Within the last 10 years, numerous R genes have been cloned from natural genetic variation in model as well as crop plants, and these have been classified according to their motifs. Some of the downstream signalling components have also been identified by artificial mutagenesis. Recently, cloning of three of these signalling genes (COI1, RAR1 and SGT1b) from Arabidopsis, barley and tobacco have helped uncover the physiological link between defence signalling and ubiquitin-mediated protein degradation. The physical association of COI1 and SGT1b with the components of ubiquitin-ligase complexes has been shown. In addition, post-transcriptional silencing of some of the subunits of the ubiquitin-ligase complex has led to a loss of resistance, indicating that protein degradation may also act as a regulatory mechanism in plant defence. Over the next few years, we should expect to see more examples of the interplay between the defence response and protein degradation in plants.


Two independent pathways of regulated necrosis mediate ischemia-reperfusion injury.
Regulated necrosis (RN) may result from cyclophilin (Cyp)D-mediated mitochondrial permeability transition (MPT) and receptor-interacting protein kinase (RIPK)1-mediated necroptosis, but it is currently unclear whether there is one common pathway in which CypD and RIPK1 act in or whether separate RN pathways exist. Here, we demonstrate that necroptosis in ischemia-reperfusion injury (IRI) in mice occurs as primary organ damage, independent of the immune system, and that mice deficient for RIPK3, the essential downstream partner of RIPK1 in necroptosis, are protected from IRI. Protection of RIPK3-knockout mice was significantly stronger than of CypD-deficient mice. Mechanistically, in vivo analysis of cisplatin-induced acute kidney injury and hyperacute TNF-shock models in mice suggested the distinctness of CypD-mediated MPT from RIPK1/RIPK3-mediated necroptosis. We, therefore, generated CypD-RIPK3 double-deficient mice that are viable and fertile without an overt phenotype and that survived prolonged IRI, which was lethal to each single knockout. Combined application of the RIPK1 inhibitor necrostatin-1 and the MPT inhibitor sanglifehrin A confirmed the results with mutant mice. The data demonstrate the pathophysiological coexistence and corelevance of two separate pathways of RN in IRI and suggest that combination therapy targeting distinct RN pathways can be beneficial in the treatment of ischemic injury.


Outcomes of Left Ventricular Assist Device Implantation in Patients With Uncommon Etiology Cardiomyopathy.
The impact of uncommon etiology cardiomyopathies on Left-ventricular assist device (LVAD)-recipient outcomes is not very well known. This study aimed to characterize patients with uncommon cardiomyopathy etiologies and examine the outcomes between uncommon and ischemic/idiopathic dilated cardiomyopathy. This observational study was conducted in 19 centers between 2006 and 2016. Baseline characteristics and outcomes of patients with uncommon etiology were compared to patients with idiopathic dilated/ischemic cardiomyopathies. Among 652 LVAD-recipients included, a total of 590 (90.5%) patients were classified as ischemic/idiopathic and 62 (9.5%) patients were classified in the "uncommon etiologies" group. Main uncommon etiologies were: hypertrophic (n = 12(19%)); cancer therapeutics-related cardiac dysfunction (CTRCD) (n = 12(19%)); myocarditis (n = 11(18%)); valvulopathy (n = 9(15%)) and others (n = 18(29%)). Patients with uncommon etiologies were significantly younger with more female and presented less co-morbidities. Additionally, patients with uncommon cardiomyopathies were less implanted as destination therapy compared with ischemic/idiopathic group (29% vs 38.8%). During a follow-up period of 9.1 months, both groups experienced similar survival. However, subgroup of hypertrophic/valvular cardiomyopathies and CTRCD had significantly higher mortality compared to the ischemic/idiopathic or myocarditis/others cardiomyopathies. Conversely, patients with myocarditis/others etiologies experienced a better survival. Indeed, the 12-months survival in the myocarditis/others; ischemic/idiopathic and hypertrophic/CTRCD/valvulopathy group were 77%; 65%, and 46% respectively. In conclusion, LVAD-recipients with hypertrophic cardiomyopathy, valvular heart disease and CTRCD experienced the higher mortality rate.


Kinetics of low-density lipoprotein receptor activity in Hep-G2 cells: derivation and validation of a Briggs-Haldane-based kinetic model for evaluating receptor-mediated endocytotic processes in which receptors recycle.
The process of receptor-mediated endocytosis for receptors that recycle to the cell surface in an active form can be considered as being kinetically analogous to that of a uni-substrate, uni-product enzyme-catalysed reaction. In this study we have derived steady-state initial-velocity rate equations for this process, based on classical Briggs-Haldane and King-Altman kinetic approaches to multi-step reactions, and have evaluated this kinetic paradigm, using as a model system the low-density lipoprotein (LDL)-receptor-mediated endocytosis of the trapped label [14C]sucrose-LDL in uninduced, steady-state Hep-G2 cells. Using the derived rate equations, together with experimentally determined values for Bmax (123 fmol/mg of cell protein), Kd (14.3 nM), the endocytotic rate constant ke (analogous to kcat; 0.163 min-1), Km (80 nM) and maximal internalization velocity (26.4 fmol/min per mg), we have calculated the ratio ke/Km (0.00204 nM-1.min-1), the bimolecular rate constant for LDL and LDL-receptor association (0. 00248 nM-1.min-1), the first-order rate constant for LDL-LDL-receptor complex dissociation (0.0354 min-1), the total cellular content of LDL receptors (154 fmol/mg of cell protein), the intracellular LDL receptor concentration (30.7 fmol/mg of cell protein) and the pseudo-first-order rate constant for LDL receptor recycling (0.0653 min-1). Based on this mathematical model, the kinetic mechanism for the receptor-mediated endocytosis of [14C]sucrose-LDL by steady-state Hep-G2 cells is one of constitutive endocytosis via independent internalization sites that follows steady-state Briggs-Haldane kinetics, such that LDL-LDL-receptor interactions are characterized by a rapid-high-affinity ligand-receptor association, followed by ligand-receptor complex internalization that is rapid relative to complex dissociation, and by receptor recycling that is more rapid than complex internalization and that serves to maintain 80% of cellular LDL receptors on the cell surface in the steady-state. The consistency with which these quantitative observations parallel previous qualitative observations regarding LDL-receptor-mediated endocytosis, together with the high correlation between theoretical internalization velocities (calculated from determined rate constants) and experimental internalization velocities, underscore the validity of considering receptor-mediated endocytotic processes for recycling receptors in catalytic terms.


Treatment of residual disease following neoadjuvant therapy in breast cancer.
Substantial advances have been made in the systemic treatment of breast cancer with residual disease following neoadjuvant therapy. We reviewed recent and ongoing studies informing the standard clinical management of residual disease by subtype: HER2+, TNBC, and HR+/HER2-, as well as strategies for BRCA+ disease. We conclude with a discussion of ongoing clinical trials and current controversies regarding the treatment of residual disease in breast cancer.


Radiation therapy with concomitant hydroxyurea and fluorouracil in stage II and III head and neck cancer.
In 1986, a multi-institutional phase II trial was begun to study the use of chemotherapy with concomitant radiation in patients with stage II and III head and neck cancer. End points were overall survival, progression-free survival, local/regional control, and toxicity in the setting of organ preservation with concomitant treatment. Eligible patients with stage II or III disease received chemotherapy and radiation on a 2-week cycle. Chemotherapy consisted of continuous infusion fluorouracil (5-FU) at 800 mg/m2/d for 5 consecutive days (days 1 to 5) and hydroxyurea (HU) at 1 g orally every 12 hours for 13 doses starting the evening before the start of irradiation. Radiation therapy was given as single 1.8- to 2.0-Gy fractions for 5 consecutive days (days 1 to 5) with chemotherapy. Each 5 days of treatment was followed by a 9-day break (days 6 to 14), during which no additional treatment was given. Treatment cycles were repeated until the completion of the planned radiation dose (six to eight cycles). Between 1989 and 1996, 60 patients were enrolled. All patients were eligible for analysis, with a median follow-up of 52 months for surviving patients and 42 months for all patients. Grade 3 to 4 mucositis occurred in 57% of patients. The 5 year-actuarial overall survival, progression-free survival, and local/regional control were 65%, 82%, and 86%, respectively. Eight patients developed local and/or regional recurrence after treatment. Surgical salvage was possible in three of these patients. Thus, the ultimate 5-year local/ regional control was 91%. Concomitant radiation and chemotherapy with 5-FU and HU is an effective regimen in patients with stage II and III head and neck cancer. Progression-free survival and local/regional control appear to be superior to expected rates in patients treated with surgery and radiation. Further testing of this regimen in a phase III setting is indicated.


The early history of glaucoma: the glaucous eye (800 BC to 1050 AD).
To the ancient Greeks, glaukos occasionally described diseased eyes, but more typically described healthy irides, which were glaucous (light blue, gray, or green). During the Hippocratic period, a pathologic glaukos pupil indicated a media opacity that was not dark. Although not emphasized by present-day ophthalmologists, the pupil in acute angle closure may appear somewhat green, as the mid-dilated pupil exposes the cataractous lens. The ancient Greeks would probably have described a (normal) green iris or (diseased) green pupil as glaukos. During the early Common Era, eye pain, a glaucous hue, pupil irregularities, and absence of light perception indicated a poor prognosis with couching. Galen associated the glaucous hue with a large, anterior, or hard crystalline lens. Medieval Arabic authors translated glaukos as zarqaa, which also commonly described light irides. Ibn Sina (otherwise known as Avicenna) wrote that the zarqaa hue could occur due to anterior prominence of the lens and could occur in an acquired manner. The disease defined by the glaucous pupil in antiquity is ultimately indeterminate, as the complete syndrome of acute angle closure was not described. Nonetheless, it is intriguing that the glaucous pupil connoted a poor prognosis, and came to be associated with a large, anterior, or hard crystalline lens.


On co-activation pattern analysis and non-stationarity of resting brain activity.
The non-stationarity of resting-state brain activity has received increasing attention in recent years. Functional connectivity (FC) analysis with short sliding windows and coactivation pattern (CAP) analysis are two widely used methods for assessing the dynamic characteristics of brain activity observed with functional magnetic resonance imaging (fMRI). However, the statistical nature of the dynamics captured by these techniques needs to be verified. In this study, we found that the results of CAP analysis were similar for real fMRI data and simulated stationary data with matching covariance structures and spectral contents. We also found that, for both the real and simulated data, CAPs were clustered into spatially heterogeneous modules. Moreover, for each of the modules in the real data, a spatially similar module was found in the simulated data. The present results suggest that care needs to be taken when interpreting observations drawn from CAP analysis as it does not necessarily reflect non-stationarity or a mixture of states in resting brain activity.


Mitigating Contemporary Trauma Impacts Using Ancient Applications.
The COVID-19 pandemic represents the most significant global challenge in a generation. Based on extant data from previous pandemics, demographic, occupational, and psychological factors have been linked to distress and for some vulnerable members of society. COVID-19 has added to the layers of grief and distress of existing trauma. Evidence-based frameworks exist to guide our individual and collective response to reduce the trauma associated with the experience of a pandemic. Pandemic and post-pandemic measures to ameliorate impacts require a multi-disciplined approach, central to which is community connectedness, resilience, and access to support. We advocate for the acceptance and broader application of Dadirri, a healing practice held by the Ngan'gikurunggurr and Ngen'giwumirri Aboriginal people of the Daly River region in the Northern Territory, Australia. This modality engages therapeutic phases that are comparable with other practiced trauma therapies. The demonstrated therapeutic outcomes from Dadirri can be attained through an individualistic or in a relational engagement context. This practice is accessible to all ages, is non-specific to gender and is suitable for people constrained in their mobility or limited by resources, pertinent in pandemic affected settings.


The Wiskott-Aldrich syndrome. Results of transfer factor therapy.
12 patients with Wiskott-Aldrich syndrome were treated with therapeutic doses of transfer factor in an attempt to induce cellular immunity. Clinical improvement was noted after transfer factor therapy in 7 of the 12 patients treated. Because this disease has a variable course and temporary spontaneous improvement can occur, the observed improvement cannot necessarily be attributed to the transfer factor. However, in two patients repeated remissions consistently followed transfer factor administration on repeated occasions. This included freedom from infections, regression of splenomegaly, and clearing of eczema. An unexpected finding was a decrease in bleeding in 3 of the 10 patients who had bleeding. Conversion of skin reactivity was obtained in all seven patients who clinically seemed to respond to transfer factor. In vitro studies performed after the administration of transfer factor demonstrated that the lymphocytes of the patients now produced migration inhibitory factor in response to appropriate test antigens, but did not undergo increased radioactive thymidine incorporation in response to the same antigens. A defect in the monocyte IgG receptors has been found in certain patients with the disease, and the current study shows that all patients with defective monocyte IgG receptors responded to transfer factor, whereas only one patient with normal receptors showed any response. This test may thus prove to be useful in predicting the results of transfer factor therapy in patients with Wiskott-Aldrich syndrome, although evaluation of a larger series of patients will be necessary to confirm this point. We conclude that cellular immunity can be induced, that there appears to be clinical benefit in certain patients with Wiskott-Aldrich syndrome by the use of transfer factor, and that this mode of therapy warrents trial in these patients and others with defects of cellular immunity.


Mortality from solid tumors among workers in formaldehyde industries: an update of the NCI cohort.
Formaldehyde, a widely used chemical, is considered a human carcinogen. We extended follow-up of the largest industrial cohort of workers in formaldehyde industries (n = 25,619) by 10 years through 2004. Standardized mortality ratios (SMRs) and rate ratios (RRs) were calculated for deaths from solid tumors using quantitative formaldehyde exposure estimates. During 998,239 person-years, 13,951 deaths occurred. With one additional death, previously observed excesses for nasopharyngeal cancer (n = 10) persisted for peak, average intensity and cumulative exposure; RRs in the highest exposure categories were 7.66 (95% CI: 0.94, 62.34), P-trend = 0.005, 11.54 (95% CI: 1.38, 96.81), P-trend = 0.09, and 2.94 (95% CI: 0.65, 13.28), P-trend = 0.06, respectively. For all cancer, solid tumors and lung cancer, SMRs among exposed workers were elevated, but internal analyses revealed no positive associations with formaldehyde exposure. Consistent with previous analyses of this cohort, this update continues to suggest a link between formaldehyde exposure and nasopharyngeal cancer.


Evaluation of Venezuelan Equine Encephalitis (VEE) replicon-based Outer surface protein A (OspA) vaccines in a tick challenge mouse model of Lyme disease.
Venezuelan Equine Encephalitis (VEE) virus replicon particles (VRPs) encoding Borrelia burgdorferi Outer surface protein A (OspA) were evaluated for their ability to induce an immune response and provide protection from tick-borne spirochetes. VRPs expressing ospA that accumulated intracellularly (VRP OspA) or that was secreted from host cells (VRP tPA-OspA) were tested. Both VRP OspA and VRP tPA-OspA expressed ospA in immunized mice. Mice vaccinated with VRPs expressing secreted OspA produced significant amounts of anti-OspA antibodies, whereas VRPs expressing intracellular OspA were less immunogenic. The VRP method of delivery induced a Th1 type immune response unlike the recombinant OspA protein in Freund's adjuvant, which induced a mixed (Th1 and Th2) immune response. The VRP tPA-OspA construct induced an immune response that reduced the bacterial load in feeding Ixodes scapularis and blocked transmission to the host. These results indicate that VRPs are capable of providing protection against tick-borne B. burgdorferi, and potentially can be used for developing improved vaccines against Lyme disease.


Relationship between isokinetic strength of the knee joint and countermovement jump performance in elite boxers.
The lower limbs play a key role to develop the linear momentum for hitting power in effective boxing. The knee extensor and flexor strength guarantees the dynamic stability of boxers. The insufficient extensor strength of the lower extremities causes compensation during flexion resulting in movement errors or damage to knee joint muscles. This study was conducted to explore the isokinetic concentric strength of the knee flexor and extensor and the relationship between isokinetic knee extensors strength and countermovement jump (CMJ) performance in elite boxers. Thirteen elite male boxers (Age: 25.15 ± 3.98 years, height 1.72 ± 0.04 m, weight 61.82 ± 10.46 kg, training years = 11.56 ± 2.67 years) performed the CMJ, and the isokinetic knee test was performed using the Biodex dynamometer. The maximal isokinetic peak torque of the knee extensor and flexor muscles was recorded at three angular velocities (60°/s, 180°/s, and 240°/s) on both sides of the legs. The relative peak value of torque in the knee extensors decreased significantly with increasing angular velocity. A difference in relative peak torque (RPT) was only seen at 60°/s in knee flexors. However, the H/Q ratio increased as the velocity increased from 60°/s to 240°/s (P < 0.05). The highest peak torque was found in the knee extensors at a velocity of 240°/s (r = 0.73, P < 0.001). The correlation between RPT and vertical jump height was the strongest at 240°/s. The strongest relationship was found between the height of the CMJ and the RPT of the deficit of knee extensors. We suggest that explosive force training of the isokinetic muscles should be optimally carried out at a speed of 240°/s. The results of this study provide a reference for boxers to improve their jump height and lower-limb explosive strength through isokinetic strength training of the knee flexor and extensor.


Dependency Change with Aging and Associated Factors in Uruguay: A Cohort Study.
To assess the heterogeneity of transitions toward dependency in older adults and to explore the robustness of results to different operationalizations of dependency. Using data from people aged 60 years and older from a national representative study in Uruguay (Encuesta Longitudinal de Protección Social, N = 5071), we fitted multinomial regressions adjusted by sociodemographic and health characteristics to model transitions into dependency and death. We used a harder operationalization with basic activities of daily living (Katz-dependency) and Comprehensive-dependency with basic, instrumental, and advanced activities. Increasing age (RRR = 1.08, CI = [1.05; 1.12], p < .001) and having comorbidities (RRR = 2.16, CI = [1.31; 3.57], p = .003) increased the risk of transition from nondependent to dependent using Katz-dependency. Women with at least two chronic conditions have increased risk of Comprehensive-dependency (RRR = 1.79, CI = [1.15; 2.80], p = .010). Inconsistencies in findings emerged when evaluating transitions into dependency with the different measures, which may have social care implications.


Developmental regulation and neuronal expression of the cellular disintegrin ADAM11 gene in mouse nervous system.
ADAM11 is the prototype member of the predominantly CNS-associated clade of the ADAM metalloprotease-disintegrins that has been implicated in neural adhesion and axon guidance. The present study describes the spatiotemporal expression pattern of the ADAM11 gene in adult and developing mouse, and identifies the cells expressing the gene. In the adult CNS, ADAM11 mRNA was present throughout the forebrain, including different cortical fields and diencephalic nuclei. In brainstem, low to moderate expression was detected in certain midbrain nuclei, while several pontine and medullary nuclei showed a very strong signal. High expression was observed in the cerebellar cortex and spinal cord. In addition, ADAM11 was expressed in ganglia of the peripheral nervous system (PNS), retinae, testes, liver, and at lower levels in epidermal and mucosal epithelia, kidney, and salivary gland. The expression was localized to neurons in all examined CNS and PNS subfields. During pre- and perinatal development, ADAM11 was differentially expressed both in the developing PNS and CNS, as well as in heart, kidney, eyes, and brown fat. The present results suggest a widespread involvement of ADAM11 in neuron-neuron or neuron-glial cell interactions during development as well as in the adult nervous system. They provide novel complementary information to recently accumulated data on CNS integrin gene expression and offer useful clues for further studies of the neural functions of ADAMs and integrins.


Resigned professionalism? Non-acute inpatients and resident education.
A growing group of inpatients on acute clinical teaching units have non-acute needs, yet require attention by the team. While anecdotally, these patients have inspired frustration and resource pressures in clinical settings, little is known about the ways in which they influence physician perceptions of the learning environment. This qualitative study explored residents' and attending physicians' perceptions of caring for these patients, including their educational value. Using constructivist grounded theory, we conducted seven homogeneous focus groups and three interviews with residents and attending physicians from neurology and general internal medicine. A constant comparative analytical approach was employed alongside data collection, using theoretical sampling to explore emergent themes. Residents consistently described non-acute patients as non-educational, uninteresting, but still in need of care. Some attending physicians echoed this view, while others described multiple learning opportunities presented by non-acute patients. Both groups described residents as engaging with non-acute patients in a professional capacity, but not as learners. This engagement in a professional capacity could be considered diligent disinterest, or resigned professionalism. A constructivist understanding of the dynamics which influence learning in the workplace was used to explore the reasons why the residents in our study did not recognize the learning opportunities presented by non-acute patients. Our results resonate with Billett's theory of workplace affordances, which offers an explanation as to why learners may not identify or take advantage of potential learning opportunities. Overall, our study assists our understanding of the sociocultural factors that influence learners' choices to engage with particular clinical learning opportunities.


Phylogeography and population differentiation in Hepatozoon canis (Apicomplexa: Hepatozoidae) reveal expansion and gene flow in world populations.
Hepatozoon canis is a protozoan transmitted to dogs and other wild carnivores by the ingestion of ticks containing mature oocysts and is considered the principal cause of canine hepatozoonosis in the world. Here, we examined ribosomal RNA 18S gene sequence variation to determine the genetic differences and phylogeographic diversity of H. canis from various geographical areas around the world. We used 550 publicly available sequences of H. canis from 46 countries to assess haplotype relationships, geographical structure, genetic diversity indices, and relationships among populations. We performed neutrality tests and pairwise comparisons of fixation index (FST) values between groups and pairwise comparisons of FST values between populations. To determine whether populations are structured, analyses of molecular variance (AMOVAs) and spatial analysis of molecular variance (SAMOVA) were performed. The dataset of H. canis yielded 76 haplotypes. Differentiation among populations indicated that there is no phylogeographical structure (GST = 0.302 ± 0.0475). Moreover, when samples were grouped by continents a significant FST was obtained, meaning that populations were genetically differentiated. The AMOVA showed that 57.4% of the genetic variation was explained by differences within populations when all locations were treated as a single group and revealed that there is no population structure when populations are grouped into two, three, and four groups (FCT, p > 0.05), suggesting that dispersal between populations is high. SAMOVA revealed significant FCT values for groups K = 5. The Tajima's D and Fu's Fs show that populations have undergone recent expansion, and the mismatch distribution analysis showed population expansion (multimodal distribution). The current molecular data confirmed that H. canis does not show phylogeographic or population structure. The haplotypes exhibit low genetic differentiation, suggesting a recent expansion due to gene flow among populations. These results provide pivotal information required for future detailed population genetic analysis or to establish control strategies of this parasite.


Cholinergic modulation of mesolimbic dopamine function and reward.
The substantial health risk posed by obesity and compulsive drug use has compelled a serious research effort to identify the neurobiological substrates that underlie the development these pathological conditions. Despite substantial progress, an understanding of the neurochemical systems that mediate the motivational aspects of drug-seeking and craving remains incomplete. Important work from the laboratory of Bart Hoebel has provided key information on neurochemical systems that interact with dopamine (DA) as potentially important components in both the development of addiction and the expression of compulsive behaviors such as binge eating. One such modulatory system appears to be cholinergic pathways that interact with DA systems at all levels of the reward circuit. Cholinergic cells in the pons project to DA-rich cell body regions in the ventral tegmental area (VTA) and substantial nigra (SN) where they modulate the activity of dopaminergic neurons and reward processing. The DA terminal region of the nucleus accumbens (NAc) contains a small but particularly important group of cholinergic interneurons, which have extensive dendritic arbors that make synapses with a vast majority of NAc neurons and afferents. Together with acetylcholine (ACh) input onto DA cell bodies, cholinergic systems could serve a vital role in gating information flow concerning the motivational value of stimuli through the mesolimbic system. In this report we highlight evidence that CNS cholinergic systems play a pivotal role in behaviors that are motivated by both natural and drug rewards. We argue that the search for underlying neurochemical substrates of compulsive behaviors, as well as attempts to identify potential pharmacotherapeutic targets to combat them, must include a consideration of central cholinergic systems.


Virus Removal and Inactivation Mechanisms during Iron Electrocoagulation: Capsid and Genome Damages and Electro-Fenton Reactions.
Virus destabilization and inactivation are critical considerations in providing safe drinking water. We demonstrate that iron electrocoagulation simultaneously removed (via sweep flocculation) and inactivated a non-enveloped virus surrogate (MS2 bacteriophage) under slightly acidic conditions, resulting in highly effective virus control (e.g., 5-logs at 20 mg Fe/L and pH 6.4 in 30 min). Electrocoagulation simultaneously generated H2O2 and Fe(II) that can potentially trigger electro-Fenton reactions to produce reactive oxygen species such as •OH and high valent oxoiron(IV) that are capable of inactivating viruses. To date, viral attenuation during water treatment has been largely probed by evaluating infective virions (as plaque forming units) or genomic damage (via the quantitative polymerase chain reaction). In addition to these existing means of assessing virus attenuation, a novel technique of correlating transmission electron micrographs of electrocoagulated MS2 with their computationally altered three-dimensional electron density maps was developed to provide direct visual evidence of capsid morphological damages during electrocoagulation. The majority of coliphages lost at least 10-60% of the capsid protein missing a minimum of one of the 5-fold and two of 3- and 2-fold regions upon electrocoagulation, revealing substantial localized capsid deformation. Attenuated total reflectance-Fourier transform infrared spectroscopy revealed potential oxidation of viral coat proteins and modification of their secondary structures that were attributed to reactive oxygen species. Iron electrocoagulation simultaneously disinfects and coagulates non-enveloped viruses (unlike conventional coagulation), adding to the robustness of multiple barriers necessary for public health protection and appears to be a promising technology for small-scale distributed water treatment.


Enriching our understanding of vulnerability through the experiences and perspectives of individuals living with mental illness.
Vulnerability is a central concept in research ethics and typically serves to identify individuals or groups whose participation in research prompts specific concerns or warrants special consideration. While theoretical discussions on vulnerability have contributed valuable insights to discussions of mental health research and care, they have not been enriched thus far by stakeholder perspectives. This oversight has important consequences for the ways in which we frame vulnerability. It misses the far-reaching insights and experiential knowledge of mental health research participants whose experiences this ethics concept ultimately seeks to capture. In view of this gap, our study combines a pragmatist ethics framework and interpretive phenomenological analysis of qualitative interviews to explore user perspectives on the notion of vulnerability through the lived experiences of mental health research participants and patient-advocates. Importantly, these perspectives offer essential clues to address the difficulties of operationalizing concerns for vulnerability in concrete and practical ways. They help refine our understanding of this key ethics concept.


Planning and orthodontic preparation for maxillary incisors reshaping.
Having a beautiful smile is the main desire of people seeking dental treatment. To achieve this goal, many variables must be considered. These include tooth alignment, color, shape and size, besides their harmonious relationship with the lips and face. An individualized diagnosis is essential to achieve the best result. Within this context, facial analysis and the characteristics of shape, size and position of maxillary incisors play a key role. This paper describes clinical situations in which interdisciplinary treatment was performed to achieve esthetic results in a conservative manner and ensuring greater stability. In most cases requiring reshaping of maxillary incisors for esthetic reasons, prior orthodontic movement is essential. The main focus of this paper is to discuss the many variables involved in these situations. The results of treatments described in this paper were obtained by means of a multidisciplinary approach, involving Orthodontics and Esthetic Dentistry, acting in harmony and recognizing their possibilities and limitations, in order to offer patients the best esthetic solution for their smile. The best treatment option is not always the easiest or fastest. The dentist, as a health professional, should consider the patients' wishes but also perform treatments with minimal intervention, and the best and most predictable esthetic result, also focusing on function and health.


Analysis of genomic damage in the mutagen-sensitive mus-201 mutant of Drosophila melanogaster by arbitrarily primed PCR (AP-PCR) fingerprinting.
DNA repair mechanisms are important to maintain the stability of the genome. In Drosophila melanogaster, the mus-201 gene is required in the excision repair process. To study the contribution of the mus-201 gene in the stability of the Drosophila genome, we have used the arbitrarily primed PCR fingerprinting method (AP-PCR). We have analysed the changes in the genomic DNA fingerprints from the progeny of wild-type males crossed with mus-201 repair-deficient or repair-proficient females. After induction of DNA damage with 2-acetylaminofluorene (2-AAF) in the wild-type parental males, quantitative and qualitative differences in the AP-PCR fingerprints were detected between the two crosses, and the estimate of the genomic damage detected by AP-PCR has clearly shown that the mus-201 repair deficiency is associated with an increase of genomic damage. The predominant type of alterations detected by AP-PCR under the mus-201 repair-deficient conditions agree with the results obtained in microsatellite PCR analysis, suggesting that the role of the mus-201 gene, necessary in excision repair, is not associated to the mismatch repair process. The work reported here demonstrates that the AP-PCR is a suitable technique to analyse genetic alterations in D. melanogaster and, consequently, can be used to compare the susceptibility to genomic damage of different DNA repair mutants.


Involvement of sigma 1 receptor in the SSRI-induced suppression of the methamphetamine-induced behavioral sensitization and rewarding effects in mice.
The abuse of methamphetamine causes abnormal behaviors which are indistinguishable from schizophrenia in humans. Recent reports have shown that selective serotonin reuptake inhibitors (SSRIs) have beneficial effects on methamphetamine-related behaviors, including behavioral sensitization and rewarding effects in animals. However, the exact mechanisms by which SSRIs affect methamphetamine-related behaviors are not yet clear. The present study was designed to investigate the effects of SSRIs on the development of methamphetamine-induced behavioral sensitization and rewarding effects in mice. Behavioral sensitization was measured by examining the locomotor activity of mice in a tilting cage after repeated injections of methamphetamine. Repeated administration of methamphetamine significantly induced a behavioral sensitization. Some SSRIs (fluoxetine and fluvoxamine), which have sigma-1 receptor agonistic activity, inhibited the development of methamphetamine-induced behavioral sensitization. Fluoxetine also dose-dependently attenuated the rewarding effects of methamphetamine as measured by the conditioned place preference paradigm. Furthermore, the sigma-1 receptor antagonist NE-100 significantly reversed the inhibitory effects of fluoxetine on methamphetamine-induced behavioral sensitization and rewarding effects. These results suggest that sigma-1 receptor agonistic activity might be involved in the attenuating effects of fluoxetine and fluvoxamine on methamphetamine-induced behavioral sensitization and rewarding effects.


Detecting Steps Walking at very Low Speeds Combining Outlier Detection, Transition Matrices and Autoencoders from Acceleration Patterns.
In this paper, we develop and validate a new algorithm to detect steps while walking at speeds between 30 and 40 steps per minute based on the data sensed from a single tri-axial accelerometer. The algorithm concatenates three consecutive phases. First, an outlier detection is performed on the sensed data based on the Mahalanobis distance to pre-detect candidate points in the acceleration time series that may contain a ground contact segment of data while walking. Second, the acceleration segment around the pre-detected point is used to calculate the transition matrix in order to capture the time dependencies. Finally, autoencoders, trained with data segments containing ground contact transition matrices from acceleration series from labeled steps are used to reconstruct the computed transition matrices at each pre-detected point. A similarity index is used to assess if the pre-selected point contains a true step in the 30-40 steps per minute speed range. Our experimental results, based on a database from three different participants performing similar activities to the target one, are able to achieve a recall = 0.88 with precision = 0.50 improving the results when directly applying the autoencoders to acceleration patterns (recall = 0.77 with precision = 0.50).


Physiology and function of platelets from patients with Alzheimer's disease.
The discovery that intact Alzheimer amyloid precursor protein is present in platelet granules, has created a great interest in the biochemistry, physiology and function of platelets of patients with Alzheimer disease (AD). In this study we monitored various biochemical and physiological parameters, such as serotonin and adenine nucleotide levels, membrane fluidity, agonist-mediated release of arachidonic acid, thromboxane formation, calcium mobilization, as well as irreversible aggregation and secretion of granule contents. Platelets of patients with AD responded poorly when stirred with weak or potent agonists on a platelet aggregometer. Although capable of agonist-mediated calcium mobilization and synthesis of thromboxanes, the aggregation response of platelets of patients with AD to thrombin and archidonate was considerably compromised. In view of the normal biochemistry and signal transduction capabilities, the compromised response of these cells to potent agonists like thrombin suggested an extrinsic defect. The present study has shown that a plasmatic factor is at least in part responsible for the functional abnormalities of AD platelets.


Alternative splicing of the human gene SYBL1 modulates protein domain architecture of Longin VAMP7/TI-VAMP, showing both non-SNARE and synaptobrevin-like isoforms.
The control of intracellular vesicle trafficking is an ideal target to weigh the role of alternative splicing in shaping genomes to make cells. Alternative splicing has been reported for several Soluble N-ethylmaleimide-sensitive factor Attachment protein REceptors of the vesicle (v-SNAREs) or of the target membrane (t-SNARES), which are crucial to intracellular membrane fusion and protein and lipid traffic in Eukaryotes. However, splicing has not yet been investigated in Longins, i.e. the most widespread v-SNAREs. Longins are essential in Eukaryotes and prototyped by VAMP7, Sec22b and Ykt6, sharing a conserved N-terminal Longin domain which regulates membrane fusion and subcellular targeting. Human VAMP7/TI-VAMP, encoded by gene SYBL1, is involved in multiple cell pathways, including control of neurite outgrowth. Alternative splicing of SYBL1 by exon skipping events results in the production of a number of VAMP7 isoforms. In-frame or frameshift coding sequence modifications modulate domain architecture of VAMP7 isoforms, which can lack whole domains or domain fragments and show variant or extra domains. Intriguingly, two main types of VAMP7 isoforms either share the inhibitory Longin domain and lack the fusion-promoting SNARE motif, or vice versa. Expression analysis in different tissues and cell lines, quantitative real time RT-PCR and confocal microscopy analysis of fluorescent protein-tagged isoforms demonstrate that VAMP7 variants have different tissue specificities and subcellular localizations. Moreover, design and use of isoform-specific antibodies provided preliminary evidence for the existence of splice variants at the protein level. Previous evidence on VAMP7 suggests inhibitory functions for the Longin domain and fusion/growth promoting activity for the Δ-longin molecule. Thus, non-SNARE isoforms with Longin domain and non-longin SNARE isoforms might have somehow opposite regulatory functions. When considering splice variants as "natural mutants", evidence on modulation of subcellular localization by variation in domain combination can shed further light on targeting determinants. Although further work will be needed to characterize identified variants, our data might open the route to unravel novel molecular partners and mechanisms, accounting for the multiplicity of functions carried out by the different members of the Longin proteins family.


Differential modulation of adherence of oral streptococci by human neutrophil myeloperoxidase.
In this study, the modulation of adherence of hydrogen peroxide (H2O2)-producing and non-H2O2-producing strains of oral streptococci by the host leukocyte enzyme myeloperoxidase (MPO) was examined. It was found that exposure to MPO decreased adherence of many strains of oral streptococci to saliva-coated hydroxyapatite beads in the presence of exogenous H2O2 and chloride. The MPO-H2O2-Cl-system increased the adherence of one strain. In the absence of exogenous H2O2, the MPO-H2O2-Cl-system decreased the adherence of H2O2-producing strains only. Glucose increased streptococcal H2O2 production and also increased the anti-adhesive activity of MPO in the absence of exogenous H2O2. We conclude that: (1) host leukocytes can modulate the adherence of oral streptococci via MPO; (2) endogenous production of H2O2 by the oral streptococci can provide sufficient substrate H2O2 to drive this system; and (3) MPO will exert differential modulatory effects on the adherence of oral streptococci, based in part upon the level of endogenous H2O2 production and in part upon the particular characteristics of the adhesins of the bacteria.


Negative thermal expansion in single-component systems with isotropic interactions.
We have devised an isotropic interaction potential that gives rise to negative thermal expansion (NTE) behavior in equilibrium many-particle systems in both two and three dimensions over a wide temperature and pressure range (including zero pressure). An optimization procedure is used in order to find a potential that yields a strong NTE effect. A key feature of the potential that gives rise to this behavior is the softened interior of its basin of attraction. Although such anomalous behavior is well-known in material systems with directional interactions (e.g., zirconium tungstate), to our knowledge, this is the first time that NTE behavior has been established to occur in single-component many-particle systems for isotropic interactions. Using constant-pressure Monte Carlo simulations, we show that as the temperature is increased, the system exhibits negative, zero, and then positive thermal expansion before melting (for both two- and three-dimensional systems). The behavior is explicitly compared to that of a Lennard-Jones system, which exhibits typical expansion upon heating for all temperatures and pressures.


Individual analysis of patients with HoFH participating in a phase 3 trial with lomitapide: The Italian cohort.
The efficacy and safety of lomitapide as adjunct treatment for adults with homozygous familial hypercholesterolaemia (HoFH) have been confirmed in a phase 3 trial. Given the small number of patients (N = 29), and variations in patient characteristics, examining individual cases provides additional details regarding patient management with lomitapide. Here, we examine the details of the Italian patient cohort in the phase 3 trial. The methodology of the multinational, single-arm, open-label, 78-week, dose-escalation, phase 3 trial has been previously reported. The current report details the Italian cohort of six patients (three males, three females) based on individual patient data, individual patient histories and narratives, and by mean data ± SD. Lomitapide was administered according to the dose-escalation protocol. At Week 78, concentrations of low-density lipoprotein-cholesterol were decreased by a mean of 42.6 ± 21.8% compared with baseline. Lomitapide was similarly well tolerated in the Italian cohort as in the entire study population. The most common adverse events were gastrointestinal symptoms. One patient showed an increase in liver transaminases >5× upper limit of normal that resolved after lomitapide treatment was reduced and maintained at a lower dose. The efficacy, safety and tolerability of lomitapide demonstrated in the Italian subgroup of patients are consistent with findings in the entire study population, and illustrate the broad applicability of lomitapide therapy across genotypes and clinical phenotypes. These data also provide an insight into the management of lomitapide use in a cohort of patients within a clinical trial protocol. Clinicaltrials.gov Identifier: NCT00730236.


[Reliability of clinical studies in the diagnosis of malaria fever in West African endemic areas].
We have studied 930 febrile cases in a savanna area of Burkina Faso. We have diagnosed malarial fevers according to the only clinical data in 333 cases and it was confirmed in 154 cases by parasitological study; for 597 cases a non malaria reason was given according to the only clinical data and it was confirmed in 507 cases by biological studies. On the whole the error rate was of 28% on the clinical examination alone and this proportion was equal or superior whatever the parasitological threshold was. The clinical examination is not allow by it self to diagnose a malaria fever case.


RNA imaging in living mice enabled by an in vivo hybridization chain reaction circuit with a tripartite DNA probe.
RNA imaging in living animals helps decipher biology and creates new theranostics for disease treatment. Due to their low delivery efficiency and high background, however, fluorescence probes for in situ RNA imaging in living mice have not been reported. We develop a new cell-targeting fluorescent probe that enables RNA imaging in living mice via an in vivo hybridization chain reaction (HCR). The minimalistic Y-shaped design of the tripartite DNA probe improves its performance in live animal studies and serves as a modular scaffold for three DNA motifs for cell-targeting and the HCR circuit. The tripartite DNA probe allows facile synthesis with a high yield and demonstrates ultrasensitive RNA detection in vitro. The probe also exhibits selective and efficient internalization into folate (FA) receptor-overexpressed cells via a caveolar-mediated endocytosis mechanism and produces fluorescence signals dynamically correlated with intracellular target expressions. Furthermore, the probe exhibits specific delivery into tumor cells and allows high-contrast imaging of miR-21 in living mice. The tripartite DNA design may open the door for intracellular RNA imaging in living animals using DNA-minimal structures and its design strategy can help future development of DNA-based multi-functional molecular probes.


Molecular Characterization and Geographic Distribution of a Mymonavirus in the Population of Botrytis cinerea.
Here, we characterized a negative single-stranded (-ss)RNA mycovirus, Botrytis cinerea mymonavirus 1 (BcMyV1), isolated from the phytopathogenic fungus Botrytis cinerea. The genome of BcMyV1 is 7863 nt in length, possessing three open reading frames (ORF1⁻3). The ORF1 encodes a large polypeptide containing a conserved mononegaviral RNA-dependent RNA polymerase (RdRp) domain showing homology to the protein L of mymonaviruses, whereas the possible functions of the remaining two ORFs are still unknown. The internal cDNA sequence (10-7829) of BcMyV1 was 97.9% identical to the full-length cDNA sequence of Sclerotinia sclerotiorum negative stranded RNA virus 7 (SsNSRV7), a virus-like contig obtained from Sclerotinia sclerotiorum metatranscriptomes, indicating BcMyV1 should be a strain of SsNSRV7. Phylogenetic analysis based on RdRp domains showed that BcMyV1 was clustered with the viruses in the family Mymonaviridae, suggesting it is a member of Mymonaviridae. BcMyV1 may be widely distributed in regions where B. cinerea occurs in China and even over the world, although it infected only 0.8% of tested B. cinerea strains.


Iron-binding proteins in adenoid cystic carcinoma of salivary glands: an immunohistochemical study.
An immunoperoxidase staining technique was used for detecting three major iron-binding proteins (transferrin, ferritin and lactoferrin) in 54 adenoid cystic carcinomas (ACCs) of major and minor salivary glands. Twenty-three normal salivary glands were investigated for comparison. Transferrin staining was detected mainly in the intercalated duct and serous acinar cells, and was found inconsistently in the myoepithelial cells surrounding normal ductules. Ferritin was always absent in the normal epithelial component of salivary gland. The presence of lactoferrin was demonstrated in the serous acinar cells and intercalated duct cells of normal salivary tissues. Five histological patterns were found in ACC, and for the cellular components of each, it was possible to establish a special immunohistochemical profile. Transferrin positivity was detected in the small angular cells of 25 cases (48%), in the duct luminal cells of 19 cases (37%) and in the hyalinized stroma of 4 cases (8%). Ferritin staining was positive in the small angular cells of 23 cases (44%) and in the duct luminal cells of 15 cases (29%). Lactoferrin was detected in only duct luminal cells of 38 ACCs (73%). The comparative immunohistochemical analysis between transferrin and ferritin showed a similar distribution in this carcinoma. On the basis of the immunohistochemical data, lactoferrin might be used as a marker of glandular differentiation.


[Differentiated prophylaxis of recurrent myocardial infarction].
149 patients stated at primarily examination as healthy had subsequently myocardial infarction (MI) at the age of 28-55 years. Preinfarction mean level of the risk factors was 3.3 per a man. For 5 years after MI the patients received differentiated therapy oriented to individual features of ischemic heart disease (IHD). The response was achieved in the majority of patients. To the end of the trial repeated MI occurred in 12.8% of the patients. 80.1% of them had angina pectoris (89.2%-angina of class I and II). 89.3% of the patients were able to work, 87.2% have lowered their mean level of risk factors to 1.5. It is demonstrated that a favourable course of postmyocardial infarction IHD was determined by adequacy of prophylactic measures rather than by health status before MI.


[Human brucellosis in Poland in 2002].
The registered 31 cases of human brucellosis in Poland comprise chronically ill professionals who in the past were involved for many years in the control of animal brucellosis. One case of acute infection was imported from Mediterranean region.


Photoinduced, copper-catalyzed alkylation of amides with unactivated secondary alkyl halides at room temperature.
The development of a mild and general method for the alkylation of amides with relatively unreactive alkyl halides (i.e., poor substrates for SN2 reactions) is an ongoing challenge in organic synthesis. We describe herein a versatile transition-metal-catalyzed approach: in particular, a photoinduced, copper-catalyzed monoalkylation of primary amides. A broad array of alkyl and aryl amides (as well as a lactam and a 2-oxazolidinone) couple with unactivated secondary (and hindered primary) alkyl bromides and iodides using a single set of comparatively simple and mild conditions: inexpensive CuI as the catalyst, no separate added ligand, and C-N bond formation at room temperature. The method is compatible with a variety of functional groups, such as an olefin, a carbamate, a thiophene, and a pyridine, and it has been applied to the synthesis of an opioid receptor antagonist. A range of mechanistic observations, including reactivity and stereochemical studies, are consistent with a coupling pathway that includes photoexcitation of a copper-amidate complex, followed by electron transfer to form an alkyl radical.


Meningeal tumor: A rare extrahepatic association in patients with polycystic liver disease enrolled for liver transplantation.
In the present study, we described a rare association of polycystic liver disease (PCLD) with intracranial meningiomas in patients included on a liver transplant list, focusing on the diagnosis, treatment and possible association with any genetic alterations. Two female patients, aged 39 and 49 years were included on a liver transplant list due to extensive PCLD, with symptoms related to an abdominal compartmental syndrome. Screening for extrahepatic manifestation revealed a right frontal meningioma in the first patient, and a parietal posterior calcified meningioma in the second patient, measuring 1 and 7×3×2 cm in diameter, respectively. Following tumor removal, the histological pattern was compatible with fibrous and transitional meningioma, respectively. Cytogenetic studies conducted following surgery did not reveal any changes in metaphase chromosomes. The postoperative follow-up for the two patients was uneventful, without complications, with the patients remaining on a liver transplant waiting list. We conclude that screening for extrahepatic manifestations of PCLD is mandatory, as certain lesions require treatment prior to liver transplantation. The lack of a genetic or familial association between these two cases show they are likely to have occurred by chance, rather than representing a previously unrecognized association between polycystic liver disease and cranial meningioma.


Hedgehog signaling patterns the tracheal branches.
The elaborate branching pattern of the Drosophila tracheal system originates from ten tracheal placodes on both sides of the embryo, each consisting of about 80 cells. Simultaneous cell migration from each tracheal pit in six different directions gives rise to the stereotyped branching pattern. Each branch contains a fixed number of cells. Previous work has shown that in the dorsoventral axis, localized activation of the Dpp, Wnt and EGF receptor (DER) pathways, subdivides the tracheal pit into distinct domains. We present the role of the Hedgehog (Hh) signaling system in patterning the tracheal branches. Hh is expressed in segmental stripes abutting the anterior border of the tracheal placodes. Induction of patched expression, which results from activation by Hh, demonstrates that cells in the anterior half of the tracheal pit are activated. In hh-mutant embryos migration of all tracheal branches is absent or stalled. These defects arise from a direct effect of Hh on tracheal cells, rather than by indirect effects on patterning of the ectoderm. Tracheal cell migration could be rescued by expressing Hh only in the tracheal cells, without rescuing the ectodermal defects. Signaling by several pathways, including the Hh pathway, thus serves to subdivide the uniform population of tracheal cells into distinct cell types that will subsequently be recruited into the different branches.


Inhibition of autophagy potentiates the antitumor effect of the multikinase inhibitor sorafenib in hepatocellular carcinoma.
Multikinase inhibitor sorafenib inhibits proliferation and angiogenesis of tumors by suppressing the Raf/MEK/ERK signaling pathway and VEGF receptor tyrosine kinase. It significantly prolongs median survival of patients with advanced hepatocellular carcinoma (HCC) but the response is disease-stabilizing and cytostatic rather than one of tumor regression. To examine the mechanisms underlying the relative resistance in HCC, we investigated the role of autophagy, an evolutionarily conserved self-digestion pathway, in hepatoma cells in vitro and in vivo. Sorafenib treatment led to accumulation of autophagosomes as evidenced by conversion from LC3-I to LC3-II observed by immunoblot in Huh7, HLF and PLC/PRF/5 cells. This induction was due to activation of autophagic flux, as there was further increase in LC3-II expression upon treatment with lysosomal inhibitors, clear decline of the autophagy substrate p62, and an mRFP-GFP-LC3 fluorescence change in sorafenib-treated hepatoma cells. Sorafenib inhibited the mammalian target of rapamycin complex 1 and its inhibition led to accumulation of LC3-II. Pharmacological inhibition of autophagic flux by chloroquine increased apoptosis and decreased cell viability in hepatoma cells. siRNA-mediated knockdown of the ATG7 gene also sensitized hepatoma cells to sorafenib. Finally, sorafenib induced autophagy in Huh7 xenograft tumors in nude mice and coadministration with chloroquine significantly suppressed tumor growth compared with sorafenib alone. In conclusion, sorafenib administration induced autophagosome formation and enhanced autophagic activity, which conferred a survival advantage to hepatoma cells. Concomitant inhibition of autophagy may be an attractive strategy for unlocking the antitumor potential of sorafenib in HCC.


Bone Organoids: Recent Advances and Future Challenges.
Bone defects stemming from tumorous growths, traumatic events, and diverse conditions present a profound conundrum in clinical practice and research. While bone has the inherent ability to regenerate, substantial bone anomalies require bone regeneration techniques. Bone organoids represent a new concept in this field, involving the three-dimensional (3D) self-assembly of bone-associated stem cells guided in vitro with or without extracellular matrix material, resulting in a tissue that mimics the structural, functional, and genetic properties of native bone tissue. Within the scientific panorama, bone organoids have ascended to an esteemed status, securing significant experimental endorsement. Through a synthesis of current literature and pioneering studies, this review offers a comprehensive survey of the bone organoid paradigm, delves into the quintessential architecture and ontogeny of bone, and highlights the latest progress in bone organoid fabrication. Furthermore, we identify existing challenges and prospective directions for future research, advocating for interdisciplinary collaboration to fully harness the potential of this burgeoning domain. Conclusively, as bone organoid technology continues to mature, its implications for both clinical and research landscapes are poised to be profound. This article is protected by copyright. All rights reserved.


Acute-phase protein α1-antitrypsin--a novel regulator of angiopoietin-like protein 4 transcription and secretion.
The angiopoietin-like protein 4 (angptl4, also known as peroxisome proliferator-activated receptor [PPAR]γ-induced angiopoietin-related protein) is a multifunctional protein associated with acute-phase response. The mechanisms accounting for the increase in angptl4 expression are largely unknown. This study shows that human α1-antitrypsin (A1AT) upregulates expression and release of angplt4 in human blood adherent mononuclear cells and in primary human lung microvascular endothelial cells in a concentration- and time-dependent manner. Mononuclear cells treated for 1 h with A1AT (from 0.1 to 4 mg/ml) increased mRNA of angptl4 from 2- to 174-fold, respectively, relative to controls. In endothelial cells, the maximal effect on angptl4 expression was achieved at 8 h with 2 mg/ml A1AT (11-fold induction versus controls). In 10 emphysema patients receiving A1AT therapy (Prolastin), plasma angptl4 levels were higher relative to patients without therapy (nanograms per milliliter, mean [95% confidence interval] 127.1 [99.5-154.6] versus 76.8 [54.8-98.8], respectively, p = 0.045) and correlated with A1AT levels. The effect of A1AT on angptl4 expression was significantly diminished in cells pretreated with a specific inhibitor of ERK1/2 activation (UO126), irreversible and selective PPARγ antagonist (GW9662), or genistein, a ligand for PPARγ. GW9662 did not alter the ability of A1AT to induce ERK1/2 phosphorylation, suggesting that PPARγ is a critical mediator in the A1AT-driven angptl4 expression. In contrast, the forced accumulation of HIF-1α, an upregulator of angptl4 expression, enhanced the effect of A1AT. Thus, acute-phase protein A1AT is a physiological regulator of angptl4, another acute-phase protein.


MethyLight droplet digital PCR for detection and absolute quantification of infrequently methylated alleles.
Aberrant DNA methylation is a common epigenetic alteration found in colorectal adenomas and cancers and plays a role in cancer initiation and progression. Aberrantly methylated DNA loci can also be found infrequently present in normal colon tissue, where they seem to have potential to be used as colorectal cancer (CRC) risk biomarkers. However, detection and precise quantification of the infrequent methylation events seen in normal colon is likely beyond the capability of commonly used PCR technologies. To determine the potential for methylated DNA loci as CRC risk biomarkers, we developed MethyLight droplet digital PCR (ddPCR) assays and compared their performance to the widely used conventional MethyLight PCR. Our analyses demonstrated the capacity of MethyLight ddPCR to detect a single methylated NTRK3 allele from among more than 3125 unmethylated alleles, 25-fold more sensitive than conventional MethyLight PCR. The MethyLight ddPCR assay detected as little as 19 and 38 haploid genome equivalents of methylated EVL and methylated NTRK3, respectively, which far exceeded conventional MethyLight PCR (379 haploid genome equivalents for both genes). When assessing methylated EVL levels in CRC tissue samples, MethyLight ddPCR reduced coefficients of variation (CV) to 6-65% of CVs seen with conventional MethyLight PCR. Importantly, we showed the ability of MethyLight ddPCR to detect infrequently methylated EVL alleles in normal colon mucosa samples that could not be detected by conventional MethyLight PCR. This study suggests that the sensitivity and precision of methylation detection by MethyLight ddPCR enhances the potential of methylated alleles for use as CRC risk biomarkers.


A perspective of nanotechnology in hypersensitivity reactions including drug allergy.
We provide an overview of the application of the concepts of nanoscience and nanotechnology as a novel scientific approach to the area of nanomedicine related to the domain of the immune system. Particular emphasis will be paid to studies on drug allergy reactions. Several well defined chemical structures arranged in the dimension of the nanoscale are currently being studied for biomedical purposes. By interacting with the immune system, some of these show promising applications as vaccines, diagnostic tools and activators/effectors of the immune response. Even a brief listing of some key applications of nanostructured materials shows how broad and intense this area of nanomedicine is. As a result of the development of nanoscience and nanotechnology applied to medicine, new approaches can be envisioned for problems related to the modulation of the immune response, as well as in immunodiagnosis, and to design new tools to solve related medical challenges. Nanoparticles offer unique advantages with which to exploit new properties and for materials to play a major role in new diagnostic techniques and therapies. Fullerene-C60 and multivalent functionalized gold nanoparticles of various sizes have led to new tools and opened up new ways to study and interact with the immune system. Some of the most versatile nanostructures are dendrimers. In their interaction with the immune system they can naturally occurring macromolecules, taking advantage of the fact that dendrimers can be synthesized into nanosized structures. Their multivalence can be successfully exploited in vaccines and diagnostic tests for allergic reactions.


[Assessment of tuberculosis treatment accessibility for patients co-infected or not with the human immunodeficiency virus].
This study aimed to evaluate accessibility to treatment for people with TB co-infected or not with HIV. This cross-sectional study addressed issues regarding accessibility to treatment in a city in the interior of São Paulo state, Brazil. The instrument Primary Care Assessment Tool was utilized with 95 people. To evaluate access to treatment, Student's t test was used. The mean scores of variables were analyzed separately and compared between two groups (people with TB co-infected with HIV and people with TB not co-infected with HIV ). Mean scores showed that HIV co-infected people presented greater difficulties in gaining access than those not co-infected. Professionals visited co-infected people more often when compared to those not co-infected; the co-infected people almost never accessed treatment for their disease in the Health Unit nearest their home. There is, therefore, the need for greater integration and communication between the programs for treatment of Tuberculosis and STD/AIDS.


HLA-F complex without peptide binds to MHC class I protein in the open conformer form.
HLA-F has low levels of polymorphism in humans and is highly conserved among primates, suggesting a conserved function in the immune response. In this study, we probed the structure of HLA-F on the surface of B lymphoblastoid cell lines and activated lymphocytes by direct measurement of peptide binding to native HLA-F. Our findings suggested that HLA-F is expressed independently of bound peptide, at least in regard to peptide complexity profiles similar to those of either HLA-E or classical MHC class I (MHC-I). As a further probe of native HLA-F structure, we used a number of complementary approaches to explore the interactions of HLA-F with other molecules, at the cell surface, intracellularly, and in direct physical biochemical measurements. This analysis demonstrated that HLA-F surface expression was coincident with MHC-I H chain (HC) expression and was downregulated upon perturbation of MHC-I HC structure. It was further possible to directly demonstrate that MHC-I would interact with HLA-F only when in the form of an open conformer free of peptide and not as a trimeric complex. This interaction was directly observed by coimmunoprecipitation and by surface plasmon resonance and indirectly on the surface of cells through coincident tetramer and MHC-I HC colocalization. These data suggest that HLA-F is expressed independently of peptide and that a physical interaction specific to MHC-I HC plays a role in the function of MHC-I HC expression in activated lymphocytes.