Clinical significance of BCR-ABL fusion gene subtypes in chronic myelogenous and acute lymphoblastic leukemias.
Some reports have suggested that chronic myeloid leukemia (CML) patients have a higher prevalence of M-bcr than acute lymphoblastic leukemia (ALL) patients, which show a higher prevalence of m-bcr. However, the relationship between BCR-ABL subtypes and progression of CML and ALL remains unclear. 354 CML chronic phase (CML-CP) patients, 26 CML blastic phase (CML-BP) patients and 72 ALL patients before treatment with BCR-ABL positive were recruited for blood routine examination and bone marrow smear cytology. Some 80 CML-CP and 32 ALL patients after imatinib (IM) treatment were followed-up for BCR-ABL relative concentrations detected after treatment for 3, 6 and 9 months and 1 year. Before treatment, CML-CP patients showed lower BCR-ABL relative concentrations with a higher proportion of M-bcr (42.7%) compared to CML-BP and ALL patients while ALL patients had a higher BCR-ABL relative concentration with high expression of m-bcr (51.4%). Patients with M-bcr demonstrated higher WBC counts than those with m-bcr and the mixed group and higher PLT counts were noted in the CML-CP and ALL groups. After imatinib (IM) treatment, patients with m-bcr showed higher BCR-ABL relative concentrations in both CML-CP and ALL groups. This study identified the BCR-ABL gene as an important factor in CML and ALL cases. The M-bcr subtype was associated more with CML while the m-bcr subtype was associated more with ALL. Patients with m-bcr seem to have a poorer response to IM in either CML or ALL patients compared to M-bcr patients.


Dopaminergic and benzodiazepine receptors studied in vivo by PET.
Using Positron Emission Tomography (PET) and specific radioligands, dopaminergic D2 (DA-D2 receptors) and benzodiazepine receptors (BZ-receptors) were studied in living animals during normal and pathological conditions. In vivo characterization of both receptors was performed using two highly specific antagonists namely: 11C-Ro 15 1788 for BZ-receptors and 76 Br-Bromospiperone for DA-D2 receptors. Changes in 11c-Ro 15 1788 specific binding to BZ-receptors were observed during convulsive seizures. After MPTP treatment, a decrease in the 76 Br-Bromospiperone striatal specific binding was observed, correlated with the establishment of a Parkinson-like syndrome.


Immunoassay on free-standing electrospun membranes.
For the purpose of immunoassay, electrospun membranes can be thought as the threadlike self-assembling of nano/microbeads. Nonwoven membranes of electrospun poly(epsilon-caprolactone) (PCL) fibers display excellent tenacity, flexibility and suitable surface energy. These PCL membranes exhibit easy handling in air, fast spreading, and wetting in aqueous solution, and rapid adsorption of protein molecules by hydrophobic interaction. After a fold-and-press process, the membrane porosity was reduced from approximately 75% to less than 10%, whereas the thickness increased from 5.3 to 280 microm. The resulting fluorescence signal from adsorbed protein increased>120x. With anti-HSA and HSA-FITC as an immunoassay model, a linear detection range from 500 ng/mL down to 1 ng/mL is obtained, with a detection of limit (LOD) of approximately 0.08 ng/mL. By comparison, conventional nitrocellulose and a 24.3 microm PCL fiber electrospun membrane displayed a much higher LOD of approximately 100 ng/mL. Immunoassay on free-standing electrospun membrane successfully combines the low-cost and simplicity of conventional membrane immunoassay, with the fast reaction speed and high sensitivity characteristic of magnetic nano/microbeads bioassays.


A novel cylindrical overlap-and-fling mechanism used by sea butterflies.
The clap-and-fling mechanism is a well-studied, unsteady lift generation mechanism widely used by flying insects and is considered obligatory for tiny insects flying at low to intermediate Reynolds numbers, Re However, some aquatic zooplankters including some pteropod (i.e. sea butterfly) and heteropod species swimming at low to intermediate Re also use the clap-and-fling mechanism. These marine snails have extremely flexible, actively deformed, muscular wings which they flap reciprocally to create propulsive force, and these wings may enable novel lift generation mechanisms not available to insects, which have less flexible, passively deformed wings. Using high-speed stereophotogrammetry and micro-particle image velocimetry, we describe a novel cylindrical overlap-and-fling mechanism used by the pteropod species Cuvierina atlantica In this maneuver, the pteropod's wingtips overlap at the end of each half-stroke to sequentially form a downward-opening cone, a cylinder and an upward-opening cone. The transition from downward-opening cone to cylinder produces a downward-directed jet at the trailing edges. Similarly, the transition from cylinder to upward-opening cone produces downward flow into the gap between the wings, a leading edge vortex ring and a corresponding sharp increase in swimming speed. The ability of this pteropod species to perform the cylindrical overlap-and-fling maneuver twice during each stroke is enabled by its slender body and highly flexible wings. The cylindrical overlap-and-fling mechanism observed here may inspire the design of new soft robotic aquatic vehicles incorporating highly flexible propulsors to take advantage of this novel lift generation technique.


New developments in the diagnosis of viral diseases.
Major technical advances have occurred, especially in the last 5 years, in the laboratory diagnosis of viral infections. Immunologic detection of immediate early antigens in specimens such as bronchoalveolar lavage fluid and blood inoculated into shell vial cell cultures, particularly for herpesvirus (cytomegalovirus, herpes simplex virus, varicella-zoster virus), has provided results 16 to 48 hours after inoculation rather than the several days required for recognition of cytopathic effects in conventional tube cell cultures. Similarly, cytomegalovirus viremia can be detected directly by immunostaining of peripheral blood leukocytes with commercially available reagents the same day the specimen is submitted to the laboratory. Single-test membrane immunoassays have provided rapid (15 minutes) detection of viral antigens (respiratory syncytial virus, rotavirus, influenza virus type A). In the near future, diagnostic virology laboratories will be expected to monitor viral strains for susceptibility to the growing list of antiviral drugs. Amplification of nucleic acid sequences of viruses from cerebrospinal fluid or tissue, which generally does not yield isolates by conventional diagnostic techniques, has added a new dimension to the laboratory diagnosis of viral infection.


[Vertical talus].
Authors differentiate four etiological groups of vertical talus. In their material the congenital and symptomatic rocker bottom foot was rarer than the paralytic one. Good result can be expected mainly from the operative treatment. From 69 cases good clinical and radiological result was reached in 58, whereas in 6 cases the treatment had no result. It was their experience that a reduction of the talus and of the navicular bone can be reached by tenotomies and capsulotomies and by these a normal form of the foot can be secured. In the conservation of the operative results the significance of the organized care is stressed.


Mesenchymal stem cell-derived factors: Immuno-modulatory effects and therapeutic potential.
Stem cell-based therapy is considered to be a new hope in transplantation medicine. Among stem cells, mesenchymal stem cells (MSCs) are, due to their differentiation and immuno-modulatory characteristics, the most commonly used as therapeutic agents in the treatment of immune-mediated diseases. MSCs migrate to the site of inflammation and modulate immune response. The capacity of MSC to alter phenotype and function of immune cells are largely due to the production of soluble factors which expression varies depending on the pathologic condition to which MSCs are exposed. Under inflammatory conditions, MSCs-derived factors suppress both innate and adaptive immunity by attenuating maturation and capacity for antigen presentation of dendritic cells, by inducing polarization of macrophages towards alternative phenotype, by inhibiting activation and proliferation of T and B lymphocytes and by reducing cytotoxicity of NK and NKT cells. In this review, we emphasized current findings regarding immuno-modulatory effects of MSC-derived factors and emphasize their potential in the therapy of immune-mediated diseases. © 2017 BioFactors, 43(5):633-644, 2017.


The novel role of IFITM1-3 in myogenic differentiation of C2C12 cells.
Interferon-induced transmembrane proteins (IFITMs 1, 2, and 3) play a critical role in preventing pathogen infection in vertebrates. They are also involved in the occurrence and prognosis of cancer. Myogenesis is a complex process regulated by several factors. This study disclosed that Ifitm1-3 were upregulated in the process of myogenic differentiation of C2C12 myoblasts on days 3, 5, and 7. This positively correlated with the expression of differentiation factors MyoD, myogenin, Mrf5, and desmin. Furthermore, knockdown of Ifitm1-3 by their individual siRNAs inhibited myogenesis of C2C12 myoblasts, with relative downregulation of MyoD, myogenin, Mrf5, and desmin. Subsequently, myotube formation and fusion percentage decreased. Co-immunoprecipitation combined with LC-MS/MS analysis uncovered the interaction proteins of IFITM1 and IFITM3 in C2C12 myoblasts. A total of 84 overlapped interaction proteins of IFITM1 and IFITM3 were identified, and one of the clusters was engaged in cytoskeletal and sarcomere proteins, including desmin, myosin, actin, vimentin, nestin, ankycorbin, and nucleolin. Hence, we hypothesize that these interacting proteins may function as scaffolds for IFITM1-3, possibly through the interaction protein desmin to initiate further interaction with other proteins to participate in myogenesis; however, the molecular mechanisms remain unclear. Our study may contribute to the development of novel therapeutics for myopathic diseases.


ProgSIO-MSA: Progressive-based single iterative optimization framework for multiple sequence alignment using an effective scoring system.
Aligning more than two biological sequences is termed multiple sequence alignment (MSA). To analyze biological sequences, MSA is one of the primary activities with potential applications in phylogenetics, homology markers, protein structure prediction, gene regulation, and drug discovery. MSA problem is considered as NP-complete. Moreover, with the advancement of Next-Generation Sequencing techniques, all the gene and protein databases are consistently loaded with a vast amount of raw sequence data which are neither analyzed nor annotated. To analyze these growing volumes of raw sequences, the need of computationally-efficient (polynomial time) models with accurate alignment is high. In this study, a progressive-based alignment model is proposed, named ProgSIO-MSA, which consists of an effective scoring system and an optimization framework. The proposed scoring system aligns sequences effectively using the combination of two scoring strategies, i.e. Look Back Ahead, that scores a residue pair dynamically based on the status information of the previous position to improve the sum-of-pair score, and Position-Residue-Specific Dynamic Gap Penalty, that dynamically penalizes a gap using mutation matrix on the basis of residue and its position information. The proposed single iterative optimization (SIO) framework identifies and optimizes the local optima trap to improve the alignment quality. The proposed model is evaluated against progressive-based state-of-the-art models on two benchmark datasets, i.e. BAliBASE and SABmark. The alignment quality (biological accuracy) of the proposed model is increased by a factor of 17.7% on BAliBASE dataset. The proposed model's efficiency is compared with state-of-the-art models using time complexity as well as runtime analysis. Wilcoxon signed-rank statistical test results concluded that the quality of the proposed model significantly outperformed progressive-based state-of-the-art models.


Auxiliary partial liver transplantation for end-stage chronic liver disease.
Auxiliary heterotopic liver transplantation is theoretically attractive because it leaves the recipient's liver in place. The surgical trauma of hepatectomy is avoided, and failure of the graft does not necessarily lead to the death of the patient or a second, emergency transplantation. Another advantage is that matching the body sizes of the donor and the recipient is not mandatory, which increases the number of possible donors. However, previous clinical results of auxiliary liver transplantation have been poor. We performed auxiliary partial liver transplantation in six consecutive patients with end-stage chronic liver disease who were not accepted for orthotopic liver transplantation because they had massive ascites, deficient clotting function, cachexia, or poor pulmonary reserve. The donor liver was transplanted to the right subhepatic region after removal of segments II and III, and it was provided with portal and arterial blood. There were no major changes in hemodynamic measurements during surgery. The mean hospital stay after transplantation was 22.7 days (range, 14 to 29). After a mean follow-up period of 14 months (range, 5 to 23), all patients were alive, with good graft function as demonstrated by scintigraphy, Doppler ultrasonography, and synthesis of clotting factors. From these observations we conclude that auxiliary partial liver transplantation is an attractive alternative to orthotopic liver transplantation in high-risk patients. Its role in other patients who need liver transplants remains to be defined.


Microsatellite diversity, population subdivision and gene flow in the Lipizzan horse.
Blood samples of 561 Lipizzan horses from subpopulations (studs) of seven European countries representing a large fraction of the breed's population were used to examine the genetic diversity, population subdivision and gene flow in the breed. DNA analysis based on 18 microsatellite loci revealed that genetic diversity (observed heterozygosity = 0.663, gene diversity = 0.675 and the mean number of alleles = 7.056) in the Lipizzan horse is similar to other horse breeds as well as to other domestic animal species. The genetic differentiation between Lipizzan horses from different studs, although moderate, was apparent (pairwise F(ST) coefficients ranged from 0.021 to 0.080). Complementary findings explaining the genetic relationship among studs were revealed by genetic distance and principal component analysis. One genetic cluster consisted of the subpopulations of Austria, Italy and Slovenia, which represent the classical pool of Lipizzan horse breeding. A second cluster was formed by the Croatian, Hungarian and Slovakian subpopulations. The Romanian subpopulation formed a separate unit. The largest genetic differentiation was found between the Romanian and Italian subpopulation. Genetic results are consistent with the known breeding history of the Lipizzan horse. Correct stud assignment was obtained for 80.9% and 92.1% of Lipizzan horses depending on the inclusion or exclusion of migrant horses, respectively. The results of the present study will be useful for the development of breeding strategies, which consider classical horse breeding as well as recent achievements of population and conservation genetics.


β-adrenergic signaling broadly contributes to LTP induction.
Long-lasting forms of long-term potentiation (LTP) represent one of the major cellular mechanisms underlying learning and memory. One of the fundamental questions in the field of LTP is why different molecules are critical for long-lasting forms of LTP induced by diverse experimental protocols. Further complexity stems from spatial aspects of signaling networks, such that some molecules function in the dendrite and some are critical in the spine. We investigated whether the diverse experimental evidence can be unified by creating a spatial, mechanistic model of multiple signaling pathways in hippocampal CA1 neurons. Our results show that the combination of activity of several key kinases can predict the occurrence of long-lasting forms of LTP for multiple experimental protocols. Specifically Ca2+/calmodulin activated kinase II, protein kinase A and exchange protein activated by cAMP (Epac) together predict the occurrence of LTP in response to strong stimulation (multiple trains of 100 Hz) or weak stimulation augmented by isoproterenol. Furthermore, our analysis suggests that activation of the β-adrenergic receptor either via canonical (Gs-coupled) or non-canonical (Gi-coupled) pathways underpins most forms of long-lasting LTP. Simulations make the experimentally testable prediction that a complete antagonist of the β-adrenergic receptor will likely block long-lasting LTP in response to strong stimulation. Collectively these results suggest that converging molecular mechanisms allow CA1 neurons to flexibly utilize signaling mechanisms best tuned to temporal pattern of synaptic input to achieve long-lasting LTP and memory storage.


Catalytic mechanism of Escherichia coli endonuclease VIII: roles of the intercalation loop and the zinc finger.
Endonuclease VIII (Nei) excises oxidatively damaged pyrimidines from DNA and shares structural and functional homology with formamidopyrimidine-DNA glycosylase. Although the structure of Escherichia coli Nei is solved [Zharkov et al. (2002) EMBO J. 21, 789-800], the functions of many of its amino acid residues involved in catalysis and substrate specificity are not known. We constructed a series of Nei mutants that interfere with eversion of the damaged base from the helix (QLY69-71AAA, DeltaQLY69-71) or perturb the conserved zinc finger (R171A, Q261A). Steady-state kinetics were measured with these mutant enzymes using substrates containing 5,6-dihydrouracil, two enantiomers of thymine glycol, 8-oxo-7,8-dihydroguanine, and an abasic site positioned opposite each of the four canonical DNA bases. To some extent, all Nei mutants were deficient in processing damaged DNA, with mutations in the zinc finger generally having a more profound effect. Wild-type Nei showed prominent opposite-base specificity (G > C approximately = T > A) when the lesion was 5,6-dihydrouracil or cis-(5S,6R)-thymine glycol but not for other lesions tested. Mutations in the Q69-Y71 loop eliminated this effect. Only wild-type Nei and Nei-Q261A mutants could be reductively cross-linked to damaged base-containing DNA. Experiments involving trapping with NaBH4 and the kinetics of DNA cleavage catalyzed by Nei-Q261A suggested that this mutant was deficient in regenerating free enzyme from the Nei-DNA covalent complex formed during the reaction. We conclude that the opposite-base specificity of Nei is primarily governed by residues in the Q69-Y71 loop and that both this loop and the zinc finger contribute significantly to the substrate specificity of Nei.


Selective hypoaldosteronism despite prolonged pre- and postoperative hyperreninemia in primary aldosteronism.
In a prospective study of 7 patients with aldosterone-producing adenoma (APA), long-term (6-72 months) preoperative stimulation of plasma renin activity (PRA) by diuretic therapy (spironolactone plus hydrochlorothiazide) did not prevent selective aldosterone deficiency postoperatively. In all patients aldosterone excretion rate (AER) fell to subnormal values (from a mean of 97 to 2.6 mug/24 h) following removal of APA, although PRA remained elevated. Generalized adrenocortical insufficiency was excluded by the demonstration of normal baseline plasma cortisol and urinary 17-OHCS and the appropriate response to ACTH stimulation. In 6 of 7 patients studied 1-3 months postoperatively, short-term (4 days) sodium deprivation evoked normal increases in PRA, but AER response was blunted (except in 1). Restudy of 3 of 6 patients after 6-12 months revealed that aldosterone production had returned to normal. These results indicate that renin deficiency is not the principal cause of postoperative selective hypoaldosteronism in these patients. On the other hand, they appear to substantiate the possibility raised by in vitro and in vivo studies that spironolactone can directly inhibit aldosterone biosynthesis.


[The music quality feeling and music perception of adult cochlear implant recipients].
Objective:To assess the music quality feeling and music perception abilities of adults with cochlear implants and to probe the correlation between the two. Method:Forty adult cochlear implant recipients recruited from the Bejing Tongren Hosipital(27 males, 13 females, average ages at 36.13±13.33) and 40 normal-hearing subjects(21 males, 19females, average ages at 26.53±6.94) participated in this study. Musical Sounds In Cochlear Implant were used to assess the pitch, melody and timbre perception for both normal-hearing subjects and adult cochlear implant recipients. To assess the self-reported music quality feeling of normal-hearing subjects and adult cochlear implant recipients from three aspects of listening music quality(Naturalness, comfort, and clarity). Using some questions about musical background and listening habits of the Munich Music questionnaire(MUMU) to assess the musical experience scores of all subjects.Result:For cochlear implant recipients, the pitch discrimination thresholds at A4 and C4 and the correction rate of melody discrimination and instrument identification test were significantly different from those for normalhearing subjects(P<0.05). The average music quality feeling scores of the cochlear implant recipients and normal-hearing subjects were 4.75±2.76 and 8.33±1.47, respectively. The differences between the two groups were significant(P<0.05). Type of CI had no significant influence on the results of musical quality feeling(P>0.05). The scores of music quality feeling was significantly related with the pitch discrimination thresholds at A4 and C4(P<0.05), but had no significant correlation with the correlation rate of melody discrimination test and instrument identification test(P>0.05).Conclusion:For adult cochlear implant recipients, the music quality feeling and the music perception ability are weaker than normal-hearing subjects. The music quality feeling is related to the pitch perception, but had no relationship with the melody and timbre perception ability.


Protein kinase C-mediated phosphorylation of HIV-I nef in human cell lines.
Stable human cell lines expressing the human immunodeficiency virus type I (HIV-I) Nef protein from inducible promoters were used to analyze the phosphorylation status of Nef in vivo. Nef phosphorylation in both HeLa and Jurkat cells was stimulated by phorbol ester treatment. Phosphoamino acid analysis revealed a predominance of phosphoserine with a small proportion of phosphothreonine. Treatment of cells with selective protein kinase inhibitors revealed that Nef phosphorylation was markedly reduced by bisindolylmaleimide, an inhibitor of protein kinase C, but was unaffected by inhibitors of mitogen-activated protein kinase kinase or cAMP-dependent kinase. These data implicate protein kinase C in Nef phosphorylation in vivo, and thus confirm and extend earlier in vitro data. Phosphorylation of a nonmyristoylated Nef mutant was impaired, suggesting that membrane targeting of Nef was required for phosphorylation. This was expected given that activated protein kinase C translocates from the cytosol to the plasma membrane. However, analysis of the subcellular localization of phosphorylated wild-type Nef revealed that both the cytosolic and membrane-associated pools of Nef were phosphorylated to an equivalent extent. Thus the significance of myristoylation for Nef function may be in influencing protein conformation, although these data could be explained by a transient and dynamic interaction between myristoylated Nef and the plasma membrane.


Multislice computed tomography coronary angiography: clinical applications.
Cardiac and coronary computed tomography (CT) is becoming increasingly common in clinical practice. Even if there is no well-established evidence, this diagnostic modality is so strong and effective and, in skilled hand, it can be readily used in clinical practice. After learning its potential and the technical limits, this tool could be used for risk stratification as well as for revascularization evaluation. In this review, we will describe the results of present literature, clinical applications at present considered suitable to CT technology (i.e. 64-slice and dual-source scanners) and future applications and innovations.


Fine Mapping of QTLs for Ascochyta Blight Resistance in Pea Using Heterogeneous Inbred Families.
Ascochyta blight (AB) is an important disease of pea which can cause severe grain yield loss under wet conditions. In our previous study, we identified two quantitative trait loci (QTLs) abIII-1 and abI-IV-2 for AB resistance and these QTLs were consistent across locations and/or years in an inter-specific pea population (PR-19) developed from a cross between Alfetta (Pisum sativum) and P651 (P. fulvum). The objectives of this study were to fine map the abIII-1 and abI-IV-2 QTLs using a high density single nucleotide polymorphism (SNP)-based genetic linkage map and analyze identified markers in heterogeneous inbred family (HIF) populations. Selective genotyping of 51 PR-19 recombinant inbred lines was performed using genotyping-by-sequencing (GBS) and the resulting high density genetic linkage map was used to identify eight new SNP markers within the abI-IV-2 QTL, whereas no additional SNPs were identified within the abIII-1 QTL. Two HIF populations HIF-224 (143 lines) and HIF-173 (126 lines) were developed from F6 RILs PR-19-224 and PR-19-173, respectively. The HIF populations evaluated under field conditions in 2015 and 2016 showed a wide range of variation for reaction to AB resistance. Lodging score had significant positive (P < 0.001) correlation with AB scores. HIFs were genotyped using SNP markers within targeted QTLs. The genotypic and phenotypic data of the HIFs were used to identify two new QTLs, abI-IV-2.1 and abI-IV-2.2 for AB resistance within the abI-IV-2 QTL. These QTLs individually explained 5.5 to 14% of the total phenotypic variation. Resistance to lodging was also associated with these two QTLs. Identified SNP markers will be useful in marker assisted selection for development of pea cultivars with improved AB resistance.


Placental labyrinth formation in mice requires endothelial FLRT2/UNC5B signaling.
The placental labyrinth is the interface for gas and nutrient exchange between the embryo and the mother; hence its proper development is essential for embryogenesis. However, the molecular mechanism underlying development of the placental labyrinth, particularly in terms of its endothelial organization, is not well understood. Here, we determined that fibronectin leucine-rich transmembrane protein 2 (FLRT2), a repulsive ligand of the UNC5 receptor family for neurons, is unexpectedly expressed in endothelial cells specifically in the placental labyrinth. Mice lacking FLRT2 in endothelial cells exhibited embryonic lethality at mid-gestation, with systemic congestion and hypoxia. Although they lacked apparent deformities in the embryonic vasculature and heart, the placental labyrinths of these embryos exhibited aberrant alignment of endothelial cells, which disturbed the feto-maternal circulation. Interestingly, this vascular deformity was related to endothelial repulsion through binding to the UNC5B receptor. Our results suggest that the proper organization of the placental labyrinth depends on coordinated inter-endothelial repulsion, which prevents uncontrolled layering of the endothelium.


Functional involvement of cerebral cortex in Duchenne muscular dystrophy.
Transcranial stimulation was performed in 4 patients with Duchenne muscular dystrophy and 4 control subjects. The patients' central motor conduction time was normal. The threshold for evoking electromyographic responses using electrical anodal stimulation was the same in both groups, but the threshold for stimulation with a circular magnetic coil at the vertex was higher in the patients. This is compatible with reduced cortical excitability that may be related to the deficiency of brain synaptic dystrophin.


[Compulsory admissions to Frederiksberg Hospital after introduction of the new psychiatric legislation compared with the period just before the change of the law].
The new Danish Act on confinement and other means of compulsion in psychiatry came into force October 1st, 1989. To investigate changes connected with the new act all involuntary commitments to the psychiatric department of the hospital of Frederiksberg, with a catchment area of 85,000 inhabitants, were prospectively registered during 23 months before, 25 months after the new legislation. Information about the commitment was obtained from the admission papers and the case records. The patients were asked to participate in an interview. Both before and after the new act there were about 35 commitments per year. There was a trend towards an increasing number of women being committed by reason of threat to the patient's health after the new act. No significant changes were observed regarding age, condition at admission, diagnoses, duration of hospitalization or condition at discharge. A little more than half of the committed patients participated in the interviews. The new act stressed the rights of the patients. In spite of this the patients did not express any increase in their understanding of or satisfaction with their hospitalization.


Volatilomics Reveals Potential Biomarkers for Identification of Renal Cell Carcinoma: An In Vitro Approach.
The identification of noninvasive biomarkers able to detect renal cell carcinoma (RCC) at an early stage remains an unmet clinical need. The recognition that altered metabolism is a core hallmark of cancer boosted metabolomic studies focused in the search for cancer biomarkers. The present work aims to evaluate the performance of the volatile metabolites present in the extracellular medium to discriminate RCC cell lines with distinct histological subtypes (clear cell and papillary) and metastatic potential from non-tumorigenic renal cells. Hence, volatile organic compounds (VOCs) and volatile carbonyl compounds (VCCs) were extracted by headspace solid-phase microextraction (HS-SPME) and analyzed by gas chromatography-mass spectrometry (GC-MS). Multivariate and univariate analysis unveiled a panel of metabolites responsible for the separation between groups, mostly belonging to ketones, alcohols, alkanes and aldehydes classes. Some metabolites were found similarly altered for all RCC cell lines compared to non-tumorigenic cells, namely 2-ethylhexanol, tetradecane, formaldehyde, acetone (increased) and cyclohexanone and acetaldehyde (decreased). Furthermore, significantly altered levels of cyclohexanol, decanal, decane, dodecane and 4-methylbenzaldehyde were observed in all metastatic RCC cell lines when compared with the non-metastatic ones. Moreover, some alterations in the volatile composition were also observed between RCC histological subtypes. Overall, our results demonstrate the potential of volatile profiling for identification of noninvasive candidate biomarkers for early RCC diagnosis.


Chemotherapy-induced peripheral neuropathy-part 2: focus on the prevention of oxaliplatin-induced neurotoxicity.
Chemotherapy-induced peripheral neuropathy (CIPN) is regarded as one of the most common dose-limiting adverse effects of several chemotherapeutic agents, such as platinum derivatives (oxaliplatin and cisplatin), taxanes, vinca alkaloids and bortezomib. CIPN affects more than 60% of patients receiving anticancer therapy and although it is a nonfatal condition, it significantly worsens patients' quality of life. The number of analgesic drugs used to relieve pain symptoms in CIPN is very limited and their efficacy in CIPN is significantly lower than that observed in other neuropathic pain types. Importantly, there are currently no recommended options for effective prevention of CIPN, and strong evidence for the utility and clinical efficacy of some previously tested preventive therapies is still limited. The present article is the second one in the two-part series of review articles focused on CIPN. It summarizes the most recent advances in the field of studies on CIPN caused by oxaliplatin, the third-generation platinum-based antitumor drug used to treat colorectal cancer. Pharmacological properties of oxaliplatin, genetic, molecular and clinical features of oxaliplatin-induced neuropathy are discussed. Available therapies, as well as results from clinical trials assessing drug candidates for the prevention of oxaliplatin-induced neuropathy are summarized. Emerging novel chemical structures-potential future preventative pharmacotherapies for CIPN caused by oxaliplatin are reported.


Cross-induction of predominant NPb idiotypic antibodies with derivatives of (4-hydroxy-3-nitrophenyl) acetyl.
Immunization of C57BL/6 mice with bovine gamma-globulin (BGG) conjugated with (4-hydroxy-3-nitrophenyl) acetyl (NP) induced a population of anti-NP antibodies that bear predominantly lambda light chain, exhibit heteroclitic affinity for heterologous NP derivatives, and share NPb idiotype. The present study analyzes the idiotypes of antibodies induced with BGG conjugated with the iodo-, bromo-, or nitro-NP derivatives (NIP, NBrP, and NNP). NIP-BGG, NBrP-BGG, and NNP-BGG, induce specific antibodies bearing NPb idiotype in C57BL/6 or C57BL/10 congenic mice, but not in many other inbred strains. Furthermore, the quantity of NPb idiotype in immune sera from various mouse strains immunized with NIP-BGG, NBrP-BGG, and NNP-BGG was similar to that in sera from mice immunized with NP-BGG. Anti-idiotypic antisera against C57BL anti-NP, anti-NIP, or anti-NBrP antibodies exhibit extensive idiotype binding of specifically purified B6 anti-NP, -NIP, -NBrP, and -NNP antibodies. These purified antibodies contain a high percentage (greater than 70%) of lambda-chain-bearing molecules. The data indicate that an extensively shared repertoire composed of predominantly lambda-bearing NPb-positive idiotypic antibodies is used in response to NP and its derivatives in C57BL mice.


Assessment of UL56 Mutations before Letermovir Therapy in Refractory Cytomegalovirus Transplant Recipients.
De novo mutations in the UL56 terminase subunit and its associated phenotypes were studied in the context of cytomegalovirus (CMV) transplant recipients clinically resistant to DNA-polymerase inhibitors, naive to letermovir. R246C was the only UL56 variant detected by standard and deep sequencing, located within the letermovir-resistance-associated region (residues 230-370). R246C emerged in 2/80 transplant recipients (1 hematopoietic and 1 heart) since first cytomegalovirus replication and responded transiently to various alternative antiviral treatments in vivo. Recombinant phenotyping showed R246C conferred an advanced viral fitness and was sensitive to ganciclovir, cidofovir, foscarnet, maribavir, and letermovir. These results demonstrate a low rate (2.5%) of natural occurring polymorphisms within the letermovir-resistant-associated region before its administration. Identification of high replicative capacity variants in patients not responding to treatment or experiencing relapses could be helpful to guide further therapy and dosing of antiviral molecules. IMPORTANCE We provide comprehensive data on the clinical correlates of both CMV genotypic follow-up by standard and deep sequencing and the clinical outcomes, as well as recombinant phenotypic results of this novel mutation. Our study emphasizes that the clinical follow-up in combination with genotypic and phenotypic studies is essential for the assessment and optimization of patients experiencing HCMV relapses or not responding to antiviral therapy. This information may be important for other researchers and clinicians working in the field to improve the care of transplant patients since drug-resistant CMV infections are an important emerging problem even with the new antiviral development.


Kinetics of N-nitrosopiperazine formation from nitrite and piperazine in CO2 capture.
Piperazine (PZ) is an efficient amine for carbon capture systems, but it can form N-nitrosopiperazine (MNPZ), a carcinogen, from nitrogen oxides (NO(x)) in flue gas from coal or natural gas combustion. The reaction of nitrite with PZ was studied in 0.1 to 5 mol/dm(3) PZ with 0.001 to 0.8 mol CO2/mol PZ at 50 to 135 °C. The reaction forming MNPZ is first order in nitrite, piperazine carbamate species, and hydronium ion. The activation energy is 84 ± 2 kJ/mol with a rate constant of 8.5 × 10(3) ± 1.4 × 10(3) dm(6) mol(-2) s(-1) at 100 °C. The proposed mechanism involves protonation of the carbamate species, nucleophilic attack of the carbamic acid, and formation of bicarbonate and MNPZ. These kinetics and mechanism will be useful in identifying inhibitors and other strategies to reduce nitrosamine accumulation in CO2 capture by scrubbing with PZ or other amines.


Hansen Neuropathy: Still a Possible Diagnosis in the Investigation of a Peripheral Neuropathy.
Leprosy is still one of the most frequent causes of peripheral neuropathy. Although regarded as eradicated in Portugal, is still documented in neuropathological study of patients with clinical peripheral neuropathy without proper diagnosis. Review of the cases of Hansen disease neuropathy diagnosed in Neuropathology Unit of Centro Hospitalar do Porto between 1978 and 2013, atending to gender, age, clinical manifestations and neuropathological findings. Twenty one patients were identified with neuropathological diagnosis of Hansenâs disease neuropathy, predominantly male. The mean age at diagnosis was 52 years, and sensory symptoms predominate as neurological manifestation of disease. Interval between symptoms and diagnosis was 1-38 years. In most nerve samples tuberculoid type of disease was identified. Bacilli were detected in skin and nerve in 44% of cases. Mononeuritis is the most common presentation of leprosy but other clinical manifestations are possible, including skin lesions. Infection with M. leprae injures myelinated and unmyelinated fibres, with replacement of nerve tissue by collagen fibrosis. The diagnosis of leprosy is only achieved by neuropathological study of skin lesions and / or peripheral nerve, supported by the identification of the bacillus. Hansen disease remains a public health problem in tropical areas and, although rare, still described in Western countries reason why should still be considered as a diagnostic possibility in the investigation of peripheral neuropathy.


Human leucocytes in asthenozoospermic patients: endothelial nitric oxide synthase expression.
In a basic study at the Andrology Unit, Department of Clinical and Molecular Sciences, Polytechnic University of Marche, Ancona, Italy, we evaluated the pattern of mRNA endothelial nitric oxide synthase (eNOS) expression in human blood leucocytes isolated from normozoospermic fertile and asthenozoospermic infertile men to elucidate any pathogenic involvement in sperm cell motility. Forty infertile men with idiopathic asthenozoospermia and 45 normozoospermic fertile donors, age-matched, were included. Semen parameters were evaluated, and expression analysis of mRNA was performed in human leucocytes using reverse transcription polymerase chain reaction. Sperm volume, count, motility and morphology were determined, and eNOS expression and Western blotting analyses were performed. A positive correlation was observed between the concentrations of NO and the percentage of immotile spermatozoa. The mRNA of eNOS was more expressed in peripheral blood leucocytes isolated from asthenozoospermic infertile men versus those of fertile normozoospermic men (7.46 ± 0.38 versus 7.06 ± 0.56, P = 0.0355). A significant up-regulation of eNOS gene in peripheral blood leucocytes was 1.52-fold higher than that of fertile donors. It is concluded that eNOS expression and activity are enhanced in blood leucocytes in men with idiopathic asthenozoospermia.


Dietary Plant Sterols Supplementation Increases In Vivo Nitrite and Nitrate Production in Healthy Adults: A Randomized, Controlled Study.
A randomized, double-blinded, placebo-controlled and crossover study was conducted to simultaneously measure the effects, 3 h after consumption and after 4-wk daily exposure to plant sterols-enriched food product, on in vivo nitrite and nitrate production in healthy adults. Eighteen healthy participants (67% female, 35.3 [mean] ± 9.5 [SD] years, mean body mass index 22.8 kg/m2 ) received 2 soy milk (20 g) treatments daily: placebo and one containing 2.0 g free plant sterols equivalent of their palmityl esters (β-sitosterol, 55%; campesterol, 29%; and stigmasterol, 23%). Nitrite and nitrate concentrations were measured in the blood plasma and urine, using stable isotope-labeled gas chromatography-mass spectrometry. L-arginine and asymmetric dimethylarginine concentrations in blood serum were measured using commercially available enzyme immunoassays. Nitrite and nitrate concentrations in blood plasma (nitrite 5.83 ± 0.50 vs. 4.52 ± 0.27; nitrate 15.78 ± 0.96 vs. 13.43 ± 0.81 μmol/L) and urine (nitrite 1.12 ± 0.22 vs. 0.92 ± 0.36, nitrate 12.23 ± 1.15 vs. 9.71 ± 2.04 μmol/L) were significantly elevated after 4-wk plant sterols supplementation Placebo and 3-h treatments did not affect the blood plasma and urinary concentrations of nitrite and nitrate. Circulating levels of L-arginine and asymmetric dimethylarginine were unchanged in the placebo and treatment arms. Total plant sterols, β-Sitosterol, campesterol, and stigmasterol concentrations were significantly elevated after 4-wk treatments compared to the placebo and 3-h treatments. Blood plasma nitrite and nitrate concentrations correlated significantly with the plasma total and specific plant sterol concentrations. Our results suggest that dietary plant sterols, in the combination used, can upregulate nitrite, and nitrate production in vivo.


Diversity of learning psychoanalysis and analytical psychology.
As the subject of this Conference is 'Diversity and its Limits: New Directions in Analytical Psychology and Psychoanalysis', I would like to investigate some aspects of education at our Institute--the East European Institute of Psychoanalysis--from the point of view of the meeting of West and East. I will look at the influence of this diversity on the formation and development of psychoanalysis and analytical psychology in Russia.


Membrane translocation of penetratin and its derivatives in different cell lines.
The third helix of the homeodomain of the Antennapedia homeoprotein can translocate through the cell membrane into the nucleus and can be used as an intracellular vehicle for the delivery of oligopeptides and oligonucleotides. A 16-amino acid-long peptide fragment, called penetratin, is internalized by the cells in a specific, non-receptor-mediated manner. For a better understanding of the mechanism of the transfer, penetratin and two analogs were synthesized:The conformation of penetratin peptides 1-3 was examined in both extracellular matrix-mimetic and membrane-mimetic environments. (1)H-NMR and CD spectroscopic measurements were performed in mixtures of TFE/water with different ratios. Peptides 1-3 were labeled by reacting their N-terminal free amino group with fluorescein isothiocyanate (FITC). Membrane translocation of the labelled peptides was studied with cell cultures [WEHI 164 murine fibrosarcoma cells (WC/1); chicken fibroblast cells (CEC-32); chicken monocytic cells (HD-11); human fibroblast (SV 80) and human monocytic cells (MonoMac-6)]. Confocal laser scanning microscopy and flow cytometry assay were used to study membrane translocation. Amphiphilicity was calculated for each peptide. In our experiments all the penetratin peptides penetrated into the cells. Helical conformation and membrane translocation ability showed little correlation: substitution of the two Trp with Phe increased the stability of helical conformation but decreased membrane translocation activity. The results of fluorescence microscopy and flow cytometry show that penetratin can be translocated into the cells by two mechanisms: endocytosis and direct transport through the cell membrane.


Slowered turnover of norepinephrine in the brain associated with an acute elevation in blood pressure of rats.
Acute responses of blood pressure and turnover of norepinephrine in the brain to intracisternally injected guanethidine were studied in rats. Systolic blood pressure in guanethidine-treated rats showed a dose-relating rise for over 9 hrs, while norepinephrine contents in cortex-cerebellum, brain stem and heart were not affected. The endogenous norepinephrine in brain samples, however, did not decrease against alpha-methyl-p-tyrosine to result in regression coefficients significantly different from those in the saline-treated group whereas norepinephrine in heart was reduced similarly in the 2 groups, indicating a slowered turnover of norepinephrine selectively in the brain. The results may indicate that the retarded norepinephrine turnover in the brain is causatively related with an acute elevation in blood pressure.


Tumour-to-tumour metastasis: breast carcinoma to an olfactory neuroblastoma.
Tumour-to-tumour metastasis is a rare phenomenon. It occurs when a primary tumour is a recipient of a separate tumour within the same individual. We present a case of a 66-year-old woman with known breast cancer who presented with one-sided nasal symptoms. Examination and imaging revealed a unilateral polyp arising from the skull base. She underwent endoscopic polypectomy with the histology demonstrating tumour-to-tumour metastasis from a breast carcinoma to an olfactory neuroblastoma, a rare sinonasal tumour. Clinicians should be cautious of distant metastases in any patient presenting with head and neck symptoms and a known primary tumour. This is the first documented case of this type.


Hirsutism as the initial presentation of malignant ovarian Leydig cell tumor: A case report.
Ovarian steroid cell tumors are a rare subtype of sex-cord stromal cell tumors. Overall, these tumors make <0.1% of all ovarian tumors. These neoplasms can be divided according to the cell of origin into stromal luteomas, Leydig cell tumors, and steroid cell tumors not otherwise specified. These tumors can be benign, malignant, or borderline, with variable presentation. We report a case of 24-year-old virgin female who was referred to our hospital after being diagnosed with steroid cell tumor-not otherwise specified. Prior to her admission, the patient had been treated unsuccessfully with oral contraceptive pills due to male-pattern facial hair growth, abdominal cramps, and irregular menstrual cycle. Lack of improvement warranted further investigations. Hormonal studies showed an elevated total testosterone, dehydroepiandrosterone sulfate, and morning fasting cortisol. Ultrasonography and computed tomography confirmed the presence of a large pelvic mass with mixed solid and cystic component. Therefore, unilateral salpingo-oophorectomy was performed. Pathological and immunohistochemical examination suggested the presence of a large ovarian steroid cell tumor-not otherwise specified with malignant behavior. The patient did not receive adjuvant therapy and developed metastatic disease. She received four cycles of BEP protocol with no improvement, so she was referred to our center to continue oncological management. Case revision confirmed the presence of steroid cell tumor, but of a different subtype: Leydig cell. She received six cycles of carboplatin-paclitaxel, but her assessment showed disease progression. We report this case with review of literature regarding the appropriate approach to these rare tumors. Although rare, ovarian steroid cell tumors should be included in the differential diagnosis of virilization in young females, especially those refractory to hormonal therapy. In our study, we aimed to present the first reported Palestinian case, which highlights the importance of detailed morphological examination in addition to the difficulties encountered to reach a proper diagnosis. We also provided a review of the existing literature regarding chemotherapeutic lines used in such cases and the response to each.


[The investigation of angiotensin converting enzyme I/D and plasminogen activator inhibitor-1 4G/5G polymorphisms in venous thromboembolism patients].
Deep venous thrombosis and pulmonary embolism, known as venous thromboembolism and seen as a fairly common multifactorial diseases. Differ between populations due to genetic factors, several polymorphisms associated with venous thromboembolism was conducted. As a result of these studies the relationship between disease development and polymorphism is not clear yet. In this study we aimed to investigate the role of angiotensin converting enzyme insersion/deletion (ACE I/D) and plasminogen activator inhibitor-1 4G/5G (PAI-1 4G/5G) polymorphism in the development of disease. In our study, DNA isolated from 80 venous thromboembolism patients and 79 control groups was used. While the classical polymerase chain reaction method used to investigate the ACE I/D polymorphism, the polymerase chain reaction based on allele-specific amplification was used for the detection of PAI-1 4G/5G polymorphism. As a result, there were no significant statistical differences for ACE I/D and PAI-1 4G/5G polymorphism among patient and control groups (p> 0.05). These findings revealed that there is no relationship between these polymorphisms and the development of venous thromboembolism, but large-scale studies are need to be done.


Cell-cycle dependent appearance of murine leukemia-sarcoma virus antigens.
Normal rat kidney (NRK) cells, NRK cells infected with Rauscher murine leukemia virus, and NRK cells infected with Kirsten murine sarcoma-leukemia virus (NRK-K) were synchronized by a double thymidine block. At intervals after release from thymidine blockage, the cells were examined for the presence of viral antigens in the cytoplasm and on the cell surface by immunofluorescent microscopy by using goat anti-Rauscher murine leukemia virus and goat anti-Moloney leukemia virus (Tween-ether disrupted) sera. Detection of viral antigens in the cytoplasm was periodic during the cell cycle. Antigens were detected first during the S phase, increased during the G2 phase, and disappeared during the M and G1 phases. A similar pattern of surface immunofluorescence was observed. Infectious virus was detected in culture fluids from synchronized cells during the M phase. Surface immunofluorescence was detected in NRK-K cells with anti-Rauscher murine leukemia virus and may represent the presence of group-specific antigens on the cell surface. Control, uninfected NRK cells, which did not normally fluoresce, showed weak immunofluorescence during the S and G2 phases after synchronization. Synchronization can be used to amplify latent oncornavirus expression.


Correlation-induced double-plasmon excitation in simple metals studied by inelastic x-ray scattering.
We report a new type of peaklike structure observed in the tail of the dynamic structure factor of simple metals, measured by inelastic x-ray scattering. Based on the momentum-transfer dependence of the energy position and the intensity of this structure, it has been unambiguously attributed to intrinsic plasmon-plasmon excitations, an electronic correlation effect that was theoretically predicted by many-body perturbation theory of the homogeneous-electron-gas model beyond the random-phase approximation. This signature appears to be largely unaffected by electron-ion interaction effects. Thus a structure that is primarily caused by correlation effects in the electron gas has been found experimentally in the dynamic structure factor of simple metals.


Interleukin-6, a reliable prognostic factor for ischemic stroke.
Interleukin-6 (IL-6) is one of the inflammatory mediators characterized by elevated levels in ischemic stroke (IS) patients. The present study set out to assess the role of IL-6, as a marker for inflammation, in the severity and prognosis of acute IS. In a cross-sectional descriptive study, 45 patients with acute IS were selected. Patients with their first day of stroke were included in the study. National Institutes of Health Stroke Scale (NIHSS) and the modified Rankin Scale (mRS) for stroke severity were evaluated on Days 1, 5, 90, and 365. Serum IL-6 level was measured by enzyme-linked immunosorbent assay (ELISA) on days 1 and 5. In the present study, 45 patients with a mean age of 77.6 ± 4.9 including 32 (71%) men and 13 (28.9%) women were studied. Death occurred in 2 (4.4%) patients before discharge from the hospital; the others, be that as it may, followed the study until Day 365 with a mortality rate of 6 (13.3%). A positive significant correlation was found between IL-6, and NIHSS and mRS of the patients from the time of admission to the end of the follow-up period (P < 0.001, r = 0.6). Moreover, there was a significant correlation between IL-6 and infarction size in brain magnetic resonance imaging (MRI) scan (P < 0.001, r = 0.7). The evidence from the present study suggests that IL-6 contributes to determination of severity of ischemic stroke. In addition, IL-6 concentrations affect clinical outcomes in ischemic stroke.


Traveling in time: a time-left analogue for humans.
Two experiments studied normal humans in an analogue of the time-left procedure of J. Gibbon and R. M. Church (1981). In Experiment 1 the "standard" alternative (S) was always half the length of the "comparison" time-left link (C), and S ranged from 4 to 8 s. Humans showed an increasing preference for the time-left alternative with increasing elapsed time in the interval, and indifference points strongly supported the idea of a linear, rather than a logarithmic, time scale. Experiment 2 used some conditions in which S was greater or less than C/2, and preference for the time-left alternative varied systematically with the S/C ratio. Data from both experiments showed reasonable superposition, suggesting underlying scalar timing processes in time left in humans.


Coagulotoxicity of Bothrops (Lancehead Pit-Vipers) Venoms from Brazil: Differential Biochemistry and Antivenom Efficacy Resulting from Prey-Driven Venom Variation.
Lancehead pit-vipers (Bothrops genus) are an extremely diverse and medically important group responsible for the greatest number of snakebite envenomations and deaths in South America. Bothrops atrox (common lancehead), responsible for majority of snakebites and related deaths within the Brazilian Amazon, is a highly adaptable and widely distributed species, whose venom variability has been related to several factors, including geographical distribution and habitat type. This study examined venoms from four B. atrox populations (Belterra and Santarém, PA; Pres. Figueiredo, AM and São Bento, MA), and two additional Bothrops species (B. jararaca and B. neuwiedi) from Southeastern region for their coagulotoxic effects upon different plasmas (human, amphibian, and avian). The results revealed inter⁻ and intraspecific variations in coagulotoxicity, including distinct activities between the three plasmas, with variations in the latter two linked to ecological niche occupied by the snakes. Also examined were the correlated biochemical mechanisms of venom action. Significant variation in the relative reliance upon the cofactors calcium and phospholipid were revealed, and the relative dependency did not significantly correlate with potency. Relative levels of Factor X or prothrombin activating toxins correlated with prey type and prey escape potential. The antivenom was shown to perform better in neutralising prothrombin activation activity than neutralising Factor X activation activity. Thus, the data reveal new information regarding the evolutionary selection pressures shaping snake venom evolution, while also having significant implications for the treatment of the envenomed patient. These results are, therefore, an intersection between evolutionary biology and clinical medicine.


Microbiological patterns in vaginitis.
To determine the incidence and microbial aetiology of vaginitis, and the usefulness of Gram staining of vaginal fluid in detecting the aetiology of vaginitis in Sri Lankan women. Prospective study with microbiological investigation of high vaginal specimens from symptomatic and healthy groups of women. A gynaecological clinic of the De Soysa Hospital for Women, and a family planning clinic of the Family Health Bureau, Colombo. 114 untreated patients with symptoms suggestive of vaginitis and 109 healthy controls. Candida species were isolated from 36% patients and 9% of controls. Gardnerella vaginalis was recovered from 8.8% of patients and 7.3% of controls. The bacterial vaginitis group consisted of several distinct subgroups, viz Gardnerella vaginalis vaginitis, anaerobic vaginitis, non-anaerobic vaginitis and polymicrobial vaginitis. The incidence of trichomoniasis was lower (4.4%) compared to other studies. Other bacteria (anaerobes, enterococci, coliforms, Group B Streptococci) were isolated in significantly higher numbers in the symptomatic group compared to the control group. Microscopy showed that the majority (73%) of the Candida vaginitis patients retained the normal Lactobacillus morphotype. In bacterial vaginitis Lactobacillus morphotype was replaced by other bacterial morphotypes. In our study 36% had candidiasis, 4.4% had trichomoniasis and 47% had bacterial vaginitis. In 12% there was no detectable pathogen. Microscopy of vaginal fluid is useful in determining the aetiology of vaginitis.


Bone marrow transplantation in the treatment of severe immunodeficiencies: possibilities and problems.
Infants and children suffering from severe primary immunodeficiencies may be cured by bone marrow transplantation from a healthy donor. Data obtained in 14 European centers show that about 60% of the patients are surviving disease-free, if they were grafted with bone marrow cells from an HLA-identical related donor. Results of transplantation of T-cell depleted bone marrow from an HLA-haploidentical related donor were also excellent in infants with severe combined immunodeficiency, with 60% recovery. This therapy is superior to transplantation of fetal tissues. HLA-haploidentical T-cell depleted marrow transplantation for other severe immunodeficiencies was less successful. This was mainly due to failure of engraftment, despite intensive conditioning of the recipient, and to infectious complications e.g. by reactivation of latently present viruses.


Putting the periodicity back into the periodic leg movement index: an alternative data-driven algorithm for the computation of this index during sleep and wakefulness.
To evaluate an alternative index for periodic leg movements during sleep (PLMS) and wakefulness (PLMW) expected to be similar to the standard index when leg movement activity is genuinely periodic, but significantly lower when periodicity is low. One-hundred-and-seven subjects with restless legs syndrome (RLS) were retrospectively identified and included (47 males, 60 females, mean age 56.9 years), along with 63 controls (33 males, 30 females, 42.2 years). Night-to-night variability was analysed in a subgroup of 17 subjects with RLS. PLMW were evaluated in a subgroup of 66 RLS subjects. Two 'alternative' PLMS/PLMW indices were calculated: one increased the lower limit of the inter-movement intervals from 5 to 10 s ('Alt1') and another additionally considered only series not interrupted by LMs with a short inter-movement interval ('Alt2'). Despite a high correlation between methods, only the Alt2 algorithm provided significantly different results, with PLMS/PLMW indices being consistently lower than those provided by the other two methods. The difference was more evident in the controls and during wakefulness, when periodicity was lower. The difference between the Alt2 and the standard PLMS index showed a significantly negative correlation with the Periodicity Index. Night-to-night variability was similar for all PLMS indices and significantly higher than the variability seen in the Periodicity Index. This methodological study introduces an alternative to the standard PLMS/PLMW indices, initiating the validation process for a new way of computing the PLMS/PLMW index, more adherent to the parameters that allows a reliable evaluation of their periodicity.


[In vitro comparative study of the protective effect of different theophyllines on the leucocytes' histamine-release induced by antigen (author's transl)].
The recent discovery of the protective action of xanthic bases towards histamine-release consecutive to the antigen-antibody reaction led the authors to study the importance of this protection induced by various theophylline derivatives. The technique used was that described by LICHTENSTEIN and OSLER: isolation of leucocytes, measurement of histamine liberated in the presence of antigen, same measurement done in the presence of antigen and theophylline. The antigens used were: house dust, Dermatophagoides pteronyssimus (DP), graminaceae pollens. The tested theophyllines were: the theophylline base, aminophylline, bamiphylline, diprophylline, thio theo, piperazine acefilinate. All these substances studied brought a reduction of the histamine-release but according to a variable frequency and intensity: the thio theo, the diprophylline and the piperazine acefilinate gave the best results. Besides the latters are clearer when the antigen was dust or DP. This study enables the confirmation of the inhibiting action of theophyllines on histamine-release induced by antigen, but complementary studies on a larger number of cases seem necessary in order to determine precisely the most efficient derivatives.


Atrial tachycardia arising from the right atrial inferoseptum masquerading as common atrial flutter.
Radiofrequency catheter ablation was performed in 2 patients with atrial tachycardia (AT). In both cases the AT originated from the inferoseptal portion of the right atrium, and the cycle length was 210 ms. The surface ECG demonstrated common counterclockwise atrial flutter, probably caused by functional block in the clockwise direction at the cavo-tricuspid isthmus and posterior right atrium with rapid activation of the origin. Although rare (2%), AT originating from the inferoseptum of the right atrium should be considered when the surface ECG exhibits common atrial flutter.


Phase Control of Nanocrystalline Inclusions in Bioprecipitated Titania with a Panel of Mutant Silica-Binding Proteins.
The biomimetic route to inorganic synthesis presents an opportunity to produce complex materials with superior properties under ambient conditions and from nontoxic precursors. While there has been significant progress in using solid-binding peptides (SBPs), proteins, and organisms to produce a variety of inorganic and hybrid structures, it has been more challenging to understand the interplay of solution conditions and solid-binding peptide (SBP) sequence, structure, and self-association on synthetic outcomes. Here, we show that fusing the Car9 silica-binding peptide-but not the silaffin-derived R5 peptide-to superfolder green fluorescent protein (sfGFP) enhances the ability of micromolar concentrations of protein to induce rapid titania (TiO2) precipitation from acidified solutions of tetrakis(di-lactato)-oxo-titanate (TiBALDH). TiO2 is produced stoichiometrically and although predominantly amorphous, contains nanosized anatase and monoclinic TiO2(B) inclusions. Remarkably, the phase of these nanocrystallites can be tuned from about 80% TiO2(B) to about 65% anatase by using Car9 mutants impaired in their ability to drive the formation of higher-order sfGFP-Car9 oligomers. Our results suggest that the presentation of multiple basic side chains in an extended plane formed by SBP self-association is critical to template the formation of monoclinic crystallites and underscore the subtle influence that single or dual substitutions in dodecameric SBPs can exert on the yield and crystallinity of biomineralized inorganics.


Prevalence of antibodies of field pseudorabies virus in pigs of herd vaccinated with live vaccine.
This trial was performed in a 350-sow farrow-to-finish pig farm in which animals had been vaccinated against Aujeszky's disease with glycoprotein gC deletion (gC(-)) vaccine. The prevalence of antibodies against field pseudorabies virus (PRV) in serum samples collected from sows and fattening pigs during the period March 1995 to May 1997 was determined by using a commercial assay kit. The frequencies with which PRV-positive sows were found were 28.9% and 25.0% in March and October of 1995, 12.5% and 2.5% in April and October of 1996, and 0% in May of 1997. The fattening pigs tested at these times had no antibody to field PRV. These results thus indicate the probability that the PRV infection at the farm was eliminated by the gC(-) vaccine.


Dietary supplementation of 25-hydroxycholecalciferol improves cardiac function and livability in broiler breeder hens-amelioration of blood pressure and vascular remodeling.
A supplement of 69 μg 25-hydroxycholecalciferol (25-OH-D3)/kg feed suppressed the mortality in feed-restricted broiler breeder hens and in hens allowed ad libitum feed intake (Ad-hens) in a feeding trial from age 26 to 60 wk. Outcomes for the mechanisms found that 25-OH-D3 relieved systemic hypoxia, pathological cardiac remodeling and arrhythmias, and hepatopathology to improve hens' livability. In the study, we further evaluated the effect of 25-OH-D3 on blood pressure and vascular remodeling relative to cardiac pathogenesis of sudden death (SD). Ad libitum feed intake increased mechanical loading and contributed to maladaptive cardiac hypertrophy as evidenced by consistently elevated peripheral arterial blood pressure in Ad-hens before SD (P < 0.05). In planned longitudinal measurements, Ad-hens also showed higher right ventricle systolic pressure and right ventricle diastolic pressure (RVDP) (P < 0.05). Supplemental 25-OH-D3 relieved peripheral hypertension and prevented time-dependent increases of RVDP in Ad-hens through the renin-angiotensin system and circulating nitric oxide availability and by regulating vascular remodeling including elastin/collagen ratio and smooth muscle cell proliferation in the pulmonary artery for improved elasticity/stiffness (P < 0.05). The antihypertensive effect via the renin-angiotensin system and nitric oxide regulation in respect to heart rate and arrhythmias by 25-OH-D3 were further confirmed in 51 week-old feed-restricted broiler breeder hens challenged with salt loading for 5 wk. Despite feed restriction, the most feed-efficient hens of feed-restricted groups also exhibited cardiac pathological hypertrophy, in conjunction with higher right ventricle systolic pressure, RVDP, plasma nitric oxide levels, and more dramatic arterial remodeling (P < 0.05). These results suggest that peripheral and pulmonary hypertension are the key drivers of SD and that 25-OH-D3 is an effective antihypertensive supplement to alleviate cardiac pathogenesis and improve livability in broiler breeder hens.


Copper-catalyzed extended Pummerer reactions of ketene dithioacetal monoxides with alkynyl sulfides and ynamides with an accompanying oxygen rearrangement.
The first examples of metal-catalyzed extended Pummerer reactions through the activation of sulfoxides are described. The copper-catalyzed reactions of ketene dithioacetal monoxides with alkynyl sulfides and ynamides provided a wide variety of γ,γ-disulfanyl-β,γ-unsaturated carbonyl compounds with an accompanying oxygen rearrangement. The products can be easily converted into 1,4-dicarbonyl compounds and substituted heteroaromatics. DFT calculations and mechanistic experiments revealed a new interesting stepwise addition/oxygen rearrangement mechanism.


Mantle cell lymphoma with t(11;14) and unmutated or mutated VH genes expresses AID and undergoes isotype switch events.
Isotype switch commonly follows onset of somatic hypermutation in the germinal center (GC), with activation-induced cytidine deaminase (AID) as a prerequisite. Mantle cell lymphoma (MCL) with t(11;14) includes a subset with unmutated (UM) and a minor subset with mutated (MUT) VH genes. Here, we investigated whether switch events and AID expression occur in MCL. In 4 of 6 UM and 4 of 7 MUT MCLs, alternative tumor-derived Cgamma,alpha,epsilon transcripts were identified. AID transcripts, including a splice variant, were common to both subsets. AID expression correlated with switch in 8 of 8 cases, but in 3 of 5 cases it occurred with switch absent. Circle transcripts (Igamma-Cmicro/Ialpha-Cmu) were identified in 5 of 7 evaluated cases. In 1 of 12 cases, 12% of tumor cells expressed immunoglobulin L-restricted surface IgA. Ongoing switch recombination events appear to be a feature of MCL, likely restricted to a minor tumor subpopulation, with occasional variant sIg expression. UM MCLs implicate origins from pre-GC B cells and reveal switch events at ectopic sites.


[Intrapericardial rupture of an aneurysm of the right anterior sinus of Valsalva: clinical case and review of the literature].
Intrapericardial ruptured aneurysm of the right anterior sinus of Valsalva. A intrapericardial ruptured aneurysm of the right anterior sinus of Valsalva is presented. An infectious origin was suspected. The aneurysm was treated by plication, which allowed to correct the preoperative insufficiency. Aneurysms of the Valsalva sinus can remain asymptomatic. When they are small, a conservative treatment is conceivable. Many complications can occur: rupture (more often in right cavities), aortic insufficiency, coronary insufficiency, cardiac failure, rhythm disturbances. This complications necessitate a surgical treatment. Aortic valvular replacement is indicated only when the structure of the valve is altered.


Selection of topical antimicrobial agents for cultured skin for burns by combined assessment of cellular cytotoxicity and antimicrobial activity.
Cultured epidermal skin has become an adjunctive therapy for treatment of major burn injuries, but its effectiveness is greatly limited due to destruction by microbial contamination. To evaluate candidate drugs for use with cultured skin, a combined cytotoxicity-antimicrobial assay system was developed for determination of toxicity to cultured human keratinocytes and fibroblasts, and to common burn wound organisms (20 bacterial and 4 fungal strains). Candidate agents including Hibiclens (n = 3), amikacin, piperacillin, norfloxacin, and nystatin were tested separately and in combination (n = 6 each) for inhibition of growth of human cells and lytic activity on microorganisms in the wet disc assay. The data showed that: (1) Hibiclens was uniformly toxic to both cultured human cells and microorganisms; (2) norfloxacin had dose-dependent toxicity to human cells and broad effectiveness against microorganisms; and (3) norfloxacin (25 micrograms/mL) plus nystatin (100 U/mL) had low toxicity to human cells and high toxicity to both Gram-positive and Gram-negative bacteria (20 of 20) and fungi (4 of 4). Selection of topical antimicrobial drugs by these assays may improve effectiveness of cultured skin for burns and may be extended to the control of other surgical wound infections.


Unusually high frequency MHC class I alleles in Mauritian origin cynomolgus macaques.
Acute shortages of Indian origin Rhesus macaques significantly hinder HIV/AIDS research. Cellular immune responses are particularly difficult to study because only a subset of animals possess MHC class I (MHC I) alleles with defined peptide-binding specificities. To expand the pool of nonhuman primates suitable for studies of cellular immunity, we defined 66 MHC I alleles in Cynomolgus macaques (Macaca fascicularis) of Chinese, Vietnamese, and Mauritian origin. Most MHC I alleles were found only in animals from a single geographic origin, suggesting that Cynomolgus macaques from different origins are not interchangeable in studies of cellular immunity. Animals from Mauritius may be particularly valuable because >50% of these Cynomolgus macaques share the MHC class I allele combination Mafa-B*430101, Mafa-B*440101, and Mafa-B*460101. The increased MHC I allele sharing of Mauritian origin Cynomolgus macaques may dramatically reduce the overall number of animals needed to study cellular immune responses in nonhuman primates while simultaneously reducing the confounding effects of genetic heterogeneity in HIV/AIDS research.


Epidemiological comparisons of codon usage patterns among HIV-1 isolates from Asia, Europe, Africa and the Americas.
To investigate the genomic properties of HIV-1, we collected 3,081 sequences from the HIV Sequence Database. The sequences were categorized according to sampling region, country, year, subtype, gene name, and sequence and were saved in a database constructed for this study. The relative synonymous codon usage (RSCU) values of matrix, capsid, and gp120 and gp41 genes were calculated using correspondence analysis. The synonymous codon usage patterns based on the geographical regions of African countries showed broad distributions; when all the other regions, including Asia, Europe, and the Americas, were taken into account, the Asian countries tended to be divided into two groups. The sequences were clustered into nine non-CRF subtypes. Among these, subtype C showed the most distinct codon usage pattern. To determine why the codon usage patterns in Asian countries were divided into two groups for four target genes, the sequences of the isolates from the Asian countries were analyzed. As a result, the synonymous codon usage patterns among Asian countries were divided into two groups, the southern Asian countries and the other Asian countries, with subtype 01_AE being the most dominant subtype in southern Asia. In summary, the synonymous codon usage patterns among the individual HIV-1 subtypes reflect genetic variations, and this bioinformatics technique may be useful in conjunction with phylogenetic methods for predicting the evolutionary patterns of pandemic viruses.


Characterization of the human ridged and non-ridged skin: a comprehensive histological, histochemical and immunohistochemical analysis.
The structure of the human skin is directly dependent on its location and the mechanical forces to which it is subjected. In the present work, we have performed a comprehensive analysis of the human ridged and non-ridged skin to identify the differences and similarities between both skin types. For this purpose, human skin samples were obtained from dorsal hand skin (DHS), palmar hand skin (PHS), dorsal foot skin (DFS) and plantar foot skin (PFS) from the same cadaveric donors. Histological, histochemical and semiquantitative and quantitative immunohistochemical analyses were carried out to evaluate the epidermis, dermis and basement membrane. Results show that the epithelial layer of ridged skin had larger cell number and size than non-ridged skin for most strata. Melanocytes and Langerhans cells were more abundant in non-ridged skin, whereas Merkel cells were preferentially found in ridged skin. The expression pattern of CK5/6 was slightly differed between non-ridged and ridged skin. Involucrin expression was slightly more intense in non-ridged skin than in ridged skin. Collagen was more abundant in foot skin dermis than in hand skin, and in ridged skin as compared to non-ridged skin. Elastic fibers were more abundant in DHS. Biglycan was more abundant in foot skin than in hand skin. No differences were found for blood and lymphatic vessels. The basement membrane laminin was preferentially found in foot skin. These results revealed important differences at the epithelial, dermal and basement membrane levels that could contribute to a better knowledge of the human skin histology.


Randomization and the analysis of variance.
This paper attempts to strengthen faith in randomization, first by defining it in general and second by showing how it is the fundamental means, in many experiments, of generating the probability space. It is defined by the natural structure of the experimental units (E.U.'s) not by a particular experimental design imposed on that structure. Three examples are given, with the same design but different E.U. structures and randomizations, to show that models developed from experimental designs are superficial, but that their randomization is fundamental.


Early embryo loss is associated with the prior expression of macrophage activation markers in the decidua.
In early embryo loss, the activation of maternal immune effector mechanisms play a critical role in determining the success or failure of a pregnancy. We have previously shown that increased nitric oxide production by decidual macrophages is involved in early embryo loss occurring at day 12 of gestation. In this study, using reverse transcription-PCR and Southern blotting, the expression of inducible nitric oxide synthases (iNOS) and TNF-alpha mRNA was determined to quantify macrophage activation in individual murine embryos in a model of spontaneous early embryo loss. At day 8 of gestation, 32 and 29% of embryos with no apparent pathology showed an increase in iNOS and TNF-alpha mRNA expression, respectively. This corresponds to the natural resorption rate seen in the mouse model. In addition, the percentage of embryos with increased iNOS and TNF-alpha mRNA expression was further augmented when pregnant mice were induced to abort at a higher rate. These results showed, for the first time, a correlation between increased iNOS and TNF-alpha expression and embryo resorption. The results provide evidence for the presence of activated macrophages at implantation sites before overt embryo damage occurs.


Deoxygenation of sulphoxides to sulphides with trichlorophosphane.
An efficient route to deoxygenation of sulphoxides to sulphides with PCl3 under mild reaction condition was developed. PCl3 was used as a reducing agent for the first time to convert sulphoxides to sulphides. The mild conditions, use of cheap and readily available reagent, and broad substrate scope render it a useful strategy for preparing sulphides.


Preclinical systematic review & meta-analysis of cyclosporine for the treatment of myocardial ischemia-reperfusion injury.
Though best known for its immunosuppressant effects, cyclosporine A (CsA) has also been studied as a treatment to mitigate ischemia-reperfusion injury (IRI) by its inhibition of the mitochondria permeability transition pore (mPTP). Despite numerous preclinical studies supporting its benefit in reducing infarct size following myocardial IRI, large randomized controlled clinical trials have been unable to show a beneficial effect. Exploring existing preclinical data can give us the opportunity to revisit some the assumptions that may have led to the failure of these studies to translate clinically. Herein, we present a systematic review of preclinical studies testing CsA to attenuate myocardial IRI (PROSPERO CRD42020159620). We conducted a systematic search of health research databases Ovid MEDLINE, Ovid EMBASE, Web of Science BIOSIS, and Scopus, as well as Cochrane and PROSPERO systematic review databases, on March 9, 2022 for non-human in vivo animal studies of myocardial IRI, using CsA as a treatment that reported clinically relevant outcomes. Bias was assessed using the Systematic Review Centre for Laboratory Animal Experimentation's risk of bias tool and a modified Collaborative Approach to Meta Analysis and Review of Animal Data from Experimental Studies checklist. Sub-group meta-analyses were conducted to identify potential factors influencing outcomes. We identified 71 studies, 59 of which were studies of coronary occlusion. Overall, 75% of studies reported a clear positive effect of CsA in mitigating myocardial IRI by some clinically relevant parameter (e.g., infarct size). A meta-analysis including 43 coronary occlusion studies showed an overall reduction in infarct size with CsA treatment (16.09%; 95% CI: -18.50% to -13.67%). Subgroup meta-analyses identified species, age, timing of administration, and duration of ischemia as factors potentially affecting the efficacy of CsA in the setting of myocardial IRI. Our systematic review and meta-analysis identifies questions that have yet to be answered by preclinical studies, highlighting important differences between these and clinical studies that should be addressed prior to proceeding with any further clinical studies using CsA to treat IRI in the heart or other organs. We also use the example of CsA to highlight general considerations for researchers attempting to translate animal studies into the clinical setting.


17-Beta estradiol administration attenuates deficits in sustained and divided attention in young ovariectomized rats and aged acyclic female rats.
Recent evidence suggests that estrogen may interact with the basal forebrain cholinergic system to influence learning. The authors examined whether the loss of estrogen following ovariectomy (Experiment 1) or the disruption to the estrogen cycle during aging (Experiment 2) impaired performance of the 5-choice serial reaction time task (5-CSRT)--a sustained and divided attention task sensitive to cholinergic challenges in rats. In Experiment 1, posttraining ovariectomy in young rats did not disrupt baseline performance but did impair performance when attention was challenged by variation in the intertrial interval (ITI) or in the intermittent presentation of a novel distracting auditory stimulus. Administration of 17-beta estradiol rescued these impairments. Through the use of a within-subjects design, Experiment 2 revealed that 17-beta estradiol did not influence the baseline performance of 21-month-old female rats trained on the 5-CSRT task from a young age but did improve performance when attention was challenged by varying the ITI or by presenting a distracting auditory cue. The results indicate that 17-beta estradiol administration can improve specific components of attention in young ovariectomized rats and gonadally intact aged female rats.


Testicular cancer in Luxembourg: incidence and outcome in relation to the different histo-pathological types (1980-2004).
Increasing incidence rates of testicular cancer have been reported worldwide over the last three decades. Trends over time in the incidence rates of germ cell tumours (GCTs) in Luxembourg (Western Europe) and the outcome, both in relation to the different histological types, were analysed. The population-based files of the Morphologic Tumour Registry collecting at a nation-wide level all testicular cancers diagnosed between 1980 and 2004 in the central department of pathology in Luxembourg were retrospectively reviewed. In addition, the presence of concomitant malignant diseases was investigated. 397 patients with GCT were evaluated. The mean age was 33.7 years (range: 1-72). Most of the patients (58.7%) were between 15 and 34 years of age. The age-standardized incidence rates rose from 4.5 per 10(5) (1980-1984) to 7.7 per 10(5) (2000-2004). Out of 275 (69.3%) pure GCTs, 218 seminomas, 48 embryonal carcinomas, 4 yolk sac tumours, 4 malignant teratomas and 1 choriocarcinoma were identified. 30.7% GCTs were of mixed type with 17 different histological variants. 5.8% of the patients had metachronous concomitant cancers, 2% bilateral GCTs and 3.8% non-testicular neoplasms. In all histological categories, with the exception of the pure seminomas, prognosis was determined within the 24 months following diagnosis; pure seminomas need long time follow-up. Ten-year observed survival rates exceeded mostly 90%. Pure embryonal carcinomas had the worst prognosis with a 10-year observed survival rate of 87.1 +/- 12%. Testicular germ cell tumours are rare, highly curable neoplasms that generally occur in young patients. In all histological categories, except for pure seminomas, prognosis was determined within the 24 months after diagnosis. Despite better observed survival rates, patients with pure seminomas, without or with metachronous concomitant non-testicular malignancies, need long time followup strategy.


CHIKV strains Brazil (wt) and Ross (lab-adapted) differ with regard to cell host range and antiviral sensitivity and show CPE in human glioblastoma cell lines U138 and U251.
Chikungunya virus (CHIKV), a (re)emerging arbovirus, is the causative agent of chikungunya fever. To date, no approved vaccine or specific antiviral therapy are available. CHIKV has repeatedly been responsible for serious economic and public health impacts in countries where CHIKV epidemics occurred. Antiviral tests in vitro are generally performed in Vero-B4 cells, a well characterised cell line derived from the kidney of an African green monkey. In this work we characterised a CHIKV patient isolate from Brazil (CHIKVBrazil) with regard to cell affinity, infectivity, propagation and cell damage and compared it with a high-passage lab strain (CHIKVRoss). Infecting various cell lines (Vero-B4, A549, Huh-7, DBTRG, U251, and U138) with both virus strains, we found distinct differences between the two viruses. CHIKVBrazil does not cause cytopathic effects (CPE) in the human hepatocarcinoma cell line Huh-7. Neither CHIKVBrazil nor CHIKVRoss caused CPE on A549 human lung epithelial cells. The human astrocyte derived glioblastoma cell lines U138 and U251 were found to be effective models for lytic infection with both virus strains and we discuss their predictive potential for neurogenic CHIKV disease. We also detected significant differences in antiviral efficacies regarding the two CHIKV strains. Generally, the antivirals ribavirin, hydroxychloroquine (HCQ) and T-1105 seem to work better against CHIKVBrazil in glioblastoma cells than in Vero-B4. Finally, full genome analyses of the CHIKV isolates were done in order to determine their lineage and possibly explain differences in tissue range and antiviral compound efficacies.


Transcriptional regulation of hormone-synthesis and signaling pathways by overexpressing cytokinin-synthesis contributes to improved drought tolerance in creeping bentgrass.
The objective of this study was to investigate transcriptomic changes and molecular factors regulated by cytokinins that may contribute to improved drought tolerance in creeping bentgrass (Agrostis stolonifera) overexpressing adenine isopentenyltransferase (ipt). Wild-type (WT) and ipt-transgenic plants were maintained well irrigated or exposed to 21 days of drought stress in growth chambers. Transcriptomic analysis conducted by RNA-seq revealed 661 and 648 upregulated and 764 and 862 downregulated drought-responsive genes (DRGs) in the WT and ipt-transgenic plants, respectively, under drought stress using adjusted P-value of 0.001 and log2 fold change. Gene ontology (GO) term classification showed that a greater number of DRGs were found in ipt-transgenic plants than in WT plants pertaining to biological functions including metabolic process, cellular process, cell structure and growth, macromolecular complex, and binding and catalytic activity, whereas fewer DRGs were found in ipt-transgenic plants than in WT plants pertaining to response to stimulus and antioxidant activity. Furthermore, plant hormone signal transduction pathway analysis revealed three downregulated transcripts [type B - Arabidopsis response regulators (B-ARR), ABA-responsive element binding factor (ABF) and pyrabactin resistance/like (PYR/PYL)] and two upregulated transcripts (BIN2 and JAZ) that were significantly differentiated between ipt-transgenic and WT plants under drought stress, which are particularly interesting for further investigation of molecular mechanisms of hormone-regulation of drought tolerance.


2D Organic Superlattice Promoted via Combined Action of π-π Stacking and Dipole-Dipole Interaction in Discotic Liquid Crystals.
A series of discotic liquid crystals based on hexapentyloxytriphenylene (HAT5) have been investigated where one out of the six ether side chains of a triphenylene core was replaced by an ester side chain and named for 5a-5h. During the process of studying these compounds, the characteristic straight line defect of ordered columnar structure was identified by polarizing optical microscopy (POM) and liquid crystal state over a wide temperature range was obtained by differential scanning calorimetry (DSC). Basic phase structure and molecular arrangement were assigned by one-dimensional wide-angle X-ray diffraction (1D WAXD), small-angle X-ray scattering (SAXS), two-dimensional wide-angle X-ray diffraction (2D WAXD), and transmission electron microscope (TEM). Combined with sharp and regular dots in 2D WAXD patterns and characteristic peaks at small angle in SAXS pattern which indicated the existence of superlattice, we proved that 2D superlattice formed from self-assembly of discotic molecules with a polar group via π-π stacking and dipole-dipole interaction. In order to verify the effect of orientation on charge carrier mobility, their electron and hole mobilities were measured by time-of-flight (TOF) device, among which the charge carrier mobility could achieve almost twice as that of HAT5. The formation of superlattice no doubt improved their electronic properties and made them more attractive in organic electronics.


MetaSpot: A General Approach for Recognizing the Reactive Atoms Undergoing Metabolic Reactions Based on the MetaQSAR Database.
The prediction of drug metabolism is attracting great interest for the possibility of discarding molecules with unfavorable ADME/Tox profile at the early stage of the drug discovery process. In this context, artificial intelligence methods can generate highly performing predictive models if they are trained by accurate metabolic data. MetaQSAR-based datasets were collected to predict the sites of metabolism for most metabolic reactions. The models were based on a set of structural, physicochemical, and stereo-electronic descriptors and were generated by the random forest algorithm. For each considered biotransformation, two types of models were developed: the first type involved all non-reactive atoms and included atom types among the descriptors, while the second type involved only non-reactive centers having the same atom type(s) of the reactive atoms. All the models of the first type revealed very high performances; the models of the second type show on average worst performances while being almost always able to recognize the reactive centers; only conjugations with glucuronic acid are unsatisfactorily predicted by the models of the second type. Feature evaluation confirms the major role of lipophilicity, self-polarizability, and H-bonding for almost all considered reactions. The obtained results emphasize the possibility of recognizing the sites of metabolism by classification models trained on MetaQSAR database. The two types of models can be synergistically combined since the first models identify which atoms can undergo a given metabolic reactions, while the second models detect the truly reactive centers. The generated models are available as scripts for the VEGA program.


Cooperative Analysis of Structural Dynamics in RNA-Protein Complexes by Single-Molecule Förster Resonance Energy Transfer Spectroscopy.
RNA-protein complexes (RNPs) are essential components in a variety of cellular processes, and oftentimes exhibit complex structures and show mechanisms that are highly dynamic in conformation and structure. However, biochemical and structural biology approaches are mostly not able to fully elucidate the structurally and especially conformationally dynamic and heterogeneous nature of these RNPs, to which end single molecule Förster resonance energy transfer (smFRET) spectroscopy can be harnessed to fill this gap. Here we summarize the advantages of strategic smFRET studies to investigate RNP dynamics, complemented by structural and biochemical data. Focusing on recent smFRET studies of three essential biological systems, we demonstrate that investigation of RNPs on a single molecule level can answer important functional questions that remained elusive with structural or biochemical approaches alone: The complex structural rearrangements throughout the splicing cycle, unwinding dynamics of the G-quadruplex (G4) helicase RHAU, and aspects in telomere maintenance regulation and synthesis.


Microtubule-targeting agents inhibit angiogenesis at subtoxic concentrations, a process associated with inhibition of Rac1 and Cdc42 activity and changes in the endothelial cytoskeleton.
Conventional anticancer agents may display antiangiogenic effects, but the underlying mechanism is poorly understood. We determined the antiangiogenic properties of cisplatin, doxorubicin, and the microtubule-targeting agents docetaxel, epothilone B, and vinblastine at concentrations not affecting cell proliferation. We also assessed tubulin and actin morphology and the activity of two key molecules in cell motility, the small Rho GTPases Cdc42 and Rac1. The highest non-toxic concentration (HNTC) of each drug was defined as the concentration inhibiting a maximum of 10% human umbilical vein endothelial cell growth on a 1-hour drug exposure, being for cisplatin 10 micromol/L, doxorubicin 100 nmol/L, docetaxel 10 nmol/L, epothilone B 1 nmol/L, and vinblastine 10 nmol/L. Comparative endothelial cell functional assays using HNTCs for an exposure time of 1 hour indicated that endothelial cell migration in the wound assay, endothelial cell invasion in a transwell invasion system, and endothelial cell formation into tubelike structures on a layer of Matrigel were significantly inhibited by docetaxel, epothilone B, and vinblastine (P < 0.05), but not by cisplatin and doxorubicin. Docetaxel was slightly more efficient in the inhibition of endothelial cell motility than epothilone B and vinblastine. Fluorescence microscopy revealed that only the microtubule-targeting agents affected the integrity of the tubulin and F-actin cytoskeleton, which showed disturbed microtubule structures, less F-actin stress fiber formation, and appearance of nuclear F-actin rings. These observations were associated with early inhibition of Rac1 and Cdc42 activity. In conclusion, HNTCs of microtubule-targeting agents efficiently reduce endothelial cell motility by interference with microtubule dynamics preventing the activation of Rac1/Cdc42 and disorganizing the actin cytoskeleton.


Primary breast diffuse large B-cell lymphoma in the rituximab era: Therapeutic strategies and patterns of failure.
Primary breast diffuse large B-cell lymphoma (PB-DLBCL) is a rare subtype of DLBCL with limited data on patterns of failure. This multicenter study aimed to define the optimum treatment strategy and patterns of failure for PB-DLBCL patients. We retrospectively reviewed data on 108 PB-DLBCL patients from 21 Chinese medical centers. Only patients with localized disease (involvement of breast and localized lymph nodes) were included. After a median follow-up of 3.2 years, 32% of patients developed progression or relapse. A continuous pattern of relapse was observed, characterized by frequent late relapses in the contralateral breast and central nervous system (CNS). Although rituximab significantly reduced the overall cumulative risk of progression or relapse (5-year cumulative risk 57% vs 24%, P = .029), it had limited effect on the reduction of breast relapse (P = .46). Consolidative radiotherapy significantly decreased the risk of breast relapse, even in the subgroup of patients treated with rituximab (5-year cumulative risk 21.2% vs 0%, P = .012). A continuous risk of CNS progression or relapse up to 8.2 years from diagnosis was observed (10-year cumulative risk 28.3%), with a median time to CNS relapse of 3.1 years. Neither rituximab nor prophylactic intrathecal chemotherapy significantly decreased the risk of CNS relapse. In summary, our study indicates that PB-DLBCL has a continuous pattern of relapse, especially with frequent late relapses in the CNS and contralateral breast. Rituximab and RT confer complementary benefit in the reduction of relapse. However, neither the addition of rituximab nor prophylactic intrathecal chemotherapy could effectively prevent CNS relapse for PB-DLBCL patients.


Plasmonic surface nanostructuring of Au-dots@SiO2 via laser-irradiation induced dewetting.
The in situ observation of Au dot formation and the self-assembly dynamics of Au nanoparticles (NPs) was successfully demonstrated via dewetting of Au thin films on SiO2 glass substrates under nano-second pulsed laser irradiation using a multi-quantum beam high-voltage electron microscope. Moreover, using electron energy-loss spectroscopy (EELS) performed in a scanning transmission electron microscope (STEM), the plasmonic properties of the formed Au/SiO2 nanostructure were analyzed to demonstrate its validity in advanced optical devices. The uniformly distributed Au NPs evolved into a dot alignment through movement and coalescence processes was demonstrated in this in situ observation. We carried out the plasmon-loss images of the plan view and the cross-section of the Au/SiO2 nanostructures were obtained at the plasmon-loss peak energy for investigate the three-dimensional distribution of surface plasmon. Furthermore, discrete-dipole approximation (DDA) calculations were used to simulate the plasmonic properties, such as the surface plasmon resonance and the surface plasmon field distribution, of isolated single Au/SiO2 nanostructures. This STEM-EELS-acquired surface plasmon map of the cross-sectional sample is in excellent agreement with the DDA calculations. This results demonstrated the influence of the contact condition between Au NP and SiO2 glass on the plasmonic properties, and may improve the technology for developing advanced optical devices.