Secondary prevention of macrovascular events in patients with type 2 diabetes in the PROactive Study (PROspective pioglitAzone Clinical Trial In macroVascular Events): a randomised controlled trial.
Patients with type 2 diabetes are at high risk of fatal and non-fatal myocardial infarction and stroke. There is indirect evidence that agonists of peroxisome proliferator-activated receptor gamma (PPAR gamma) could reduce macrovascular complications. Our aim, therefore, was to ascertain whether pioglitazone reduces macrovascular morbidity and mortality in high-risk patients with type 2 diabetes. We did a prospective, randomised controlled trial in 5238 patients with type 2 diabetes who had evidence of macrovascular disease. We recruited patients from primary-care practices and hospitals. We assigned patients to oral pioglitazone titrated from 15 mg to 45 mg (n=2605) or matching placebo (n=2633), to be taken in addition to their glucose-lowering drugs and other medications. Our primary endpoint was the composite of all-cause mortality, non fatal myocardial infarction (including silent myocardial infarction), stroke, acute coronary syndrome, endovascular or surgical intervention in the coronary or leg arteries, and amputation above the ankle. Analysis was by intention to treat. This study is registered as an International Standard Randomised Controlled Trial, number ISRCTN NCT00174993. Two patients were lost to follow-up, but were included in analyses. The average time of observation was 34.5 months. 514 of 2605 patients in the pioglitazone group and 572 of 2633 patients in the placebo group had at least one event in the primary composite endpoint (HR 0.90, 95% CI 0.80-1.02, p=0.095). The main secondary endpoint was the composite of all-cause mortality, non-fatal myocardial infarction, and stroke. 301 patients in the pioglitazone group and 358 in the placebo group reached this endpoint (0.84, 0.72-0.98, p=0.027). Overall safety and tolerability was good with no change in the safety profile of pioglitazone identified. 6% (149 of 2065) and 4% (108 of 2633) of those in the pioglitazone and placebo groups, respectively, were admitted to hospital with heart failure; mortality rates from heart failure did not differ between groups. Pioglitazone reduces the composite of all-cause mortality, non-fatal myocardial infarction, and stroke in patients with type 2 diabetes who have a high risk of macrovascular events.


A Dosimetric Comparison between Conventional Fractionated and Hypofractionated Image-guided Radiation Therapies for Localized Prostate Cancer.
Image-guided radiation therapy (IGRT) is the preferred method for curative treatment of localized prostate cancer, which could improve disease outcome and reduce normal tissue toxicity reaction. IGRT using cone-beam computed tomography (CBCT) in combination with volumetric-modulated arc therapy (VMAT) potentially allows smaller treatment margins and dose escalation to the prostate. The aim of this study was to compare the difference of dosimetric diffusion in conventional IGRT using 7-field, step-and-shoot intensity-modulated radiation therapy (IMRT) and hypofractionated IGRT using VMAT for patients with localized prostate cancer. We studied 24 patients who received 78 Gy in 39 daily fractions or 70 Gy in 28 daily fractions to their prostate with/without the seminal vesicles using IMRT (n = 12) or VMAT (n = 12) for prostate cancer between November 2013 and October 2015. Image guidance was performed using kilovoltage CBCT scans equipped on the linear accelerator. Offline planning was performed using the daily treatment images registered with simulation computed tomography (CT) images. A total of 212 IMRT plans in conventional cohort and 292 VMAT plans in hypofractionated cohort were enrolled in the study. Dose distributions were recalculated on CBCT images registered with the planning CT scanner. Compared with 7-field, step-and-shoot IMRT, VMAT plans resulted in improved planning target volume (PTV) D95% (7663.17 ± 69.57 cGy vs. 7789.17 ± 131.76 cGy, P < 0.001). VMAT reduced the rectal D25 (P < 0.001), D35 (P < 0.001), and D50 (P < 0.001), bladder V50 (P < 0.001), D25 (P = 0.002), D35 (P = 0.028), and D50 (P = 0.029). However, VMAT did not statistically significantly reduce the rectal V50, compared with 7-field, step-and-shoot IMRT (25.02 ± 5.54% vs. 27.43 ± 8.79%, P = 0.087). To deliver the hypofractionated radiotherapy in prostate cancer, VMAT significantly increased PTV D95% dose and decreased the dose of radiation delivered to adjacent normal tissues comparing to 7-field, step-and-shoot IMRT. Daily online image-guidance and better management of bladder and rectum could make a more precise treatment delivery.


Multiple Salmonella-pathogenicity island 2 effectors are required to facilitate bacterial establishment of its intracellular niche and virulence.
The pathogenesis of Salmonella Typhimurium depends on the bacterium's ability to survive and replicate within host cells. The formation and maintenance of a unique membrane-bound compartment, termed the Salmonella-containing vacuole (SCV), is essential for S. Typhimurium pathogenesis. SCV-bound S. Typhimurium induces formation of filamentous tubules that radiate outwards from the SCV, termed Salmonella-induced filaments (SIFs). SIF formation is concomitant with the onset of replication within host epithelial cells. SIF biogenesis, formation and maintenance of the SCV, and the intracellular positioning of the SCV within the host cell requires translocation of bacterial proteins (effectors) into the host cell. Effectors secreted by the type III secretion system encoded on Salmonella pathogenicity island 2 (T3SS2) function to interfere with host cellular processes and promote both intracellular survival and replication of S. Typhimurium. Seven T3SS2-secreted effectors, SifA, SopD2, PipB2, SteA, SseJ, SseF, and SseG have previously been implicated to play complementary, redundant, and/or antagonistic roles with respect to SIF biogenesis, intracellular positioning of the SCV, and SCV membrane dynamics modulation during infection. We undertook a systematic study to delineate the contribution of each effector to these processes by (i) deleting all seven of these effectors in a single S. Typhimurium strain; and (ii) deleting combinations of multiple effectors based on putative effector function. Using this deletion mutant library, we show that each of SIF biogenesis, intracellular SCV localization, intramacrophage replication, colonization, and virulence depends on the activities of multiple effectors. Together, our data demonstrates the complex interplay between these seven effectors and highlights the necessity to study T3SS2-secreted effectors as groups, rather than studies of individual effectors.


Sodium-dependent net urea transport in rat initial inner medullary collecting ducts.
We reported that feeding rats 8% protein for 3 wk induces net urea transport and morphologic changes in initial inner medullary collecting ducts (IMCDs) which are not present in rats fed 18% protein. In this study, we measured net urea transport in microperfused initial IMCDs from rats fed 8% protein for > or = 3 wk and tested the effect of inhibiting Na+/K(+)-ATPase activity and found that adding 1 mM ouabain to the bath reversibly inhibited net urea transport from 14 +/- 3 to 6 +/- 2 pmol/mm per min (P < 0.01), and that replacing potassium (with sodium) in the bath reversibly inhibited net urea transport from 18 +/- 3 to 5 +/- 0 pmol/mm per min (P < 0.01). Replacing perfusate sodium with N-methyl-D-glucamine reversibly inhibited net urea transport from 12 +/- 2 to 0 +/- 1 pmol/mm per min (P < 0.01), whereas replacing bath sodium had no significant effect on net urea transport. Adding 10 nM vasopressin to the bath exerted no significant effect on net urea transport. Finally, we measured Na+/K(+)-ATPase activity in initial and terminal IMCDs from rats fed 18% or 8% protein and found no significant difference in either subsegment. Thus, net urea transport in initial IMCDs from rats fed 8% protein for > or = 3 wk requires sodium in the lumen, is reduced by inhibiting Na+/K(+)-ATPase, and is unchanged by vasopressin or phloretin. These results suggest that net urea transport may occur via a novel, secondary active, sodium-urea cotransporter.


Revealing radiotherapy- and chemoradiation-induced pathway dynamics in glioblastoma by analyzing multiple differential networks.
The progression of glioblastoma (GBM) is driven by dynamic alterations in the activity and connectivity of gene pathways. Revealing these dynamic events is necessary in order to understand the pathological mechanisms of, and develop effective treatments for, GBM. The present study aimed to investigate dynamic alterations in pathway activity and connectivity across radiotherapy and chemoradiation conditions in GBM, and to give system‑level insights into molecular mechanisms for GBM therapy. A total of two differential co‑expression networks (DCNs) were constructed using Pearson correlation coefficient analysis and one sided t‑tests, based on gene expression profiles and protein‑protein interaction networks, one for each condition. Subsequently, shared differential modules across DCNs were detected via significance analysis for candidate modules, which were obtained according to seed selection, module search by seed expansion and refinement of searched modules. As condition‑specific differential modules mediate differential biological processes, the module connectivity dynamic score (MCDS) was implemented to explore dynamic alterations among them. Based on DCNs with 287 nodes and 1,052 edges, a total of 28 seed genes and seven candidate modules were identified. Following significance analysis, five shared differential modules were identified in total. Dynamic alterations among these differential modules were identified using the MCDS, and one module with significant dynamic alterations was identified, termed the dynamic module. The present study revealed the dynamic alterations of shared differential modules, identified one dynamic module between the radiotherapy and chemoradiation conditions, and demonstrated that pathway dynamics may applied to the study of the pathogenesis and therapy of GBM.


[Molecular genetics of epithelial ovarian neoplasms: correlations with phenotype and biological behavior].
The phenotypic variability of epithelial ovarian neoplasms correlates with a diversity of changes on the molecular level. Invasive serous and undifferentiated ovarian carcinomas are characterized by p53 mutations, extensive loss of genetic material of chromosome 17 and complex changes on many other chromosomes. These alterations are seen only in a minority of mucinous and endometrioid carcinomas, mainly in advanced stages. Overexpression of bcl-2 is seen most frequently in endometrioid carcinomas (ca. 90% of cases), which in addition show microsatellite instability in around a third of cases, as has been described in endometrioid endometrial carcinomas. KRAS mutations are characteristic for mucinous LMP tumors (borderline tumors) and mucinous carcinomas (40-50% of cases). Furthermore, KRAS mutations have been described in around a third of serous LMP tumors, which in addition show microsatellite instability in up to 40% of cases. Serous LMP tumors never harbour complex chromosomal aberrations.


The SL1-SL2 (stem-loop) domain is the primary determinant for stability of the gamma retroviral genomic RNA dimer.
Retroviral genomes are assembled from two sense-strand RNAs by noncovalent interactions at their 5' ends, forming a dimer. The RNA dimerization domain is a potential target for antiretroviral therapy and represents a compelling RNA folding problem. The fundamental dimerization unit for the Moloney murine sarcoma gamma retrovirus spans a 170-nucleotide minimal dimerization active sequence. In the dimer, two self-complementary sequences, PAL1 and PAL2, form intermolecular duplexes, and an SL1-SL2 (stem-loop) domain forms loop-loop base pairs, mediated by GACG tetraloops, and extensive tertiary interactions. To develop a framework for assembly of the retroviral RNA dimer, we quantified the stability of and established nucleotide resolution secondary structure models for sequence variants in which each motif was compromised. Base pairing and tertiary interactions between SL1-SL2 domains contribute a large free energy increment of -10 kcal/mol. In contrast, even though the PAL1 and PAL2 intermolecular duplexes span 10 and 16 bp in the dimer, respectively, they contribute only -2.5 kcal/mol to stability, roughly equal to a single new base pair. First, these results emphasize that the energetic costs for disrupting interactions in the monomer state nearly balance the PAL1 and PAL2 base pairing interactions that form in the dimer. Second, intermolecular duplex formation plays a biological role distinct from simply stabilizing the structure of the retroviral genomic RNA dimer.


Contribution of bone morphogenetic protein-2 to aortic valve calcification in aged rat.
Although aging is well established as an important risk factor for aortic stenosis, the mechanism of age-related aortic valve calcification is yet unknown. Here, we investigated this mechanism in tissue and cellular levels using middle-aged rats. Aortic valve specimens were obtained by dissecting from 9-week-old (young) and 30-week-old (aged) male Wistar rats. In the aged rats, the main risk factors for aortic stenosis in plasma were still in the normal range; however, their number of calcified specimens was significantly increased in comparison with the young rats. Aortic valve interstitial cells (AVICs) obtained from explants of aortic valve specimens were cultured for 14 days after reaching confluence. Spontaneous calcification, the expressions of calcigenic genes, that is, BMP-2, alkaline phosphatase (ALP), and osterix (osteogenic transcription factor) and ALP enzyme activity in AVICs from aged rats were enhanced in comparison with those from young rats. However, neither typical calcification inducing reagents (dexamethasone, β-glycerophosphate, and high concentration of phosphate) nor tumor necrosis factor-α (an inflammatory cytokine) accelerated the spontaneous calcification of AVICs from aged rats. These results suggest that aortic valve calcification progresses with age partly through an activation of the BMP-2 pathway.


Neonatal thyroxine, maternal thyroid function, and child cognition.
Thyroid hormone is essential for normal brain development. Limited data are available regarding whether thyroid function in neonates influences later cognitive development. Our objective was to study associations of newborn T4 levels with maternal thyroid function and childhood cognition. We studied participants in Project Viva, a cohort study in Massachusetts. We studied a total of 500 children born 1999--2003 at 34 wk or more. We determined cognitive test scores at ages 6 months and 3 yr. Mean newborn T4 at a mean age of 1.94 d was 17.6 (sd 4.0) microg/dl, and levels were higher in girls [1.07 microg/dl; 95% confidence interval (CI) 0.38, 1.76] and infants born after longer gestation (0.42 microg/dl; 95% CI 0.17, 0.67 per wk). Newborn T4 levels were not associated with maternal T4, TSH, or thyroid peroxidase antibody levels. On multivariable linear regression analysis, adjusting for maternal and child characteristics, higher newborn T4 was unexpectedly associated with poorer scores on the visual recognition memory test among infants at age 6 months (-0.5; 95% CI -0.9, -0.2), but not with scores at age 3 yr on either the Peabody Picture Vocabulary Test (0.2; 95% CI -0.1, 0.5) or the Wide Range Assessment of Visual Motor Abilities (0.1; 95% CI -0.2, 0.3). Maternal thyroid function test results were not associated with child cognitive test scores. Newborn T4 concentrations within a normal physiological reference range are not associated with maternal thyroid function and do not predict cognitive outcome in a population living in an iodine-sufficient area.


Pre-Columbian Alloy Objects from Guerrero, Mexico.
Fifteen metal artifacts from coastal Guerrero were analyzed. Gold-, silver-, and copper-base alloys were found, and a deliberate manufacture of bronze is indicated. Metallographic studies show a high degree of sophistication in the art of cold-working metal.


Environmental RNA as a Tool for Marine Community Biodiversity Assessments.
Microscopic organisms are often overlooked in traditional diversity assessments due to the difficulty of identifying them based on morphology. Metabarcoding is a method for rapidly identifying organisms where Environmental DNA (eDNA) is used as a template. However, legacy DNA is problematically detected from organisms no longer in the environment during sampling. Environmental RNA (eRNA), which is only produced by living organisms, can also be collected from environmental samples and used for metabarcoding. The aim of this study was to determine differences in community composition and diversity between eRNA and eDNA templates for metabarcoding. Using mesocosms containing field-collected communities from an estuary, RNA and DNA were co-extracted from sediment, libraries were prepared for two loci (18S and COI), and sequenced using an Illumina MiSeq. Results show a higher number of unique sequences detected from eRNA in both markers and higher α-diversity compared to eDNA. Significant differences between eRNA and eDNA for all β-diversity metrics were also detected. This study is the first to demonstrate community differences detected with eRNA compared to eDNA from an estuarine system and illustrates the broad applications of eRNA as a tool for assessing benthic community diversity, particularly for environmental conservation and management applications.


Systemic mastocytosis: case report and literature review.
A case of systemic mastocytosis is reported and the literature reviewed. Although the dermatological manifestation of this rare disease, in the form of urticaria pigmentosa, is well recognized and goes along with the osteosclerotic changes that can be seen with systemic mastocytosis, it is imperative to rule out malignancy as a cause for a new onset of ascites and newly discovered sclerotic osseous changes.


Effect of Downstream Sequence on the Cleavage of Envelop Protein 1 Signal Sequence in Hepatitis C Virus.
The RNA genome of hepatitis C virus encodes a polyprotein of 3 000 amino acids, which is processed into 10 viral proteins by proteases provided by host cells and virus itself. Multiple precursors are produced due to inefficient processing. Here, the study of E1 signal sequence (C/E1 site) processing in eukaryotic vaccinia virus/T7 system is reported. Differently truncated HCV structural proteins were expressed in this system. It was found that the efficient cleavage of E1 signal sequence was affected by downstream envelope protein sequences. When the lacZ gene encoding a product with similar size was engineered downstream to the E1 signal sequence, the inefficient cleavage of signal sequence was also observed, suggesting that the effect of downstream sequence on the cleavage was due to the presence of the envelop protein sequences. Computer-aided analysis clearly showed that E1 signal sequences was a typical signal sequence. The influence of downstream sequences to signal sequence cleavage demonstrated here was uncommon. To date, similar observations were only reported for the processing of IL-12 signal sequence and the C/prM site of flavivirus. As both flavivirus and HCV are classified into the same Flaviviridae family, this downstream-sequence-related cleavage of signal sequence worths further studying.


[Role of Anammox Bacteria in Removal of Nitrogen Compounds from Wastewater].
The review deals with the unique microbial group responsible for anaerobic ammonium oxidation with nitrite (anammox), and with the role of this process in development of the biotechnology for removal of nitrogen compounds from wastewater. The history of the study of this process is briefly related. Up-to date knowledge on the intracellular organization, energy metabolism, growth stoichiometry, and physiology of anammox bacteria is described, and the main methods for cultivation of these microorganisms are characterized. Special attention is paid to the problems associated with practical application of anammox bacteria, which result from their extremely slow growth, the absence of pure cultures, and the interaction with other microbial groups.


Conversion of lysine 91 to methionine or glutamic acid in human choriogonadotropin alpha results in the loss of cAMP inducibility.
Human choriogonadotropin (hCG) is a heterodimeric glycoprotein hormone. The alpha subunit comprises 92 amino acids, of which 6 are Lys residues (Morgan, F.G., Birken, S., and Canfield, R.E. (1975) J. Biol. Chem. 250, 5247-5258). Our photoaffinity-labeling studies indicate that several of these Lys residues make contact with the lutropin receptor and are covalently cross-linked to the receptor. Lys-91 of the alpha subunit is of interest because deletion of the two alpha C-terminal residues, Lys-91 and Ser-92, results in a significant reduction in the bioactivity of lutropin and thyrotropin (Cheng, K.-W., Glazer, A.N., and Pierce, J.G. (1973) J. Biol. Chem. 248, 7930-7937). To determine the importance of Lys-alpha 91, we substituted it with Arg, Met, or Glu. The resulting mutant alpha cDNA constructs were co-transfected into CHO cells with the wild type hCG beta cDNA construct. Secreted hCG dimers were assayed for binding to receptors on porcine granulosa cells and stimulation of cAMP synthesis in a murine Leydig tumor cell line. The natural hCG, wild type hCG, and all mutant hCGs recognized the receptor, although with somewhat divergent affinities. However, there was a striking difference in the ability of cAMP induction. The natural hCG, wild type hCG, and Lys-91----Arg mutant hCG induced cAMP synthesis, whereas the Lys-91----Met and Lys-91----Glu mutants did not. These results demonstrate that Lys-91 is important for receptor modulation in the stimulation of cAMP synthesis.


Flexible conductive-bridging random-access-memory cell vertically stacked with top Ag electrode, PEO, PVK, and bottom Pt electrode.
Flexible conductive-bridging random-access-memory (RAM) cells were fabricated with a cross-bar memory cell stacked with a top Ag electrode, conductive polymer (poly(n-vinylcarbazole): PVK), electrolyte (polyethylene oxide: PEO), bottom Pt electrode, and flexible substrate (polyethersulfone: PES), exhibiting the bipolar switching behavior of resistive random access memory (ReRAM). The cell also exhibited bending-fatigue-free nonvolatile memory characteristics: i.e., a set voltage of 1.0 V, a reset voltage of -1.6 V, retention time of >1 × 10(5) s with a memory margin of 9.2 × 10(5), program/erase endurance cycles of >10(2) with a memory margin of 8.4 × 10(5), and bending-fatigue-free cycles of ∼1 × 10(3) with a memory margin (I(on)/I(off)) of 3.3 × 10(5).


Polycomblike protein PHF1b: a transcriptional sensor for GABA receptor activity.
The γ-aminobutyric acid (GABA) type A receptor (GABA(A)R) contains the recognition sites for a variety of agents used in the treatment of brain disorders, including anxiety and epilepsy. A better understanding of how receptor expression is regulated in individual neurons may provide novel opportunities for therapeutic intervention. Towards this goal we have studied transcription of a GABA(A)R subunit gene (GABRB1) whose activity is autologously regulated by GABA via a 10 base pair initiator-like element (β(1)-INR). By screening a human cDNA brain library with a yeast one-hybrid assay, the Polycomblike (PCL) gene product PHD finger protein transcript b (PHF1b) was identified as a β(1)-INR associated protein. Promoter/reporter assays in primary rat cortical cells demonstrate that PHF1b is an activator at GABRB1, and chromatin immunoprecipitation assays reveal that presence of PHF1 at endogenous Gabrb1 is regulated by GABA(A)R activation. PCL is a member of the Polycomb group required for correct spatial expression of homeotic genes in Drosophila. We now show that PHF1b recognition of β(1)-INR is dependent on a plant homeodomain, an adjacent helix-loop-helix, and short glycine rich motif. In neurons, it co-immunoprecipitates with SUZ12, a key component of the Polycomb Repressive Complex 2 (PRC2) that regulates a number of important cellular processes, including gene silencing via histone H3 lysine 27 trimethylation (H3K27me3). The observation that chronic exposure to GABA reduces PHF1 binding and H3K27 monomethylation, which is associated with transcriptional activation, strongly suggests that PHF1b may be a molecular transducer of GABA(A)R function and thus GABA-mediated neurotransmission in the central nervous system.


A large-scale method for the selective depletion of alphabeta T lymphocytes from PBSC for allogeneic transplantation.
We sought to develop a method for the clinical large-scale depletion of alphabeta T lymphocytes from mobilized peripheral stem cells, which would allow the allogeneic transplantation of a graft enriched for stem cells, natural killer (NK) cells and gammadelta T lymphocytes. Therefore, we obtained mononuclear cells from either mobilized or non-mobilized healthy adult volunteer donors and incubated the cells with a biotinylated anti-alphabeta T-cell Ab and subsequently with an anti-biotin Ab conjugated with magnetic microbeads. The depletion was then performed using a CliniMACS device. The median T-cell depletion was 3.9 log (range 3.5-4.1 log). The recovery of the gammadelta and NK cells was 92% and 80%, respectively. The recovery of CD34+ stem cells from the mobilized donors was 66%. This method had no negative influence on the in vitro colony formation of stem cells, and transplantation of alphabeta-depleted cells into NOD-SCID IL-2 common gamma chain knockout (NOD-scid IL2r (null)) mice resulted in a rapid engraftment of human myeloid and lymphoid cells. This method will allow large-scale depletion of alphabeta T cells from mobilized peripheral blood in clinical trials.


A Novel Automated Peak Frequency Annotation Algorithm for Identifying Deceleration Zones and Ventricular Tachycardia Ablation Sites.
Current annotation of local fractionated signals during ventricular electroanatomical mapping (EAM) requires manual input subject to variability and error. To evaluate a novel peak frequency (PF) annotation software for its ability to automatically detect late potentials (LP) and local abnormal ventricular activity (LAVA), determine an optimal range for display, and assess its impact on isochronal late activation mapping (ILAM). EAM data from 25 patients who underwent ventricular tachycardia (VT) ablation were retrospectively analyzed. Samplings of electrogram peak frequencies from areas of normal bipolar voltage, areas of low voltage, and areas of low voltage with fractioned signals were performed. An optimal range of frequency display was identified from these patients and applied to a validation cohort of 10 prospective patients to assess high PF within scar as a predictor of VT ablation target sites, in particular deceleration zones (DZs) identified by ILAM, LP, and LAVA. Voltage and PF ranges of normal endocardial tissue varied widely. Using 220 Hz as a frequency cut-off value in areas of low bipolar voltage, areas of high fractionation were identified with a sensitivity of 91% and a specificity of 85% There was no significant reduction in targeted DZ surface areas and colocalization with DZs was observed in all cases. Applied to the prospective cohort, PF predicted fractionated areas and DZ in 9/10 patients. A PF annotation algorithm with a cut off of 220 Hz accurately identifies areas of fractioned signals and accurately predicts DZs during ILAM.


Expression of aromatase and 5-alpha-reductase genes in endometrial adenocarcinoma.
The aim of this study was to investigate the expression of genes coding aromatase and 5-alpha-reductase type 1 and type 2 in endometrial adenocarcinoma, with parallel analysis of testosterone concentrations in serum isolated from blood of basilic and ovarian vein. A significant difference between the copy numbers of mRNA for 5-alpha-reductase type 1 and type 2 was disclosed. The content of the mRNA copies was higher for 5-alpha-reductase type 2 than for type 1. We noted a positive correlation between the mRNA copy number for aromatase and 5-alpha-reductase type 1 as well as between the mRNA copy number for aromatase and 5-alpha-reductase type 2. A positive correlation was disclosed between the mRNA copy number for 5-alpha-reductase type 1 and type 2. We concluded that in tissues of endometrial adenocarcinoma, there is expression of aromatase and 5-alpha-reductase type 1 and 2 genes, and that the expression of aromatase and 5-alpha-reductase genes in tissues of endometrial adenocarcinoma is not controlled by testosterone.


Cyclic AMP mediates serotonin-induced synaptic enhancement of lateral giant interneuron of the crayfish.
The lateral giant (LG)-mediated escape behavior of the crayfish habituates readily on repetitive sensory stimulation. Recent studies suggested that the biogenic amines serotonin and octopamine modulate the time course of recovery and/or re-depression of the LG response after habituation. However, little is known of how serotonin and octopamine effect LG habituation and what second-messenger cascades they may activate. To investigate the effect of biogenic amines on LG habituation, serotonin and octopamine were superfused before presenting repetitive sensory stimulation. Serotonin and octopamine increased the number of stimuli needed to habituate the LG response. Their effects were mimicked by mixed application of a cAMP analogue [8-(4-chlorophenylthio)-cAMP (CPT-cAMP)] and a phosphodiesterase inhibitor [3-isobutyl-1-methylxanthine (IBMX)] but not by a cGMP analogue (8-bromoguanosine 3',5'-cyclic monophosphate). Perfusion of the adenylate cyclase inhibitor (SQ22536) abolished the effect of serotonin but not that of octopamine. To investigate the site of action of each biogenic amines in the neural circuit meditating LG escape, the effect of drugs on directly and indirectly elicited postsynaptic potentials in LG was investigated. Serotonin, octopamine, and a mixture of CPT-cAMP and IBMX increased both the direct and indirect synaptic inputs. Simultaneous application of SQ22536 abolished the effect of serotonin on both inputs but did not block the effect of octopamine. Direct injection of the cAMP analogue (Sp-isomer of adenosine-3',5'-cyclic monophosphorothioate) into LG increased both the direct and indirect inputs to LG. These results indicate that serotonin mediates an increase in cAMP levels in LG, but octopamine acts independently of cAMP and cGMP.


Development of intramolecular vicinal diamination of alkenes: from palladium to bromine catalysis.
Palladium catalysis has been instrumental in the development of the intramolecular diamination of alkenes. Reagent combinations of a palladium catalyst and iodosobenzene diacetate or copper(II) salts, respectively, represent the broad applicability and mechanistic variation. Recent work has established alternative copper and bromine catalysts. The occupation with this reaction has also contributed to the development of high oxidation state metal catalysis in alkene difunctionalization and significantly broadened the spectrum of Pd-catalyzed C-N bond-forming reactions in general.


Establishing Recovery Colleges in mental healthcare services: A Royal Commission recommendation.
Recovery Colleges are an innovative education-based approach to support mental health recovery that, following the recent Royal Commission, will have to be established in every area mental health service within the state of Victoria. This paper describes the rationale, benefits and some of the key considerations to successfully establish Recovery Colleges. The establishment of Recovery Colleges has the potential to drive culture change within mental health services and embed recovery orientation within service provision as well as engaging service users in their own recovery journey. There are significant challenges, however, in implementing the collaborative, co-produced model within the constraints of a publicly funded mental health clinical service. This paper considers some of the practice implications for public mental health services in developing and integrating Recovery Colleges. The paper, like everything we do at the Recovery College, is co-produced and co-authored - in this case, by a lived experience expert, a medically trained expert and a research/writing expert.


Rhabdomyolysis after infection and taking a cold medicine in a patient who was susceptible to malignant hyperthermia.
A case of rhabdomyolysis after a possible viral infection and the use of a cold medication is reported. A 41-year-old man who presented with dysarthria, dysphagia, progressive weakness of his muscles and a high grade fever was admitted. He suffered from massive rhabdomyolysis, acute renal failure, and bronchopneumonia. Hemodialysis, antibiotics, and hydration therapy were effective in the treatment of his illness. Although the cause of the rhabdomyolysis was not completely clear, he was subsequently shown to be susceptible to malignant hyperthermia (MH) based on the results of a caffeine-halothane contracture test. When a mild recurrence occurred during a follow-up muscle biopsy, intravenous dantrolene sodium was administered and he improved immediately. This case suggests that MH should be considered in patients with rhabdomyolysis when the cause is unclear. The caffeine-halothane contracture test may also be helpful in the diagnosis.


Sudden mono-ocular blindness with recurrent transient diplopia and ptosis in a middle-aged woman.
Sudden painless loss of vision is an ophthalmologic and a medical emergency resulting from various causes such as occlusion of retinal artery or vein, macular or vitreous haemorrhages, retinal detachment, and anterior and posterior ischaemic optic neuropathy. We report a 48-year-old woman presenting with right monocular blindness due to branch retinal artery occlusion whose vision recovered due to timely paracentesis coupled with treatment with adequate antiplatelet agents and anticoagulants. The patient had transient diplopia and ptosis despite adequate antiplatelet agents and anticoagulants. Thorough search for aetiology revealed the underlying cause to be aortoarteritis. Aortoarteritis is a rare disease, and ocular involvement occurs late in the disease. We review ophthalmologic manifestation of aortoarteritis and diagnostic utilities of various modalities for aortoarteritis.


A tale of 141 municipalities: the spatial distribution of dengue in Mato Grosso, Brazil.
In recent years, the state of Mato Grosso has presented one of the highest dengue incidence rates in Brazil. The meeting of the Amazon, Cerrado and Pantanal biomes results in a large variation of rainfall and temperature across different regions of the state. In addition, Mato Grosso has been undergoing intense urban growth since the 1970s, mainly due to the colonization of the Mid-North and North regions. We analyzed factors involved in dengue incidence in Mato Grosso from 2008 to 2019. The Moran Global Index was used to assess spatial autocorrelation of dengue incidence using explanatory variables such as temperature, precipitation, deforestation, population density and municipal development index. Areas at risk of dengue were grouped by the Local Moran Indicator. We noticed that areas at risk of dengue expanded from the Mid-North region to the North; the same pattern occurred from the Southeast to the Northeast; the South region remained at low-risk levels. The increase in incidence was influenced by precipitation, deforestation and the municipal development index. The identification of risk areas for dengue in space and time enables public health authorities to focus their control and prevention efforts, reducing infestation and the potential impact of dengue in the human population.


The effect of nanoparticles on soil and rhizosphere bacteria and plant growth in lettuce seedlings.
Nanomaterials are increasingly being considered for use in agricultural applications, where they have been suggested for a range of uses including fertilizer and pesticide applications. Among nanomaterial applications, agricultural use has a particularly high likelihood of introducing significant quantities of nanomaterials to the environment. The focus of this work was on conducting preliminary experiments examining how nanomaterials might influence rhizosphere bacteria, and in turn influence plant growth. For this work, buttercrunch lettuce seeds were grown in the presence of suspensions of three different nanoparticles. Two of the studied nanomaterials, amine-modified polystyrene nanospheres and titanium dioxide nanoparticles, caused significant decreases in both rhizosphere bacterial counts and plant root and stem growth. In contrast, sulfate-modified polystyrene nanospheres actually increased rhizosphere bacterial counts, but had no significant impact on growth. Only the amine-modified polystyrene nanospheres were found to attach to root surfaces, suggesting that nanomaterial attachment to root surfaces is not a requirement for hindered plant growth. It was hypothesized that attachment of amine-modified polystyrene and TiO2 nanomaterials to bacteria themselves could be changing the bacteria surface properties, and ultimately reducing bacterial affinity for root surfaces.


A biochemical and histologic rationale for the treatment of hydrofluoric acid burns with calcium gluconate.
Hydrofluoric acid has unique properties that make it attractive for a variety of industrial and household uses. Exposure to dilute and concentrated solutions of hydrofluoric acid can lead to severe pain and tissue necrosis. Local treatment with topical calcium gluconate and subdermal injections of 0.5 ml 10% solution of calcium gluconate per cm2 of affected tissue has been advocated but frequently fails to relieve the patient of pain. Intraarterial infusion of calcium gluconate has been advocated for these patients, but several reports have been made of massive soft-tissue loss associated with such therapy. The purpose of this study was to show what, if any, were the microscopic effects on the distal arterial tree of intraarterial infusion of calcium gluconate. By studying 1 micron-thick cuts of distal rat aortas after proximal infusion of concentrated (10%) and dilute (2%) calcium gluconate, we were able to show that the incidence of microperforations in the intima and media of the rat aorta increased with the concentration of calcium gluconate. We conclude that intraarterial infusions should be reserved for only the most severe cases of hydrofluoric acid burns unresponsive to local therapy.


Effect of Plastic Deformation on the Structure and Mechanical Properties of the Zn-4Ag-1Cu Zinc Alloy.
It is known that zinc biodegradable alloys are a promising material for producing biomedical implants for orthopedics and vascular stents. Among them, the Zn-Ag-Cu zinc alloy is of special interest due to the antibacterial and antimicrobial properties of Ag and Cu. To improve the mechanical properties of the Zn-4Ag-1Cu zinc alloy, the effect of equal-channel angular pressing (ECAP) on the microstructure and strength has been investigated. The ECAP conditions for the Zn-4Ag-1Cu alloy were chosen by modeling in the Deform 3 D program (temperature and strain rate). The microstructure was analyzed using transmission electron microscopy, scanning electron microscopy and X-ray diffraction analysis. The study of strength was carried out by measuring the microhardness and tensile tests of small samples with a gauge dimension of 0.8 × 1 × 4 mm3. The microstructure after ECAP was characterized by equiaxed grains ranging in a size from 1.5 µm to 4 µm with particles in a size from 200 nm to 1 µm uniformly distributed along the boundaries. The ECAP samples showed a high strength of 348 MPa and good ductility of up to 30%, demonstrating their great potential as promising materials for producing medical stents.


Orthodontic postgraduate training in Germany--have we anything to learn?
In 1996, the authors were able to visit the University Orthodontic Unit in Marburg, Germany. Author FL was able to undertake the visit as part of a discretionary secondment for specialist training and this article describes the author's (FL) experience of orthodontic, postgraduate training there, and highlights differences between training in the U.K. and Germany. In the current climate of change with regard to specialist training, it seemed timely to investigate how training occurs elsewhere in Europe.


LDR brachytherapy: can low dose rate hypersensitivity from the "inverse" dose rate effect cause excessive cell killing to peripherial connective tissues and organs?
Examined here are the possible effects of the "inverse" dose rate effect (IDRE) on low dose rate (LDR) brachytherapy. The hyper-radiosensitivity and induced radioresistance (HRS/IRR) effect benefits cell killing in radiotherapy, and IDRE and HRS/IRR seem to be generated from the same radioprotective mechanisms. We have computed the IDRE excess cell killing experienced in LDR brachytherapy using permanent seed implants. We conclude, firstly, that IDRE is a dose rate-dependent manifestation of HRS/IRR. Secondly, the presence of HRS/IRR or IDRE in a cell species or tissue must be determined by direct dose-response measurements. Thirdly, a reasonable estimate is that 50-80% of human adjoining connective and organ tissues experience IDRE from permanent implanted LDR brachytherapy. If IDRE occurs for tissues at point A for cervical cancer, the excess cell killing will be about a factor of 3.5-4.0 if the initial dose rate is 50-70 cGy h(-1). It is greater for adjacent tissues at lower dose rates and higher for lower initial dose rates at point A. Finally, higher post-treatment complications are observed in LDR brachytherapy, often for unknown reasons. Some of these are probably a result of IDRE excess cell killing. Measurements of IDRE need be performed for connective and adjacent organ tissues, i.e. bladder, rectum, urinary tract and small bowels. The measured dose rate-dependent dose responses should extended to <10 cGy h(-1) and involve multiple patients to detect patient variability. Results may suggest a preference for high dose rate brachytherapy or LDR brachytherapy without permanent retention of the implant seeds (hence the dose rates in peripheral tissues and organs remain above IDRE thresholds).


The Human Genome Project in the United States: a perspective on the commercial, ethical, legislative and health care issues.
The Human Genome Project represents a government supported effort to map and sequence the human genome. Governmental support for the Project should include increased emphasis on grants and contracts to industry. This is particularly true for small highly innovative biotechnology companies that can rapidly integrate and use technology as a base for product development. Private industry must be integrated as a partner into the Project, as it will be in Japan and Europe. There is a consensus in industry that the Genome Project is, at this stage at least, a science research project funded by government and not, at present, a commercial project. It is not seriously expected to have any substantial widespread commercial impact in the near future. The ultimate commercial benefits of the Human Genome Project, in terms of definable health care products, may not be realized until well into the next century. Yet, there are a few companies for which the Genome Project affords immediate commercial opportunities in certain niche areas. The Genome Project can be expected to have a significant impact upon medical knowledge and treatment. Though the Genome Project is just beginning, much of the type of medical knowledge expected to be gained from the Project is already present and even being exploited, albeit on a small scale. The application of genetic understanding to practical applications raises ethical, medical and legal issues central to the Genome Project. Unfortunately, emotion and sensationalism sometimes dominate and prevent a constructive discussion of ethical and social issues pertaining to genetics. To answer public concerns about human gene transfer experiments, the medical and biotechnology communities must constructively discuss the medical realities, the benefits to human health and the adequacy of the current governmental oversights. These presentations must be understandable by the lay public and must address their fears. Failure to assuage public fears and concerns, no matter how abstract or unrealistic they may be to the medical/scientific community, will lead to increased governmental controls and regulatory burdens.


Behaviour in biotin-deficient rats of thymic peptides controlling DNA transcription.
Biotin-deficient rats show a slowing down of the growth and an involution of the thymus. The amount of the thymic peptides controlling DNA template, if referred to the thymus weight is higher in deficient than in control rats; no significant difference is noticed among the contents of the active peptides when evaluated per rat. The inhibiting activity on RNA synthesis is the same for the peptides extracted from normal and from biotin-deficient rat thymus.


Study of inhalation toxicity of performic, peracetic and perpropionic acid in mice.
The acute and subacute inhalation toxicity of laboratory performic, peracetic and stabilized aqueous peracetic acid solution. The differences in the toxicity of the studied of the commercial preparation Persteril for mice was studied. Persteril is a 40% peracids were not statistically significant. The histological findings in the lung tissue of intoxicated mice show that the effet of the given peracids is qualitatively the same. Although their toxicity is not very high, in repared inhalation they can induce temporary pathomorphological lesions in the lung parenchyma and have a negative effect on the development of mice in concentrations of about only 70 mg/m3 air. As regards their inhalation toxicity, performic and perpropionic acid have no advantages over peracetic acid for practical use in the air disinfection. The findings show that it is necessary for persons carrying out spatial disinfection with one of the studied peracids to wear an protective mask.


Volar radiocarpal dislocation with ulnar translocation.
The first known reported case of volar radiocarpal dislocation with ulnar translocation is described with a review of previous literature on the isolated components of these injuries. Radiographic criteria for their diagnosis are presented with recommendations for early operative intervention to minimize the risk of subsequent carpal instability and provide an optimum functional result. An awareness of the possible coexistence of these injuries leading to early diagnosis and expedient treatment is essential to a good functional result.


Cutaneous Tuberculosis: An Update.
Among extra pulmonary tuberculosis cutaneous tuberculosis occurs rarely, despite a high and increasing prevalence of tuberculosis worldwide particularly in developing countries. Cutaneous tuberculosis can be acquired exogenously or endogenously and present as a diverse of differing clinical morphologies. Diagnosis of these lesions can be difficult, as they resemble many other dermatological conditions that are often primarily considered. Moreover, microbiological confirmation is poor, despite scientific advances, such as the more frequent use of polymerase chain reaction. Furthermore, treatment of cutaneous tuberculosis is also challenging in many cases.


Dopaminergic circuitry and risk/reward decision making: implications for schizophrenia.
Abnormal reinforcement learning and representations of reward value are present in schizophrenia, and these impairments can manifest as deficits in risk/reward decision making. These abnormalities may be due in part to dopaminergic dysfunction within cortico-limbic-striatal circuitry. Evidence from studies with laboratory animal have revealed that normal DA activity within different nodes of these circuits is critical for mediating dissociable processes that can refine decision biases. Moreover, both phasic and tonic dopamine transmission appear to play separate yet complementary roles in these processes. Tonic dopamine release within the prefrontal cortex and nucleus accumbens, serves as a "running rate-meter" of reward and reflects contextual information such as reward uncertainty and overt choice behavior. On the other hand, manipulations of outcome-related phasic dopamine bursts and dips suggest these signals provide rapid feedback to allow for quick adjustments in choice as reward contingencies change. The lateral habenula is a key input to the DA system that phasic signals is necessary for expressing subjective decision biases; as suppression of activity within this nucleus leads to catastrophic impairments in decision making and random patterns of choice behavior. As schizophrenia is characterized by impairments in using positive and negative feedback to appropriately guide decision making, these findings suggest that these deficits in these processes may be mediated, at least in part, by abnormalities in both tonic and phasic dopamine transmission.


Reflections on academic supervision.
Academic supervision is vital for students who are developing academic careers. New lecturers have little training to prepare them for the role of supervisor and may not feel adequately prepared to support the needs of students. This article discusses academic supervision from the viewpoint of a nurse lecturer, examines the nature of supervision and makes recommendations for practice.


FER kinase promotes breast cancer metastasis by regulating α6- and β1-integrin-dependent cell adhesion and anoikis resistance.
Metastatic breast cancer cannot be treated successfully. Currently, the targeted therapies for metastatic disease are limited to human epidermal growth factor receptor 2 and hormone receptor antagonists. Understanding the mechanisms of breast cancer growth and metastasis is therefore crucial for the development of new intervention strategies. Here, we show that FER kinase (FER) controls migration and metastasis of invasive human breast cancer cell lines by regulating α6- and β1-integrin-dependent adhesion. Conversely, the overexpression of FER in non-metastatic breast cancer cells induces pro-invasive features. FER drives anoikis resistance, regulates tumour growth and is necessary for metastasis in a mouse model of human breast cancer. In human invasive breast cancer, high FER expression is an independent prognostic factor that correlates with high-grade basal/triple-negative tumours and worse overall survival, especially in lymph node-negative patients. These findings establish FER as a promising target for the prevention and inhibition of metastatic breast cancer.


A mixed antiglobulin test with kidney cells in suspension for IgG antibody in human allograft recipients.
A modification of the mixed antiglobulin test for the detection of IgG antibodies directed against human kidney cells has been devised which uses a suspension of individual kidney cells rather than a monolayer culture as antigen so that it may be enployed clinically as a prospective crossmatch for organ transplantation. The antiglobulin reagent is added to the sensitized kidney cells rather than to the indicator erythrocytes, which reduces the background of nonspecific reations almost to zero. It is reproducible. The mixed antiglobulin test detected antibody in the sera of patients on chronic dialysis two to four times more frequently than did either the immune adherence test or the most sensitive modification of the microlymphocytotoxicity test which was utilized. It detected the development of antibody specific for donor kidney cells in the sera of 5 of 10 allograft receipients during periods of good to moderate renal transplant function several months before rejection.


Ultrasonography for Noninvasive Assessment of Portal Hypertension.
Portal hypertension is a major pathophysiology in patients with cirrhosis. Portal pressure is the gold standard to evaluate the severity of portal hypertension, and radiological intervention is the only procedure for pressure measurement. Ultrasound (US) is a simple and noninvasive imaging modality available worldwide. B-mode imaging allows broad applications for patients to detect and characterize chronic liver diseases and focal hepatic lesions. The Doppler technique offers real-time observation of blood flow with qualitative and quantitative assessments, and the application of microbubble-based contrast agents has improved the detectability of peripheral blood flow. In addition, elastography for the liver and spleen covers a wider field beyond the original purpose of fibrosis assessment. These developments enhance the practical use of US in the evaluation of portal hemodynamic abnormalities. This article reviews the recent progress of US in the assessment of portal hypertension.


Prolonged P wave duration predicts stroke mortality among type 2 diabetic patients with prevalent non-major macrovascular disease.
Prolonged P wave duration is a marker of delayed inter-atrial conduction which may predict cardiovascular disease (CVD). Type 2 diabetes is a risk factor for all atherosclerotic manifestations including stroke. We evaluated the prognostic significance of prolonged P wave duration among middle-aged Finnish type 2 diabetes patients with and without prevalent non-major macrovascular disease (PNMMVD) with respect to total and stroke mortality. We followed up for 18 years 739 type 2 diabetic patients without previous major CVD event at baseline. Participants were stratified according to P wave duration (<114 or ≥ 114 ms) and PNMMVD (i.e. coronary heart disease defined as ischaemic ECG changes and typical symptoms of angina pectoris, or claudication; yes or no). The Cox proportional hazards model was used to estimate the joint association between P wave duration, PNMMVD and the mortality risk. During the follow-up, 509 patients died, and 59 of them died from stroke. Those who had prolonged P wave duration had 2.45 (95% confidence interval: 1.11-5.37) increased stroke mortality among PNMMVD patients. In patients without PNMMVD, there was no relationship between P wave duration and stroke mortality. As an easily measurable factor P wave duration merits further studies with higher number of patients to evaluate its importance in the estimation of stroke risk in type 2 diabetic patients with PNMMVD.


HLA-DP and susceptibility to insulin-dependent diabetes mellitus in Japanese.
Human leukocyte antigen (HLA) genes are candidates for susceptibility genes in insulin-dependent diabetes mellitus (IDDM). Recently, the association of DR and DQ with IDDM has been reported, but the role of HLA-DP genes remains uncertain. To address the question, we analyzed the DPB1 gene of 20 Japanese IDDM patients and 30 control subjects using a combination of polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analysis (PCR-RFLP method). DPB1*0501 was the most frequent allele both in Japanese patients and control subjects. There was no appreciable association between IDDM and the DPB1 allele in Japanese. The absence of association between IDDM and DP, in spite of the known association between this disease and both DR and DQ, suggests that the HLA locus (loci) telomeric to DP encodes susceptibility to IDDM.


Understanding the shapes of bacteria just got more complicated.
The paper by Briegel et al. in this issue of Molecular Microbiology uses advanced cryo-transmission electron microscopy (cryoTEM) techniques to reveal four separate locations of cytoplasmic filament bundles in Caulobacter crescentus. Intuitively, these filaments should be rather rigid protein structures and composed of previously identified shape-forming proteins, such as crescentin or MreB. Yet, deletion mutants lacking these proteins still possessed filaments and still possessed wild-type morphology. These results suggest that a complex combination of protein structures, including those of crescentin, MreB and these newly identified bundles, in combination with the cell envelope help maintain the complicated shape of C. crescentus. Other bacteria might have similar architectural proteins to assist in maintaining the cell contours during growth and division.


Influence of grape density and harvest date on changes in phenolic composition, phenol extractability indices, and instrumental texture properties during ripening.
Changes in the phenolic composition, phenol extractability indices, and mechanical properties occur in grape berries during the ripening process, but the heterogeneity of the grapes harvested at different ripening stages affects the reliability of the results obtained. In this work, these changes were studied in Nebbiolo grapes harvested during five consecutive weeks and then separated according to three density classes. The changes observed in chemical and mechanical parameters through the ripening process are more related to berry density than harvest date. Therefore, the winemaker has to select the flotation density according to the objective quality properties of the wine to be elaborated. On the other hand, the stiffer grapes were associated with a higher accumulation of proanthocyanidins. The harder grapes provided the higher concentration and extractability of flavanols reactive to vanillin, whereas the thicker ones facilitated the extraction of proanthocyanidins.


Axial elongation measured by long scan depth optical coherence tomography during pilocarpine-induced accommodation in intraocular lens-implanted eyes.
We used an ultra-long scan depth optical coherence tomography (UL-OCT) system to investigate changes in axial biometry of pseudophakic eyes during pilocarpine- induced accommodation. The right eyes from 25 healthy subjects (age range 49 to 84 years) with an intraocular lens (IOL) were imaged twice in the non-accommodative and the accommodative states. A custom-built UL-OCT instrument imaged the whole eye. Then accommodation was induced by two drops of 0.5% pilocarpine hydrochloride separated by a 5-minute interval. Following the same protocol, images were acquired again 30 minutes after the first drop. The central corneal thickness (CCT), anterior chamber depth (ACD), IOL thickness (IOLT), and vitreous length (VL) were obtained using custom automated software. The axial length (AL) was calculated by summing the CCT, ACD, IOLT, and VL. With accommodation, ACD increased by +0.08 ± 0.09 mm, while the VL decreased by -0.04 ± 0.09 mm (paired t-test each, P<0.05). CCT and IOLT remained constant during accommodation (P > 0.05). The non-accommodative AL was 23.47 ± 0.93 mm, and it increased by +0.04 ± 0.04 mm after accommodation (P<0.01). The AL increased and the IOL moved backward during pilocarpine-induced accommodation in pseudophakic eyes.


Genotyping NAT2 with only two SNPs (rs1041983 and rs1801280) outperforms the tagging SNP rs1495741 and is equivalent to the conventional 7-SNP NAT2 genotype.
Genotyping N-acetyltransferase 2 (NAT2) is of high relevance for individualized dosing of antituberculosis drugs and bladder cancer epidemiology. In this study we compared a recently published tagging single nucleotide polymorphism (SNP) (rs1495741) to the conventional 7-SNP genotype (G191A, C282T, T341C, C481T, G590A, A803G and G857A haplotype pairs) and systematically analysed if novel SNP combinations outperform the latter. For this purpose, we studied 3177 individuals by PCR and phenotyped 344 individuals by the caffeine test. Although the tagSNP and the 7-SNP genotype showed a high degree of correlation (R=0.933, P<0.0001) the 7-SNP genotype nevertheless outperformed the tagging SNP with respect to specificity (1.0 vs. 0.9444, P=0.0065). Considering all possible SNP combinations in a receiver operating characteristic analysis we identified a 2-SNP genotype (C282T, T341C) that outperformed the tagging SNP and was equivalent to the 7-SNP genotype. The 2-SNP genotype predicted the correct phenotype with a sensitivity of 0.8643 and a specificity of 1.0. In addition, it predicted the 7-SNP genotype with sensitivity and specificity of 0.9993 and 0.9880, respectively. The prediction of the NAT2 genotype by the 2-SNP genotype performed similar in populations of Caucasian, Venezuelan and Pakistani background. A 2-SNP genotype predicts NAT2 phenotypes with similar sensitivity and specificity as the conventional 7-SNP genotype. This procedure represents a facilitation in individualized dosing of NAT2 substrates without losing sensitivity or specificity.


High school to college transition: a profile of the stressors, physical and psychological health issues that affect the first-year on-campus college student.
The purpose of this article is to profile identified stressors, physical and psychological health issues that affect first-year campus college students as they transition from high school to college. The Health Behaviors, Self-Rated Health and Quality of Life (QOL) questionnaire was administered to 514 university college students. Results from this study determined that there were significant differences among student life stressors and physical and psychological health status between first-year on-campus and first-year off-campus college students. Most importantly this study documented compelling information regarding selection of roommate, poor housing, chronic and temporary diseases, injury and prescription medicine among college students attending a university in North Carolina. Implications for university health administrators, student affairs personnel, counselors and faculty are discussed.


A Good Day for Helminths: how parasite-derived GDH suppresses inflammatory responses.
Parasitic helminths are often associated with immunoregulation, which allows them to survive in their hosts in the face of type 2 immune responses. They achieve this feat through the secretion of multiple immunomodulatory factors. In this issue of EMBO Reports, Prodjinotho et al show that the parasitic cestode Taenia solium induces regulatory T-cell responses in mice and humans through the release of the metabolic enzyme Glutamate dehydrogenase (GDH), which may be a conserved pathway of immunoregulation in many helminths (Prodjinotho et al, 2022).


Redox-dependent expression of cyclin D1 and cell proliferation by Nox1 in mouse lung epithelial cells.
NADPH oxidases produce reactive oxygen species (ROS) that serve as co-stimulatory signals for cell proliferation. In mouse lung epithelial cells that express Nox1, Nox2, Nox4, p22(phox), p47(phox), p67(phox), and Noxo1, overexpression of Nox1 delayed cell cycle withdrawal by maintaining AP-1-dependent expression of cyclin D1 in low serum conditions. In cycling cells, the effects of Nox1 were dose dependent: levels of Nox1 that induced 3- to 10-fold increases in ROS promoted phosphorylation of ERK1/2 and expression of cyclin D1, whereas expression of Nox1 with Noxo1 and Noxa1 (or expression of Nox4 alone) that induced substantial increases in intracellular ROS inhibited cyclin D1 and proliferation. Catalase reversed the effects of Nox1 on cyclin D1 and cell proliferation. Diphenylene iodonium, an inhibitor of NADPH oxidase activity, did not affect dosedependent responses of ERK1/2 or Akt to serum, but markedly inhibited the sequential expression of c-Fos and Fra-1 required for induction of cyclin D1 during cell cycle re-entry. These results indicate that Nox1 stimulates cell proliferation in actively cycling cells by reducing the requirement for growth factors to maintain expression of cyclin D1, whereas during cell cycle re-entry, NADPH oxidase activity is required for transcriptional activation of Fos family genes during the immediate early gene response.


Factors impacting the access and use of formal health and social services by caregivers of stroke survivors: an interpretive description study.
Evidence has shown that family and friend caregivers of stroke survivors are significantly and negatively impacted by caregiving. The negative effects of caregiving may persist over time suggesting that caregivers might benefit from ongoing engagement with supportive services. However, little is known about caregivers' use of formally funded health and social services, or the factors influencing their access to and use of these services. The aim of this study is to increase understanding of the factors that influence stroke caregivers' access and use of formal health and social services, from the perspective of stroke caregivers and healthcare providers. A qualitative study was conducted with stroke caregivers and health providers in Ontario, Canada using interpretive description. In-depth interviews were conducted with caregivers of survivors who experienced a stroke between six months to five years previous and healthcare providers who support caregivers and stroke survivors. All participants provided written informed consent. Interview data were analyzed using constant comparison to identify codes and develop key thematic constructs. A total of 40 interviews were conducted with 22 stroke caregivers at an average 30-months post-stroke and 18 health providers. Factors that influenced stroke caregivers' access and use of services included: finances and transportation; challenges caregivers faced in caring for their health; trust that they could leave their family member and trust in health providers; limited information pertaining to services and a lack of suitable services; and the response of their social networks to their caregiving situation. Stroke caregivers experience significant challenges in accessing and using formal health and social services. These challenges could be addressed by increasing availability of subsidized community-based supports such as respite and counselling tailored to meet the ongoing needs of caregivers. Systemic change is needed by the health system that readily includes and supports caregivers throughout the stroke recovery continuum, particularly in the community setting.


Cardiovascular damage due to COVID-19: what do we need to know?
Severe Acute Respiratory Syndrome Coronavirus 2 is part of the Cononaviridae family and is the causative agent of the 2019 (Covid-19) Coronavirus pandemic declared by the World Health Organization in March, 2020. This virus has a high rate of transmission, affecting several individuals, and has caused thousands of deaths. The clinical manifestations of Severe Acute Respiratory Syndrome Coronavirus 2 infection are not restricted only to the respiratory tract, and there is an express involvement of the cardiovascular system with a higher risk of death in this group. In such patients there is an overactivation of renin-angiotensin-aldosterone system, which promotes an increase in the expression of angiotensin-converting enzyme - 2 that acts as a receptor for the SPIKE protein expressed by the virus and enables the interaction between the host cell and Severe Acute Respiratory Syndrome Coronavirus 2. This process of infection causes a hyperinflammatory state that increases the inflammatory markers of cardiac injury. Hence, an adequate understanding and clinical guidance regarding the monitoring, and controlling the damage in these patients is essential to avoid worsening of their clinical condition and to prevent death.


Adherence to medication in patients with chronic kidney disease: a systematic review of qualitative research.
Non-adherence to multipharmacological treatment increases the risk of morbidity, mortality and hospitalization. We know little about the perspective of patients with chronic kidney disease regarding factors influencing medicine taking. This study aimed to synthesize findings from qualitative studies of patients' experiences of factors that facilitate and hinder adherence to medication. A systematic review of qualitative studies adhering to the Enhancing transparency in reporting the synthesis of qualitative research (ENTREQ) framework. Systematic searches were conducted in several databases. We used thematic synthesis and the Confidence in the Evidence from Reviews of Qualitative Research (CERQual) approach to assess the confidence of the evidence. Nineteen studies involving 381 patients with chronic kidney disease were included. We identified three analytical themes; logistics, benchmarking the need for medication; and the quality of the patient-physician relationship, with seven descriptive sub-themes as factors influencing patients' adherence to medications. Helping patients to map their everyday activities and motivating them to associate medications with everyday activities may facilitate adherence to medications. Addressing patient beliefs about medications, supporting patients in coping with side effects of medications and eliciting patients' wishes for involvement in treatment decisions may also facilitate adherence. Barriers to adherence were the costs of buying medications, and lacking understanding of the indications and effects of medications. The findings in this synthesis resonate with previous research and extend the known literature by synthesizing and formally assessing confidence in the evidence.


Physiology of Fetal Heart Rate Monitoring.
Fetal heart tracings (FHTs) are useful as a window into the oxygenation status of the fetal brain. Patterns in the FHT reflect the oxygen status of the fetal brain. Fetal adaptive response to progressive hypoxemia and acidosis are detectable and produce recognizable patterns in the fetal heart rate. The basic physiology and adaptive responses that regulate the fetal heart rate and physiological fetal adaptations to stress as reflected in the FHTs are described. Mechanisms of oxygen delivery to the fetus including ways in which those mechanisms can be disrupted are reviewed.


[Changes in total body potassium and ion distribution compartments in general surgery and intensive care patients].
Measurements of total body potassium in a whole body counter revealed significant total body potassium depletion in patients suffering from high output enteric fistulas, short bowel syndrome and Crohn's disease. The clinical importance of this finding is derived from the fact that the degree of potassium depletion was found to be related to mortality, and that the introduction of an infusional regimen for preparation for surgery in Crohn's disease which effected intracellular potassium refilling resulted in a decreased surgical mortality and complication rate. In conclusion specific nutritional support as an essential measure in preparing such patients for elective surgery is particularly recommended. Significant cellular potassium depletion was also found in critically ill patients dying from sepsis. In contrast to the aforementioned diseases with malnutrition, where an increased fecal loss of potassium is the most likely cause of potassium deficiency, a different mechanism for cellular potassium loss is suggested in this condition from additional indicator dilution studies. Rather here the finding of increased distribution volumes for ions physiologically predominantly restricted to the extracellular space (82Br and 24Na) indicates a change of cell membran permeability suggestive of so called sick cell syndrome.


[Gangrene of the hand due to accidental intra-arterial injection of floxacilline: about a case].
Floxacilline is a beta-lactam antibiotic of the penicillin class. In our context it is used to fight against infections caused by gram-positive bacteria, including staphylococcus aureus. However, floxacilline should be administered with great caution due to its possible side-effects. Our study reports the case of a 6-year old boy who underwent surgery for the treatment of a humeral fracture. The child was treated with injectable floxacilline following a suspected orthopaedic device infection 2 months after surgery. The day after starting antibiotic treatment, the child presented with acute ischemia of the right hand. Then, he was referred to us. Explorations objectified occlusion of the radial artery. The patient underwent relieving fasciotomy and received postoperative heparin. The evolution was marked by gangrene affecting the whole hand. The aim of our study was to educate caregivers about the risk of occurrence of this disastrous complication and about the measures to prevent it.


Investigation on some biological aspects of Chrysoperla lucasina (Chrysopidae: Neuroptera) on Bemisia tabaci in laboratory conditions.
Bemisia tabaci is one of the most important key pests of many types of cultivated plants. Lacewings (Chrysopidae: Neuroptera) are predatory insects, widely used in biological control programs. Between them green lacewing is a promising biological control agent of pests in green houses and crop fields. In this study, gravid females of the green lacewing Chrysoperla lucasina (Lacroix) were captured from Sarepolzahab ( altitude 540m, latitude 34 degrees ,14' N 46 degrees, 9' E) in western part of Iran. Collected insects were reared in a growth chamber, under experimental conditions (25 +/- 1 degrees C, 70 +/- 5% RH and a photoperiod of 16:8 L: D). Different diets were offered to larvae which consisted of a whitefly species B. tabaci, an aphid Myzus persica and also lyophilized powder of drone honeybee (Apis melifera). As different foods were used to nurish larvae, so for each diet, mean larval period were calculated, and finally means were compared to each other. Anova in MSTAT-C was used for analysis of variance, and Duncan multiple range test (DMRT) to compare between means. The results showed that larvae had maximum duration of 27 +/- 0.33 days when fed on honeybee lyophilized powder and the minimum value was 17.9 +/- 0.3 days for B. tabaci. 25 +/- 0.27 day recorded for M. persicae. Food preference of the 3rd instar larvae of green lacewing was surveyed, they showed a food preference to M. persicae, to compare with B. tabaci, as the former has a bigger body size, so more easily to be captured by the predator larvae. The 3rd instar larvae of lacewing were more voracious on preys, than the 1st or the 2nd instar larvae. Statistically speaking, there were a significantly difference when mean of different preys consumed by predator larvae were compared. We found, that when the predator larvae have fed on B. tabaci, their development time was shorter, and when arrived to adult stage, the adults showed, an improved fertility. The results indicated that the suitable prey not only can increase the rate of through accelerating developmental stages of the predator and by means of an increase in its pupal body weight consequently promoting the fecundity of resulting adults, but also can alter predators population density in relation to own production numbers.


Convergence of afferent impulses from individual semicircular canals of pontine reticular units.
The effect of afferent impulses on single pontine RF units was recorded by means of tungsten microelectrodes in 22 curarized guinea pigs followed thermic stimulation of individual semicircular canals. The labyrinthine stimulation consisted of short-lasting circumscribed heating (45 degrees C) of the canals through a silver thermode. Unitary response to labyrinthine stimulation was characterized by excitation and inhibition of rest discharge rate. Out of 86 units, 66 received bilateral labyrinthine influences responding in greatest number (20 units) to four semicircular canals. Only two units were unaffected. No significant difference in RF responses was noticed between concordant and antagonistic canals. Reticular formation units showed great degree of facilitation of response and convergence of impulses from the single six semicircular canals.


Partial rescue of mucopolysaccharidosis type VII mice with a lifelong engraftment of allogeneic stem cells in utero.
In utero hematopoietic cell transplantation (IUHCT) has been performed in Mucopolysaccharidosis Type VII (MPSVII) mice, but a lifelong engraftment of allogeneic donor cells has not been achieved. In this study, we sought to confirm a lifelong engraftment of allogeneic donor cells immunologically matched to the mother and to achieve partial rescue of phenotypes in the original MPSVII strain through IUHCT by intravenous injection. We performed in vitro fertilization in a MPSVII murine model and transferred affected embryos to ICR/B6-GFP surrogate mothers in cases where fetuses receiving IUHCT were all homozygous. Lineage-depleted cells from ICR/B6-GFP mice were injected intravenously at E14.5. Chimerism was confirmed by flow cytometry at 4 weeks after birth, and β-glucuronidase activity in serum and several phenotypes were assessed at 8 weeks of age or later. Donor cells in chimeric mice from ICR/B6-GFP mothers were detected at death, and were confirmed in several tissues including the brains of sacrificed chimeric mice. Although the serum enzyme activity of chimeric mice was extremely low, the engraftment rate of donor cells correlated with enzyme activity. Furthermore, improvement of bone structure and rescue of reproductive ability were confirmed in our limited preclinical study. We confirmed the lifelong engraftment of donor cells in an original immunocompetent MPSVII murine model using intravenous IUHCT with cells immunologically matched to the mother without myeloablation, and the improvement of several phenotypes.


Long non-coding RNA NEAT1 facilitates the growth, migration, and invasion of ovarian cancer cells via the let-7 g/MEST/ATGL axis.
Growing evidence indicates a significant role of long non-coding RNA (lncRNA) nuclear-enriched abundant transcript 1 (NEAT1) in ovarian cancer, a frequently occurring malignant tumor in women; however, the possible effects of an interplay of NEAT1 with microRNA (miRNA or miR) let-7 g in ovarian cancer are not known. The current study aimed to investigate the role of the NEAT1/let-7 g axis in the growth, migration, and invasion of ovarian cancer cells and explore underlying mechanisms. NEAT1 expression levels were examined in clinical tissue samples and cell lines. The relationships between NEAT1, let-7 g, and MEST were then analyzed. Gain- or loss-of-function approaches were used to manipulate NEAT1 and let-7 g. The effects of NEAT1 on cell proliferation, migration, invasion, and apoptosis were evaluated. Mouse xenograft models of ovarian cancer cells were established to verify the function of NEAT1 in vivo. NEAT1 expression was elevated while let-7 g was decreased in ovarian cancer clinical tissue samples and cell lines. A negative correlation existed between NEAT1 and let-7 g, whereby NEAT1 competitively bound to let-7 g and consequently down-regulate let-7 g expression. By this mechanism, the growth, migration, and invasion of ovarian cancer cells were stimulated. In addition, let-7 g targeted mesoderm specific transcript (MEST) and inhibited its expression, leading to promotion of adipose triglyceride lipase (ATGL) expression and inhibition of ovarian cancer cell growth, migration, and invasion. However, the effect of let-7 g was abolished by overexpression of MEST. Furthermore, silencing of NEAT1 decreased the xenograft tumor growth by decreasing MEST while up-regulating let-7 g and ATGL. Cumulatively, the findings demonstrated that NEAT1 could promote malignant phenotypes of ovarian cancer cells by regulating the let-7 g/MEST/ATGL signaling axis. Therefore, NEAT1 can be regarded as an important molecular target and biomarker for ovarian cancer.


Switching behaviour of modulated ferroelectrics: the kinetics of the field induced lock-in transition in K₂SeO₄.
The field induced switching process across the ferroelectric lock-in transition in K(2)SeO(4) has been studied on a millisecond timescale using stroboscopic neutron diffraction. The time evolution of both the first and the third order satellites was examined. The time constants are found to vary with temperature between 0.2 and 1.2 ms. This is an order of magnitude faster than in the isostructural Rb(2)ZnCl(4). Moreover, the time dependence of the satellite's linewidth provides information about the evolution of the coherence of the modulated structure.


How will global change affect plant reproduction? A framework for mast seeding trends.
Forest ecology traditionally focuses on plant growth and survival, leaving seed production as a major demographic process lacking a framework for how it will be affected by global change. Understanding plant reproductive responses to changing climate is complicated by masting, the annually variable seed production synchronized within populations. Predicting trends in masting is crucial, because masting impacts seed predation and pollination enough to override simple trends in mean seed production. Proximate mechanisms of seed production patterns in perennial plants are gathered to identify processes through which masting may be affected by a changing environment. Predicting trends in masting will require understanding the mechanisms that cause predictable seed failure after high-seed years, and the stochastic mechanisms that synchronize individuals in high-seed years.


[Acute coronary syndrome with and without ST elevation].
Coronary artery disease accounts for most deaths in western communities. The acute coronary syndrome subsidizes ST elevation myocardial infarction, non-ST elevation myocardial infarction, and unstable angina pectoris. They are characterized by an acute onset of chest pain. The high number of acute coronary syndromes of more than 400,000 per year in Germany demonstrates the necessity of guidelines. Such guidelines are available from different cardiac societies. The implementation of the guidelines of the German Cardiac Society and the European Society of Cardiology in the daily clinical practice are demonstrated in this review by means of two case presentations. Special attention has been given to diagnostic measures, risk stratification, and different therapeutic options. For the diagnostic work-up in the acute phase, the ECG and the assessment of cardiac biomarkers play the central role. For patients with ST elevation myocardial infarction, primary interventional diagnostics and therapy are the first choice. For patients presenting with acute coronary syndromes without ST elevation, a risk-adapted therapeutic approach should be chosen. High-risk patients (elevated troponins, clinical, rhythmologic, and hemodynamic instability, ST depression, or diabetes mellitus) should be treated with an early invasive approach within 48-72 h. Low-risk patients can be treated primarily conservatively. For all patients who undergo interventional treatment, administration of an aggressive antiaggregatory therapy, including acetylsalicylic acid, clopidogrel, glycoprotein IIb/IIIa receptor antagonists, and heparin, is indicated in the acute phase. In the chronic phase, an adequate treatment of cardiovascular risk factors is of paramount importance.


Brain structure, function, and genetics revealed by studies of the eye and face.
Understanding the structure and function of the human brain is intrinsically interesting and fundamental to improved diagnosis, treatment, and prevention of human neurological diseases, which constitute an increasing global burden. The intimate connections between brain and face, and brain and eye, have been utilized to access brain structure and function. Concepts and recent progress are reviewed here. Continued work on biological links between brain and eye or face has uncovered further genetic abnormalities causing facial or eye anomalies, which in either case may clearly indicate changes in the underlying brain. Cause and effect can be difficult to disentangle, but the use of conditional animal models can help establish whether brain changes are the result of face or eye changes or the result of a parallel influence on brain and eye or face. The application of newer methods and technologies such as parameterization of facial characteristics and comparative genomic hybridization has led to new discoveries and insights. Further interdisciplinary studies into brain structure and function through the windows of the face and the eye, with the application of genome-wide studies in larger cohorts, will potentially enable more discovery and critically may reveal unsuspected therapeutic targets in human disease.


Features of a novel protein, rusticalin, from the ascidian Styela rustica reveal ancestral horizontal gene transfer event.
The transfer of genetic material from non-parent organisms is called horizontal gene transfer (HGT). One of the most conclusive cases of HGT in metazoans was previously described for the cellulose synthase gene in ascidians. In this study we identified a new protein, rusticalin, from the ascidian Styela rustica and presented evidence for its likely origin by HGT. Discernible homologues of rusticalin were found in placozoans, coral, and basal Chordates. Rusticalin was predicted to consist of two distinct regions, an N-terminal domain and a C-terminal domain. The N-terminal domain comprises two cysteine-rich repeats and shows remote similarity to the tick carboxypeptidase inhibitor. The C-terminal domain shares significant sequence similarity with bacterial MD peptidases and bacteriophage A500 L-alanyl-D-glutamate peptidase. A possible transfer of the C-terminal domain by bacteriophage was confirmed by an analysis of noncoding sequences of C. intestinalis rusticalin-like gene, which was found to contain a sequence similar to the bacteriophage A500 recombination site. Moreover, a sequence similar to the bacteriophage recombination site was found to be adjacent to the cellulose synthase catalytic subunit gene in the genome of Streptomices sp., the donor of ascidian cellulose synthase. The C-terminal domain of rusticalin and rusticalin-like proteins is likely to be horizontally transferred by the bacteriophage A500. A common mechanism involving bacteriophage mediated gene transfer can be proposed for at least two HGT events in ascidians.


Synthesis, structure, spectral and electrochemical properties, and catalytic use of cobalt(III)-oxo cubane clusters.
Olive-green cobalt(III) complexes having the general formula Co4O4(O2CMe)4L4 (1) where L = py (1a), 4-Mepy (1b), 4-Etpy (1c), and 4-CNpy (1d) have been prepared by the H2O2 oxidation of a mixture of Co2+, MeCO2-, and pyridine or 4-substituted pyridines in a 1:2:1 molar ratio in methanol. Spectroscopic and X-ray crystallographic studies show that these complexes contain a tetrameric cubane-like core [Co4(mu3-O)4]4+ where the four cobalt atoms form an approximate tetrahedron with edge lengths of approximately 2.75 A. Each cobalt in the crystallographically determined structure of Co4(mu3-O)4(mu-O2CMe)4L4 in 1a.NaClO4.3.5H2O and 1b.3H2O is hexacoordinate. Infrared spectra of the complexes show characteristic bands near 700, 635, and 580 cm-1 due to the central cubane-like core. 1H NMR spectra of the complexes show that the dissolved species are essentially diamagnetic and also that the complexes maintain their integrity in solution. UV-vis spectra of the green solutions have been interpreted in terms of ligand-field and charge-transfer bands. The electrochemical behavior of the complexes studied by cyclic and differential pulse voltammetric techniques indicates that the [(CoIII)4(mu3-O)4]4+ core present in the complexes undergoes a reversible one-electron oxidation to the [(CoIII)3CoIV(mu3-O)4]5+ core with an E1/2 value below 1 V. This suggests that these complexes of cobalt may be suitable as catalysts for the oxidation of organic compounds. Preliminary investigations indicate that 1a has a role to play in the cobalt-catalyzed aerobic oxidation of neat ethylbenzene and p-xylene.


Processing determinants required for in vitro cleavage of the poliovirus P1 precursor to capsid proteins.
We generated defined alterations in poliovirus protein-processing substrates and assayed the effects of these alterations with an in vitro expression system. A complete cDNA copy of the poliovirus genome was inserted into a bacteriophage T7 transcription vector. Using this expression template, we produced RNA transcripts containing defined regions of the poliovirus capsid precursor polypeptide (P1) and RNA transcripts containing mutations in the P1 and P2 regions. In vitro translation of P1-derived transcripts allowed us to characterize the 3C-mediated cleavage of P1 to capsid proteins. We demonstrated that, for either posttranslational or cotranslational cleavage at any of the Q-G amino acid pairs within P1, almost the entire P1 precursor is required. We also demonstrated that minimal sequences 3' to the 2A coding sequence are required to generate active 2A proteinase in vitro and that two specific four-amino-acid insertions in protein 2C do not alter 2A- or 3C-mediated processing of the poliovirus polyprotein. In addition, we demonstrated that substantial deletion of P1 sequences does not alter 2A-mediated cleavage of the Y-G site at the P1-P2 junction. These results allowed us to compare the P1 sequences required for 2A- versus 3C-mediated processing of the capsid precursor, and we discuss these results in the context of the three-dimensional structure of the capsid proteins.


Chondroptosis: a variant of apoptotic cell death in chondrocytes?
Evidence has accumulated in recent years that programmed cell death (PCD) is not necessarily synonymous with the classical apoptosis, as defined by Kerr and Wyllie, but that cells use a variety of pathways to undergo cell death, which are reflected by different morphologies. Although chondrocytes with the hallmark features of classical apoptosis have been demonstrated in culture, such cells are extremely rare in vivo. The present review focuses on the morphological differences between dying chondrocytes and classical apoptotic cells. We propose the term 'chondroptosis' to reflect the fact that such cells are undergoing apoptosis in a non-classical manner that appears to be typical of programmed chondrocyte death in vivo. Unlike classical apoptosis, chondroptosis involves an initial increase in the endoplasmic reticulum and Golgi apparatus, reflecting an increase in protein synthesis. The increased ER membranes also segment the cytoplasm and provide compartments within which cytoplasm and organelles are digested. In addition, destruction occurs within autophagic vacuoles and cell remnants are blebbed into the lacunae. Together these processes lead to complete self-destruction of the chondrocyte as evidenced by the presence of empty lacunae. It is speculated that the endoplasmic reticulum pathway of apoptosis plays a greater role in chondroptosis than receptor-mediated or mitochondrial pathways and that lysosomal proteases are at least as important as caspases. Because chondroptosis does not depend on phagocytosis, it may be more advantageous in vivo, where chondrocytes are isolated within their lacunae. At present the initiation factors or the molecular pathways involved in chondroptosis remain unclear.


The carboxyl terminus of Rtt109 functions in chaperone control of histone acetylation.
Rtt109 is a fungal histone acetyltransferase (HAT) that catalyzes histone H3 acetylation functionally associated with chromatin assembly. Rtt109-mediated H3 acetylation involves two histone chaperones, Asf1 and Vps75. In vivo, Rtt109 requires both chaperones for histone H3 lysine 9 acetylation (H3K9ac) but only Asf1 for full H3K56ac. In vitro, Rtt109-Vps75 catalyzes both H3K9ac and H3K56ac, whereas Rtt109-Asf1 catalyzes only H3K56ac. In this study, we extend the in vitro chaperone-associated substrate specificity of Rtt109 by showing that it acetylates vertebrate linker histone in the presence of Vps75 but not Asf1. In addition, we demonstrate that in Saccharomyces cerevisiae a short basic sequence at the carboxyl terminus of Rtt109 (Rtt109C) is required for H3K9ac in vivo. Furthermore, through in vitro and in vivo studies, we demonstrate that Rtt109C is required for optimal H3K56ac by the HAT in the presence of full-length Asf1. When Rtt109C is absent, Vps75 becomes important for H3K56ac by Rtt109 in vivo. In addition, we show that lysine 290 (K290) in Rtt109 is required in vivo for Vps75 to enhance the activity of the HAT. This is the first in vivo evidence for a role for Vps75 in H3K56ac. Taken together, our results contribute to a better understanding of chaperone control of Rtt109-mediated H3 acetylation.


Inflammatory myoblastic tumor of the uterus and interleukin-6.
We report a patient with inflammatory myoblastic tumor of the uterus with constitutional symptoms. Surgical excision was curative and brought a rapid normalization in the elevated serum level of interleukin-6. We revealed an overproduction of interleukin-6 by a detection of messenger RNA by use of reverse transcriptase-polymerase chain reaction and immunohistochemistry.


Frequency and factors associated with unexpected death in an acute palliative care unit: expect the unexpected.
Few studies have examined the frequency of unexpected death and its associated factors in a palliative care setting. To determine the frequency of unexpected death in two acute palliative care units (APCUs); to compare the frequency of signs of impending death between expected and unexpected deaths; and to determine the predictors associated with unexpected death. In this prospective, longitudinal, observational study, consecutive patients admitted to two APCUs were enrolled and physical signs of impending death were documented twice daily until discharge or death. Physicians were asked to complete a survey within 24 hours of APCU death. The death was considered unexpected if the physician answered "yes" to the question "Were you surprised by the timing of the death?" In total, 193 of 203 after-death assessments (95%) were collected for analysis. Nineteen of 193 patients died unexpectedly (10%). Signs of impending death, including non-reactive pupils, inability to close eyelids, decreased response to verbal stimuli, drooping of nasolabial folds, peripheral cyanosis, pulselessness of the radial artery, and respiration with mandibular movement, were documented more frequently in expected deaths than unexpected deaths (P < 0.05). Longer disease duration was associated with unexpected death (33 months vs. 12 months, P = 0.009). Unexpected death occurred in an unexpectedly high proportion of patients in the APCU setting and was associated with fewer signs of impending death. Our findings highlight the need for palliative care teams to be prepared for the unexpected.


1H-MR Spectroscopy of the Early Developmental Brain, Neonatal Encephalopathies, and Neurometabolic Disorders.
MRI interpretations of the pediatric brain are often challenging for general radiologists and clinicians because MR signals and morphology are continuously changing in the developing brain. Furthermore, the developing brain reacts differently to injuries, resulting in imaging characteristics that differ from those of the mature brain. Proton magnetic resonance spectroscopy (1H-MRS) is a non-invasive method for assessing neurological abnormalities at the microscopic level and measures in vivo brain metabolites using a clinical MR machine. In MR examinations of the pediatric brain, 1H-MRS demonstrates its powerful diagnostic capability when MRI is insufficient for diagnostic features. MRI and 1H-MRS may be complementary tools for diagnosing and monitoring diseases. However, there is currently no consensus on how to include 1H-MRS in clinical MR examinations. An overview of the clinical implementation of 1H-MRS for the assessment of early pediatric developmental brains as well as the diagnosis, prognostification, and disease monitoring of various non-neoplastic brain disorders, including neonatal encephalopathies and neurometabolic/neurodegenerative diseases, was provided herein. Qualitative and quantitative 1H-MRS is a powerful non-invasive tool for accessing various brain metabolites to confirm age appropriate peaks and detect abnormal peaks or deficient or reduced peaks, which may facilitate the identification of metabolic and neurodegenerative disorders as well as damage associated with hypoxic-ischemic encephalopathy (HIE). Moreover, 1H-MRS has potential as a biomarker for monitoring therapeutic efficacy in metabolic diseases and neonatal HIE. It also provides insights into the pathophysiologies of various disorders, which may facilitate the use of novel therapeutic approaches. Therefore, 1H-MRS needs to be included more frequently in routine clinical MR examinations of pediatric patients with neurological disorders.


[A case of 5-year survival with combination chemotherapy for advanced colonic cancer with multiple lung and liver metastases and peritoneal dissemination].
A 74-year-old man was admitted to the hospital because of abdominal fullness and constipation. Endoscopy and abdominal computed tomography (CT) scans showed an advanced sigmoid colonic cancer with multiple lung and liver metastases and peritoneal dissemination. Sigmoid resection was performed to relieve ileus. Postoperative chemotherapy was administered for metastatic lesions. We administered systemic chemotherapy with tegafur-uraci(l UFT)/UZEL( 10 months), oxaliplatin with fluorouracil (5-FU) and folinic acid (FOLFOX; 11 courses), irinotecan with 5-FU and folinic acid (FOLFIRI; 13 courses), irinotecan plus S-1 (IRIS; 23 courses), bevacizumab (32 courses), cetuximab (29 courses), and peritoneal infusion of paclitaxel( 65 courses). The patient survived for 5 years after the initial treatment.


Chromosome evolution in new world monkeys (Platyrrhini).
During the last decades, New World monkey (NWM, Platyrrhini, Anthropoideae) comparative cytogenetics has shed light on many fundamental aspects of genome organisation and evolution in this fascinating, but also highly endangered group of neotropical primates. In this review, we first provide an overview about the evolutionary origin of the inferred ancestral NWM karyotype of 2n = 54 chromosomes and about the lineage-specific chromosome rearrangements resulting in the highly divergent karyotypes of extant NWM species, ranging from 2n = 16 in a titi monkey to 2n = 62 in a woolly monkey. Next, we discuss the available data on the chromosome phylogeny of NWM in the context of recent molecular phylogenetic analyses. In the last part, we highlight some recent research on the molecular mechanisms responsible for the large-scale evolutionary genomic changes in platyrrhine monkeys.