[Study of the Mg2+-ATPase reaction of myosin using the NMR-31P method. Detection of adenylate kinase activity in a purified myosin subfragment I].
Isolated myosin heads (HMM-S1) were concentrated by ultrafiltration in an "Amicon" cell to a concentration of 120 mg/ml. Incubation of HMM-S1 with ATP in the presence of Mg2+ produced an AMP peak in the spectrum of NMR-31P whose size was increasing linearly with the reaction time. It was demonstrated that after heating of the sample the appearance of AMP in the incubation mixture was due to the presence of a small amount of adenylate kinase (or myokinase) in the purified preparation of HMM-S1.


Genetic diversity of Gerbich alleles in Brazilians reveals an unexpected prevalence of the GE:-2,-3,4 phenotype.
Gerbich (GE) blood group system carries high-frequency antigens and the absence of them leads to rare phenotypes: GE:-2,3,4, GE:-2,-3,4 and GE:-2,-3,-4. Their serological differentiation is limited and misclassification of Gerbich phenotypes may occur, but this can be avoided by molecular characterization. This study aimed to characterize the molecular background responsible for rare Gerbich phenotypes in Brazilian population. We selected eight samples from patients with anti-Ge, six from their relatives and nine samples with normal expression of Gerbich antigens. Serological tests were performed in gel and red blood cells (RBCs) were tested with anti-Ge2 and anti-Ge3. Monocyte monolayer assay (MMA) was performed. Molecular investigation was performed with allele-specific polymerase chain reaction and DNA sequencing. Patient plasma samples reacted with all commercial RBCs. Patient RBCs showed negative results with anti-Ge2 and anti-Ge3. Using MMA two of eight antibodies were clinically significant. Exon 3 was not amplified in any of the patient samples and in two samples from relatives, suggesting the presence of GE*01.-03/GE*01.-03. By sequencing, we identified the genetic variability that interferes with the definition of deletion breakpoints, thus two options of genetic structure were suggested to be responsible for the GE:-2,-3,4 phenotype. This study showed for the first time the genetic diversity of GYPC alleles for carriers of Gerbich-negative phenotypes in a Brazilian population and showed an unexpected prevalence of the GE:-2,-3,4 phenotype. It also demonstrated the importance of using molecular tools to correctly classify Gerbich phenotypes for selection of variants in antigen-matched transfusions.


The relationship between cognitive impairment, cognitive fatigue, and visual evoked potential latency in people with multiple sclerosis.
Fatigue in people with multiple sclerosis (PwMS) can impact physical, cognitive, and psychosocial domains of daily life. The experience of fatigue in PwMS is thought to originate from the central nervous system, particularly for the domain of cognitive fatigue. Here, we tested the hypothesis that fatigue scores in PwMS would be significantly associated with an index of neural activity - the latency of the P100 of the visual evoked potential (VEP) - in line with the notion of a centralized origin of fatigue. We predicted that prolonged VEP latency would be associated with greater fatigue, and that this relationship would be the most pronounced within the domain of cognitive fatigue. PwMS (n=249) completed the Modified Fatigue Impact Scale (Global, Physical, Cognitive, and Psychosocial scales of the MFIS) and Fatigue Severity Scale. VEP latency was obtained using an alternating checkerboard paradigm. We also examined the influence of depression (Beck Depression Inventory, second edition, BDI-II) and cognitive functioning (NeuroTrax testing battery) on the VEP/fatigue relationship. Surprisingly, we observed that earlier (not later) VEP latency was significantly associated with higher MFIS Cognitive score. The negative relationship between VEP latency and cognitive fatigue was evident in PwMS that had poor cognitive performance as measured by a latent variable that reflected attention, executive function, information processing speed, and motor skills; but a significant relationship was not observed in PwMS that exhibited good performance on this measure. These findings can be interpreted within a metacognitive framework - greater fatigue may be perceived when neural performance and the level of mental effort expended does not translate to efficient cognitive performance. Cognitive fatigue may be particularly salient in PwMS when neural resources are unable to compensate for cognitive difficulties. The mismatch between the expectation of what ought to occur and what does occur during cognitive performance may be a key feature of the experience of cognitive fatigue for some PwMS.


microRNA-27a-3p delivered by extracellular vesicles from glioblastoma cells induces M2 macrophage polarization via the EZH1/KDM3A/CTGF axis.
Glioblastoma (GBM) cell-derived extracellular vesicles (EVs) have been demonstrated to modulate tumor microenvironment. In the present study, we attempted to discuss the role of hsa-microRNA-27a-3p (miR-27a-3p) delivered by GBM-EVs in M2 macrophage polarization. The isolated GBM-EVs were co-cultured with macrophages. After co-culture under normoxia/hypoxia, the effect of EV-derived hsa-miR-27a-3p on GBM cell biological processes was analyzed. Additionally, the target genes of hsa-miR-27a-3p were predicted. Moreover, the binding of enhancer of zeste homologue 1 (EZH1) to lysine-specific demethylase 3A (KDM3A) promoter region and the interaction between KDM3A and connective tissue growth factor (CTGF) were analyzed. GBM mouse models were established to verify the functions of EV-derived hsa-miR-27a-3p in vivo. We found increased hsa-miR-27a-3p in GBM tissues as well as GBM-EVs, which induced M2 polarization, thus promoting proliferative, migrative and invasive potentials of GBM cells. hsa-miR-27a-3p targeted EZH1 and promoted KDM3A expression to elevate the CTGF expression. GBM-EV-delivered hsa-miR-27a-3p promoted the KDM3A-upregulated CTGF by downregulating EZH1, thereby promoting M2 macrophage polarization and development of GBM in vivo. We demonstrated that EV-derived hsa-miR-27a-3p may promote M2 macrophage polarization to induce GBM.


Effects of the ABCDE bundle on the prevention of post-intensive care syndrome: A retrospective study.
To identify the effects of each domain of the early and modified ABCDE bundle on post-intensive care syndrome (PICS). This is a retrospective study. We analysed the data from electronic medical records of 91 intensive care patients who received therapeutic interventions in stages, based on the early ABCDE bundle (admitted to the intensive care unit [ICU] from June - August 2013) and 94 patients who received interventions using a modified ABCDE bundle developed through continuous quality improvement activities (admitted to the ICU from June to August 2014). In the ABC domain, the percentage of patients showing sedation levels of alertness and calmness increased significantly from 58.2% using the early ABCDE bundle to 72.4% using the modified ABCDE bundle. Coma prevalence decreased significantly from 45.1% using the early ABCDE bundle to 28.7% using the modified ABCDE bundle. In the E domain, the percentage of patients receiving early mobility interventions increased significantly from 11% using the early ABCDE bundle to 54.3% using the modified ABCDE bundle. The ABCDE bundle in the ICU helped prevent PICS by reducing deep sedation and immobilization among intensive care patients. To effectively use the ABCDE bundle, it is necessary for institutions to develop suitable protocols for each constituent element and to test their effectiveness. The ABCDE bundle was a suitable tool to support evidence-based practice in intensive care patients, including oversedation and immobilization, which is related to the prevention of PICS. Individual institutions will need to actively use the ABCDE bundle in the ICU, by developing protocols and testing their effectiveness.


Plight of CORMs: The unreliability of four commercially available CO-releasing molecules, CORM-2, CORM-3, CORM-A1, and CORM-401, in studying CO biology.
Carbon monoxide (CO) is an endogenously produced gaseous signaling molecule with demonstrated pharmacological effects. In studying CO biology, three delivery forms have been used: CO gas, CO in solution, and CO donors of various types. Among the CO donors, four carbonyl complexes with either a transition metal ion or borane (BH3) (termed CO-releasing molecules or CORMs) have played the most prominent roles appearing in over 650 publications. These are CORM-2, CORM-3, CORM-A1, and CORM-401. Intriguingly, there have been unique biology findings that were only observed with these CORMs, but not CO gas; yet these properties were often attributed to CO, raising puzzling questions as to why CO source would make such a fundamental difference in terms of CO biology. Recent years have seen a large number of reports of chemical reactivity (e.g., catalase-like activity, reaction with thiol, and reduction of NAD(P)+) and demonstrated CO-independent biological activity for these four CORMs. Further, CORM-A1 releases CO in an idiosyncratic fashion; CO release from CORM-401 is strongly influenced or even dependent on reaction with an oxidant and/or a nucleophile; CORM-2 mostly releases CO2, not CO, after a water-gas shift reaction except in the presence of a strong nucleophile; and CORM-3 does not release CO except in the presence of a strong nucleophile. All these beg the question as to what constitutes an appropriate CO donor for studying CO biology. This review critically summarizes literature findings related to these aspects, with the aim of helping result interpretation when using these CORMs and development of essential criteria for an appropriate donor for studying CO biology.


The human kallikrein protein 5 (hK5) is enzymatically active, glycosylated and forms complexes with two protease inhibitors in ovarian cancer fluids.
The kallikrein family is a group of 15 serine protease genes clustered on chromosome 19q13.4. Binding of kallikreins to protease inhibitors is an important mechanism for regulating their enzymatic activity and may have potential clinical applications. Human kallikrein gene 5 (KLK5) is a member of this family and encodes for a secreted serine protease (hK5). This kallikrein was shown to be differentially expressed at the mRNA and protein levels in diverse malignancies. Our objective was to study the enzymatic activity and the interaction of recombinant hK5 protein with protease inhibitors. Recombinant hK5 protein was produced in yeast and mammalian expression systems and purified by chromatography. HPLC fractionation, followed by ELISA-type assays, immunoblotting and radiolabeling experiments were performed to detect the possible interactions between hK5 and proteinase inhibitors in serum. Enzymatic deglycosylation was performed to examine the glycosylation pattern of the protein. The enzymatic activity of hK5 was tested using trypsin and chymotrypsin-specific synthetic fluorogenic substrates. In serum and ascites fluid, in addition to the free ( approximately 40 kDa) form, hK5 forms complexes with alpha(1)-antitrypsin and alpha(2)-macroglobulin. These complexes were detected by hybrid ELISA-type assays using hK5-specific coating antibodies and inhibitor detection antibodies. The ability of hK5 to bind to these inhibitors was further verified in vitro. Spiking of serum samples with 125I-labeled hK5 results in the distribution of the protein in two higher molecular mass (bound) forms, in addition to the unbound form. The hK5 mature enzyme is active and shows trypsin, but not chymotrypsin-like, activity. The pro-form of hK5 is not active. Recombinant hK5 shows a higher than predicted molecular mass due to glycosylation. hK5 is partially complexed with alpha(1)-antitrypsin and alpha(2)-macroglobulin in serum and ascites fluid of ovarian cancer patients. The recombinant protein is glycosylated and its mature form shows trypsin-like activity.


Hippocampal activation during processing of previously seen visual stimulus pairs.
Activity in the hippocampus is modulated by novelty detection, and by the processing of conjunctions between two stimuli. We investigated whether the hippocampus is activated by discrimination of stimulus-stimulus relationships in novel versus familiar pairs of visual stimuli in 15 healthy subjects using functional magnetic resonance imaging. Subjects were asked to recognize the previously rewarded stimulus in each case. We found significantly greater activation of the right hippocampus when discriminating previously seen compared with novel pairs of visual stimuli. This activation was evident in individual subjects and was not related to stimulus novelty, reward contingency, or task instruction. Right hippocampal activation during discrimination of previously seen pairs of objects was correlated with activity in the anteromedial thalamus, cingulate cortex, and contralateral hippocampus.


Nitric therapy in preterm infants: rationalised approach based on functional neonatal echocardiography.
Use of inhaled nitric oxide (iNO) in preterm infants is not supported by current evidence. In 2013, in Australia and New Zealand, 14% infants' ≤25 weeks of gestations were administered iNO. Within the cohort administered iNO, we aimed to identify subgroups where it may be more efficacious and compared characteristics before and after the set-up of the functional echocardiography (fEcho) programme. A retrospective audit for the period 2000-2013 involving preterm infants administered iNO in the first four weeks of life was performed. Comparisons were made between the two time epochs: up to 2007 and post-2007. Eighty-five infants fulfilled the inclusion criteria; 62 (73%) were ≤28 weeks of gestation; 51 (60%) survived. Amongst survivors, gestation and birthweight were higher and oxygenation index (OI) was lower. Fourteen (16.5%) infants weighed small for gestation age; survival was lower in this subgroup (6/14, 43%, p = 0.0005). The fEcho programme increased prenitric assessments for a definitive diagnosis and monitoring; iNO was started earlier, at a lower OI with a trend towards reduced usage (hours). Characteristics of subgroups (within the cohort of infants ≤34 weeks of gestation) more likely to benefit from iNO therapy were identified. Use of fEcho could rationalise usage.


The dying patient.
The dying patient experiences extraordinary emotional stress, as do the family and physician. Their distress is compounded by the fact that members of our society are poorly prepared to deal with death. The defenses that individuals mobilize to deal with their death-anxiety are often maladaptive. The psychiatrist, through the use of understanding of psycho-dynamics, can help herself or himself and then others in responding adaptively to the dying patient. Most medical schools all but ignore death education, despite the clear relevance of such teaching to their students. Psychiatric educators have a role to play in developing and implementing the psychological study of terminal illness in medical schools.


High Risk or Risky Highs: Understanding the Links Between Alcohol and Cannabis Use on the Transition From Suicidal Ideation to Attempts in Australian Men.
Alcohol and cannabis use are consistently associated with greater risk of suicide, particularly among men and in higher-income countries (e.g., Australia). Adult data (n = 7,464) from waves 1 and 2 of Ten to Men: The Australian Longitudinal Study on Male Health were used to explore whether alcohol and/or cannabis use increased the longitudinal risk of a suicide attempt among suicidal ideators. Cannabis use was associated with increased risk of transitioning from suicidal ideation to making a suicide attempt; no association was found for alcohol. Broadly, these findings indicate that greater cannabis but not alcohol use may increase risk of transitioning to making a suicide attempt among those who are thinking about suicide.


The keyhole concept in neurosurgery.
Improvements in preoperative diagnostic imaging as well as in microsurgical techniques significantly advanced the development of transcranial neurosurgery, allowing the treatment of complicated diseases through smaller and more specific approaches. In this article, authors overviewed their experience in transcranial endoscope-assisted microsurgery, using limited-sized keyhole craniotomies. Over a 10-year period, authors treated more than 3000 patients according to the transcranial endoscope-assisted microsurgery concept, advanced by the pioneer of minimally invasive neurosurgery, Axel Perneczky. In all cases, meticulous preoperative planning was done for determining the site, size, and optimal placement of the craniotomy as well as the trajectory toward the surgical target. Most importantly, the surgical approach was performed either completely or at least under permanent presence of the responsible senior surgeon from the moment of patient positioning until wound closure. The minimally invasive keyhole approaches allowed safe intraoperative control and adequate dealing with intracranial lesions. Essential preconditions for keyhole surgery were 1) careful selection of cases, 2) accurate preoperative planning, 3) placement of the craniotomy tailored to the individual case, and 4) intraoperative use of transcranial endoscope-assisted microsurgery techniques. Advantages of intraoperative endoscopic visualization were increased light intensity, extended viewing angle, and clear depiction of details even in hidden parts of the surgical field.


Interplay between epithelial and mesenchymal cells unveils essential proinflammatory and pro-resolutive mediators modulated by photobiomodulation therapy at 660 nm.
Photobiomodulation therapy (PBMT) has been widely used to promote tissue repair. However, PBMT's critical roles in the epithelial and mesenchymal tissues interactions are still barely known. Herein, we investigated light parameters on challenged keratinocytes (KC)-i.e., cultivated under oxidative stress-solely or associated with fibroblasts (FB) in a co-culture system. Cells were treated with PBMT at the wavelength of 660 nm, at 20 mW and 0.71 W/cm2 . Three different energy densities were primarily evaluated on KC: 1 (1.4 s), 5 (7 s), and 50 J/cm2 (70 s). Next, KC and FB were co-cultured and assessed at 5 J/cm2 . This energy density was also tested in ex vivo murine skin samples. Our main data suggest that PBMT can increase cellular proliferation at low doses and cell migration in a biphasic mode (1 and 50 J/cm2 ), both further confirmed by the epidermal growth factor receptor ligand-amphiregulin-upregulation. IL-1RA mRNA-the IL-1β (interleukin-1β) receptor antagonist recognized to fasten wound repair-was upregulated in the co-culture system. Upon PBMT, the ex vivo findings showed a progressive increase in the epidermal thickness, although presenting qualitatively less differentiated epithelium than the control group. In conclusion, PBMT effects are dependent on the cellular interactions with the surrounding microenvironment. Ultimately, PBMT is anti-inflammatory and contributes to the expression of critical mediators of wound repair.


Prophylaxis and management of postoperative nausea and vomiting in enhanced recovery protocols: Expert Opinion statement from the American Society for Enhanced Recovery (ASER).
International experience and evidence-based practices have shown that reduction in variability through use of protocolized perioperative care improves surgical outcomes and reduces costs to patients and healthcare systems. In this series of Expert Opinions, we provide consensus recommendations for the various components of perioperative care to aid with the development of enhanced recovery after surgery protocols.


Successful Treatment of Prolonged, Severe Coronavirus Disease 2019 Lower Respiratory Tract Disease in a B cell Acute Lymphoblastic Leukemia Patient With an Extended Course of Remdesivir and Nirmatrelvir/Ritonavir.
A patient with B-cell acute lymphoblastic leukemia (ALL) and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) had persistent, progressive pneumonia with viremia after 5 months of infection despite monoclonal antibodies, intravenous (IV) remdesivir and prolonged oral steroids. Twenty days of nirmatrelvir/ritonavir and 10 days of IV remdesivir led to full recovery.


Identification of candidate genes in rice for resistance to sheath blight disease by whole genome sequencing.
Recent advances in whole genome sequencing (WGS) have allowed identification of genes for disease susceptibility in humans. The objective of our research was to exploit whole genome sequences of 13 rice (Oryza sativa L.) inbred lines to identify non-synonymous SNPs (nsSNPs) and candidate genes for resistance to sheath blight, a disease of worldwide significance. WGS by the Illumina GA IIx platform produced an average 5× coverage with ~700 K variants detected per line when compared to the Nipponbare reference genome. Two filtering strategies were developed to identify nsSNPs between two groups of known resistant and susceptible lines. A total of 333 nsSNPs detected in the resistant lines were absent in the susceptible group. Selected variants associated with resistance were found in 11 of 12 chromosomes. More than 200 genes with selected nsSNPs were assigned to 42 categories based on gene family/gene ontology. Several candidate genes belonged to families reported in previous studies, and three new regions with novel candidates were also identified. A subset of 24 nsSNPs detected in 23 genes was selected for further study. Individual alleles of the 24 nsSNPs were evaluated by PCR whose presence or absence corresponded to known resistant or susceptible phenotypes of nine additional lines. Sanger sequencing confirmed presence of 12 selected nsSNPs in two lines. "Resistant" nsSNP alleles were detected in two accessions of O. nivara that suggests sources for resistance occur in additional Oryza sp. Results from this study provide a foundation for future basic research and marker-assisted breeding of rice for sheath blight resistance.


A murine model of HIV encephalitis: xenotransplantation of HIV-infected human neuroglia into SCID mouse brain.
The degree of neuronal damage in HIV encephalitis (HIVE) and the mechanisms leading to it are not known. Post-mortem human studies provide limited information concerning pathogenesis, and there are few animal models of HIVE. We have developed a murine model of HIVE based on HIV-infected human brain tissues grafted within the host CNS milieu. HIV-infected blood-derived macrophages were cocultured with and incorporated within second trimester human fetal brain neuroglia. Mixed neuronal-glial aggregates were injected into the striatum of SCID mice where they survived for more than 6 months. Cellular proliferation and differentiation were determined by immunohistochemical staining for proliferating cell nuclear antigen and cellular markers. Synapse formation was seen by immunocytochemistry for synapsin and by electron microscopy. Virus was detected by immunohistochemical staining for HIV gp41. Based on the long-term survival of human neuroglial xenografts containing HIV infected macrophages, we believe that this model will support the study of chronic HIV-associated neurodegeneration and the testing of various CNS targeted therapeutic interventions.


Subunit-specific phosphorylation of pyruvate kinase in medullary thyroid carcinomas of the rat.
Pyruvate kinase from anaplastic medullary thyroid carcinomas contains predominantly K-type subunits, whereas pyruvate kinase from differentiated medullary thyroid carcinomas consist of M- and K-type subunits in about equal proportion. In order to analyse the incorporation of phosphate in the respective isozymes after endogenous phosphorylation of cytosolic extracts with [32P]ATP, homotetrameric isozymes as well as heterotetrameric hybrids of differentiated tumors were resolved by affinity chromatography on Blue-Sepharose CL-6B and, if necessary, further purified by immunoprecipitation. SDS-polyacrylamide gel electrophoresis of purified isozymes and subsequent autoradiography showed the incorporation of phosphate in the K4-type isozymes, but not in the other isozymes. The phosphorylation appeared to be cAMP-independent and occurred on a serine residue.


Cortisol Concentrations in Well-Regulated Dogs with Hyperadrenocorticism Treated with Trilostane.
There are no clear treatment guidelines for dogs with clinically well-regulated hyperadrenocorticism in which serum cortisol concentrations before and after an ACTH stimulation test performed 3-6 hours after trilostane administration are < 2.0 μg/dL. To determine if serum cortisol concentrations measured before (Pre1) and after (Post1) ACTH stimulation at 3-6 hours after trilostane administration are significantly lower than cortisol concentrations measured before (Pre2) and after (Post2) ACTH stimulation 9-12 hours after trilostane administration, in a specific population of dogs with clinically well-regulated hyperadrenocorticism and Pre1 and Post1 <2 μg/dL. Thirteen client-owned dogs with clinically well-regulated hyperadrenocorticism and Pre1 and Post1 serum cortisol concentrations <2.0 μg/dL 3-6 hours after trilostane administration. Prospective study. Dogs had a second ACTH stimulation test performed 9-12 hours after trilostane administration, on the same day of the first ACTH stimulation test. Cortisol concentrations before and after ACTH stimulation were compared using a paired t-test. Cortisol concentrations before (1.4 ± 0.3 μg/dL) and after the first stimulation (1.5 ± 0.3 μg/dL, mean ± SD) were significantly lower than cortisol concentration before the second stimulation (3.3 ± 1.6 μg/dL, P = .0012 each). Cortisol concentration before the first stimulation was also significantly lower than cortisol concentration after the second stimulation (5.3 ± 2.4 μg/dL, P = .0001). In dogs with clinically well-regulated, trilostane-treated, hyperadrenocorticism, and cortisol concentrations <2 μg/dL before and after the first stimulation, a second ACTH stimulation test performed 9-12 hours after treatment can result in higher cortisol concentrations that could support continued trilostane treatment.


Conjunctival incisions for trabeculectomy and their relationship to the type of bleb formation--a preliminary study.
The bleb which develops following trabeculectomy may be diffuse or cystic in nature. Cystic blebs can be complicated by infection, leakage of aqueous, astigmatism, and problems related to contact lens wear, whereas diffuse blebs are not associated with such problems. We report a preliminary prospective photographic evaluation of bleb formation following each of three types of conjunctival incision: (a) fornix based (b) straight limbus based and (c) curved limbus based. The vascular pattern and morphology of the drainage bleb formed was dependent upon the type of conjunctival incision made. The fornix based flap was found most likely to give rise to a diffuse bleb with a normal vascular pattern.


[Characteristics of the evoked electrical reactions of the nucleus lateralis posterior in the rabbit thalamus and their dependence on cortical and reticular formation function].
The electrical responses of n. lateralis posterior of the thalamus (nLP) to light stimuli of different intensity were recorded in alert rabbits, and their interrelationships with the reactions of retina, superior colliculi and visual cortex were analysed. The retina rather than superior colliculi appeared to play a major role in afferent supply of nLP in rabbit. The organized mechanism of facilitating reticular control of the nLP function was shown, with the adrenoreactive mechanism playing the most important role in the reticular influence. The inhibitory influence of visual and sensomotor cortical areas upon neuronal mechanism of nLP forming the responses to photostimuli, was revealed. The effects of visual cortex were the most expressive and stable.


The impact of intraoperative residual mild regurgitation after repair of degenerative mitral regurgitation.
During degenerative mitral repair, surgeons must decide if further repair is warranted for residual mild mitral regurgitation. We examined the incidence of mild mitral regurgitation, late echocardiographic and clinical outcomes, and influence of surgical experience in decision making. From April 2004 to June 2018, 1155 of 1195 patients with pure degenerative disease underwent repair (97% repair rate). Propensity score matching was performed between patients with trace/no mitral regurgitation and patients with mild residual mitral regurgitation. Late echocardiographic outcome and freedom from reoperation were compared using competing-risks models. A comparison of outcomes of the referent surgeon (89.8% of repairs) with all other surgeons was performed. Mild mitral regurgitation was present in 73 patients (6%). Propensity score-matched analyses compared 69 patients with mild mitral regurgitation with 198 patients without mitral regurgitation. Late moderate or greater mitral regurgitation was higher in those with mild mitral regurgitation than in those with no mitral regurgitation (17% vs 7%, P = .033), as was late moderate-severe or greater mitral regurgitation (6% vs 1%, P = .016). Ten-year freedom from reoperation was low in both groups (99.5% no vs 96.9% mild; P = .10). The referent surgeon had fewer patients with mild residual mitral regurgitation (6% vs 11%, P = .027) and less progression of mitral regurgitation compared with other surgeons (late moderate or greater mitral regurgitation 6% vs 15%, P = .002). Residual mild mitral regurgitation was uncommon, and late progression to moderate or greater mitral regurgitation was rare and never led to late mitral reoperation. Experienced surgeons may be better able to determine repairs likely to remain stable, and most mild residual mitral regurgitation does not require re-repair.


Novel modes of oestrogen receptor agonism and antagonism by hydroxylated and chlorinated biphenyls, revealed by conformation-specific peptide recognition patterns.
Because of the concern about environmental chemicals with oestrogenic and anti-oestrogenic effects, there is a need to construct biosensors for classifying such chemicals according to their effect on oestrogen receptor conformation. The conformation of the ligand-binding domains (LBD) of oestrogen receptor-alpha and -beta determine their transcription regulation activity. Some ligands, i.e., the natural oestrogen oestradiol, induce an active conformation allowing interaction with co-activators. In contrast, antagonists like ICI 182, 780, because of their bulky side chains, do not allow an alpha-helix 12 positioning compatible with co-activator binding. Another type of oestrogen receptor-ligand interactions, termed "passive antagonism", was first defined by X-ray crystal structure analysis of receptors in complex with the side chain-less 5,11-cis-diethyl-5,6,11,12-tetrahydrochrysene-2,8-diol (THC). We have now used the ability of peptides selected from phage-displayed peptide libraries to bind conformation specifically to oestrogen receptor-alpha and -beta LBDs to analyse conformations induced by THC and a group of chlorinated biphenyls and their aryl-hydroxylated metabolites, suspected of being environmental chemical disruptors. In oestrogen receptor-beta, THC defined a "passive antagonist" peptide recognition pattern, which was also induced by several antagonistic hydroxylated biphenyls, while a clearly different peptide recognition pattern was induced by their chlorinated agonistic counterparts. In oestrogen receptor-alpha, THC induced a conformation similar to that induced by oestriol and other oestrogen receptor-alpha agonists, which, as evaluated by site-directed mutagenesis, have a functionally important interaction with oestrogen receptor-alpha residue His524. We conclude that the peptide recognition pattern can be used to classify suspected environmental endocrine disruptors according the oestrogen receptor-alpha and -beta conformations they induce.


Flaviviridae viruses use a common molecular mechanism to escape nucleoside analogue inhibitors.
The RNA-dependent RNA polymerases of Flaviviridae viruses are crucial for replication. The Flaviviridae polymerase is organized into structural motifs (A-G), with motifs F, A, C and E containing interrogating, priming and catalytic substrate-interacting sites. Modified nucleoside analogues act as antiviral drugs by targeting Flaviviridae polymerases and integrating into the synthesized product causing premature termination. A threonine mutation of a conserved serine residue in motif B of Flaviviridae polymerases renders resistance to 2'-C-methylated nucleoside analogues. The mechanism how this single mutation causes Flaviviridae viruses to escape nucleoside analogues is not yet known. Given the pivotal position of the serine residue in motif B that supports motif F, we hypothesized the threonine mutation causes alterations in nucleoside exploration within the entry tunnel. Implementing a stochastic molecular software showed the all-atom 2'-C-methylated analogue reaction within the active sites of wild type and serine-threonine mutant polymerases from Hepacivirus and Flavivirus. Compared with the wild type, the serine-threonine mutant polymerases caused a significant decrease of analogue contacts with conserved interrogating residues in motif F and a displacement of metal ion cofactors. The simulations significantly showed that during the analogue exploration of the active site the hydrophobic methyl group in the serine-threonine mutant repels water-mediated hydrogen bonds with the 2'-C-methylated analogue, causing a concentration of water-mediated bonds at the substrate-interacting sites. Collectively, the data are an insight into a molecular escape mechanism by Flaviviridae viruses from 2'-C-methylated nucleoside analogue inhibitors.


Possibility of a dark matter interpretation for the excess in isotropic radio emission reported by ARCADE.
The ARCADE 2 Collaboration has recently measured an isotropic radio emission which is significantly brighter than the expected contributions from known extra-galactic sources. The simplest explanation of such excess involves a "new" population of unresolved sources which become the most numerous at very low (observationally unreached) brightness. We investigate this scenario in terms of synchrotron radiation induced by weakly interacting massive particle (WIMP) annihilations or decays in extra-galactic halos. Intriguingly, for light-mass WIMPs with a thermal annihilation cross section, the level of expected radio emission matches the ARCADE observations.


Airway constrictor effects of leukotriene D4 in dogs with hyperreactive airways.
Leukotriene D4 (5 micrograms/ml) aerosol constricts airways of dogs with nonspecific airway hyperreactivity but not of mongrel dogs which lack nonspecific airway hyperreactivity. RL increased 200 +/- 25% and Cdyn decreased to 77 +/- 5% of the prechallenge value. LTD4 (10 micrograms/ml) produced no further increase. Atropine (0.2 mg/kg) prevented the increase in RL and decrease in Cdyn, suggesting that part of the effect of LTD4 on airways is neurally mediated.


Cariogenicity of soluble starch in oral in vitro biofilm and experimental rat caries studies: a comparison.
Common belief suggests that starch is less cariogenic than sugar; however, the related literature is quite controversial. We aimed to compare cariogenic and microbiological effects of soluble starch in both a standard animal model and an oral biofilm system, and to assess the possible substitution of the animal model. Six-species biofilms were grown anaerobically on enamel discs in saliva and medium with glucose/sucrose, starch (average molecular weight of 5000, average polymerization grade of 31), or mixtures thereof. After 64.5 h of biofilm formation, the microbiota were quantitated by cultivation and demineralization was measured by quantitative light-induced fluorescence. To assess caries incidence in rats, the same microbiota as in the biofilm experiments were applied. The animals were fed diets containing either glucose, glucose/sucrose, glucose/sucrose/starch or starch alone. Results with both models show that demineralization was significantly smaller with starch than sucrose. The data demonstrate that soluble starch is substantially less cariogenic than glucose/sucrose. By leading to the same scientific evidence as its in vivo counterpart, the described in vitro biofilm system provides an interesting and valuable tool in the quest to reduce experimentation with animals.


Healthcare utilization and mental health outcomes among nonfatal shooting assault victims.
Victims of nonfatal shooting (NFS) assaults suffer from emotional and physical trauma; however, little is understood about clinical care utilization patterns among victims. This study examines the healthcare utilization and mental health outcomes before and after an index NFS victimization. A longitudinal dataset of police and clinical data were linked at the individual level to define a cohort of NFS victims with one or more clinical encounter in the 24-months preceding an index NFS injury (N = 2,681) in Indianapolis, Indiana between 2005 and 2018. Mental health was defined using ICD diagnosis codes from any emergency department, inpatient, or outpatient encounter and clinical care utilization was the number of unique encounters within the 24-months preceding and following an index NFS injury. Multivariable logistic regression was conducted to examine factors associated with a mental health diagnosis in the post injury period. Analyses were conducted in October 2021-March 2022. Overall clinical care utilization (Mean: pre = 277.7 (SD 235.3) vs. post = 333.9 (SD 255.1), p < 0.001) and mental health prevalence (14.4% pre vs. 18.8% post, p < 0.001) increased in the 24-months following an index NFS compared to the prior 24-months. Preinjury mental health utilization increased the odds of receiving a mental health diagnosis in the 24-months following an index NFS injury - particularly for Black victims (Odds Ratio 1.69, 95% CI 1.01, 2.85). The findings indicate missed opportunities within the healthcare system to connect NFS victims with needed mental health services, as well as the importance of premorbid connection to mental health care.


Skull burns resulting in calvarial defects: cognitive and affective outcomes.
To determine the extent of cognitive and affective difficulties experienced in children with full thickness burns of the calvaria caused by flame and electrical sources. Medical records were examined retrospectively to identify patients with full thickness burns of the calvaria. Data were reviewed from the acute period and long term for these patients and their matched controls. Fourteen children were identified who sustained full thickness burns. The average age of injury was 7.1+/-7.3 years and average percent TBSA was 39.7+/-26.3. Of these 14 patients, 8 were under the age of 4 years at the time of injury and suffered flame injuries. Six children were over 11 years old at injury and suffered electrical burns. Imaging was performed on 10 patients acutely and revealed 5 with major brain injury. The youngest patients suffering flame injuries experienced significant difficulty with developmental functioning acutely and a trend towards long-term cognitive difficulty. Older children did not differ from their matched controls at any time. Children with burns to the calvarium can be at risk for affective and cognitive sequalae. Therefore, we recommend comprehensive neuropsychological evaluations at multiple time points to adequately assess functioning across time.


MicroRNA signature of small-cell lung cancer after treatment failure: impact on oncogenic targets by miR-30a-3p control.
Small-cell lung cancer (SCLC) is associated with a high mortality rate and limited treatment efficacy. We created a microRNA (miRNA) expression signature by RNA sequencing using specimens from patients with SCLC who had failed treatment. Forty-nine miRNAs were downregulated in SCLC tissues and were candidate tumor-suppressive miRNAs. In this signature, both guide and passenger strands were downregulated for five miRNAs (miR-30a, miR-34b, miR-34c, miR-223, and miR-4529). Recent studies have revealed that passenger strands of miRNAs are involved in the molecular pathogenesis of human cancer. Although miR-30a-5p (the guide strand) has been shown to be a tumor-suppressive miRNA in various types of cancers, miR-30a-3p (the passenger strand) function is not well characterized in SCLC cells. We investigated the functional significance of miR-30a-3p and oncogenic genes regulated by miR-30a-3p in SCLC cells. Ectopic expression assays showed that miR-30a-3p expression inhibited cell proliferation and induced cell cycle arrest and apoptosis in two SCLC cell lines. Furthermore, in silico database searches and gene expression assays identified 25 genes as putative targets of miR-30a-3p in SCLC cells. Luciferase reporter assays revealed that downstream neighbor of SON (DONSON) was directly regulated by miR-30a-3p in SCLC cells. Knockdown of DONSON induced cell cycle arrest in SCLC cells and DONSON overexpression were detected in SCLC clinical samples. Analyzing the regulatory networks of tumor-suppressive miRNAs may lead to the identification of therapeutic targets in SCLC.


Poor outcome in bacterial peritonitis is associated with dysregulated microRNAs and an increased inflammatory response.
Peritonitis is a common cause of surgical sepsis. The failure of the host to mount an appropriate immune response contributes to persistence of the infection. We investigated the role microRNAs may play in this failed immune response. Klebsiella pneumoniae was injected intraperitoneally in mice. Weight loss was used to predict clinical outcome. Peritoneal exudate cells (PECs) and supernatant were collected. RNA from PECs was run on screening microRNA array cards to determine gene expression, and validated by single assay analysis. Cytokine levels in supernatant were assayed by enzyme-linked immunosorbent assay. Despite similar bacterial levels, PEC counts were higher in the predicted death group. The predicted deaths had higher levels of proinflammatory tumor necrosis factor-α/IL-6 and significantly lower levels of interleukin-10. MiR-221 was up-regulated in both the predicted death and predicted survivor groups. Five miRNAs were up-regulated in the predicted survivor group compared with normal controls. Higher PEC counts and proinflammatory cytokines in the predicted death group indicates an exaggerated inflammatory response, with lower IL-10 levels despite similar bacterial counts. There were two dysregulated miRNAs with transcriptional targets that may explain our results. A more balanced immune response with an appropriate counter inflammatory response may be important for improving survival.


Weighted Kaplan-Meier statistics: a class of distance tests for censored survival data.
A class of statistics based on the integrated weighted difference in Kaplan-Meier estimators is introduced for the two-sample censored data problem. With positive weight functions these statistics are intuitive for and sensitive against the alternative of stochastic ordering. The standard weighted log-rank statistics are not always sensitive against this alternative, particularly if the hazard functions cross. Qualitative comparisons are made between the weighted log-rank statistics and these weighted Kaplan-Meier (WKM) statistics. A statement of null asymptotic distribution theory is given and the choice of weight function is discussed in some detail. Results from small-sample simulation studies indicate that these statistics compare favorably with the log-rank procedure even under the proportional hazards alternative, and may perform better than it under the crossing hazards alternative.


Detection of Dopamine Receptors Using Nanoscale Dendrimer for Potential Application in Targeted Delivery and Whole-Body Imaging: Synthesis and In Vivo Organ Distribution.
Dopamine is one of the most important neurotransmitters released by neurons in the central nervous system, and a variety of neurological illnesses and mental disorders are associated with impairments in the secretion and functionality of dopamine. Dopamine, depending on the type of receptors, can act as a stimulant or an inhibitor. In this study, dendrimer-conjugated dopamine was utilized as a chelating agent for Technetium-99m to investigate the organ distribution of this compound in vivo using the single-photon emission computed tomography (SPECT) technique. For this purpose, dendrimers were synthesized using polyethylene glycol diacid and citric acid precursors, and dopamine was conjugated to the dendrimer using EDC/NHS cross-linker. The results showed no sign of toxicity of the dopamine-functionalized dendrimers on HEK-293 cell lines. The optimization of labeling conditions was conducted using the experimental design method (i.e., conjugate value, pH, and the amount of reducing agent), and then labeling efficiency was evaluated by thin-layer chromatography (TLC). Finally, the study of organ distribution in normal mice using SPECT imaging and comparing it with gene expression in different organs revealed that dopamine D1 receptors exhibited the highest accumulation in the liver and that the drug retained its specificity.


Balancing Early Aggression Against Risk of Progression in Multiple Sclerosis.
Multiple sclerosis is a chronic demyelinating disease characterized by focal and diffuse inflammation of the central nervous system resulting in significant physical and cognitive disabilities. Disease-modifying therapies targeting the dysfunctional immune response are most effective in the first few years after disease onset, indicating that there is a limited time window for therapy to influence the disease course. No evidence of disease activity is emerging as a new standard for treatment response and may be associated with improved long-term disability outcomes. An aggressive management strategy, including earlier use of more potent immunomodulatory agents and close monitoring of the clinical and radiologic response to treatment, is recommended to minimize early brain volume loss and slow the progression of physical and cognitive impairments in patients with relapsing-remitting multiple sclerosis.


Abnormalities in glucose tolerance are common in children with fanconi anemia and associated with impaired insulin secretion.
To determine prevalence of abnormal glucose metabolism in Fanconi Anemia (FA). Thirty-nine children with FA underwent 2-hr oral glucose tolerance test (OGTT). Reference lean adolescents (REF) were older than FA patients (mean +/- SD: FA 8.6 +/- 3.9 years, REF 19.8 +/- 0.3 years, P < 0.001), but comparable in BMI Z-scores (FA 1.25 +/- 0.58, REF -0.02 +/- 0.24; P = 0.24). Patients had normal glucose tolerance (NGT) or abnormal glucose metabolism (AGM) by American Diabetes Association Criteria. Insulinogenic index estimated beta-cell function. Insulin resistance estimation used homeostatic model assessment (HOMA-IR). Insulin secretion estimation relative to insulin sensitivity used disposition index (DI). Among FA patients, 46% had AGM. Compared to REF, there were significant differences in glycemic responses (area under curve: FA-NGT 344 +/- 42, FA-AGM 596 +/- 35, REF 208 +/- 25 mM, P < 0.0001) and insulinogenic index (FA-NGT 105 +/- 29, FA-AGM 44 +/- 8, and REF 173 +/- 41 pM/mM, P < 0.05). Insulin sensitivity did not differ among NGT, AGM, and REF (HOMA-IR: FA-NGT 1.9 +/- 0.4, FA-AGM 2.2 +/- 0.5, REF 1.3 +/- 0.2, P = NS). However, DI was significantly lower in both FA groups than REF [NGT 63.6 +/- 16.5 vs. AGM 26.4 +/- 3.5 (P < 0.048); REF 132.6 +/- 24.5 (NGT and AGM vs. REF, both P < 0.0002)]. Abnormalities in glucose metabolism are frequent in young FA patients without prior diagnosis of diabetes, and are associated with marked defects in insulin secretion.


Epidemiology of bloodstream infections and time to detection of positive blood cultures: an evaluation of the automated BacT/Alert and BACTEC 9240 systems.
Data of 3,097 blood culture sets processed with the BacT/Alert system in 1997 were compared to those of 3,158 blood culture sets processed with BACTEC 9240 in 1999. Agents responsible for bloodstream infections (BSI) were detected in 15.9% and 20.0% of blood cultures in 1997 and 1999, respectively. The incidence of BSI was 9.3 (1997) vs. 11.3 (1999) per 1,000 admissions. In both years, S. aureus was the most frequent isolate, followed by E. coli. Overall, the mean detection time (MDT) obtained with the BACTEC 9240 was significantly shorter than that of the BacT/Alert. Significant MDT differences were found for all organisms, except for Enterobacteriaceae (12.7 vs. 10.6 h). With both systems, over 95% positive samples were detected within 3 days, indicating that a 4-day incubation protocol may disclose most BSI agents. Thus, the added speed of the BACTEC 9240 allowed a particularly fast clinical management of septic patients.


Aeromonads in acute diarrhoea and asymptomatic infections in Nigerian children.
Stool samples of 616 asymptomatic and 296 diarrhoeic school children were compared for the recovery rate of Aeromonas spp. on ampicillin (10 micrograms/ml) sheep blood agar. Culture filtrates of isolates were tested for haemolysin production with 1% freshly washed rabbit erythrocytes. Stools of 9 (3.0%) diarrhoeic children yielded five strains of A. hydrophila and four of A. veronii (two each of biotypes sobria and veronii), compared to 12 (1.9%) (p > 0.01) asymptomatic children who harbored seven A. hydrophila and five A. caviae strains. Isolates from-diarrhoeic stools were exclusively from children < or = 5 years, while all infected asymptomatic children were > or = 6 years. Culture filtrates of all nine diarrhoeic strains were uniformly enterotoxigenic (intestinal weight ratio > 0.083) and produced haemolysin titres > 128. These phenotypes where variable in carriage strains of A. hydrophila but were not detected in A. caviae. The recovery of A. hydrophila, and A. veronii biotypes from diarrhoeic stools of children < or = 5 years may suggest their involvement in diarrhoea causation in the absence of other diarrhoeagenic agents.


Adherence to time and temperature standards and food acceptability.
The objective of this study was to examine time and temperature of food under actual operating conditions during product flow, from procurement through service, and to seek relationships to the amounts of food consumed during the noon meal by children in Head Start early childhood developmental centers. A process flow model was developed for a conventional foodservice system such as that used in Head Start. The preparation and service of the lunch menus served to 101 3- to 5-year-old children in three centers were studied during a 3-week period for adherence to time and temperature standards. Weight of food consumed was measured and evaluated by a preference ratings panel for acceptability. Food was well accepted, was consumed by the children, and received very good adult preference panel ratings. The largest variations from the time standard were in processing-heating hot vegetables, while the greatest temperature variations were across all process steps for entrées. Deviations from recommended time and temperature standards for handling of milk were significantly correlated with lower consumption of milk.


Plant signaling: The sugar-coated story of root growth.
A new study draws attention to photosynthetically produced sucrose as a major shoot-derived and auxin-dependent regulator of root growth and development in plants.


[Photoreactivation of cells and phages injuried by ultraviolet radiation in the ecological long-wave band].
Photoreactivation (PR) was measured after inactivation by far (254 nm), middle (300-315 nm) and near (315-400 nm) UV radiation of Paramecium caudatum and 8 strains of Escherichia coli differing in PR and dark repair capability. PR volume was high and practically the same after irradiation by far and middle UV, but PR was not observed in near UV-inactivated cells of all the strains. It is proposed that pyrimidine dimers are not significant in near UV lethal lesions in cells, as near UV-irradiated phages (T7 and lambdacI 857) are not photoreactivated in undamaged host bacterial cells.


Caloric content of a meal affects duration but not contractile pattern of duodenal motility in man.
The variability of the fasted duodenal contractile pattern and the patterns of contraction during the fed phase was examined in normal volunteers. Prolonged recordings from the duodenum and proximal jejunum were achieved using a series of transducers mounted on a 2.3-mm catheter. A total of 58 interMMC intervals and the response to 18 meals was examined. There was marked inter- and intrasubject variability in the fasted state, even within one study. The phase II pattern was examined in detail and propagated single peaks, propagated clusters, and repeated propagated clusters are described. Single peaks could be propagated as rapidly as 16 cm/sec. Single peaks were propagated more rapidly than propagated multiple peaks. During phase III, duodenal contractions occurred at 11.3 +/- 0.09/min and jejunal contractions at 10.73 +/- 0.15/min. The rate of progression of the onset of phase III was 0.145 +/- 0.015 cm/sec. The effect of the caloric content of the meal was examined by determining the effect of 150-kcal, 300-kcal, and 600-kcal meals on the fed pattern. Increasing caloric content increased the duration of the fed pattern but had no effect on the total or normalized motility index or on the change in motility index over time during the fed pattern. The types of contractions seen during the fed pattern are described. Propagated clusters over at least 16 cm are common during the fed phase in normals, with 10% of all contractions seen during the fed phase being propagated over 28 cm. No difference in the patterns of contractions or their propagation was seen with the different caloric contents of the meals.(ABSTRACT TRUNCATED AT 250 WORDS)


A new isotope framework to decipher leaf-root nitrogen allocation and assimilation among plants in a tropical invaded ecosystem.
Exotic plant invasion is an urgent issue occurring in the biosphere, which can be stimulated by environmental nitrogen (N) loading. However, the allocation and assimilation of soil N sources between leaves and roots remain unclear for plants in invaded ecosystems, which hampers the understanding of mechanisms behind the expansion of invasive plants and the co-existence of native plants. This work established a new framework to use N concentrations and isotopes of soils, roots, and leaves to quantitatively decipher intra-plant N allocation and assimilation among plant species under no invasion and under the invasion of Chromolaena odorata and Ageratina adenophora in a tropical ecosystem of SW China. We found that the assimilation of N derived from both soil ammonium (NH4+) and nitrate (NO3-) were higher in leaves than in roots for invasive plants, leading to higher leaf N levels than native plants. Compared with the same species under no invasion, most native plants under invasion showed higher N concentrations and NH4+ assimilations in both leaves and roots, and increases in leaf N were higher than in root N for native plants under invasion. These results inform that preferential N allocation, dominated by NH4+-derived N, to leaves over roots as an important N-use strategy for plant invasion and co-existence in the studied tropical ecosystem.


Material density iodine images in dual-energy CT: Detection and characterization of hypervascular liver lesions compared to magnetic resonance imaging.
To determine the diagnostic potential of Material Density (MD) iodine images in dual-energy CT (DECT) for the detection and characterization of hypervascular liver lesions compared to monenergetic 65keV images, using MRI as the standard. The study complied with HIPAA guidelines and was approved by the institutional review board. Fifty-two patients (36 men, 16 women; age range, 29-87 years) with 236 hypervascular liver lesions (benign, n=31; malignant, n=205; mean diameter, 29.4mm; range: 6-90.6mm) were included. All of them underwent both contrast-enhanced single-source DECT and contrast-enhanced abdominal MRI within three months. Late arterial phase CT imaging was performed with dual energies of 140 and 80kVp. Protocol A showed monoenergetic 65keV images, and protocol B presented MD-iodine images. Three radiologists qualitatively evaluated randomized images, and lesion detection, characterization, and reader confidence were recorded. Liver-to-lesion ratio (LLR) and contrast-to-noise ratio (CNR) were assessed on protocol A, protocol B, and MRI. Paired t-tests were used to compare LLR, CNR, and the number of detected lesions. LLR was significantly increased in protocol B (2.8±2.33) compared to protocol A (0.77±0.55) and MRI (0.61±0.66). CNR was significantly higher in protocol B (0.08±0.04) compared to protocol A (0.01±0.01) and MRI (0.01±0.01). All three observers correctly identified more liver lesions using protocol B vs protocol A: 83.13% vs 63.64%, 84.57% vs 68.09%, and 79.37% vs 65.52%. There was no significant difference between the three observers in classification of a lesion as benign or malignant. However, higher diagnostic confidence was reported more frequently by the experienced radiologist when using protocol B vs protocol A (84.6% vs 75%). MD-iodine images in DECT help to increase the conspicuity and detection of hypervascular liver lesions.


The effects of obesity on longitudinal ventricular and atrial strain in a rural population of Spanish children and adolescents, evaluated by a new strain software.
Research on the relationship between body mass index (BMI) and strain values in children and adolescents is limited. Our aim was to analyse the relationship between BMI and strain values of both ventricles and left atrium in children and adolescents. Both ventricles and left atrial strain values were compared among different BMI categories in children and adolescents from a town in the South of Spain. Of the 198 subjects, aged 6-17 years, 53% were of normal weight, 26% were overweight and 21% had obesity. Lower absolute values of left ventricular global longitudinal strain (25.9 ± 2.0% vs. 26.9 ± 2.2%, p = 0.002) and right ventricular free wall longitudinal strain (29.5 ± 4.2% vs. 30.8 ± 4.5%, p = 0.04) were found in subjects with obesity and overweight versus subjects with normal weight. A lower right ventricular four-chamber longitudinal strain was also observed in males with obesity and overweight (24.8 ± 3.3% vs. 26.4 ± 3.6%, p = 0.03). Statistically significant negative correlations of BMI were found for all ventricular, but not atrial, strain values in univariate analysis. This association turned non-significant for right ventricular four-chamber longitudinal strain in multivariate analysis. Utilizing this new strain software, children and adolescents with high BMI were associated with significantly lower values for left and right ventricular free wall longitudinal strain, without impact in left atrial strain.


Reading ability and phonological awareness in Japanese children with dyslexia.
In alphabetic languages, the deficit of the phonological awareness is considered as the core deficit in developmental dyslexia. However, the role of phonological awareness in the acquisition of reading Japanese kana, the transparent, mora-based phonogram, has not been understood completely. We examine the abilities of Japanese dyslexic children on different types of Japanese phonological tasks, and discussed which tasks significantly account for each aspect of reading ability. Fifteen dyslexic children (9.53+/-1.52 years old), and 15 children with normal reading ability (9.17+/-0.90 years old) participated in this study. They performed three types of phonological awareness tasks. The mora counting task and the mora reversal task of words require phonological awareness at the mora level. The letter rhyming task, which resembles the task in English language, requires phonological awareness at the phoneme level. We evaluated the reading ability by the reading speed, the reading errors, and the number of pauses while reading sentences aloud. The task performances of the dyslexic group on all three phonological awareness tasks were significantly lower than those of the control group. Stepwise multiple regression analysis revealed that the mora counting task and the rhyming letter task most significantly explained the reading speed and number of reading pauses. The mora reversal task of words, together with the antegraded digit span, significantly explained the reading errors. Japanese dyslexics showed deficits of phonological awareness both at the mora and the phoneme levels. Phonological awareness must be crucial for acquiring the ability of decoding phonograms, including Japanese kana.


Cardia carcinoma considered as a distinct clinical entity.
From 1 January 1969 until 31 December 1985, gastric resections were performed in 937 patients with carcinoma of the stomach and 422 patients with carcinoma of the cardia. During this period, the general frequency of cardia carcinoma increased continuously from 6.6 per cent (1969-71) to 37.5 per cent (1984-85) and a corresponding decrease of carcinomas in other sites was observed. Early carcinomas (pT1) of the oesophageal margin (n = 10, 2.4 per cent) were diagnosed less often than early carcinomas in other sites (n = 177, 18.9 per cent). Whereas 51.4 per cent of all resected carcinomas of the stomach showed no lymph node metastases (pN0), only 14.1 per cent of cardia carcinomas belonged to this category. With respect to operative treatment, Laurén's classification which defines the required margin of clearance according to diffuse type (frequency in cardia carcinoma 43 per cent, in other stomach sites 51 per cent), is of great significance. We believe that extensive oesophageal resection is necessary for patients with such difficult tumours. The pattern of metastasis in cardia carcinoma is similar to that of cancer in other stomach sites, the preferred area is the regional lymph nodes on the lesser curve (83 per cent). Histological classification and metastasis pattern show that cardia carcinoma is a type of carcinoma of the stomach which must be treated according to the criteria of stomach cancer surgery.


Underlying issues in bioaccessibility and bioavailability: experimental methods.
This article presents experimental designs focusing on assessing of the bioavailability of metals in aquatic organisms, soil organisms, microorganisms, plants, birds, and mammals. Standardized test systems receive the greatest emphasis. With regard to microorganisms, animals, and plants, the study concentrates on toxicity as an indicator for bioavailability. In respective test procedures, results are usually calculated for total chemical concentrations; chemical analyses are not commonly in routine assessments. For soil organisms chiefly exposed by the water pathway, the bioavailable fraction of contaminants can be roughly determined by chemical analyses in aqueous soil extracts simulating soil pore water concentrations. Human toxicity can be determined using adequate in vitro test designs. In addition to experimental designs, results from the literature dealing with specific problems of bioavailability are presented.


Impact of interprofessional in situ simulations on acute pediatric burn management: Combining technical and non-technical burn team skills.
This study aimed to evaluate the impact of interprofessional in situ simulations on the technical and non-technical skills of pediatric burn teams in acute burn management. This quasi-experimental study consisted of a one-group pre- and post-test design conducted in a pediatric burn center in Turkey. The sample consisted of nine interprofessional burn team members. Data collection tools consisted of the following: descriptive data form, burn technical skills checklists, simulation evaluation form, and Anesthesiologists' non-technical skills in Denmark rating form. We found no statistically significant difference between the pre- and post-test scores for technical (p = 0.285) and non-technical skill (p = 0.180) scores. Burn team members evaluated the highest score in almost all criteria for in situ simulations. The interprofessional in situ simulations did not improve the burn teams' acute burn management; however, according to a self-report, burn team members were satisfied with the interprofessional in situ simulation experiences and achieved their own gains.


An increased risk of pulmonary hypertension in patients with combined pulmonary fibrosis and emphysema: a meta-analysis.
Pulmonary hypertension (PH) is a common complication of combined pulmonary fibrosis and emphysema (CPFE). Whether the incidence of PH is increased in CPFE compared with pure pulmonary fibrosis or emphysema remains unclear. This meta-analysis aimed to evaluate the risk of PH in patients with CPFE compared to those with IPF or COPD/emphysema. We searched the PubMed, Embase, Cochrane Library, and CNKI databases for relevant studies focusing on the incidence of PH in patients with CPFE and IPF or emphysema. Pooled odds ratios (ORs) and standard mean differences (SMD) with 95% confidence intervals (95% CIs) were used to evaluate the differences in the clinical characteristics presence and severity of PH between patients with CPFE, IPF, or emphysema. The survival impact of PH in patients with CPFE was assessed using hazard ratios (HRs). A total of 13 eligible studies were included in the meta-analysis, involving 560, 720, and 316 patients with CPFE, IPF, and emphysema, respectively. Patients with CPFE had an increased PH risk with a higher frequency of pulmonary hypertension and higher estimated systolic pulmonary artery pressure (esPAP), compared with those with IPF (OR: 2.66; 95% CI: 1.55-4.57; P < 0.01; SMD: 0.86; 95% CI: 0.52-1.19; P < 0.01) or emphysema (OR: 3.19; 95% CI: 1.42-7.14; P < 0.01; SMD: 0.73; 95% CI: 0.50-0.96; P < 0.01). In addition, the patients with CPFE combined with PH had a poor prognosis than patients with CPFE without PH (HR: 6.16; 95% CI: 2.53-15.03; P < 0.01). Our meta-analysis showed that patients with CPFE were associated with a significantly higher risk of PH compared with those with IPF or emphysema alone. The presence of PH was a poor predictor of mortality.


The links between microclimatic and particulate matter concentration in a multi-storey car parking: a case study iran.
Multi-storey cars increasing with population growth have excellent security and temporary parking for cars in big cities, which isn't suitable for parking in the streets. The goals of this study are (1) to determine PM concentrations in the ZGP and (2) to investigate the effect of temperature and humidity on PM concentration in ZGP. This study measured the levels of emitted PM1, PM2.5, and PM10 by GRIMM EDM 107 laser dust monitor in a busy multi-storey parking garage located in Qom. Moreover, the relationship between microclimatic parameters and the contaminants mentioned above was investigated. Samples were collected in two stages in different spatiotemporal conditions, namely, the summer and autumn of 2017. The results indicate that during the sampling period, the daily mean ± standard deviation of PM10, PM2.5, and PM1 were 120.9 ± 90.6, 28.5 ± 10.4, and 10.8 ± 3.8 µg/m3, respectively. A decrease in pollution level was observed during the measurement period. During rush hours, the levels of particulate matter increased. Also, a significant positive relationship between indoor humidity and particle level was observed, while there was a meaningful, inverse relationship between temperature and particle level. The high PM concentration in the parking garage indicates the necessity of proper management and planning.


Prevalence of papillary muscle hypertrophy in fabry disease.
Fabry disease (FD) is an X-linked genetic lysosomal disease, in which a deficit in the alpha-galactosidase A enzyme results in lysosomal build-up of globotriaosylceramide in several organs, causing cardiac, renal and cerebrovascular complications. The aim of this study was to assess the prevalence of papillary muscle hypertrophy (PMH) in patients with FD. A group of 63 patients with FD and a positive genetic diagnosis were studied and were divided into two groups: one included 24 patients with FD and LVH and another group included 39 patients with FD and without LVH. Papillary muscles were measured from the left parasternal short axis view, defining PMH as a diastolic thickness greater than 11 mm in any diameter. Patients with FD and LVH had a high prevalence of anterolateral PMH (66.6%), and such prevalence was lower for the posteromedial PMH (33.3%). However, patients who had not yet developed LVH had a high prevalence of anterolateral PMH (33.3%). Patients with FD in the pre-clinical stage (without LVH) have a high prevalence of PMH, especially involving the anterolateral papillary muscle. This finding could be an early marker for the development of LVH, allowing to suspect the disease during its early stages, and begin enzyme replacement therapy in the appropriate patients.


Brn-4 transcription factor expression targeted to the early developing mouse pancreas induces ectopic glucagon gene expression in insulin-producing beta cells.
The endocrine pancreas is comprised of beta and alpha cells producing the glucostatic hormones insulin and glucagon, respectively, and arises during development by the differentiation of stem/progenitor cells in the foregut programmed by the beta cell lineage-specific homeodomain protein Idx-1. Brain-4 (Brn-4) is expressed in the pancreatic anlaga of the mouse foregut at e10 in the alpha cells and transactivates glucagon gene expression. We expressed Brn-4 in pancreatic precursors or beta cell lineage in transgenic mice by placing it under either Idx-1 or insulin promoter (rat insulin II promoter) control, respectively. Idx-1 expression occurs at developmental day e8.5, and insulin expression occurs at e9.5, respectively. Misexpression of Brn-4 by the Idx-1 promoter results in ectopic expression of the proglucagon gene in insulin-expressing pancreatic beta cells, whereas misexpression by rat insulin II promoter did not. The early developmental expression of Brn-4 appears to be a dominant regulator of the glucagon expressing alpha cell lineage, even in the context of the beta cell lineage.


Allogeneic bone marrow transplantation versus intensification chemotherapy for acute myelogenous leukaemia in first remission: a prospective controlled trial.
In 1982 we began a prospective controlled trial to assess the effectiveness of allogeneic bone marrow transplantation and intensive post-remission chemotherapy for patients with acute myelogenous leukaemia in first complete remission. Fourteen patients, 3-45 years of age, who had an HLA-identical sibling donor, received bone marrow transplantation. Twenty-five patients who either lacked an HLA-identical sibling or were over 45 years of age received intensive consolidation chemotherapy including high-dose cytosine arabinoside with or without adriamycin. The actuarial rate of continued complete remission (CCR) at 3 years was significantly higher in the transplantation group than in the chemotherapy group: 70% (95% confidence interval 35-91%) compared with 10% (95% confidence interval 2-30%); P = 0.01. However, the actuarial rate of CCR was not significantly different between the transplantation group and patients under 45 years in the chemotherapy group: 70% (95% confidence interval 35-91%) compared with 17% (95% confidence interval 4-45%), 0.1 greater than P greater than 0.05. The actuarial probability of leukaemia relapse was significantly lower in the transplantation group than in the chemotherapy group: 10% (95% confidence interval 4-21%) compared with 88% (95% confidence interval 70-96%), 0.005 greater than P greater than 0.001. There was no significant difference between both groups if we compare only the patients who died of non-leukaemic causes: 22% (95% confidence interval 9-42%) versus 25% (95% confidence interval 7-59%), P = NS. In summary, this study shows that allogeneic bone marrow transplantation is a better anti-leukaemic treatment than is intensive consolidation chemotherapy in patients with AML in first complete remission.


A new FGFR inhibitor disrupts the TGF-β1-induced fibrotic process.
Pulmonary fibrosis (PF) is chronic and irreversible damage to the lung characterized by fibroblast activation and matrix deposition. Although recently approved novel anti-fibrotic agents can improve the lung function and survival of patients with PF, the overall outcomes remain poor. In this study, a novel imidazopurine compound, 3-(2-chloro-6-fluorobenzyl)-1,6,7-trimethyl-1H-imidazo[2,1-f]purine-2,4(3H,8H)-dione (IM-1918), markedly inhibited transforming growth factor (TGF)-β-stimulated reporter activity and reduced the expression of representative fibrotic markers, such as connective tissue growth factor, fibronectin, collagen and α-smooth muscle actin, on human lung fibroblasts. However, IM-1918 neither decreased Smad-2 and Smad-3 nor affected p38MAPK and JNK. Instead, IM-1918 reduced Akt and extracellular signal-regulated kinase 1/2 phosphorylation increased by TGF-β. Additionally, IM-1918 inhibited the phosphorylation of fibroblast growth factor receptors 1 and 3. In a bleomycin-induced murine lung fibrosis model, IM-1918 profoundly reduced fibrotic areas and decreased collagen and α-smooth muscle actin accumulation. These results suggest that IM-1918 can be applied to treat lung fibrosis.


Baseline health care for refugees in The Netherlands.
In the Netherlands there are some 20,000 refugees from different parts of the world (e.g. Vietnam, Latin America). Most of them have experienced a form of organized violence. The somatic and psychosocial complaints of the refugees are comparable to those of Dutch war victims. They are mostly of an aspecific kind and making a diagnosis can be difficult because of the culturally different presentation of ill being. In order to help general practitioners in making a diagnosis the Refugee Health Centre (CGV) has made a classification of complaints according to whether or not they have a specific cause. It is clear from the literature that there are different opinions about the causes of the somatic complaints. As far as the psychic complaints are concerned it is remarkable that in our pilot study (n = 135) only 6% of the examined refugees suffer from a classical picture of the post-traumatic stress disorder; in a number of cases the picture is limited to some components only. Psychosocial complaints of refugees are subdivided and described. The philosophy of the CGV-treatment is to give assistance as much as possible in the refugee's neighbourhood; so that the clinician(s) will become part of the refugee's new social network. Another very important aspect of the assistance given is preventing medicalization of psychological problems. The basis of help is a recognition of the problems and complaints of the refugee. The structure of the Dutch health care, built up in 'lines', is very often very confusing for a refugee; this confusion can cause communication difficulties between refugee and clinician. To develop methods of treatment, definition and registration of complaints and problems can be a first step.


Effects of fluoxetine on the 5-HT1A receptor and recovery of cognitive function after traumatic brain injury in rats.
This study examined the effects of chronic administration of fluoxetine, a selective serotonin reuptake inhibitor, on cognitive performance and 5-HT1A receptor immunoreactivity following traumatic brain injury. Rats received a moderate severity of lateral fluid percussive injury or sham injury 24 hr after surgical preparation. Fluoxetine or vehicle was administered chronically on postinjury days 1-15. Motor performance and Morris water maze performance were assessed on postinjury days 1-5 and 11-15, respectively. Results indicated that chronic fluoxetine treatment did not affect motor or maze performance. Injured groups showed significantly higher 5-HT1A receptor immunoreactivity on postinjury day 15 than sham-injured rats, and fluoxetine treatment did not alter 5-HT1A receptor immunoreactivity. These results indicate that chronic postinjury fluoxetine administration did not influence the recovery of motor or Morris water maze performance following lateral fluid percussive injury. They also indicate that injury-induced changes in the 5-HT1A receptor may contribute to traumatic brain injury-induced cognitive deficits.


A spectral kurtosis based blind deconvolution approach for spur gear fault diagnosis.
Unanticipated background noises often convolute fault information in the gearboxes' vibration response. The Blind Deconvolution strategy has been extensively applied for fault-impulse enhancement to aid gear fault detection. The existing deconvolution strategies involve designing an optimum filter applied in the time domain. Gear tooth wear leads to the excitation of Gear Mesh Frequency harmonics. Hence, spectral analysis is typically used for gearbox fault detection. As such, feature enhancement in the order domain is more practical than existing blind deconvolution approaches. This study proposes a Spectral Kurtosis-based blind deconvolution strategy with filtering done in the order domain, to aid gear fault detection. Experimental analyses show 109.76% and 64.48% better performance for constant and real-world speed operation, respectively, for the proposed method to aid spectral analysis-based fault detection.


Genetic Profiling of Cell-Free DNA in Liquid Biopsies: A Complementary Tool for the Diagnosis of B-Cell Lymphomas and the Surveillance of Measurable Residual Disease.
To assess the potential value of LiqBio as a complementary tool for diagnosis and surveillance of BCL. This prospective multi-center study included 78 patients (25 follicular lymphomas (FL) and 53 large B-cell lymphomas (LBCL)). We performed next-generation sequencing (NGS) of cfDNA LiqBio and paired gDNA tissue biopsies at diagnosis and compared the mutational statuses. Also, through NGS of LiqBio, we identified MRD biomarkers and compared this novel LiqBio-MRD method with PET/CT in detecting MRD at follow-up. We identified mutations in 71% of LiqBio and 95% of tissue biopsies, and found a correlation between variant allele frequency of somatic mutations. Additionally, we identified mutations in 73% of LiqBio from patients with no available tissue samples or no mutations in them. Regarding the utility of LiqBio-MRD as a dynamic monitoring tool, when compared with the PET/CT method, a lower sensitivity was observed for LiqBio-MRD at 92.3% (vs. 100% for PET/CT), but a higher specificity of 91.3% (vs. 86.9% for PET/CT). Genetic profiling of tumor cfDNA in plasma LiqBio is a complementary tool for BCL diagnosis and MRD surveillance.


The vasculature and limb development.
The developing vascular pattern of the embryonic chick limb results from a combination of two properties: the intrinsic self-assembly and branching properties of the vascular cells and the extrinsic information associated with the expanding mitotic population of mesenchymal cells; and the inhibitory factors which restrict the entrance of vessels into particular domains and/or decrease the branching frequency of such vessels. It is hypothesized that an important component of limb pattern formation is the interplay between the dividing population of mesenchymal cells and the intrinsic properties of the vascular cells. It is further asserted that the presence of particular vascular elements may, indeed, be 'positional information'. Two examples are cited involving aspects of limb duplication to support this possibility; it is suggested that vascular vessel size of a host limb may dictate the polarity of duplication events. The presented hypothesis emphasizes that the interplay between the intrinsic properties of self-assembly into tissues and extrinsic factors which establish boundaries and morphologies is involved in both vascular and limb pattern formation.


Association of heart failure duration with clinical outcomes after transcatheter mitral valve repair for functional mitral regurgitation.
Little is known about the association of heart failure (HF) chronicity with clinical outcomes after transcatheter mitral valve repair (TMVR) for functional mitral regurgitation (MR). From January 2011 to March 2019, consecutive patients with functional MR who underwent a MitraClip procedure were analyzed. The patients were divided into two groups according to HF duration-those with duration ≤18 months and those with >18 months. The primary outcome measure was a composite of all-cause mortality and re-HF rehospitalization within 1 year after the procedure. These outcomes were also assessed separately. A Cox proportional hazard model was conducted for investigating the association of HF duration with the primary outcome. A total of 208 patients were analyzed. Patients with HF duration >18 months had a higher rate of the primary outcome compared to those with HF duration ≤18 months (38.1 vs. 19.0%, log-rank p = .003). A longer duration of HF was associated with an increased risk of the primary outcomes (adjusted-HR of >18 months, 2.12 95% CI, 1.14-4.19; p = .03; adjusted-HR (hazard ratios) for 1 year increase, 1.05; 95% CI, 1.02-1.09; p = .004). The association of HF duration with the primary outcomes showed a steep rise during the first 2 years of HF duration and progressive increase after 5 years. A longer HF duration before TMVR was associated with an increased risk of all-cause mortality or HF rehospitalization. HF duration can be used for the risk stratification marker in patients undergoing TMVR for functional MR.


Tourette's syndrome in Taiwan: an epidemiological study of tic disorders in an elementary school at Taipei County.
We have done an epidemiological study in an elementary school with 2000 Taiwanese children aged from 6 to 12 years, and found 11 of them with Tourette's syndrome. The prevalence rate of Tourette's syndrome was around 0.56%. The ratio of male to female was 9 to 2. The comorbid rate of attention-deficit/hyperactivity disorder was 36%, self-injurious behaviors 27%, and obsessive-compulsive behaviors 18%. The familial rate of Tourette's syndrome was 27%. We also found that another 98 children had transient tic disorders. For 73% of patients with mild tics, understanding and acceptance from family, teachers, and friends are the most important things. When tics are so severe that medication is necessary, haloperidol is no longer the first or only choice. We tried clonidine, atypical neuroleptics such as risperidone or olanzapine, or pergolide. The first support group was established in 1999 for children with tics in Taiwan and transformed to Taiwan Tourette Family association in June 2001 to provide further service for Tourette's syndrome.


Desmopressin and bleeding risk after percutaneous kidney biopsy.
Desmopressin is used to reduce bleeding after kidney biopsy but evidence supporting its use is weak, especially in patients with elevated creatinine. The present study was undertaken to evaluate efficacy of desmopressin in reducing bleeding after percutaneous kidney biopsy. Retrospective cohort study. 269 of 322 patients undergoing percutaneous kidney biopsy between January 1, 2014 and January 31, 2018 were included. Patients had normal bleeding time, platelet count and coagulation profile. Primary outcome was defined as composite of hemoglobin drop ≥1 g/dL, hematoma on post biopsy ultrasound, gross hematuria, erythrocyte transfusion or angiography to stop bleeding. Association of desmopressin with outcomes was assessed using linear (for continuous variables) and logistic (for binary variables) regression models. Propensity score was used to minimize potential confounding. Desmopressin was administered to 100/269 (37.17%) patients. After propensity score adjustment patients who received desmopressin had increased odds of post biopsy bleeding [OR 3.88 (1.95-7.74), p < 0.001]. Creatinine at time of biopsy influenced bleeding risk; gender, emergent vs elective biopsy, obesity, AKI, diabetes, hypertension or bleeding time did not influence bleeding risk. Administration of desmopressin to patients with serum creatinine ≥1.8 mg/dL decreased bleeding risk [OR 2.11 (95% CI 0.87-5.11), p = 0.09] but increased bleeding risk when serum creatinine was < 1.8 mg/dL (OR 9.72 (95% CI 2.95-31.96), p < 0.001). Desmopressin should not be used routinely prior to percutaneous kidney biopsy in patients at low risk for bleeding but should be reserved for patients who are at high risk for bleeding.


Comparison of lithium disilicate-reinforced glass ceramic surface treatment with hydrofluoric acid, Nd:YAG, and CO2 lasers on shear bond strength of metal brackets.
To evaluate and compare the effects of different surface conditioning methods of lithium disilicate-reinforced ceramic on shear bond strength (SBS) of metallic brackets. Thirty-six lithium disilicate ceramic blocks mounted in acrylic resin blocks were assigned to 3 groups (n = 12): 9.6% hydrofluoric acid (HF); neodymium-doped yttrium aluminium garnet (Nd:YAG) laser; and carbon dioxide (CO2) laser. The glass ceramic surfaces were primed with a silane, and the brackets were bonded using a light-cured composite resin. SBS test was carried out in a universal testing machine at 0.5 mm/min crosshead speed until the brackets were debonded. The remaining adhesive was evaluated under a stereomicroscope in terms of the adhesive remnant index (ARI). The surface hardness was determined with a 100-gr force using a microhardness tester. Glass ceramic surface changes were evaluated using the scanning electron microscope. One-way ANOVA and post hoc Tamhane tests were used to compare microhardness values, and Kruskal-Wallis and Mann-Whitney U tests were used to analyze SBS values and ARI. The median and interquartile range of SBS values in 3 groups were 6.48 (1.56-15.18), 1.26 (0.83-1.67), and 0.99 MPa (0.70-2.10), respectively. Microhardness analysis revealed significant differences between the CO2 laser and intact porcelain groups (P = 0.003), without significant differences between the other groups. Group 1 exhibited the highest ARI. Neither CO2 nor Nd:YAG lasers resulted in adequate surface changes for bonding of brackets on ceramics compared with the samples conditioned with HF. CO2 laser decreased the microhardness of ceramics. Surface conditioning with HF resulted in clinically acceptable SBS values.


Mechanisms of motor vehicle accidents that predict major injury.
To assess whether prehospital triage guidelines, based on mechanistic criteria alone, accurately identify victims of motor vehicle accidents (MVA) with major injury. Retrospective analysis of the Royal Melbourne Hospital trauma database. Mechanisms analysed were those outlined by the American College of Surgeons Committee on Trauma and Advanced Trauma Life Support/Early Management of Severe Trauma prehospital triage guidelines. There were 621 MVA analysed, 253 with major injury (40.7%). Multivariate logistic regression indicated prolonged extrication time (P < 0.0001), cabin intrusion (P = 0.047), high speed (P = 0.003) and ejection from vehicle (P = 0.04) were statistically associated with major injury. Vehicle rollover and fatality in the same vehicle were not statistically associated with major injury. These data suggest that existing guidelines for the prehospital triage of MVA victims, based on mechanistic criteria alone may need revision.


Promoter engineering for efficient production of sucrose phosphorylase in Bacillus subtilis and its application in enzymatic synthesis of 2-O-α-D-glucopyranosyl-L-ascorbic acid.
2-O-α-D-glucopyranosyl-L-ascorbic acid (AA-2G), a stable glucoside derivative of L-ascorbic acid (L-AA), can be one-step synthesized by sucrose phosphorylase (SPase). In this study, we attempted to produce extracellular SPase in Bacillus subtilis WB800 for the food-grade production of AA-2G. The results showed that the secretion of SPases did not require signal peptide. Promoter and its compatibility to target SPase gene were proved to be the key factors for high-level secretion. The strong promoter P43 and synthetic SPase gene derived from Bifidobacterium longum (BloSPase) were selected due to generate a relatively high extracellular activity (0.94 U/mL) for L-AA glycosylation. A highly active dual-promoter system PsigH-100-P43 was further constructed, which produced the highest extracellular and intracellular activity were 5.53 U/mL and 6.85 U/mL in fed-batch fermentation, respectively. Up to 113.58 g/L of AA-2G could be achieved by the supernatant of fermentation broth and a higher yield of 146.42 g/L was obtained by whole-cells biotransformation. Therefore, the optimal dual-promoter system in B. subtilis is suitable for the food-grade scale-up production of AA-2G.


Imaging CXCR4 Expression with (99m)Tc-Radiolabeled Small-Interference RNA in Experimental Human Breast Cancer Xenografts.
Noninvasive quantification of chemokine receptor 4 (CXCR4) expression could serve as a prognostic indicator and may be of value for the design of personalized therapies and posttreatment monitoring. The objective of the present study was to assess the use of (99m)Tc-radiolabeled small-interference RNA (siRNA) targeting CXCR4 to detect CXCR4 expression in vivo. CXCR4 siRNAs were radiolabeled with (99m)Tc using the bifunctional chelator hydrazinonicotinamide (HYNIC), and the labeling efficiency, specific activity and radiochemical purity were determined. The stability of the probe in serum was assessed by measuring its radiochemical purity and inhibitory activity by RT-PCR and western blotting. Biodistribution studies and static imaging were performed in MDA-MB-231 tumor-bearing mice. Radiochemical purity remained highly stable in PBS and fresh human serum at room temperature and at 37 °C. Radiolabeled siRNA1 showed strong inhibitory effects similar to those of unlabeled siRNA1 on both CXCR4 messenger RNA (mRNA) and protein in vitro. The excretion of the probe occurred mainly through the liver and kidneys. Tumors were clearly visualized at 1-10 h after injection of the probe, but not after injection of the control probe. (99m)Tc-labeled CXCR4 siRNA1 shows tumor-specific accumulation and could be a promising strategy for the visualization of CXCR4 expression in human breast cancer.


Cervicogenic headache.
The connection between headache and the cervical spine has been a theme of debate for decades. Cervicogenic headache is a headache related to the cervical spine that often is misdiagnosed and treated inadequately because of confusing and varying terminology. In this article, we discuss our experience in diagnosing and treating cervicogenic headache.


Rho-regulatory proteins in breast cancer cell motility and invasion.
The importance of the Rho-GTPases in cancer progression, particularly in the area of metastasis, is becoming increasingly evident. This review will provide an overview of the role of the Rho-regulatory proteins in breast cancer metastatis.


Incidence of cancer in patients with ulcerative colitis 30 years after diagnosis (the IBSEN study).
Patients with ulcerative colitis (UC) have shown an increased risk for colorectal cancer, hepatobiliary, hematologic, and skin cancers, but updated long-term data is needed. This study aimed to estimate the risk of cancer in patients with UC compared to the general Norwegian population, in a population-based cohort (the IBSEN study), 30 years after diagnosis; and to identify possible risk factors associated with cancer. The IBSEN cohort prospectively included all incident patients between 1990 and 1993. Cancer incidence data were obtained from the Cancer Registry of Norway. The overall and cancer-specific hazard ratios (HR) were modelled using Cox regression. Standardized incidence ratios were estimated compared to the general population. In total, the cohort included 519 patients, and 83 cases were diagnosed with cancer. There was no statistically significant difference in the overall cancer risk (HR = 1.01, 95% CI: [0.79-1.29]) and colorectal cancer risk (HR = 1.37, 95% CI: [0.75-2.47]) between patients and controls. The incidence of biliary tract cancer was higher than expected (SIR = 9.84, 95%CI: [3.19-20.15]), especially when UC patients suffered from primary sclerosing cholangitis. Male UC patients were also more at risk of being diagnosed with hematologic malignancies (HR = 3.48, 95% CI: [1.55-7.82]). Being prescribed thiopurines was associated with a higher risk of cancer (HR = 2.03, 95% CI: [1.02-4.01]). At 30 years after diagnosis, the risk of all cancer in patients with UC was not significantly increased compared with the general population. However, the risks of biliary tract cancer and hematologic cancers were increased, particularly in male patients.


Origin of the 3-methylglutaryl moiety in caprazamycin biosynthesis.
Caprazamycins are liponucleoside antibiotics showing bioactivity against Gram-positive bacteria including clinically relevant Mycobacterium tuberculosis by targeting the bacterial MraY-translocase. Their chemical structure contains a unique 3-methylglutaryl moiety which they only share with the closely related liposidomycins. Although the biosynthesis of caprazamycin is understood to some extent, the origin of 3-methylglutaryl-CoA for caprazamycin biosynthesis remains elusive. In this work, we demonstrate two pathways of the heterologous producer Streptomyces coelicolor M1154 capable of supplying 3-methylglutaryl-CoA: One is encoded by the caprazamycin gene cluster itself including the 3-hydroxy-3-methylglutaryl-CoA synthase Cpz5. The second pathway is part of primary metabolism of the host cell and encodes for the leucine/isovalerate utilization pathway (Liu-pathway). We could identify the liu cluster in S. coelicolor M1154 and gene deletions showed that the intermediate 3-methylglutaconyl-CoA is used for 3-methylglutaryl-CoA biosynthesis. This is the first report of this intermediate being hijacked for secondary metabolite biosynthesis. Furthermore, Cpz20 and Cpz25 from the caprazamycin gene cluster were found to be part of a common route after both individual pathways are merged together. The unique 3-methylglutaryl moiety in caprazamycin originates both from the caprazamycin gene cluster and the leucine/isovalerate utilization pathway of the heterologous host. Our study enhanced the knowledge on the caprazamycin biosynthesis and points out the importance of primary metabolism of the host cell for biosynthesis of natural products.


Bacterial pneumonia and subsequent dementia risk: A nationwide cohort study.
Bacterial pneumonia is associated with an increased risk of dementia. However, the association between different pathogens of bacterial pneumonia and the risk of dementia remains unclear. Using the Taiwan National Health Insurance Research Database, we recruited 11,712 patients with bacterial pneumonia and 11,120 controls between 1997 and 2012 and followed them up until the end of 2013. A diagnosis of dementia, Alzheimer's disease (AD), vascular dementia (VaD), and unspecified dementia were identified during the follow-up period. Cox regression analyses were performed with adjustments for confounders. Sensitivity analysis was conducted to exclude patients with prodromal dementia. Patients with bacterial pneumonia were more likely to develop dementia (hazard ratio [HR]: 2.83, 95% confidence interval [CI]: 2.53-3.18), AD (HR: 2.44, 95% CI: 1.65-3.61), VaD (HR: 4.15, 95% CI: 3.20-5.38), and unspecified dementia (HR: 2.62, 95% CI: 2.29-3.00) compared with controls after adjusting for potential confounders. Subgroup pathogen analyses showed that the HR of AD was 3.85 (1.66-8.96) for Hemophilus, and the HR of VaD was 5.40 for Staphylococcus. The risks of dementia and VaD were associated with repeated hospitalization due to bacterial pneumonia in a dose-dependent manner. Sensitivity analyses after exclusion of the first three years or first five years of observation and after exclusion case enrollment before 2010 or 2008 showed consistent findings. Different pathogens are associated with different risks of AD, VaD, and unspecified dementia. Further studies are necessary to investigate the underlying mechanisms of bacterial pneumonia and dementia.


Lumbar spondylolisthesis with isthmic defects.
The importance of spondylolisthesis, aside from the obvious malalignment and malformation of the affected vertebrae, rests mainly on the possibility and the extent of impairment of activity. In this discussion the authors are concerned mainly with the myelographic signs associated with spondylolisthesis in patients with isthmic changes.


Characterization and outcomes of optic nerve gliomas: a population-based analysis.
Optic nerve gliomas (ONG) are rare astrocytic neoplasms. A paucity of literature exists on the epidemiology and outcomes of ONG. Here, we present a series of 445 cases of ONG obtained from the Surveillance, epidemiology and end results (SEER) database. Data on patient and tumor characteristics as well as initial treatment with surgery or radiation were extracted from the SEER Database. Survival rates were calculated using the Kaplan-Meier method. A multivariate analysis was performed to determine independent prognostic factors predicting mortality hazard ratios (HRs) using Cox proportional hazards modeling. The median age range at diagnosis was 5-9 years. Twenty percent of patients were over the age of 20 years. Amongst patients with information available on tumor grade (n = 131), 83% had a low-grade tumors and 17% had a high-grade tumors. Sixteen percent of patients received radiation therapy and 18.4% of patient underwent a sub- or gross total resection. The 5 year overall survival was 96% and 20% for patients with low- and high-grade tumors, respectively. In a multivariate analysis, grade was the only significant predictor of overall survival (HR 29.3, CI: 4.3, 205.4, P < 0.001). Age at diagnosis, receipt of radiation therapy, and extent of surgical resection were not significantly correlated with overall survival. In conclusion, ONG are rare tumors seen predominantly in children. The overall prognosis of high-grade tumors remains poor in all age groups despite multi-modality treatment.