Amplitude difference between pattern-evoked potentials after left and right hemifield stimulation in normal subjects.
The amplitude of visual evoked potentials (VEPs) to pattern hemifield stimulation was measured in 14 normal right-handed subjects. The paired t test was used to determine if the values differed between right and left hemifield stimulation. The amplitude of transient VEPs from a midoccipital electrode and a lateral occipital electrode ipsilateral to the hemifield stimulated was significantly greater with right hemifield stimulation. This result is compatible with neuroanatomic asymmetries of human striate cortex. Amplitude distribution over the scalp was markedly different between transient and steady-state VEPs that probably reflect activities of different populations of cortical neurons.


Cystatin C and cystatin SN as possible soluble tumor markers in malignant uveal melanoma.
The aim of the study was to determine the concentration of endogenous cystatin C and cystatin SN, as potential tumor biomarkers, in the serum and biological fluids of the eye in both healthy controls and patients with uveal melanoma. The concentration of both cystatins was determined in the intraocular fluid (IOF), tear fluid, and serum of patients with uveal melanoma and compared to baseline measurements in IOF, tears, serum, cerebral spinal fluid, saliva and urine of healthy controls. The concentration of cystatin C in all the biological matrices obtained from healthy controls significantly exceeded the concentration of cystatin SN and was independent of gender. Cystatin C concentrations in the tear fluid of patients with uveal melanoma (both the eye with the malignancy, as well as the contralateral, non-affected eye), were significantly greater than cystatin C concentrations in the tear fluid of healthy controls and was independent of tumor size. The concentration of cystatin SN in IOF of patients with uveal melanoma was significantly less than the corresponding concentration of cystatin SN in healthy controls. The ratio of cystatins (CysC:CysSN) in both the serum and tear fluid, as well as the concentration of cystatin SN in IOF, would appear to strongly suggest the presence of uveal melanoma. It is further suggested that multiple diagnostic criteria be utilized if a patient is suspected of having uveal melanoma, such as determination of the cystatin C and cystatin SN concentrations in serum, tears, and IOF, ocular fundus and ultrasound imaging, and biopsy with histopathological evaluation.


Impact of junctional ectopic tachycardia on postoperative morbidity following repair of congenital heart defects.
To determine the incidence of postoperative junctional ectopic tachycardia (JET), we reviewed 343 consecutive patients undergoing surgery between 1997 and 1999. The impact of this arrhythmia on in-hospital morbidity and our protocol for treatment were assessed. We reviewed the postoperative course of patients undergoing surgery for ventricular septal defect (VSD; n=161), tetralogy of Fallot (TOF; n=114), atrioventricular septal defect (AVSD; n=58) and common arterial trunk (n=10). All patients with JET received treatment, in a stepwise manner, beginning with surface cooling, continuous intravenous amiodarone, and/or atrial pacing if the haemodynamics proved unstable. A linear regression model assessed the effect of these treatments upon hours of mechanical ventilation, and stay on the cardiac intensive care unit (CICU). Overall mortality was 2.9% (n=10), with three of these patients having JET and TOF. JET occurred in 37 patients (10.8%), most frequently after TOF repair (21.9%), followed by AVSD (10.3%), VSD (3.7%), and with no occurrence after repair of common arterial trunk. Mean ventilation time increased from 83 to 187 h amongst patients without and with JET patients (P<0.0001). Accordingly, CICU stay increased from 107 to 210 h when JET occurred (P<0.0001). Surface cooling was associated with a prolongation of ventilation and CICU stay, by 74 and 81 h, respectively (P<0.02; P<0.02). Amiodarone prolonged ventilation and CICU stay, respectively, by 274 and 275 h (P<0.05; P<0.06). Postoperative JET adds considerably to morbidity after congenital cardiac surgery, and is particularly frequent after TOF repair. Aggressive treatment with cooling and/or amiodarone is mandatory, but correlates with increased mechanical ventilation time and CICU stay. Better understanding of the mechanism underlying JET is required to achieve prevention, faster arrhythmic conversion, and reduction of associated in-hospital morbidity.


Hormone ontogeny in the ovine fetus. IV. Serum somatomedin activity in the fetal and neonatal lamb and pregnant ewe: correlation with maternal and fetal growth hormone, prolactin, and chorionic somatomammotropin.
Somatomedin (SM) activity was measured by a placental membrane receptor assay using 125I-labeled somatomedin A, as radioligand in serum samples obtained from 33 ovine fetuses, 14 neonatal lambs, 8 pregnant, and 3 postpartum ewes. The mean serum concentration of SM activity in eight adult rams was 2.06 +/- 0.12 U/ml. In fetal sheep, SM activity was detected at 66 days gestation (term 147 days), in the youngest fetus studied. Before 100 days of gestation, SM was lower (P less than 0.001) in fetal sheep (1.08 +/- 0.18 U/ml) than in adult rams. In fetuses between 101 and 125 days, SM rose (P less than 0.001) to 2.64 +/- 0.32 U/ml. In late gestation fetal serum SM fell but during the neonatal period it rose to 3.38 +/- 0.3 U/ml, higher (P less than 0.01) than that in adult rams. Serum SM activity in the pregnant ewe prior to 100 days was 1.01 +/- 0.11 U/ml, increased (P less than 0.05) to 1.75 +/- 0.21 U/ml between 125 days and term, and rose further to 2.56 +/- 0.32 U/ml in the postpartum period. Maternal concentrations of serum SM in late gestation were significantly less than in the fetus. Gel chromatography of fetal, maternal, and neonatal sera indicated that over 90% of SM activity circulated in high-molecular weight form. The rise in SM activity in fetal serum between 100 and 125 days parallels the rise in fetal growth hormone and prolactin concentrations; however, in maternal serum the increase in SM activity is associated with rising maternal chorionic somatomammotropin concentrations.


Prolyl Carboxypeptidase Maintains Receptor Tyrosine Kinase Signaling and Is a Potential Therapeutic Target in Triple Negative Breast Cancer.
TNBC is an aggressive cancer sub-type with limited treatment options and poor prognosis. New therapeutic targets are needed to improve outcomes in TNBC patients. PRCP is a lysosomal serine protease that cleaves peptide substrates when the penultimate amino acid is proline. A role for PRCP in TNBC or other cancers, and its potential as a therapy target has not yet been tested. In the current study, we found high tumor expression of PRCP associates with worse outcome and earlier recurrence in TNBC patients. Knockdown of PRCP or treatment with a small molecule PRCP inhibitor blocked proliferation and survival in TNBC cell lines and inhibited growth of TNBC tumors in mice. Mechanistically, we found PRCP maintains signaling from multiple receptor tyrosine kinases (RTKs), potentially by promoting crosstalk between RTKs and G-protein coupled receptors (GPCRs). Lastly, we found that the PRCP inhibitor caused synergistic killing of TNBC cells when combined with the EGFR and ErbB2 inhibitor lapatinib. Our results suggest that PRCP is potential prognostic marker for TNBC patient outcome and a novel therapeutic target for TNBC treatment.


Clastogenicity of the fungicide afugan in cultured human lymphocytes.
The genotoxic effects of the fungicide afugan were analysed by measuring chromosomal aberrations (CAs), sister chromatid exchange (SCE) and micronuclei (MN) in cultured human peripheral lymphocytes. Concentrations of 2.5, 5, 10 and 20 microg/ml of afugan were used during 24 and 48 h. Afugan significantly increased the frequency of CAs at 5, 10 and 20 microg/ml concentrations during a 48 h treatment period. A significant increase was observed for induction of SCE and MN at all treatments compared with the negative control. A significant dose-response correlation was found in all tests. Afugan did not affect the replicative index (RI), however it significantly decreased the mitotic index (MI) at all treatment concentrations except 2.5 microg/ml, and at both treatment times. The present results indicate that afugan is clastogenic and cytotoxic to cultured human lymphocytes.


Aggressive Care at the End of Life; Where Are We?
Although, efforts to encourage palliative care only for terminal patients, aggressive end-of-life care (EOL) care still common for those probably to die shortly. Multicenter experiences to investigate where did we stand in this era? A retrospective study included patients with advanced solid tumors. The presence of one or more of the following indicators in the last month of life (LM) referred to aggressive EOL care: emergency department (ED) visits ≥ twice, admission to the hospital through ED, death in critical care units (CCUs), and palliative chemotherapy (PC) at the past 2 weeks before death. A total of 435 patients, 51.5% were men with a median age of 62 years (range: 17-108), were included in the study. Most of the patients (89.2%) belonged to Group II; they had attended ED at least twice (60%), approximately 53% admitted to the hospital through ED, 31% received PC-LM with 41% of them had at the past 2 weeks before death, 13% died in the CCUs, and more than half of them (53%) survived <2 weeks. Kaplan-Meier estimator revealed that median survival was 30 days in Group I versus 13 days in Group II (odds ratio: 1.63; 95% confidence interval: 1.20-2.21; P = 0.002). The median survival was statistically significantly associated with PC-LM ≥14 days and the admission mode. There was no statistically significant association with age, sex, and primary cancer sites. The majority of our patients continue with anticancer treatments they possibly do not need and associated with poor survival.


The correlation between the adsorption of adhesive proteins and cell behaviour on hydroxyl-methyl mixed self-assembled monolayers.
The objective of this study was to compare the biological effects of two key cell-adhesive proteins, fibronectin (FN) and vitronectin (VN), upon adsorption onto molecularly-designed model surfaces. Single-component and mixed self-assembled monolayers (SAMs) of alkanethiols on gold with OH and CH(3) terminal groups were prepared at 100%, 65%, 36% and 0% of OH at the surface, to generate a range of surfaces with a simple chemistry and a wettability gradient. FN and VN were adsorbed under non-competitive (single-protein solutions) and competitive (multi-protein solutions) conditions, and compared at different levels: adsorbed amount (radiolabelling), elution, functional presentation of cell-binding domains (ELISA), and role in mediating cell adhesion (antibody-based assay). The observed trends were related to mesenchymal stem cell response in terms of adhesion and overall cell morphology. Under non-competitive conditions, adsorption of both proteins increased with surface hydrophobicity. The presence of competitive proteins significantly decreased the adsorbed amounts, although both proteins were still detected in all SAMs. Adsorption of FN followed a trend similar to that of non-competitive conditions, while adsorption of VN was higher on 100%OH-SAMs. Concerning elution, retention of adsorbed VN was always higher than that of FN. For both proteins, functional presentation of cell-binding domains was more effective on the more hydrophilic 100%OH-SAMs. This fact, coupled to the ability of this type of SAMs to selectively recruit and retain VN in the presence of competitive serum proteins, seems to correlate with the better cell response observed on these surfaces, as compared with hydrophobic 0%OH(100%CH(3))-SAMs.


Therapeutic drug monitoring of atomoxetine in children and adolescents with attention-deficit/ hyperactivity disorder: a naturalistic study.
The selective norepinephrine reuptake inhibitor atomoxetine is potentially among the first-line pharmacotherapy options for ADHD. Therapeutic drug monitoring (TDM) with the quantification and interpretation of atomoxetine serum concentrations is used to determine an individual dose followed by an optimal effectiveness and minimal side effects. The aim of this retrospective pharmacokinetic-pharmacodynamic analysis was to derive age-appropriate recommendations for the implementation of TDM to improve the efficacy and tolerability of atomoxetine in children and adolescents. Using the analytical method of high-performance liquid chromatography with UV detection, 94 serum concentrations of 74 patients between 6 and 21 years of age were determined. Therapeutic effectiveness and side effects were evaluated according to the categories "low", "moderate", and "significant". As part of TDM, a time interval with maximum concentrations of 1-3 h after the administration of atomoxetine was determined for blood sampling. In this time interval, a significant correlation between the weight-normalized dose and the serum concentrations was found. The efficacy as well as the tolerability proved to be mainly moderate or significant. A preliminary therapeutic reference range was between 100 and 400 ng/ml. Naturalistic studies have limitations. Therefore, and due to a limited study population, the results have to be regarded as preliminary observations that must be confirmed in further studies. The preliminary therapeutic reference range for children and adolescents proved to be narrower than the reference range for adult patients. However, due to good efficacy and tolerability an exact reference range remained difficult to determine.


Implication of brevetoxin B1 and PbTx-3 in neurotoxic shellfish poisoning in New Zealand by isolation and quantitative determination with liquid chromatography-tandem mass spectrometry.
Brevetoxin B1 (BTX-B1) was isolated from Austrovenus stutchburyi following the 1992-1993 outbreak of neurotoxic shellfish poisoning (NSP) in New Zealand. We report here the first isolation of PbTx-3 from the same shellfish and the development of a procedure for quantitative determination of PbTx-3 and BTX-B1. PbTx-3 was isolated by chromatography on columns of SiO2, ODS, and LH-20, followed by reverse-phase HPLCs. In mass spectrometry (MS) with an electrospray ionization (ESI) interface operating in the positive or negative ion mode, the abundant protonated ion [M+H]+ of PbTx-3 (m/z 897) and the de-sodiated ion [M-Na]- of BTX-B1 (m/z 1016) were generated, respectively. These served as precursor ions for collision-induced dissociation, and the product ions of m/z 725 from PbTx-3 and m/z 80 from BTX-B1 were identified, allowing unambiguous confirmation of these toxins by selected reaction monitoring liquid chromatography-tandem mass spectrometry (SRM LC-MS/MS) analysis. The determination limits were 0.4 and 2 ng/g for BTX-B1 and PbTx-3 at a signal-to-noise ratio of five, respectively. This LC-MS/MS method was successfully applied to determine BTX-B1 and PbTx-3 in the NSP-associated toxic shellfish. BTX-B1 was found in both A. stutchburyi and Perna canaliculus, but not in Crassostrea gigas, while PbTx-3 was found in all three.


Xyn10A, a thermostable endoxylanase from Acidothermus cellulolyticus 11B.
We cloned and purified the major family 10 xylanase (Xyn10A) from Acidothermus cellulolyticus 11B. Xyn10A was active on oat spelt and birchwood xylans between 60°C and 100°C and between pH 4 and pH 8. The optimal activity was at 90°C and pH 6; specific activity and K(m) for oat spelt xylan were 350 μmol xylose produced min⁻¹ mg of protein⁻¹ and 0.53 mg ml⁻¹, respectively. Based on xylan cleavage patterns, Xyn10A is an endoxylanase, and its half-life at 90°C was approximately 1.5 h in the presence of xylan.


Control of sterol synthesis and of hydroxymethylglutaryl CoA reductase in skin fibroblasts grown from patients with homozygous type II hyperlipoproteinemia.
In skin fibroblasts grown from four children with a homozygous form of type II hyperlipoproteinemia, the feedback control of sterol synthesis and the inhibitory effect on hydroxymethylglutaryl (HMG) CoA reductase activity by serum or low density lipoprotein were present, though diminished compared with the effects in normal fibroblasts. Stimulation of HMG CoA reductase by insulin and inhibition of acetyl CoA carboxylase by serum lipids were not impaired in these type II cells, indicating a degree of specificity in the abnormal response of the reductase. A rapid and convenient method for isolation of mevalonolactone in the course of the assay of HMG CoA reductase is described.


Dieth-yl(μ3-2-methyl-4-oxo-4H-pyran-3-olato-κ(4) O (3),O (4):O (3):O (3))tris-(μ2-2-methyl-4-oxo-4H-pyran-3-olato-κ(3) O (3),O (4):O (3))trizinc toluene disolvate.
The title compound, [Zn3(C2H5)2(C6H5O3)4]·2C7H8, crystallizes with one complex mol-ecule solvated by two mol-ecules of toluene in the asymmetric unit. The Zn(II) ions are coordinated by two terminal ethyl (Et) groups and four maltolate ligands, which act as μ3- and μ2-bridges. The metal atoms are arranged in an incomplete cubane Zn3O4 core structure, derived from one EtZnO3 tetra-hedron, one EtZnO4 bipyramid and one ZnO6 octa-hedron, sharing common corners. The structure is stabilized by weak C-H⋯O and C-H⋯π inter-actions.


Emotion regulation and touch in infants: the role of cholecystokinin and opioids.
Behavioral-pharmacological research in infant rats supports the role of cholecystokinin (CCK) and opioid peptides in mediating early learning of new associations with aspects of the nest and dam, such as maternal odor, milk, and contact. The current paper reviews research that examines the hypothesis that these neuropeptide systems are further involved in mediating emotion regulation in infants, thus playing a role in the emergence of stress-reactivity and other motivational systems. The beneficial effects of maternal proximity, handling, and touch on the development of emotion regulation have been demonstrated in both human and animal models. Interventions that promote tactile stimulation of the infant ("touch therapy") and infant-mother contact ("skin-to-skin contact" or "kangaroo care") have been shown to improve the infant's ability to self-regulate, and to moderate the effects of some risk factors. Theoretical perspectives and empirical findings regarding emotion regulation in infants are first discussed. This is followed by a review of work providing evidence in animal models (and suggestive evidence in humans) for the importance of CCK and opioid neuropeptides in affecting infant emotion regulation and the impact of touch-based interventions, in particular in the context of infant-mother attraction, contact, separation, and attachment.


Elevation of serum erythropoietin after subtotal parathyroidectomy in chronic haemodialysis patients.
After successful subtotal parathyroidectomy (PTX) in 10 chronic haemodialysis patients, significant elevation of Epo was observed, from 48.4 +/- 17.8 mU/ml(M +/- SEM) at preoperative state to 103.3 +/- 34.7 mU/ml at 6 h and 163.4 +/- 50.2 mU/ml at 12 h after PTX. Significant reductions in both PTH-m and ionized calcium (iCa) were confirmed. Since Epo did not increase in the cases with an inadequate PTX and ovariectomy, an abrupt reduction in PTH with a decrease in iCa may play some role in the elevation of Epo.


Conservation of the heterodimeric glycoprotein hormone subunit family proteins and the LGR signaling system from nematodes to humans.
Glycoprotein hormones, follicle-stimulating hormones (FSHs), luteinizing hormones (LHs), thyroid-stimulating hormones (TSHs), and chorionic gonadotropin (CG) are key endocrine hormones secreted from the pituitary gonadotrophs and thyrotrophs and the placenta in primates. These hormones, consisting of a common alpha subunit and a specific beta subunit, act through the FSH receptor (FSHR), the LH receptor (LHR), and the TSH receptor (TSHR) that are highly specific for their cognate hormones. These glycoprotein hormones are structurally and functionally conserved in various vertebrates and have been identified in most lineages of actinopterygians (bony fish) and sarcopterygians (tetra-pods). Of interest, recent genomic studies showed that vertebrate glycoprotein hormone receptors belong to an ancient subfamily of G protein-coupled receptors (GPCRs) named as leucine-rich repeat-containing GPCRs (LGRs). These findings have prompted the hypothesis that there could be additional glycoprotein hormones in vertebrate genomes. Indeed, searches of vertebrate genomes have led to the identification of two novel glycoprotein hormone subunits, glycoprotein alpha 2 (GPA2) and glycoprotein beta 5 (GPB5), as well as their homologs in invertebrates. Subsequently, it was demonstrated that GPA2 and GPB5 form a heterodimeric hormone, thyrostimulin/OGH, capable of activating TSHR in vivoand the thyroid axis in transgenic mice. However, the exact role of this novel glycoprotein hormone and its homolog in invertebrates is not clear. To gain a better understanding of the physiological role of the novel glycoprotein hormone subunits and their evolution, it is imperative to carry out systematic studies of these genes in representative model species. In the present report, we summarize our findings based on studies of genomes of model organisms from sea anemones to humans. We found that GPA2 and GPB5 represent the ancient forms of glycoprotein hormone alpha and beta subunits, respectively, and that vertebrate and invertebrate glycoprotein hormone subunit proteins shared common ancestors that evolved during early metazoan evolution. It is important to note that glycoprotein hormone alpha and beta subunit proteins from invertebrates formed a heterodimer with structural functional characteristics similar to that of vertebrate glycoprotein hormones. Taken together, both glycoprotein hormone alpha and beta subunits evolved before the evolution of nematodes, arthropods, and vertebrates.


Integrating many co-splicing networks to reconstruct splicing regulatory modules.
Alternative splicing is a ubiquitous gene regulatory mechanism that dramatically increases the complexity of the proteome. However, the mechanism for regulating alternative splicing is poorly understood, and study of coordinated splicing regulation has been limited to individual cases. To study genome-wide splicing regulation, we integrate many human RNA-seq datasets to identify splicing module, which we define as a set of cassette exons co-regulated by the same splicing factors. We have designed a tensor-based approach to identify co-splicing clusters that appear frequently across multiple conditions, thus very likely to represent splicing modules - a unit in the splicing regulatory network. In particular, we model each RNA-seq dataset as a co-splicing network, where the nodes represent exons and the edges are weighted by the correlations between exon inclusion rate profiles. We apply our tensor-based method to the 38 co-splicing networks derived from human RNA-seq datasets and indentify an atlas of frequent co-splicing clusters. We demonstrate that these identified clusters represent potential splicing modules by validating against four biological knowledge databases. The likelihood that a frequent co-splicing cluster is biologically meaningful increases with its recurrence across multiple datasets, highlighting the importance of the integrative approach. Co-splicing clusters reveal novel functional groups which cannot be identified by co-expression clusters, particularly they can grant new insights into functions associated with post-transcriptional regulation, and the same exons can dynamically participate in different pathways depending on different conditions and different other exons that are co-spliced. We propose that by identifying splicing module, a unit in the splicing regulatory network can serve as an important step to decipher the splicing code.


The distressed (type D) personality is independently associated with impaired health status and increased depressive symptoms in chronic heart failure.
Chronic heart failure (CHF) is a serious condition that is associated with impaired health status and a high prevalence of depressive symptoms. To date, little is known about the determinants of health status and depressive symptoms in CHF. Therefore, the aim of this study was to assess whether Type D personality is associated with impaired health status and increased depressive symptoms in heart failure patients, independent of disease characteristics. Eighty-four patients (63 men and 21 women, mean age=65.9+/-12.1 years) with systolic CHF completed four questionnaires to assess Type D personality (14-item Type D Personality Scale [DS14]), health status (Minnesota Living with Heart Failure Questionnaire [MLWHFQ]), depressive symptoms (Center for Epidemiological Studies Depression Scale [CES-D]) and mood status (Global Mood Scale [GMS]) when visiting an outpatient heart failure clinic. Information on clinical variables was obtained from patients' medical records. Type D patients were more likely to experience impairment in health status (18/38=47%) as compared to non-Type Ds (11/46=24%), P=0.027. They also more often reported symptoms of depression; namely 18 of 38=47% versus 6 of 46=13%, P=0.001. When controlling for severity and etiology of CHF, age and gender, Type D remained a significant associate of impaired health status [odds ratio (OR) 3.0, 95% confidence interval (CI) 1.12-7.78] and depressive symptoms (OR 6.3, 95% CI 2.08-19.12). Type D was associated with impaired health status and increased depressive symptoms in CHF patients. These preliminary findings demonstrate the value of including personality factors in CHF research.


Outcome of in vitro fertilization following stimulation with highly purified hMG or recombinant FSH in downregulated women of advanced reproductive age: a prospective, randomized and controlled trial.
To compare highly purified human menopausal gonadotropin (HP-hMG) with recombinant follicle-stimulating hormone (rFSH) on ovarian response and pregnancy outcome in downregulated women of advanced reproductive age. A prospective, randomized and controlled study of 127 consecutive normogonadotropic infertile women ≥ 35 years old undergoing their first in vitro fertilization/intracytoplasmic sperm injection cycles received ovarian stimulation with HP-hMG (n = 63) or with rFSH (n = 64) in a long gonadotropin-releasing hormone agonist protocol. More leading (≥ 18 mm) follicles and oocytes were obtained in rFSH group (p = 0.008 and p < 0.001, respectively). The proportion of top-quality embryo from oocyte retrieval and live birth rate per started cycle trended towards improvement with HP-hMG (OR 1.3, 95% CI 0.9-1.8; OR 1.9, 95% CI 0.9-3.9; respectively), although they were not significant difference between two groups. At end of stimulation, higher serum progesterone level was found in rFSH group (p < 0.001). Following downregulated women of advanced reproductive age, superiority of HP-hMG over rFSH in live birth rate could not be concluded from this study, but noninferiority was established. Pharmacodynamic differences in follicular development, oocyte/embryo quality and endocrine response exist between HP-hMG and rFSH, which may be relevant to treatment outcome.


Polyglutamation of a novel antifolate, MX-68, is not necessary for its anti-arthritic effect.
N-[[4-[(2,4-diaminopteridin-6-yl)methyl]-3,4-dihydro-2H-1,4-benzothiazin-7-yl]-carbonyl]-L-homoglutamic acid (MX-68), a derivative of methotrexate, was chemically designed to resist polyglutamation and to have a high affinity for dihydrofolate reductase, in an attempt to reduce the side effects of methotrexate. We confirmed that MX-68 did not undergo polyglutamation and investigated the pharmacological activities of MX-68 compared with methotrexate. (1) In vitro: MX-68 inhibited the activity of dihydrofolate reductase to the same degree as methotrexate-tetraglutamate. MX-68 treatment produced a similar anti-proliferative effect to that of methotrexate. However, the intracellular concentration of MX-68 was much lower than the sum of the levels of methotrexate and its polyglutamate, and the effects of MX-68 disappeared when it was removed from the culture medium. (2) In vivo: Oral administration of MX-68 suppressed the development of collagen-induced arthritis in mice and adjuvant-induced arthritis in rats, in a similar fashion to that of methotrexate. These results indicate that polyglutamation is not essential for the anti-arthritic effect of antifolates.


Carcinoma of the urinary bladder after treatment with cyclophosphamide for non-Hodgkin's lymphoma.
We observed nine cases of transitional-cell carcinoma of the urinary bladder among patients who had had long-term treatment of other cancers with cyclophosphamide. Seven of the bladder carcinomas occurred within a cohort of 471 patients treated for non-Hodgkin's lymphomas. In this cohort the relative risk of bladder cancer was 6.8 (95 percent confidence interval, 3.2 to 14.2). The cumulative risk (mean +/- SE) was 3.5 +/- 1.8 percent 8 years after the start of treatment with cyclophosphamide and 10.7 +/- 4.9 percent after 12 years. Three of the nine patients were 50 years of age or younger; seven died with progressive bladder cancer. Subsequently, an additional patient had acute nonlymphocytic leukemia. Hemorrhagic cystitis was observed in 33 patients (cumulative risk, 11.8 +/- 2.1 percent after five years). Development of carcinoma of the urinary bladder was not related to previous hemorrhagic cystitis. The results caution against long-term treatment with cyclophosphamide for diseases with a favorable prognosis.


Transthyretin and retinol-binding protein as discriminators of diabetic retinopathy in type 1 diabetes mellitus.
Diabetic retinopathy (DR) is one of the most common complications of diabetes mellitus (DM), which is still a major reason for blindness. Transthyretin (TTR) and retinol-binding protein (RBP) are thought to be related to the pathogenesis both in T2DM and T1DM. We aimed to investigate the association between serum levels of TTR, RBP, RBP/TTR ratio, and DR. This retrospective study involved 188 T1DM inpatients divided into two groups: patients with DR (n = 95) and patients without DR (n = 93). Data of serum levels on lipids and inflammation were collected. Multiple logistic regression analysis was performed to research the association between TTR, RBP, RBP/TTR, and diabetic retinopathy in T1DM. Compared with patients without DR, those with DR have a higher level of TTR (207 versus 195 mg/L, p = 0.034) and RBP4 (36.85 versus 25.68 mg/L, p < 0.001). Significant differences were also observed between two groups with respect to body mass index (BMI), blood pressure (BP), total cholesterol (TC), triglycerides (TG), low-density lipoprotein (LDL), homocysteine, apolipoprotein B (APOB), leucocyte, monocyte, neutrophil, and uric acid (p < 0.05 for all). TTR, RBP, and RBP/TTR were positively correlated with BP, BMI, TG, LDL, homocysteine, APOB, and uric acid. A multivariate logistic regression model revealed individuals with RBP4 level in the highest quartile had 58.95 times higher risk of developing diabetic retinopathy than those in the lowest quartile. In conclusion, TTR, RBP, and RBP/TTR ratio are risk factors of DR in T1DM. They are potential markers and targets for diagnosis and treatment of DR.


Systems analysis and improvement approach to improve naloxone distribution within syringe service programs: study protocol of a randomized controlled trial.
More than half a million Americans died of an opioid-related overdose between 1999 and 2020, the majority occurring between 2015 and 2020. The opioid overdose mortality epidemic disproportionately impacts Black, Indigenous, and people of color (BIPOC): since 2015, overdose mortality rates have increased substantially more among Black (114%) and Latinx (97%) populations compared with White populations (32%). This is in part due to disparities in access to naloxone, an opioid antagonist that can effectively reverse opioid overdose to prevent death. Our recent pilot work determined that many barriers to naloxone access can be identified and addressed by syringe service programs (SSPs) using the Systems Analysis and Improvement Approach to Naloxone distribution (SAIA-Naloxone). This randomized controlled trial will test SAIA-Naloxone's ability to improve naloxone distribution in general and among BIPOC specifically. We will conduct a trial with 32 SSPs across California, randomly assigning 16 to the SAIA-Naloxone arm and 16 to receive implementation as usual. SAIA-Naloxone is a multifaceted, multilevel implementation strategy through which trained facilitators work closely with SSPs to (1) assess organization-level barriers, (2) prioritize barriers for improvement, and (3) test solutions through iterative change cycles until achieving and sustaining improvements. SSPs receiving SAIA-Naloxone will work with a trained facilitator for a period of 12 months. We will test SAIA-Naloxone's ability to improve SSPs' naloxone distribution using an interrupted time series approach. Data collection will take place during a 3-month lead-in period, the 12-month active period, and for an additional 6 months afterward to determine whether impacts are sustained. We will use a structured approach to specify SAIA-Naloxone to ensure strategy activities are clearly defined and to assess SAIA-Naloxone fidelity to aid in interpreting study results. We will also assess the costs associated with SAIA-Naloxone and its cost-effectiveness. This trial takes a novel approach to improving equitable distribution of naloxone amid the ongoing epidemic and associated racial disparities. If successful, SAIA-Naloxone represents an important organizational-level solution to the multifaceted and multilevel barriers to equitable naloxone distribution.


IL-15 as memory T-cell adjuvant for topical HIV-1 DermaVir vaccine.
IL-7 and IL-15 are key cytokines involved in the generation and maintenance of memory CD8+ T-cells. We evaluated these cytokines as molecular adjuvants for topical HIV-1 DermaVir vaccine. We found that mice receiving DermaVir formulated with HIV-1 Gag plasmid in the presence of IL-7- or IL-15-encoding plasmid significantly enhanced Gag-specific central memory T-cells, as measured by a peptide-based cultured IFN-gamma ELISPOT. Additionally, IL-15 significantly improved DermaVir-induced Gag-specific effector memory CD8+ T-cell responses, measured by standard IFN-gamma ELISPOT. In a DermaVir prime/vaccinia vector boost regimen, the inclusion of IL-15 together with DermaVir significantly improved Gag-specific effector memory T-cell responses. Our study demonstrates that IL-15 is more potent than IL-7 in enhancing HIV-1-specific central memory T-cells induced by topical DermaVir. IL-15 adjuvanted DermaVir might be an alternative prime in a prophylactic vaccine regimen.


Evaluating bias and variability in diagnostic test reports.
Diagnostic testing is an important component of modern medical care. Unfortunately, many diagnostic tests are not rigorously evaluated before general application. Studies examining test characteristics often have methodologic flaws that impair their ability to provide reliable information on test performance. These flaws can introduce systematic nonrandom errors (biases) that distort measures of test accuracy. Other design errors can make it difficult to generalize the results of individual studies. These problems may enhance the apparent performance of poor tests while obscuring the performance of good tests, and they may result in the widespread use of tests with uncertain or limited efficacy. This article explores the ways in which studies of diagnostic test efficacy can be affected by bias and variability.


VRCTC-310--a novel compound of purified animal toxins separates antitumor efficacy from neurotoxicity.
Two purified animal venom toxins, crotoxin and cardiotoxin, have been combined to produce a unique natural product (VRCTC-310) currently under investigation as an antitumor agent by the National Cancer Institute. In vitro, it has demonstrated cytotoxic disease specificity and a unique mechanism of action when submitted to COMPARE analysis. In vivo, tolerance was developed to the neurotoxic properties of crotoxin which allowed comparison of several schedules of fixed and escalating daily i.m. doses to mice bearing s.c. Lewis Lung carcinoma. An 83% inhibition of tumor growth was achieved using an escalating dose schedule starting at 1.8 mg/kg and reaching 6.3 mg/kg/day on day 20. Although some irritation around the sites of i.m. injection was noted, animal weight loss was negligible and there were no other signs of adverse toxicity. This natural product represents a new, membrane interactive anticancer agent which produces a unique spectrum of cytotoxicity in vitro and which has demonstrated interesting in vivo antitumor efficacy.


Validation of a monoclonal antibody directed against the human sphingosine 1-phosphate receptor type 1.
The sphingosine 1-phosphate receptor type 1 (S1PR1) has several important functions, including stabilizing endothelial barrier and maintaining lymphocyte circulation. These functions are critically dependent on the regulation of S1PR1 cell surface expression. Currently available antibodies against human S1PR1 are not able to pick up cell surface expression on living cells by flow cytometry due to intracellular epitopes or unspecific binding. Here we describe the generation of a mouse monoclonal antibody specific for the N-terminal region of human S1PR1. It has an immunoglobulin M (IgM) kappa isotype and detects cell surface expression of recombinant human S1PR1 on overexpressing cells. Due to unspecific intracellular cell staining, it cannot be used for staining of dead cells and tissue slides or in microscopic analyses. It is also not suitable for Western blot analysis and immunoprecipitation. However, the antibody can stain for endogenous S1PR1 on human endothelial cell lines and primary human umbilical vein endothelial cells (HUVEC). Incubation of these cells with various S1PR1 agonists revealed potent S1PR1 internalization, which was not the case with the specific antagonist W146. Surprisingly, human T and B cells isolated from blood and palatine tonsils did not show specific staining, demonstrating significantly lower endogenous S1PR1 surface expression on lymphocytes than on endothelial cells.


A Case Series of Dentigerous Cyst in Paediatric Patients at Our Tertiary Institution.
Dentigerous cysts involves erupted or developing teeth in decreasing order of frequency as mandibular third molars, the maxillary canines, the maxillary third molars with rare involvement of the central incisors & are usually asymptomatic but becomes symptomatic with symptoms such as swelling, mild sensitivity, tooth mobility and displacement when it reaches size > 2 cm in diameter. Dentigerous cysts are seen most commonly in 20-30 years age group with relative low frequency in children with proportion of 4-9% of these cysts occur in the first 10 years after birth. Hospital based study conducted in the Department of ENT at Ashwini Rural medical college & Hospital, Solapur in which we have reported a series of three cases of dentigerous cyst in paediatric patients. Dentigerous Cysts in our study were associated with maxillary permanent lateral incisor, mandibular permanent premolar, Mandibular second molar tooth in pediatric age group. Provisional diagnosis of Dentigerous cyst based on clinical, radiological & biochemical study but Histopathological examination confirms the diagnosis. Dental orthopentamogram aids in diagnosing and localizing the Dentigerous cyst and ct face in all views helps to see bony effects and complications due to expansion of Dentigerous cyst. Enucleation of the cystic lesion followed by extraction of the offending teeth is a suitable treatment modality for Dentigerous cyst associated with maxilla and mandible giving adequate balance between complete removal of cyst & giving chance for eruption of unerupted teeth.


Compliance calibration for fracture testing of equine cortical bone.
An experimental compliance calibration method for measuring crack length in fracture toughness tests of cortical bone was developed. Calibration tests were conducted on twenty compact type fracture specimens machined from the mid-diaphysis of five pairs of equine third metacarpal bones. Specimens were oriented for crack propagation in a direction transverse to the longitudinal axis of the bone. Specimen compliance was determined from the load vs. crack opening displacement record over a range of crack lengths from 0.48 to 0.75 times the specimen width. The results demonstrate that the compliance calibration method developed for isotropic materials can be used to determine crack length in bone, which is transversely isotropic. However, specimens from lateral and dorsal regions exhibited significantly different compliance calibrations even after differences in elastic modulus were taken into account in the normalized compliance.


The oxygen reactions of reduced cytochrome c oxidase. Position of a form with an unusual EPR signal in the sequence of early intermediates.
The order of appearance of intermediates in the reoxidation of reduced cytochrome c oxidase by oxygen has been examined. Particular emphasis was placed on determining where the intermediate with the EPR signal at g = 5, 1.78, 1.69 (Shaw, R.W., Hansen, R.E. and Beinert, H. (1978) J. Biol. Chem. 253, 6637--6640) appears in the sequence of events during reoxidation. Flash photolysis of reduced, CO-complexed samples of cytochrome c oxidase in the presence of oxygen in a buffer containing 30% (v/v) ethylene glycol at 77 K and 195 K has been used to generate states of partial reoxidation. The intermediate with the EPR signal at g = 5, 1.78, and 1.69 can be detected as a product of the photolysis and subsequent oxidation but does not appear until the photolyzed sample is incubated at temperatures well above 196 K. In the course of the reoxidation, the intermediate characterized by the g = 5, 1.78, 1.69 signal occurs in the reaction sequence after the states referred to as 'Compound A' and 'Compound B' (Chance, B., Saronio, C., and Leigh, J.S. (1975) J. Biol. Chem. 250, 9226--9237). Its appearance is within the time range reported for the formation of 'oxygenated' cytochrome c oxidase (Orii, Y. (1979) in Cytochrome Oxidase (King, T.E., Orii, Y., Chance, B. and Okunuki, K., eds.), pp. 331--340, Elsevier/North-Holland Biomedical Press, Amsterdam).


Studies on the specific dynamic effect of proteins of different nutritional value.
Using the respiratory method the rise of metabolic rate (specific dynamic effect SDE) was determined in laboratory rats after ingestion of gluten, peas, cottage cheese and ham. The SDE was greatest after ham ingestion, which is a product of high nutritional value, while after gluten ingestion which has a low protein value, the SDE was lowest.


Impact of Fast-Acting Insulin Aspart on Glycemic Control in Patients with Type 1 Diabetes Using Intermittent-Scanning Continuous Glucose Monitoring Within a Real-World Setting: The GoBolus Study.
Background: The GoBolus study investigated the real-world effectiveness of faster aspart in patients with type 1 diabetes (T1D) using intermittent-scanning continuous glucose monitoring (iscCGM) systems. Methods: This 24-week, multicenter, single-arm, noninterventional study investigated adults with T1D (HbA1c, 7.5%-9.5%) receiving multiple daily injections (MDI) of insulin and using iscCGM within local healthcare settings for ≥6 months before switching to faster aspart at study start (week 0; baseline). Primary endpoint was HbA1c change from baseline to week 24. Exploratory endpoint was change in iscCGM metrics from baseline to week 24. Results: Overall, 243 patients were included (55.6% male), with mean age/diabetes duration, 49.9/18.8 years; mean HbA1c, 8.1%. By week 24, HbA1c had decreased by 0.19% (-2.1 mmol/mol, P < 0.0001) with no mean change in insulin doses or basal/bolus insulin ratios. For patients with sufficient available iscCGM data (n = 92): "time in range" (TIR; 3.9-10.0 mmol/L) increased from 46.9% to 50.1% (P = 0.01), corresponding to an increase of 46.1 min/day; time in hyperglycemia decreased from 49.1% to 46.1% (>10.0 mmol/L, P = 0.026) and 20.4% to 17.9% (>13.9 mmol/L, P = 0.013), corresponding to 43.5 (P = 0.024) and 35.6 (P = 0.015) fewer minutes per day on average spent in these ranges, respectively; no change for time in hypoglycemia (<3.9 and <3.0 mmol/L). Mean interstitial and postprandial glucose improved from 10.4 to 10.1 mmol/L (P = 0.035) and 11.9 to 11.0 mmol/L (P = 0.002), respectively. Conclusion: Real-world switching to faster aspart in adults with T1D on MDI improved HbA1c, increased TIR, and decreased time in hyperglycemia without affecting time in hypoglycemia. The GoBolus study: NCT03450863.


Leptin deficiency, not obesity, protects mice from Con A-induced hepatitis.
Leptin-deficient ob/ob mice are protected from Con A-induced hepatitis. However, it is unclear whether leptin deficiency or obesity itself is responsible for this protection. To address this question, wild-type (WT) obese mice with high serum leptin levels were generated by injection of gold thioglucose (WT GTG). Both Con A-injected WT and WT GTG mice developed hepatitis, whereas no hepatic damage was observed in ob/ob mice. Moreover, TNF-alpha and IFN-gamma levels as well as expression of the activation marker CD69 were elevated in liver mononuclear cells of WT and WT GTG mice, but not in ob/ob mice following administration of Con A. The liver of WT and WT GTG mice had the same percentage of NK T cells, a lymphocyte population involved in Con A-induced hepatitis. This population decreased equally in both WT and WT GTG mice after Con A injection. In contrast, the liver of ob/ob mice contained 50% less NK T cells compared to WT and WT GTG mice. Furthermore, no decrease in NK T cells was observed in Con A-injected ob/ob mice. We conclude that leptin-deficiency, not obesity, is responsible for protection from Con A-induced hepatitis.


Determining therapeutic approaches in the elderly with rectal cancer.
To evaluate the toxicity and feasibility of pelvic radiotherapy (RT) and/or surgery in elderly patients with locally advanced low-lying rectal cancer. From November 1999 to November 2005, 51 patients aged >or=70 years who underwent RT for locally advanced low-lying rectal cancer were retrospectively examined. Variables considered were age, co-morbidities (evaluated according to the Charlson score and the Cumulative Illness Rating Scale-Geriatric [CIRS-G] score) and surgery versus no surgery. The median age was 80 years (range 70-94 years) and the male : female ratio was 33 : 18. A total of 5.9% of patients were considered 'fit', 72.5% had one or more CIRS-G grade 1 or 2 co-morbidities and 21.6% had one or more CIRS-G grade 3 co-morbidities. 54.9% of patients underwent surgery and 45.1% underwent RT. Only 9 of 21 (42.8%) patients who underwent radical resection received the full course of adjuvant RT and only seven (50%) of all patients treated with RT alone received the full dose of therapy. Patients with one or more CIRS-G grade 3 co-morbidities reported similar numbers of grade 1-2 toxicities as patients with one or more CIRS-G grade 2 co-morbidities. Notwithstanding the small number of patients analysed, the findings of this study indicate that elderly patients with rectal cancer and mild co-morbidities could probably receive the same treatment as fit elderly patients, given that tolerability appeared to be similar in both categories of patients. Neither age nor co-morbidities should be considered reasons to deny the patient the possible benefits of receiving complete treatment. Moreover, Multidimensional Geriatric Assessment should always be undertaken to help clinicians make better decisions about treatment. Further prospective trials are needed to confirm these results.


Interobserver variability between cytopathologists and cytotechnologists upon application and characterization of the indeterminate category in the Milan System for Reporting Salivary Gland Cytopathology.
The indeterminate categories in the Milan System for Reporting Salivary Gland Cytopathology (MSRSGC) are diagnostically challenging because of inherent heterogeneity and complexity, with wide interobserver variability (IOV). Herein, the authors explore the concordance rate (CR) between cytopathologists (CPs) and cytotechnologists (CTs) in interpreting indeterminate salivary gland lesions using the MSRSGC. Between 2011 and 2016, 86 indeterminate fine-needle aspirations had slides available for review, of which 48 had follow-up. Four CPs and 2 CTs performed an independent, blinded review of these slides and categorized them according to the MSRSGC. The CRs between CTs and CPs with the final sign-out cytopathologist (FCP) were assessed, and interobserver agreement was categorized into uniform, majority, divided, minimal, or no agreement. The overall CR with the FCP ranged from 48.8% to 60.5% for CPs and from 22.1% to 36% for CTs. IOV κ scores for the entire group were 0.314 and, with the FCP as the reference, ranged from 0.403 to 0.539 for CPs and from 0.091 to 0.254 for CTs. Uniform, majority, divided, minimal, and no agreement was noted in 12.8%, 31.4%, 38.4%, 10.5%, and 6.9%, respectively, of all cases and in 16.7%, 35.4%, 31.3%, 8.3%, and 6.3%, respectively, of the cases with follow-up. Diagnostic challenges included distinguishing lymphoma from a reactive process and distinguishing mucin from mucin-like material. CPs had modestly higher CRs compared with CTs; and, although the variable CRs highlight indeterminate IOV, the MSRSGC enables reproducibility. Characterizing larger cohorts in the indeterminate categories will further improve MSRSGC criteria. Moreover, education on the MSRSGC should include CTs and CPs to improve overall diagnostic accuracy.


Genetic susceptibility to multiple sclerosis in Sardinians: an immunological study.
We studied the in vitro production of variably MS-related cytokines from Sardinian MS and healthy donors bearing the two "Sardinian" MS-associated HLA-DR alleles: DR3 and DR4, with the purpose to evidentiate possible differences in their immune response. ELISA were used for detection of cellular products by mitogen-activated peripheral blood mononuclear cells. PHA-activated HLA-DR4+/DR3- mononuclear cells produce significantly higher amounts of TNF-alpha compared with the DR3+/DR4-. In addition, homozygous HLA-DR3+ mononuclear cells from MS patients produce significantly lower amounts of IL-10 than those from homozygous HLA-DR3+ healthy donors. The abnormal production of detrimental or regulatory cytokines may account for the genetic susceptibility to MS in different HLA-subgroups of Sardinian MS patients.


A diagnostic test for carpal tunnel syndrome using ultrasound.
The paper describes the details of an ultrasound test for carpal tunnel syndrome and compares the results of this with E.M.G. studies in a series of 19 patients. The equipment used was a standard therapeutic ultrasound unit and the tests were performed by a physiotherapist. The results were encouraging, with a 90% correlation with E.M.G. results.


Survival rates after serious immersion accidents in childhood.
A study of childhood survival rates, after loss of consciousness in fresh water, has been undertaken. Age-specific, sex-specific and site-specific survival rates for childhood fresh water immersion accidents are reported for the first time. The overall survival rate, after loss of consciousness in the water was 0.49; swimming pool and domestic bath tub serious immersion accidents have a survival rate of 0.60 compared with a rate of 0.05 for similar immersion accidents in creeks and rivers. Young male schoolboys have the lowest potential survival (0.20 or less) of any group. Survival rates were significantly higher during the winter (0.86) than during the warmer months (survival rate of 0.49). This gives a quantitative expression to the protection from cerebral anoxia afforded by body chilling which is not extreme. Survival rates have increased significantly over the 5 year period 1971--1975; it is considered that this is due to public education campaigns of the potential danger of water to children. The use of survival rates to measure factors which modify the pathophysiology of human drowning and near-drowning is discussed.


Structural differences between apo- and holoenzyme of horse liver alcohol dehydrogenase.
The three-dimensional structure of a ternary complex of horse liver alcohol dehydrogenase with reduced nicotinamide adenine dinucleotide and the inhibitor dimethyl sulfoxide has been determined to 4.5 A resolution independently of the apoenzyme structure. The electron density maps of both structures have been compared. The two coenzyme binding domains which form the center of the dimer molecular have retained their conformation and orientation within the molecule whereas the catalytic domains rotate and narrow the cleft between the domains. The active site becomes shielded from the solution by a combination of this rotation, local movements of a loop from residues 53 to 57 and coenzyme and substrate binding. Both subunits bind coenzyme and inhibitor to the same extent. The nicotinamide ring of the coenzyme is positioned close to the active zinc atom and the inhibitor is bound to this zinc atom. The difference between the two crystallographically independent subunits is small. The proposed mechanisms of action for the enzyme based on the apoenzyme structure are confirmed by the present investigation.


Sustained remission of type 2 diabetes in rodents by centrally administered fibroblast growth factor 4.
Type 2 diabetes (T2D) is a major health and economic burden worldwide. Despite the availability of multiple drugs for short-term management, sustained remission of T2D is currently not achievable pharmacologically. Intracerebroventricular administration of fibroblast growth factor 1 (icvFGF1) induces sustained remission in T2D rodents, propelling intense research efforts to understand its mechanism of action. Whether other FGFs possess similar therapeutic benefits is currently unknown. Here, we show that icvFGF4 also elicits a sustained antidiabetic effect in both male db/db mice and diet-induced obese mice by activating FGF receptor 1 (FGFR1) expressed in glucose-sensing neurons within the mediobasal hypothalamus. Specifically, FGF4 excites glucose-excited (GE) neurons while inhibiting glucose-inhibited (GI) neurons. Moreover, icvFGF4 restores the percentage of GI neurons in db/db mice. Importantly, intranasal delivery of FGF4 alleviates hyperglycemia in db/db mice, paving the way for non-invasive therapy. We conclude that icvFGF4 holds significant therapeutic potential for achieving sustained remission of T2D.


Can we measure the intraocular pressure when the eyeball is against the pillow in the lateral decubitus position?
To evaluate the amount of intraocular pressure (IOP) change in the eye against the pillow in the lateral decubitus position (LDP). Thirty eyes from 15 healthy volunteers (12 men and three women) aged 29 ± 3 (range 25-37) years participated in this study. Using the rebound tonometer (Icare PRO, Icare Finland Oy, Helsinki, Finland), the IOP of both eyes was checked in sitting, supine, right and left LDPs. In the LDP, the additional IOP measurements were taken with the lower eyeball against the latex pillow. Baseline IOP in the sitting position was 12.7 ± 1.9 mmHg in the right eye and 12.8 ± 2.2 mmHg in the left eye. Ten minutes after shifting from the sitting to the supine position, IOP increased significantly (right eye: +1.4 ± 1.4 mmHg, p = 0.006; left eye: +1.8 ± 1.5 mmHg, p = 0.001). Changing from the supine to the right and left LDP increased significantly the IOP of dependent eye (right eye: +2.3 ± 1.8 mmHg, p = 0.001; left eye: +1.5 ± 1.8 mmHg, p = 0.011). When the dependent eye was compressed against the pillow in the LDP, the IOP of the dependent eyes increased significantly after 10 min (right eye in the right LDP: +4.1 ± 4.9 mmHg, p = 0.011; left eye in the left LDP: +3.4 ± 3.7 mmHg, p = 0.006). The IOP was significantly elevated when the eyeball was against the pillow in the LDP.


The Impact of Balanced Counseling on Contraceptive Method Choice and Determinants of Long Acting and Reversible Contraceptive Continuation in Nepal.
Introduction Long-acting reversible contraceptives (LARCs) reduce rates of unintended pregnancies and repeat abortion. Uptake and continuation rates of LARCs are very low in Nepal, despite free provision from most health facilities. We sought to establish the effectiveness of a new approach to LARC promotion in Nepal. Methods We examined change in contraceptive method mix in Nepal using service data resulting from introduction of a balanced counseling (BC) approach to family planning (FP). All staff located at nine randomly selected FP sites were trained and began applying BC in April and May 2014. Women who accepted LARCs from a participating facility were re-contacted at 1, 3, 6 and 12 months. We estimated the LARC continuation rate and assessed determinants of continuation using descriptive analysis, Kaplan-Meier survival curves and univariate and multivariate Cox proportional hazard analysis. Results A total of 5744 women received BC between April and July 2014. 1580 women (27.5 %) took up LARCs, raising its contribution to contraceptive method mix at [organization] to 40 %, significantly higher than the 15 % recorded in 2013. 913 women were followed-up, and the LARC continuation rate at 12 months was 82 %. Women's reported satisfaction with LARC [AHR 0.23; 95 % CI 0.14-0.39, p = 0.000] was the single strongest determinant of LARC continuation after adjusting for all background characteristics. Discussion The findings suggest BC is an effective approach for increasing LARC uptake in Nepal. The rate of LARC continuation and its determinants are important inputs to strategies for improved delivery of FP services.


Sox4 functions as a positive regulator of β-catenin signaling through upregulation of TCF4 during morular differentiation of endometrial carcinomas.
Sox factors function as either activators or repressors of β-catenin/TCF transcription depending on the cellular context and associated interacting proteins. Our previous study provided evidence that alteration in β-catenin signaling is an essential event during transdifferentiation toward the morular phenotype of endometrial carcinomas (Em Cas). Here, we focused on related functional roles of Sox factors. Of eight Sox factors investigated, Sox4 could enhance β-catenin/TCF4 transcription, through upregulation of TCF4 at the transcription level, without any direct β-catenin association. Cells stably overexpressing Sox4 showed significant decreases in proliferation rate, along with increases in expression of p21(WAF1), as well as TCF4, in contrast to increased cell growth observed with knockdown. Of these factors, only Sox7 could transcriptionally upregulate Sox4 expression, but it also resulted in not only inhibition of Sox4-meditated activation of β-catenin/TCF4-driven transcription, but also repression of its own promoter activity, indicating the existence of very complex feedback loop for Sox-mediated signal cascades. Finally, Sox4 immunoreactivity was frequently pronounced in morular lesions of Em Cas, the expression being positively correlated with status of β-catenin, TCF4, and Sox7, and inversely with cell proliferation. These data therefore suggest that Sox4 may serve as a positive regulator of β-catenin signaling through alteration in TCF4 expression during morular differentiation of Em Ca cells, leading to inhibition of cell proliferation. In addition, Sox7 may also participate in the process, having complex roles in modulation of signaling.


Soluble interleukin-2 receptor and soluble CD8 antigen in active rheumatoid arthritis.
Concentrations of soluble interleukin-2 receptor (sIL-2R) and of soluble CD8 antigen (sCD8) in sera and in supernatants of phytohemagglutinin (PHA)-stimulated peripheral blood mononuclear cells (PBMC) derived from patients with active rheumatoid arthritis (RA) were studied. sIL-2R concentrations in sera derived from patients with RA (1484 +/- 382 U/ml) were significantly higher than in sera derived from healthy controls (380 +/- 110 U/ml; P less than 0.0005). In contrast, supernatants of PHA-stimulated PBMC derived from patients with RA contained similar amounts of sIL-2R (727 +/- 467 U/ml) as those derived from healthy control individuals (833 +/- 508 U/ml; P greater than 0.1). When investigated for the presence of sCD8 antigen, sera derived from patients with RA contained significantly lower amounts (30 +/- 28 U/ml) than sera derived from healthy controls (405 +/- 136 U/ml; P less than 0.0005). Similarly, PHA stimulation of PBMC derived from patients with RA resulted in a significantly lower production of sCD8 (35 +/- 46 U/ml) as compared to the one obtained by PHA stimulation of PBMC derived from healthy controls (177 +/- 59 U/ml; P less than 0.0005). This difference could not be explained by a lower proliferative response to PHA by PBMC derived from patients with RA (21,474 +/- 14,022 cpm) as compared to healthy controls (29,549 +/- 11,188 cpm; P greater than 0.05). Our data demonstrate that PBMC derived from patients with active RA differ from PBMC derived from healthy individuals concerning their ability to produce sIL-2R and sCD8.


Dual-functional porous and cisplatin-loaded polymethylmethacrylate cement for reconstruction of load-bearing bone defect kills bone tumor cells.
Malignant bone tumors are usually treated by resection of tumor tissue followed by filling of the bone defect with bone graft substitutes. Polymethylmethacrylate (PMMA) cement is the most commonly used bone substitute in clinical orthopedics in view of its reliability. However, the dense nature of PMMA renders this biomaterial unsuitable for local delivery of chemotherapeutic drugs to limit the recurrence of bone tumors. Here, we introduce porosity into PMMA cement by adding carboxymethylcellulose (CMC) to facilitate such local delivery of chemotherapeutic drugs, while retaining sufficient mechanical properties for bone reconstruction in load-bearing sites. Our results show that the mechanical strength of PMMA-based cements gradually decreases with increasing CMC content. Upon incorporation of ≥3% CMC, the PMMA-based cements released up to 18% of the loaded cisplatin, in contrast to cements containing lower amounts of CMC which only released less than 2% of the cisplatin over 28 days. This release of cisplatin efficiently killed osteosarcoma cells in vitro and the fraction of dead cells increased to 91.3% at day 7, which confirms the retained chemotherapeutic activity of released cisplatin from these PMMA-based cements. Additionally, tibias filled with PMMA-based cements containing up to 3% of CMC exhibit comparable compressive strengths as compared to intact tibias. In conclusion, we demonstrate that PMMA cements can be rendered therapeutically active by introducing porosity using CMC to allow for release of cisplatin without compromising mechanical properties beyond critical levels. As such, these data suggest that our dual-functional PMMA-based cements represent a viable treatment option for filling bone defects after bone tumor resection in load-bearing sites.


Near-infrared spectroscopy in the medical management of infants.
Near-infrared spectroscopy (NIRS) is a technology that is easy to use and can provide helpful information about organ oxygenation and perfusion by measuring regional tissue oxygen saturation (rSO2) with near-infrared light. The sensors can be placed in different anatomical locations to monitor rSO2 levels in several organs. While NIRS is not without limitations, this equipment is now becoming increasingly integrated into modern healthcare practice with the goal of achieving better outcomes for patients. It can be particularly applicable in the monitoring of pediatric patients because of their size, and especially so in infant patients. Infants are ideal for NIRS monitoring as nearly all of their vital organs lie near the skin surface which near-infrared light penetrates through. In addition, infants are a difficult population to evaluate with traditional invasive monitoring techniques that normally rely on the use of larger catheters and maintaining vascular access. Pediatric clinicians can observe rSO2 values in order to gain insight about tissue perfusion, oxygenation, and the metabolic status of their patients. In this way, NIRS can be used in a non-invasive manner to either continuously or periodically check rSO2. Because of these attributes and capabilities, NIRS can be used in various pediatric inpatient settings and on a variety of patients who require monitoring. The primary objective of this review is to provide pediatric clinicians with a general understanding of how NIRS works, to discuss how it currently is being studied and employed, and how NIRS could be increasingly used in the near future, all with a focus on infant management.


Subtypes of muscarinic receptors in cultured explants of the hippocampus of the rat.
Using the tritiated, muscarinic antagonist quinuclidinyl benzilate ([3H]QNB) as a ligand, muscarinic receptors have been identified and characterized in intact, cultured explants of the hippocampus of the rat. Competition studies with scopolamine and oxotremorine indicated a certain heterogeneity in the population of muscarinic receptors, whereas atropine and pirenzepine competed with [3H]QNB in a manner consistent with only one binding site for these substances. Thus, the observed heterogenity does not fit in with the M1/M2 receptor concept. Extended studies, with the aim of determining to what extent these putative subtypes of receptors are functional, would be of interest.


Targeting a helix-in-groove interaction between E1 and E2 blocks ubiquitin transfer.
The ubiquitin-proteasome system (UPS) is a highly regulated protein disposal process critical to cell survival. Inhibiting the pathway induces proteotoxic stress and can be an effective cancer treatment. The therapeutic window observed upon proteasomal blockade has motivated multiple UPS-targeting strategies, including preventing ubiquitination altogether. E1 initiates the cascade by transferring ubiquitin to E2 enzymes. A small molecule that engages the E1 ATP-binding site and derivatizes ubiquitin disrupts enzymatic activity and kills cancer cells. However, binding-site mutations cause resistance, motivating alternative approaches to block this promising target. We identified an interaction between the E2 N-terminal alpha-1 helix and a pocket within the E1 ubiquitin-fold domain as a potentially druggable site. Stapled peptides modeled after the E2 alpha-1 helix bound to the E1 groove, induced a consequential conformational change and inhibited E1 ubiquitin thiotransfer, disrupting E2 ubiquitin charging and ubiquitination of cellular proteins. Thus, we provide a blueprint for a distinct E1-targeting strategy to treat cancer.


Recombinant thrombomodulin domain 1 rescues pathological angiogenesis by inhibition of HIF-1α-VEGF pathway.
Pathological angiogenesis (PA) contributes to various ocular diseases, including age-related macular degeneration, diabetic retinopathy, and retinopathy of prematurity, which are major causes of blindness over the world. Current treatments focus on anti-vascular endothelial growth factor (VEGF) therapy, but persistent avascular retina, recurrent intravitreal neovascularization, and general adverse effects are reported. We have previously found that recombinant thrombomodulin domain 1 (rTMD1) can suppress vascular inflammation. However, the function of rTMD1 in VEGF-induced PA remains unknown. In this study, we found that rTMD1 inhibited VEGF-induced angiogenesis in vitro. In an oxygen induced retinopathy (OIR) animal model, rTMD1 treatment significantly decreased retinal neovascularization but spared normal physiological vessel growth. Furthermore, loss of TMD1 significantly promoted PA in OIR. Meanwhile, hypoxia-inducible factor-1α, the transcription factor that upregulates VEGF, was suppressed after rTMD1 treatment. The levels of interleukin-6, and intercellular adhesion molecule-1 were also significantly suppressed. In conclusion, our results indicate that rTMD1 not only has dual effects to suppress PA and inflammation in OIR, but also can be a potential HIF-1α inhibitor for clinical use. These data bring forth the possibility of rTMD1 as a novel therapeutic agent for PA.


Determination of the main risk factors for benzodiazepine dependence using a multivariate and multidimensional approach.
The aim of this study was to identify risk factors for benzodiazepine (BZD) dependence, such as sociodemographic variables, characteristics of BZD use, and psychiatric parameters, which to date have been found to relate inconsistently to indicators of BZD dependence such as chronic BZD use and BZD withdrawal symptoms. The Benzodiazepine Dependence Self-Report Questionnaire (Bendep-SRQ), Schedules for Clinical Assessment in Neuropsychiatry (SCAN), and Symptom Checklist-90 (SCL-90) were administered to 599 outpatients using BZDs. Regression analyses were conducted using BZD dependence diagnoses and severity scales as dependent variables. BZD dependence diagnoses were only predicted by being a self-help patient and long BZD elimination half-life (for only the DSM-III-R). The main predictors of BZD dependence severity, as measured by the ICD-10, DSM-III-R scales, and Bendep-SRQ Rasch scales, were in decreasing order: (1) being a self-help patient; (2) higher BZD dose, longer duration of BZD use, younger age; and (3) non-native cultural origin, lower level of education, being in outpatient treatment for alcohol and/or drug dependence, and the interaction of BZD dose with duration of BZD use. We conclude that a limited number of recognizable risk factors appear to predict the severity of BZD dependence. Additional administration of a specific BZD dependence instrument is recommended to confirm suspected BZD dependence and guide further clinical decision-making.


Changes in the structure and function of the kidney of rats chronically exposed to cadmium. II. Histoenzymatic studies.
Early effects of cadmium (Cd) on the structure and function of the kidney were studied in an experimental model using rats intoxicated with Cd at the levels of 5 and 50 mg Cd/l drinking water. The effect of Cd was evaluated histopathologically and biochemically. Damage to the cellular structures was assessed on the basis of histoenzymatic analyses of the activity and localization of indicator enzymes (succinate dehydrogenase, lactate dehydrogenase, glucose-6-phosphatase, Mg(2+)-dependent adenosine triphosphatase and acid phosphatase). The histochemical observations indicate that Cd causes damage to the organization and function of the nephron. Several structures, i.e. endoplasmic reticulum, mitochondrion, lysosome, cellular and intracellular membrane, as well as their biological functions, i.e. aerobic and anaerobic respiration, transport functions and biochemical processes taking place in the endoplasmic reticulum, were affected. The cytotoxic action of Cd occurs mainly in the tubules and partially also in the glomeruli. The results clearly indicate that Cd damages kidney structurally and functionally even at a relatively low level (5 mg/l) corresponding to human environmental exposure, and they confirm our previous hypothesis that the threshold for the kidney effects of Cd is less than 4.08+/-0.33 microg/g kidney wet weight and higher than 2.40+/-0.15 microg/g. The target for Cd action in the kidney is the tubules (proximal convoluted tubules and straight tubules), and disturbance in their function is the main toxic effect of Cd. Renal glomeruli are also injured, but only partially, whereas in other parts of the nephron the damage is slight. The results, together with observations reported in the first paper of the study, incline us to conclude that humans environmentally exposed to Cd are at risk of tubular damage.


The patient-centered medical home, practice patterns, and functional outcomes for children with attention deficit/hyperactivity disorder.
To determine whether children with attention deficit/hyperactivity disorder (ADHD) receive care in a patient-centered medical home (PCMH) and how that relates to their ADHD treatment and functional outcomes. Cross-sectional analysis of the 2007 National Survey for Children's Health, a nationally representative survey of 91,642 parents. This analysis covers 5169 children with parent-reported ADHD ages 6-17. The independent variable is receiving care in a PCMH. Main outcome measures are receiving ADHD medication, mental health specialist involvement, and functional outcomes (difficulties with participation in activities, attending school, making friends; having problem behaviors; missed school days; and number of times parents contacted by school). Only 44% of children with ADHD received care in a PCMH. Children with ADHD receiving care in a PCMH compared with those who did not were more likely to receive medication for ADHD (odds ratio [OR], 1.4; 95% confidence interval [CI], 1.1-1.9); less likely to have mental health specialist involvement (OR, 0.6; 95% CI, 0.4-0.7); less likely to have difficulties participating in activities (OR, 0.6; 95% CI 0.4-0.8), making friends (OR, 0.6; 95% CI, 0.5-0.9), and attending school (OR, 0.4; 95% CI, 0.3-06); less likely to have problem behaviors (OR 0.6; 95% CI 0.5-0.9); had fewer missed school days (β = -1.5, 95% CI -2.4 to -0.5); and parents were contacted by school less frequently (β = -0.2, 95% CI -0.3 to -0.1). For children with ADHD, receiving care in a PCMH is associated with practice pattern change and better outcomes. The PCMH may represent a promising opportunity to improve quality of care and outcomes for children with ADHD.


Trapped sweat in basketball uniforms and the effect on sweat loss estimates.
The aims of this study were to determine: (1) trapped sweat (TS) in basketball uniforms and the effect on sweat loss (SL) estimates during a laboratory-based basketball simulation protocol; (2) the impact of exercise intensity, body mass, age, and SL on TS; and (3) TS during on-court training to assess the ecological validity of the laboratory-based results. Twenty-four recreational/competitive male basketball players (23 ± 10 years, 77.0 ± 16.7 kg) completed three randomized laboratory-based trials (Low, Moderate, and High intensity) consisting of 150-min intermittent exercise. Eighteen elite male players (23 ± 4 years, 92.0 ± 20.6 kg) were observed during coach-led, on-court training. Nude and clothed body mass were measured pre and postexercise to determine TS. Data are mean ± SD. There was a significant effect of intensity on SL and TS (P < 0.001, Low<Moderate<High, ANOVA). During Low, subjects lost 1.10 ± 0.59 kg sweat and TS was 0.11 ± 0.15 kg (8.0 ± 5.1% SL). During Moderate, subjects lost 1.60 ± 0.56 kg sweat and TS was 0.21 ± 0.21 kg (11.6 ± 6.3% SL). During High, subjects lost 2.12 ± 0.66 kg sweat and TS was 0.38 ± 0.28 kg (16.0 ± 7.4% SL). Multiple regression and partial correlation analysis suggested TS was significantly related to SL (P < 0.0001; partial r = 0.81-0.89), whereas the contributions of body mass (P = 0.22-0.92) and age (P = 0.29-0.44) were not significant. TS during on-court training was 0.35 ± 0.36 kg, which was associated with a 14.1 ± 11.5% underestimation in SL, and was not statistically different than laboratory-based results (P = 0.59). Clothed body mass measurements should be used with caution, as TS is highly variable and can cause a significant underestimation in SL in athletes with high sweating rates.


Heavy metal toxicity and the aetiology of glaucoma.
Despite recent advances, our understanding of the aetiological mechanisms underlying glaucoma remains incomplete. Heavy metals toxicity has been linked to the development of neurodegenerative diseases and various ocular pathologies. Given the similarities in pathophysiology between glaucoma and some neurodegenerative disorders, it is plausible that heavy metal toxicity may play a role in the development of glaucoma. Heavy metal exposure may be occupational, or through water or dietary contamination. In this report, we review mechanisms for systemic and neurotoxicity for arsenic, cadmium, chromium, cobalt, lead, mercury, and manganese, and weigh the evidence for an association between glaucoma and the accumulation of heavy metals either in ocular tissues or in the central nervous system.


Heterotrimeric and small molecular mass GTP-binding proteins of rat brain neuronal and glial nuclei.
Heterotrimeric and small molecular mass guanine nucleotide binding (GTP-binding) proteins were found in neuronal and glial nuclei isolated from rat brain. Neuronal nuclei bound 0.213 +/- 0.055 pmoles of GTP/microg protein (n = 8) and glial nuclei bound 0.145 +/- 0.038 pmoles of GTP/microg protein (n = 8). The intrinsic GTPase activity of neuronal and glial nuclei was 0.0019 +/- 0.0005 pmoles GTP hydrolyzed/min/microg protein (n = 10) and 0.0022 +/- 0.0006 pmoles GTP hydrolyzed/min/microg protein (n = 10), respectively. Western blot analysis was carried out using a peptide-specific antibody that recognizes a common sequence in the alpha-subunit of the various heterotrimeric G-proteins. The antibody revealed the presence of a polypeptide of molecular mass of 40 kDa only in neuronal nuclei. Small molecular mass GTP-binding proteins were detected by incubating nitrocellulose blots with [alpha-32P]GTP. The results demonstrated the presence of 25-26 kDa GTP-binding proteins in both populations of nuclei. However, the binding of [alpha-32P]GTP to neuronal nuclei was approximately 3-fold greater than to the glial nuclei. Further analysis by two-dimensional polyacrylamide gel electrophoresis resolved the neuronal nuclei 26 kDa protein into three forms (a-c) with the most acidic form (c) being the major species. The neuronal 25 kDa protein was resolved into two forms that were present in approximately equal concentration. In glial nuclei, only the 26 kDa (c) and a small amount of the 25 kDa proteins were detected. However, both populations of nuclei contained the small molecular mass GTP-binding protein, ran. Differential association of non-ran small molecular mass GTP-binding proteins and heterotrimeric G-proteins with neuronal nuclei suggests a potential role for these guanine nucleotide binding proteins in the function of this cell type.


The impact of a "narrative interview" intervention in oncology. A study protocol for a feasibility study.
Oncological diagnosis determines a biographical breakdown and requires the person to adapt to the disease. If patients, after diagnosis, ask professionals for 'compassionate care', research on these issues is still underdeveloped. There are currently no studies that use the narrative interview as an intervention tool. The objectives of the study are to evaluate: (1) the feasibility of the narrative interview intervention on cancer patients in the first diagnosis; (2) the impact of the narrative medicine intervention on the patient's self-perception, his psychological distress and adaptation to the disease. It is a mixed-method study, with an intervention (narrative interview) and quantitative evaluation before/after intervention and qualitative evaluation post-intervention (reflective writing). The analysis will use the Psychological Distress Inventory scale for the assessment of psychological distress and the Mini-Mental Adjustment to Cancer Scale for the assessment of disease adaptation. Adult patients, with oncological pathology will be recruited one month after the communication of the diagnosis, regardless of the type of tumor. The Wilcoxon test for paired data will be used to verify pre-post-intervention differences. The 'reflective writings' will be subjected to thematic analysis. The study evaluates the feasibility of the narrative interview intervention as a primary outcome. Secondly, the impact of the intervention is assessed in relation to: a) identification of risk or protective factors on psychological distress and adaptation to the disease; b) re-elaboration of the patient's experiences and experiences related to his/her own illness.


Study of the influence of meteorological factors on HFMD and prediction based on the LSTM algorithm in Fuzhou, China.
This study adopted complete meteorological indicators, including eight items, to explore their impact on hand, foot, and mouth disease (HFMD) in Fuzhou, and predict the incidence of HFMD through the long short-term memory (LSTM) neural network algorithm of artificial intelligence. A distributed lag nonlinear model (DLNM) was used to analyse the influence of meteorological factors on HFMD in Fuzhou from 2010 to 2021. Then, the numbers of HFMD cases in 2019, 2020 and 2021 were predicted using the LSTM model through multifactor single-step and multistep rolling methods. The root mean square error (RMSE), mean absolute error (MAE), mean absolute percentage error (MAPE) and symmetric mean absolute percentage error (SMAPE) were used to evaluate the accuracy of the model predictions. Overall, the effect of daily precipitation on HFMD was not significant. Low (4 hPa) and high (≥ 21 hPa) daily air pressure difference (PRSD) and low (< 7 °C) and high (> 12 °C) daily air temperature difference (TEMD) were risk factors for HFMD. The RMSE, MAE, MAPE and SMAPE of using the weekly multifactor data to predict the cases of HFMD on the following day, from 2019 to 2021, were lower than those of using the daily multifactor data to predict the cases of HFMD on the following day. In particular, the RMSE, MAE, MAPE and SMAPE of using weekly multifactor data to predict the following week's daily average cases of HFMD were much lower, and similar results were also found in urban and rural areas, which indicating that this approach was more accurate. This study's LSTM models combined with meteorological factors (excluding PRE) can be used to accurately predict HFMD in Fuzhou, especially the method of predicting the daily average cases of HFMD in the following week using weekly multifactor data.


Lmod2 piggyBac mutant mice exhibit dilated cardiomyopathy.
Leiomodin proteins, Lmod1, Lmod2 and Lmod3, are key regulators of the thin filament length in muscles. While Lmod1 is specifically expressed in smooth muscles, both Lmod2 and Lmod3 are expressed in striated muscles including both cardiac and skeletal muscles. We and others have previously shown that Lmod3 mainly function in skeletal muscles and the mutant mice display disorganized sarcomere. Lmod2 protein has been found to act as an actin filament nucleator in both cell-free assays and in cultured rat and chicken cardiomyocytes. To better understand the function of Lmod2 in vivo, we have identified and characterized a piggyBac (PB) insertional mouse mutant. Our analysis revealed that the PB transposon inserts in the first exon of the Lmod2 gene and severely disrupts its expression. We found that Lmod2 (PB/PB) mice exhibit typical dilated cardiomyopathy (DCM) with ventricular arrhythmias and postnatal lethality. Electron microscope reveals that the Lmod2 (PB/PB) hearts carry disordered sarcomere, disarrayed thin filaments, and distorted intercalated discs (ICDs). Those ICDs display not only decreased convolutions, but also reduced electron-dense staining, indicating less ICDs component proteins in Lmod2 (PB/PB) hearts. Consistent with the phenotype, the expression of the ICD component genes, β-catenin and Connexin43, are down-regulated. Taken together, our data reveal that Lmod2 is required in heart thin filaments for integrity of sarcomere and ICD and deficient mice exhibit DCM with ventricular arrhythmias and postnatal lethality. The Lmod2 (PB/PB) mutant offers a valuable resource for interrogation of pathogenesis and development of therapeutics for DCM.


References ranges and sources of variation for indices of pulsed Doppler flow velocity waveforms from the uteroplacental and fetal circulation.
Variation in indices used to characterize the maximum frequency outline of waveforms obtained from the uteroplacental and fetal circulations by duplex Doppler ultrasound was studied between observers, over a 24-h period and before and after a meal. No significant differences were found. Reference ranges were derived for three indices, the A/B ratio, the resistance index and the pulsatility index, for waveforms from the fetal descending aorta, umbilical artery, and the placental and maternal sides of the uteroplacental circulation. Indices from waveforms from the uteroplacental circulation demonstrated a fall from 16 to 24 weeks gestation with no change thereafter. The umbilical artery indices demonstrated a continuing fall with increasing gestation but the A/B ratio and resistance index from the fetal descending aorta did not change with gestation. The pulsatility index from the aorta demonstrated a gentle rise.


Hydrothermal synthesis and photoluminescent properties of stacked indium sulfide superstructures.
Unusual hierarchical stacked superstructures of cubic beta-In2S3 were fabricated via a facile hydrothermal process in the presence of a surfactant cetyltrimethylammonium bromide CTAB; the 3D superstructures were developed by helical propagation of surface steps from microflakes of 10-20 nm thickness.


Amyotrophic lateral sclerosis with a sudden-onset history.
We report on two patients with amyotrophic lateral sclerosis (ALS) complaining of sudden-onset difficulty in finger elevation. A 65-year-old man (the first patient) and a 66-year-old man (the second patient) suddenly became aware of difficulty in finger elevation of one hand. They were not aware of any other symptoms prior to the onset. In the first patient, cerebral infarction at the precentral gyrus was initially suspected. In the second patient, cervical spondylosis was initially suspected, and cervical spine surgery was planned. However, needle EMG revealed widespread neurogenic changes and abundant fasciculation potentials for both patients. Widespread weakness emerged in time and relentlessly progressed, and finally the diagnosis of ALS was made. In both cases, notable weakness in the extensor digitorum (ED) muscle with relatively mild weakness in the other muscles in the affected limb was a characteristic finding. Loss of one motor unit in ED that has already enlarged due to reinnervation must have caused sudden awareness of the weakness. Clinicians should recognize the presence of ALS patients with a sudden-onset history because the risk of initial misdiagnosis is high for such patients.


Retained subretinal perfluorocarbon liquid in microincision 23-gauge versus traditional 20-gauge vitrectomy for retinal detachment repair.
To assess the rate of retained subretinal perfluorocarbon liquid (PFCL) in patients undergoing rhegmatogenous retinal detachment (RRD) repair with sutureless 23-gauge vitrectomy versus traditional 20-gauge vitrectomy. A retrospective, consecutive, interventional comparative case series. All patients with a diagnosis of RRD who underwent pars plana vitrectomy with PFCL for RRD repair from November 1, 2005 through October 31, 2008 were included. A total of 234 RRD repairs were performed during the study period by one surgeon. Subretinal PFCL occurred in 4 of 176 eyes (2.3%) who underwent sutured 20-gauge pars plana vitrectomy and in 6 of 58 eyes (10.3%) who underwent sutureless 23-gauge pars plana vitrectomy for repair of retinal detachment (P = 0.0167, Fisher exact test, 2-tailed). There is a statistically significant 4.5-fold increased incidence of retained subretinal PFCL in patients undergoing RRD repair with sutureless 23-gauge vitrectomy versus traditional 20-gauge vitrectomy. This may be because of higher fluid flow through open 23-gauge cannulas, which causes disruption of the PFCL surface tension resulting in formation of small PFCL bubbles that can enter the subretinal space. Reduction of fluid flow may help prevent this complication.


Health disparities in epilepsy: how patient-oriented outcomes in women differ from men.
Epilepsy is a chronic disorder with multiple effects on biological, social, and psychological health. Many of these effects differ between men and women, but only sparse research has specifically addressed the relevance and importance of the differences. Available evidence suggests that men and women with epilepsy have differing rates of employment and driving, and women with epilepsy have increased risk for specific mood disorders such as post-partum depression. National surveys of physicians indicate that many physicians providing care for women with epilepsy have limited knowledge of fundamental concerns such as interactions between antiepileptic drugs and oral contraceptives, and their potential teratogenic effects. Further research and clinical implementation of improved gender-specific care is needed to optimize outcomes for women with epilepsy.


Mixed micelles of PEG(2000)-DSPE and vitamin-E TPGS for concurrent delivery of paclitaxel and parthenolide: enhanced chemosenstization and antitumor efficacy against non-small cell lung cancer (NSCLC) cell lines.
Concurrent combination of chemotherapeutic drugs is a promising alternative to single-agent therapies in cancer. In the present study, paclitaxel and parthenolide were loaded into mixed micelles and tested against taxol sensitive (A549) and resistant (A549-T24) NSCLC cell lines. Combination chemotherapy was further evaluated by isobologram analyses and combination index calculations. Drugs were loaded into micelles by the film casting method using PEG(2000)-DSPE and vitamin E-TPGS. Micelle characterization studies included the determination of particle size, encapsulation efficiency, in vitro release kinetics, as well as 1H NMR analysis. The in vitro release of both drugs was slower from the mixed micelles, which maintained an encapsulation efficiency >95% and chemical stability over a storage period of 45 days. The IC50 of paclitaxel and parthenolide determined by MTT assay were 108.6nM and 21μM, respectively, while the combination had an IC50 of 64.15nM in A549 cells. In the taxol resistant cell lines, the IC50 values of paclitaxel and parthenolide were 233nM and 32μM, respectively, while the combination had an IC(50) of 128nM. The efficacy of paclitaxel and parthenolide against both cell lines significantly increased when the drugs were coencapsulted in mixed micelles. Mixed micelles caused 79% cell death, which was significantly higher than the 46% cell death caused by the drugs in solution against taxol sensitive cell lines. In taxol resistant cell lines, the cell death caused by mixed micelles was 70% as compared to 45% cell death caused by un-encapsulated drugs. Co-encapsulation of parthenolide with paclitaxel in mixed micelles increased the anticancer activity of paclitaxel against resistant and sensitive lung cancer cell lines.


High alkaline phosphatase activity in 3p-induced human renal cancer cells.
Recent studies have suggested that a tumor suppressor gene is located on the short arm of chromosome 3 (3p) for tumorigenesis of sporadic renal cell carcinoma. In this study, we introduced 3p from normal human fibroblasts into the renal cancer cell line YCR and this introduction clearly resulted in inhibited growth in vitro and in vivo. Furthermore, we investigated several enzyme activities of original YCR cells and 3p-induced clones (YCR-151 and YCR-152) and found that only alkaline phosphatase activity in 3p-induced clones was significantly increased compared to that of the original YCR cells.


Aurovertin E, a new polyene pyrone from the basidiomycete Albatrellus confluens.
A new polyene pyrone, aurovertin E (2), was isolated along with aurovertin B (1) from the culture mycelia of the basidiomycete Albatrellus confluens. Their structures were elucidated on the basis of spectroscopic studies including 2D NMR experiments. This is the first example of the occurrence of aurovertins in basidiomycetes.


Hyperoxia and moderate hypoxia fail to affect inspiratory muscle fatigue in humans.
Normal human subjects (n = 7) breathing 21% O2 (normoxia), 13% O2 (hypoxia), or 100% O2 (hyperoxia) performed repeated maximal inspiratory maneuvers (inspiratory duration = 1.5 s, total breath duration = 3.5 s) on an "isoflow" system, which delivered a constant mouth flow (1.25 or 1 l/s) while maintaining normocapnia (5.5% end-tidal CO2). Respective mean arterial O2 saturation values (ear lobe oximetry) were 98 +/- 1, 91 +/- 4 (P less than or equal to 0.01), and 99 +/- 1% (NS). Maximal mouth pressure (Pm) was measured during inspirations at rest and during a 10-min fatigue trial, and the Pm measurements obtained during the fatigue trials were fit to an exponential equation. The parameters of the equation included the time constant (tau), which describes the rate of decay of Pm from the initial pressure (Pi) to the asymptote, or "sustainable" pressure (Ps). The mean fraction of Pm remaining at the end of the fatigue trials (Ps/Pi) was 63 +/- 5%. No significant differences in Pi, Ps, or tau were observed between O2 treatments. This suggests that fatigue of the inspiratory muscles in normal humans occurs by a mechanism that is insensitive to changes in blood O2 content that occur during inspiration of O2 in the range of 13-100%.


Finalizing the properties of porous scaffolds of aliphatic polyesters through radiation sterilization.
Porous scaffolds made of various L,L-lactide (LLA), 1,5-dioxepane-2-one (DXO) and epsilon-caprolactone (CL) copolymers were sterilized by EB- and gamma-irradiation. Differences in the comonomers, composition and the microstructure of the starting materials were used to influence the degradation mechanism and susceptibility towards irradiation and by this means to achieve sterilized scaffolds with predicted end-properties. The chemical changes and the formation of low-molecular-weight products were determined by SEC, 1H nuclear magnetic resonance (NMR), 13C NMR and gas chromatography-mass spectrometry (GC-MS). The degradation mechanism changed from random chain scission to cross-linking depending on the choice of monomers, the copolymer composition and the monomer sequences. Copolymerization of LLA with small amounts of CL or DXO increased the stability compared to that of the LLA homopolymer. Changing DXO to CL in a LLA copolymer also increased the stability. The type of radiation and the microstructure of the copolymer chains determined which of the monomer sequences were more prone to degrade. The most abundant low-molecular-weight product identified after sterilization was DXO monomer. Traces of LLA and CL monomers were also identified. Modification of the copolyester microstructure changed the degradation mechanism and the susceptibility towards irradiation. This allows the use of radiation sterilization to finalize the scaffold properties.


The effect of routine endoscopy on the detection rate of T1 gastric cancer (early gastric cancer) in Birmingham.
The introduction of routine endoscopy facilities at the General Hospital, Birmingham has made no significant impact on the proportion of stage T1 or 'early' gastric cancers detected. Prior to this, T1 cancers comprised 2.8% of all tumors diagnosed, or 4.2% of resected cases. Following establishment of the service in 1974, 3.2% of all cases and 6.3% of those resected were stage T1 lesions. The clinical presentation reflected the high proportion of tumors associated with ulceration (67%). The accuracy rate for pre-operative diagnosis of a malignant lesion was barium meal 67%, cytology alone 73%, biopsy alone 80%, and cytology and biopsy in combination, repeated if suspicious, 100%. The crude survival rate at 5 years was 83.3% (age-adjusted 96%), and at 10 years 53.8%).


Use of 3-D Contrast-Enhanced Ultrasound to Evaluate Tumor Microvasculature After Nanoparticle-Mediated Modulation.
A cost-effective method for serial in vivo imaging of tumor microvasculature has been developed. We evaluated acoustic angiography (AA) for visualizing and assessing non-small cell lung tumor (A549) microvasculature in mice before and after tumor vascular disruption by vascular-targeted gold nanoparticles and radiotherapy. Standard B-mode and microbubble-enhanced AA images were acquired at pre- and post-treatment time points. Using these modes, a new metric, 50% vessel penetration depth, was developed to characterize the 3-D spatial heterogeneity of microvascular networks. We observed an increase in tumor perfusion after radiation-induced vascular disruption, relative to control animals. This was also visualized in vessel morphology mode, which revealed a loss in vessel integrity. We found that tumors with poorly perfused vasculature at day 0 exhibited a reduced growth rate over time. This suggested a new method to reduce in-group treatment response variability using pre-treatment microvessel maps to objectively identify animals for study removal.


Biochemical and morphological characterization of spontaneously hypertensive hyperlipidaemic rats.
1. To develop and characterize a new animal model of hypertension and hyperlipidaemia, we cross-bred spontaneously hypertensive rats (SHR) with spontaneously hyperlipidaemic rats (HLR). 2. A new strain of spontaneously hypertensive hyperlipidaemic rats (SHHR) was established at generation 10 through selective mating of brothers and sisters (systolic blood pressure > 150 mmHg, plasma cholesterol > 150 mg/dL). Cross-bred Wistar-Kyoto (WKY) rats and Sprague-Dawley (SD) rats (SDWKY rats) were used as a control. 3. Adrenaline and noradrenaline levels in the plasma and adrenal medulla of male SHHR were significantly higher than those of male SDWKY rats. The tyrosine hydroxylase activity in the adrenal medulla of male SHHR was significantly higher than that of male SDWKY rats. Low-density lipoprotein expression was found in the plasma of male and female SHHR and HLR. Cholesterol 7alpha-hydroxylase mRNA expression in the liver of male SHHR was lower than that of male SDWKY rats. Endothelium lesions and lipid deposition under the endothelium were observed in the aorta of 24-month-old SHHR, especially female SHHR, but not in age-matched HLR and SDWKY rats. 4. The hypertension of this new animal model of hypertension and hyperlipidaemia may be related to increased catecholamine activity and the hyperlipidaemia may be related to changes in the expression of cholesterol 7alpha-hydroxylase mRNA and lipoprotein profiles. The SHHR may be valuable in the study of mechanisms of atherosclerosis and the evaluation of anti-atherosclerosis drugs as a new strain of cardiovascular disease.