Pair copula construction for longitudinal data with zero-inflated power series marginal distributions.
Assessing the temporal dependency among outcomes under investigation is critical in many fields. One complication in the modeling process of the discrete longitudinal data is the presence of excess zeros. We propose a framework for modeling count repeated measurements using members of power series family of distributions. The framework accommodates count outcomes having extra zeros. The longitudinal observations of response variable is modeled using pair copula constructions with a D-vine structure. The maximum likelihood estimates of parameters are obtained using a two-stage approach. Some simulation studies are performed for illustration of the proposed methods, for comparing its performance with that of a generalized linear mixed effects (GLME) model and for assessing the robustness of D-vine and GLME models with respect to the distribution of random effects. In the empirical analysis, the proposed method is applied for analysing a real data set of a kidney allograft rejection study.


Posthypnotic Amnesia in Hypnotizability Assessment: Validation of a New Scoring System for the Hypnotic Induction Profile.
The Hypnotic Induction Profile (HIP) is a standardized assessment of hypnotizability featuring a validated 0-10 scoring system, that does not factor in posthypnotic amnesia. Using confirmatory factor analyses (CFA), we compared the 10-point scoring system with a new 12-point system that includes the posthypnotic amnesia item in independent samples of individuals with fibromyalgia (n = 98) and healthy adults (n = 97). Additionally, we explored associations of the two scoring systems with measures of hypnotic phenomena. CFA results indicate that the 12-point scoring system is a good fit for the 1-factor model of hypnotizability. Posthypnotic amnesia loaded highly on the model in the fibromyalgia sample, and moderately on the model in healthy adults. Furthermore, the 12-point scoring system correlated significantly with measures of hypnotic phenomena. We conclude that the 12-point scoring system is psychometrically equivalent yet conceptually more comprehensive than the 10-point scoring system.


The importance of early parenting in at-risk families and children's social-emotional adaptation to school.
The aim of this study was to determine the specific aspects of early parenting in psychosocially at-risk families most strongly related to children's social-emotional adaptation to school. A cohort study of families (N = 318) identified as at risk for maltreatment of their newborns was conducted. Quality of early parenting was observed in the home when the child was 1 year old. Social-emotional adaptation to school was reported by teachers in first grade. Multivariable models assessed the independent influence of early parenting variables on social-emotional adaptation. Early parenting and social-emotional adaptation to school varied greatly across families. Parental warmth was associated with lower teacher ratings of shyness, concentration problems, and peer rejection. Parental lack of hostility was associated with decreased teacher ratings of concentration problems and peer rejection. Parental encouragement of developmental advance was associated with lower ratings of aggression and peer rejection. Provision of materials to promote learning and literacy was associated with lower ratings of concentration problems. In this sample of families with multiple psychosocial risks for child maltreatment, specific aspects of early parenting were associated with better social-emotional adaptation to school in the first grade in theoretically predicted ways. Improving parental knowledge about positive parenting via anticipatory guidance should be a focus of well-child visits. Well-child visit-based interventions to improve the quality of early parenting, especially among at-risk families, should be studied for their impact on parenting behavior and on children's successful social-emotional adaptation to school. Primary care providers should reinforce complementary services, such as home visiting, that seek to promote positive parenting.


External exposure estimates for individuals near the Nevada Test Site.
Individuals living near the Nevada Test Site were exposed to both beta and gamma radiations from fission products and activation products resulting from the atmospheric testing of nuclear devices. These exposures, and the resultant doses, were functions of the amount of material deposited, the time of arrival of the debris, and the amount of shielding afforded by structures. Results are presented for each of nine generic life-styles. These are representative of the living patterns of people residing in the area. For each event at each location for which data exist, a representative of each life-style was closely followed for 30 d. The results of these detailed calculations were then extrapolated to the present, employing a stochastic model. Results displayed are the geometric means and standard deviations derived from 25 independent determinations of the various quantities shown. For each determination, required parameters were randomly selected from appropriate distributions. Calculations yielded estimates for: 1) whole-body and skin dose due to gamma rays from material on the ground; 2) skin dose due to beta particles from material deposited directly on the skin; and 3) skin dose due to beta particles from material deposited on the ground. Organ dose estimates were established from the whole-body dose using appropriate conversion factors. For the homemaker life-style, the uterus dose was also calculated as a function of time for 9 mo. This assisted in estimating fetal dose as a function of gestation.


Cognitive functions in patients with phenylketonuria in long-term treatment with tetrahydrobiopterin.
Cognitive functions were assessed in 9 patients with mild to moderate phenylketonuria (PKU) ranging from 6 to 18 years of age, who were in long-term treatment (>5 years) with 5-9 mg/kg/day tetrahydrobiopterin (BH4) on compassionate use, provided by Schircks Inc. An extensive study of cognitive functions (intelligence quotient (IQ), visuospatial, visual memory, fine motor, executive and attentional functions) was conducted, and behavior was assessed using the ADHD Rating Scale and the Behavior Rating Inventory of Executive Function (BRIEF). All patients had normal IQ (M=107, SD=10). The most notable area of impairment was fine motor function, but no significant difference was found between the PKU patients in BH4 treatment who participated in the current study and PKU patients in dietary treatment who participated in a previous study. These results, however, should be interpreted with caution. It is necessary to conduct further studies with a larger number of patients, using more sensitive tests of motor function and using the formulation of BH4 that is currently available.


Cervical spine injuries in patients with head injuries.
It is generally believed that significant head injury after a traffic accident or fall is associated with a high incidence of concurrent cervical spine injury. This study prospectively examined 260 victims of traffic accidents or falls with significant head injury. The incidence of associated cervical spinal injury was only 3.5 per cent. There was no association between the severity of head injury and the incidence of cervical spine injury. The risk of concurrent spinal injury in head injury patients is not higher than the reported risk in patients without head injury. Although care should be taken to protect the cervical spine in patients with head injury, the results of the present investigation show that the risk of cervical spine injury is much less than previously reported.


RecA-Mediated gene conversion and aminoglycoside resistance in strains heterozygous for rRNA.
Clinical resistance to aminoglycosides in general is due to enzymatic drug modification. Mutational alterations of the small ribosomal subunit rRNA have recently been found to mediate acquired resistance in bacterial pathogens in vivo. In this study we investigated the effect of 16S rRNA heterozygosity (wild-type [wt] and mutant [mut] operons at position 1408 [1408wt/1408mut]) on aminoglycoside resistance. Using an integrative vector, we introduced a single copy of a mutated rRNA operon (1408 A-->G) into Mycobacterium smegmatis, which carries two chromosomal wild-type rRNA operons; the resultant transformants exhibited an aminoglycoside-sensitive phenotype. In contrast, introduction of the mutated rRNA operon into an M. smegmatis rrnB knockout strain carrying a single functional chromosomal wild-type rRNA operon resulted in aminoglycoside-resistant transformants. Subsequent analysis by DNA sequencing and RNase protection assays unexpectedly demonstrated a homozygous mutant genotype, rRNAmut/rRNAmut, in the resistant transformants. To investigate whether RecA-mediated gene conversion was responsible for the aminoglycoside-resistant phenotype in the rRNAwt/rRNAmut strains, recA mutant strains were generated by allelic exchange techniques. Transformation of the recA rrnB M. smegmatis mutant strains with an integrative vector expressing a mutated rRNA operon (Escherichia coli position 1408 A-->G) resulted in transformants with an aminoglycoside-sensitive phenotype. Subsequent analysis showed stable heterozygosity at 16S rRNA position 1408 with a single wild-type allele and a single resistant allele. These results demonstrate that rRNA-mediated mutational resistance to aminoglycosides is recessive.


Characterizing complexes with pnicogen bonds involving sp2 hybridized phosphorus atoms: (H2C═PX)2 with X = F, Cl, OH, CN, NC, CCH, H, CH3, and BH2.
Ab initio MP2/aug'-cc-pVTZ searches of the potential surfaces of (H2C═PX)2 complexes, with X = F, Cl, OH, CN, NC, CCH, H, CH3, and BH2, have been carried out to identify and characterize the properties of complexes with P···P pnicogen bonds. All (H2C═PX)2 form equilibrium conformation A dimers with C(2h) symmetry in which A-P···P-A approaches a linear alignment, with A the atom of X directly bonded to P. Conformation A dimers containing the more electronegative substituents are stabilized by a P···P pnicogen bond, have shorter P-P distances, and have binding energies which correlate with the P-P distance. Dimers stabilized by a P···P pnicogen bond and two P···H(b) interactions consist of those with the more electropositive substituents, have shorter P-H(b) distances, and have binding energies which are too high for their P-P distances. Conformation A complexes with P···H(b) interactions in addition to the P···P bond are more stable than the corresponding (PH2X)2 complexes, while with only one exception, complexes stabilized by only a P···P bond are less stable than the corresponding (PH2X)2 complexes. In the region of the potential surfaces with C-P···P-C approaching linearity (conformation B), the only planar equilibrium complex is (H2C═POH)2, which is stabilized primarily by two O-H···P hydrogen bonds. The remaining (H2C═PX)2 complexes are not stabilized by pnicogen bonds, but by π interactions between the two H2C═PX monomers which are in parallel planes. When A-P···P-C approaches linearity, two types of equilibrium structures with P···P bonds exist. Of the conformation C dimers, (H2C═POH)2 is planar and the most stable, with a P···P pnicogen bond and an O-H···P hydrogen bond. (H2C═PH)2 and (H2C═PCH3)2 are also planar, and stabilized by a P···P pnicogen bond and a P···H(b) interaction. The absence of a P···H(b) interaction results in nonplanar C' conformations with structures in which the monomers essentially retain their symmetry plane, but the plane of one molecule is rotated about the P···P bond relative to the other. C and C' dimers are less stable than the corresponding A dimers, except for (H2C═PCH3)2. (31)P chemical shielding patterns are consistent with the changing nature of the interactions which stabilize (H2C═PX)2 complexes. EOM-CCSD (31)P-(31)P spin-spin coupling constants increase quadratically as the P-P distance decreases.


Digital holographic three-dimensional Mueller matrix imaging.
A digital holographic Mueller matrix imaging method is proposed. A Mueller matrix is obtained from Stokes vectors that are calculated from the complex amplitude of the two orthogonal polarized lights. The complex amplitude is obtained by digital holography. In the proposed method, three-dimensional Mueller matrix imaging is achieved by 12 recordings. Optical experimental results confirm the feasibility of the proposed method by measuring the Mueller matrices of four linear polarizers, a quarter-wave plate, and two linear polarizers placed at different positions.


A Material Showing Colossal Positive and Negative Volumetric Thermal Expansion with Hysteretic Magnetic Transition.
It is an ongoing challenge to design and synthesize magnetic materials that undergo colossal thermal expansion and that possess potential applications as microscale or nanoscale actuators with magnetic functionality. A paramagnetic metallocyanate building block was used to construct a cyanide-bridged Fe-Co complex featuring both positive and negative colossal volumetric thermal-expansion behavior. A detailed study revealed that metal-to-metal charge transfer between 180 and 240 K induced a volumetric thermal expansion coefficient of 1498 MK-1 accompanied with hysteretic spin transition. Rotation of the magnetic building blocks induced change of π⋅⋅⋅π interactions, resulting in a negative volume expansion coefficient of -489 MK-1 , and another hysteretic magnetic transition between 300 and 350 K. This work presents a strategy for incorporating both colossal positive and negative volumetric thermal expansion with shape and magnetic memory effects in a material.


Prognostic Role of Molecular Forms of B-Type Natriuretic Peptide in Acute Heart Failure.
B-type natriuretic peptide (BNP) molecular forms 5-32, 4-32, and 3-32 are known to be present in the circulation of heart failure (HF) patients. This study investigated the prognostic role of circulating BNP molecular forms on risk prediction for patients with acute HF. BNP molecular forms were measured in plasma using an immunocapture MALDI-TOF-mass spectrometry (MS) method. Associations of molecular BNP forms with adverse outcome of all-cause mortality (death) and a composite of all-cause mortality and rehospitalization due to HF (death/HF) at 6 months and 1 year were investigated. BNP molecular forms 5-32, 4-32, and 3-32 were detected in 838 out of 904 patient samples. BNP molecular forms were all able to independently predict death and death/HF at 6 months and 1 year. BNP 5-32 was the superior form with strongest predictive qualities for death at 6 months [adjusted hazard ratio (HR) 1.31, P = 0.005] and 1 year (adjusted HR 1.29, P = 0.002) and death/HF at 1 year (adjusted HR 1.18, P = 0.011). BNP 5-32, 4-32, and 3-32 showed decreased survival rates across increasing tertiles of circulating concentrations (P ≤ 0.004). BNP molecular forms showed prognostic ability comparable with conventional BNP measurements across all end points (P = 0.002-0.032 vs P = 0.014-0.039, respectively) and reduced associations with renal dysfunction (blood urea; Spearman correlation rs = 0.187-0.246 vs rs = 0.369, respectively). BNP molecular forms, notably BNP 5-32, showed association with poor prognosis at 6 months and 1 year in patients with acute HF. This is the first study reporting the prognostic ability of molecular BNP forms in HF patients and demonstrated comparable qualities to conventional BNP measurements.


Maternal pregnancy weight gain and the risk of placental abruption.
To evaluate the relationship between pregnancy weight gain and placental abruption, Missouri's population-based, maternally linked, longitudinal dataset (1989-2005, n = 1,146,935) was assessed. Regardless of baseline body mass index, women who gained less than the optimal amount recommended by the Institute of Medicine had a 67% increased likelihood of placental abruption (adjusted odds ratio [AOR] for placental abruption = 1.673; 95%CI = 1.588-1.762) compared with those who gained an optimal amount of weight, while those who gained more than the recommended optimal amount of weight had a 30% reduced AOR for placental abruption (AOR = 0.695, 95%CI = 0.660-0.731). These findings underscore the importance of maternal weight management as part of preconception care to improve pregnancy outcomes.


Reduced growth of human sarcoma xenografts in hosts homozygous for the lit mutation.
Prior studies have shown that sarcoma growth can be stimulated by insulin-like growth factor-I (IGF-I). To extend this line of research, we carried out in vivo growth studies of primary human sarcoma in immunosuppressed control and IGF-I-deficient mice. Human sarcoma specimens (one osteosarcoma and seven soft tissue sarcomas) were harvested in the operating room and implanted in immunosuppressed mice. Second-generation sarcomas were transplanted to control (GH replete lit/+ mice) and to experimental (GH/IGF-I-deficient lit/lit) animals. When tumors reached 1,000 mm(3) in one group, average tumor size was compared in the two groups. IGF-I receptor expression was measured by RT-PCR and IGF-I receptor binding sites were assayed by radiolabeled IGF-I. Five of eight sarcomas demonstrated reduced growth in the GH/IGF-I-deficient lit/lit animals. In four of the five sarcomas that demonstrated growth inhibition, IGF-R was elevated relative to placenta or a positive control cell line (MCF-7, which is known to be responsive to IGF-I in vitro and in vivo). In three of the five sarcomas that demonstrated growth suppression, IGF-R was elevated twofold after implantation in the experimental IGF-I-deficient animals. The GH-IGF axis may be an important stimulator of tumor growth in sarcomas. These experiments suggest that IGF suppression may inhibit sarcoma growth in vivo.


Diagnostic evaluation of the substantia nigra.
An improved method for the histopathological evaluation of the substantia nigra for diagnostic purposes is proposed. This method takes advantage of the natural architecture of the constituent cell clusters of the substantia nigra by combining standardized sections in the horizontal plane and simple quantification. The use of this standardized technique should increase the diagnostic reliability and sensitivity of patterns of cell loss within the substantia nigra.


[Impact of high remnant-like lipoproteins on the function of peripheral arterial endothelium in patients with hyperlipidemia and normolipidemia: estimation using Doppler flow analysis].
Cholesterol levels of remnant-like lipoprotein (RLP-cholesterol: RLP-C), which reflect remnant lipoproteins, are associated with coronary arterial endothelial functions and cardiovascular events. The influence of RLP-C on peripheral vascular endothelial functions in patients with hyperlipidemia and normolipidemia was evaluated based on the reactivity of the antecubital artery to acetylcholine (Ach) and isosorbide dinitrate (ISDN) using a Doppler guidewire. Protocol 1: Five patients were selected, and the dose-response of the antecubital artery was evaluated by administering Ach (0.5, 5, and 50 micrograms/30 sec) or ISDN (0.025, 0.25, and 2.5 mg/30 sec). An index of vascular reactivity (brachial artery response: BR) was determined by dividing the maximal blood flow velocity after the administration of Ach or ISDN by resting blood flow velocity. Protocol 2: BR was evaluated in 48 patients after administering Ach 50 micrograms or ISDN 2.5 mg. Subsequently, these patients were divided into the following three groups based on early morning RLP-C levels: Group L (n = 11), RLP-C < 2.0 mg/dl (minimal detectable level); Group M (n = 21), 2.0 mg/dl < or = RLP-C < 5.0 mg/dl; and Group H (n = 16), 5.0 mg/dl < or = RLP-C. The factors that regulate BR-Ach 50 micrograms were also evaluated in 34 normolipidemic patients. Protocol 1: BR dose-dependently increased after the administration of Ach and ISDN. Protocol 2: BR-Ach 50 micrograms was significantly less in Group H (3.1 +/- 0.8) than in Groups M (3.8 +/- 0.9, p < 0.03) and L (4.2 +/- 0.9, p < 0.01). However, there were no significant differences in BR-ISDN 2.5 mg between the three groups. Univariate analysis in normolipidemic patients revealed that BR-Ach 50 micrograms was correlated with age (r = -0.355, p < 0.05), RLP-C (r = -0.488, p < 0.01), low-density lipoprotein cholesterol (r = -0.382, p < 0.03), systolic blood pressure (r = -0.354, p < 0.05), and diastolic blood pressure (r = -0.406, p < 0.02). Multivariate analysis using these five factors as independent variables revealed that age, RLP-C, and low-density lipoprotein cholesterol regulated BR-Ach 50 micrograms. Peripheral vascular endothelial dysfunction may occur in patients with high levels of RLP-C. RLP-C is an independent lipid factor that regulates peripheral vascular endothelial functions even in normolipidemic patients.


[HPLC-MS/MS for determination of astragalin in rat plasma and its pharmacokinetics].
To develop a LC-MS/MS method for determination of astragalin in rat plasma and study its pharmacokinetics in rats. The analytical column was packed with ZORBAX SB-C18, and the mobile phase was methanol-water containing 10 mmol/L ammonium acetate-formic acid (80:20:0.15, V/V/V). Quercetin was used as the internal standard (IS). Multiple reaction monitoring (MRM) mode was employed, and the transition of m/z was m/z 449.1→m/z 287.1 for astragalin, and m/z 301.1→m/z 151.1 for IS. A good liner relationship was obtained within the range of 1.00-1000 ng/ml (r(2)=0.9929), and the lower limit of quantification of astragalin was 1.00 ng/ml in rat plasma. The extraction recoveries were all above 93%. After oral administration of astragalin, the maximum plasma concentration of astragalin was 231.1∓67.3 ng/ml and the time to reach this value was 0.5∓0.1 h, with a half-life of 3.9∓1.3 h and an AUC of 782.6∓152.8 ng·h/ml. The method is highly sensitive, selective and rapid for determination of the concentration of astragalin in rat plasma to facilitates the study of its pharmacokinetics.


Transgenic overexpression of amphiregulin induces a mitogenic response selectively in pancreatic duct cells.
The epidermal growth factor (EGF) receptor family and the corresponding ligands are frequently overexpressed in pancreatic cancer. To compare the biological effects of transforming growth factor (TGF)-alpha and amphiregulin (AR) on growth and differentiation of the exocrine pancreas, we have generated transgenic mice overexpressing AR under control of the elastase promoter. Two independently generated transgenic mouse lines overexpress 50-, 43-, 28-, 26-, and 16-kilodalton AR forms in the pancreas. Morphologic and immunohistochemical examinations suggest that small intralobular duct and centro-acinar cells proliferate in response to AR in these mice. AR transgenic mice display increased Ras, Erk1/2, cyclin D/CDK4, and cyclin E/CDK2 activity and G1/S progression in pancreatic duct cells. In contrast to TGF-alpha transgenic mice, AR neither induced tubular complex formation nor elicited a strong fibrogenic response. AR induced a slight induction of ErbB2 on duct cells, whereas TGF-alpha resulted in overexpression of the EGF receptor in cells within tubular complexes. Furthermore, AR and TGF-alpha displayed different effects on differentiation of isolated acini in vitro comparable to the situation in vivo. These data suggest that AR induces a mitogenic response selectively in small duct cells through activation of Ras, CDK2, and CDK4, respectively. The closely related EGF receptor ligands, AR and TGF-alpha, display different biological effects when overexpressed in the exocrine pancreas in vivo.


Integrating a relaxation response-based curriculum into a public high school in Massachusetts.
Academic and societal pressures result in U.S. high school students feeling stressed. Stress management and relaxation interventions may help students increase resiliency to stress and overall well-being. The objectives of this study were to examine the feasibility (enrollment, participation and acceptability) and potential effectiveness (changes in perceived stress, anxiety, self-esteem, health-promoting behaviors, and locus of control) of a relaxation response (RR)-based curriculum integrated into the school day for high school students. The curriculum included didactic instruction, relaxation exercises, positive psychology, and cognitive restructuring. The intervention group showed significantly greater improvements in levels of perceived stress, state anxiety, and health-promoting behaviors when compared to the wait list control group. The intervention appeared most useful for girls in the intervention group. The results suggest that several modifications may increase the feasibility of using this potentially effective intervention in high schools.


Nonparametric estimates of drift and diffusion profiles via Fokker-Planck algebra.
Diffusion processes superimposed upon deterministic motion play a key role in understanding and controlling the transport of matter, energy, momentum, and even information in physics, chemistry, material science, biology, and communications technology. Given functions defining these random and deterministic components, the Fokker-Planck (FP) equation is often used to model these diffusive systems. Many methods exist for estimating the drift and diffusion profiles from one or more identifiable diffusive trajectories; however, when many identical entities diffuse simultaneously, it may not be possible to identify individual trajectories. Here we present a method capable of simultaneously providing nonparametric estimates for both drift and diffusion profiles from evolving density profiles, requiring only the validity of Langevin/FP dynamics. This algebraic FP manipulation provides a flexible and robust framework for estimating stationary drift and diffusion coefficient profiles, is not based on fluctuation theory or solved diffusion equations, and may facilitate predictions for many experimental systems. We illustrate this approach on experimental data obtained from a model lipid bilayer system exhibiting free diffusion and electric field induced drift. The wide range over which this approach provides accurate estimates for drift and diffusion profiles is demonstrated through simulation.


Changing incidence of out-of-hospital ventricular fibrillation, 1980-2000.
Recent reports from 2 European cities and an earlier observation from Seattle, Wash, suggest that the number of patients treated for out-of-hospital ventricular fibrillation (VF) has declined. To analyze the incidence of cardiac arrest and to examine relationships among incidence, sex, race, age, and first identified cardiac rhythm in Seattle. Population-based study of all cardiac arrest cases with presumed cardiac etiology who received advanced life support from Seattle Fire Department emergency medical services during specified periods between 1979 and 2000. United States Census data for Seattle in 1980, 1990, and 2000 were used to determine incidence rates for treated cardiac arrest with adjustments for age and sex. Changes in incidence of cardiac arrest and initial recorded cardiac rhythm. The adjusted annual incidence of cardiac arrest with VF as the first identified rhythm decreased by about 56% from 1980 to 2000 (from 0.85 to 0.38 per 1000; relative risk [RR], 0.44; 95% confidence interval [CI], 0.37-0.53). Similar reductions occurred in blacks (54%; RR, 0.45; 95% CI, 0.26-0.79) and whites (53%; RR, 0.47; 95% CI, 0.38-0.58) and was most evident in men (57%; RR, 0.43; 95% CI, 0.35-0.53), in whom the baseline incidence was relatively high. When all treated arrests with presumed cardiac etiology were considered, that incidence decreased by 43% (RR, 0.58; 95% CI, 0.49-0.67) in men but negligibly in women, for whom a relatively low incidence of VF also declined but was offset by more cases with asystole or pulseless electrical activity. We observed a major decline in the incidence of out-of-hospital VF and in all cases of treated cardiac arrest presumably due to heart disease in Seattle. These changes likely reflect the national decline in coronary heart disease mortality.


Centrally produced neuronal nitric oxide in the control of baroreceptor reflex sensitivity and blood pressure in normotensive and spontaneously hypertensive rats.
We studied the effect of chronic nitric oxide synthase (NOS) blockade in the brain on mean arterial pressure [MAP (mmHg)], heart rate [HR (bpm)] and baroreceptor reflex sensitivity [BRS (mean slope: bpm/mmHg)] in Wistar-Kyoto (WKY) and spontaneously hypertensive rats (SHR). Intracerebroventricular (i.c.v.) infusion of the nonselective NOS inhibitor N-Nitro-L-arginine-methylester (L-NAME) (50 microg/kg per day, 11-12 days) increased MAP in WKY and SHR (125+/-2.1 vs 118+/-1.1 controls, P<0.01 and 179+/-3.59 vs 156+/-4.0 controls, P<0.001, respectively) without affecting HR. In L-NAME-treated WKY, BRS to bradycardia was suppressed (-0.79+/-0.09 vs -1.76+/-0.17 controls, P=0.001), whereas in SHR, L-NAME did not affect BRS to bradycardia. BRS to tachycardia remained unaffected in either strain. In WKY, 7-nitroindazole (7-NI x Na+) (34 microg i.c.v./kg per day, 11-12 days), a selective nNOS inhibitor, did not affect MAP or HR, but BRS to bradycardia and tachycardia was decreased (-0.37+/-0.20 vs -0.97+/-0.41 controls, P<0.01 and -1.78+/-0.20 vs -2.52+/-0.40 controls, P=0.05, respectively). In SHR, the same dose of 7-NI x Na+ increased resting MAP (171+/-5.00 vs 150+/-7.00 controls, P<0.05) without affecting HR or BRS to bradycardia or tachycardia. Thus in WKY, BRS to acute changes in systemic blood pressure (BP) is regulated by NO produced by nNOS in the brain, serving as a neurotransmitter in sympathetic and parasympathetic efferent pathways. In SHR, systemic BP is regulated in part by NO released by the type I NOS isoenzyme in the brain.


Chemical Nature of Electrode and the Switching Response of RF-Sputtered NbOx Films.
In this study, the dominant role of the top electrode is presented for Nb2O5-based devices to demonstrate either the resistive switching or threshold characteristics. These Nb2O5-based devices may exhibit different characteristics depending on the selection of electrode. The use of the inert electrode (Au) initiates resistive switching characteristics in the Au/Nb2O5/Pt device. Alternatively, threshold characteristics are induced by using reactive electrodes (W and Nb). The X-ray photoelectron spectroscopy analysis confirms the presence of oxide layers of WOy and NbOx at interfaces for W and Nb as top electrodes. However, no interface layer between the top electrode and active layer is detected in X-ray photoelectron spectroscopy for Au as the top electrode. Moreover, the dominant phase is Nb2O5 for Au and NbO2 for W and Nb. The threshold characteristics are attributed to the reduction of Nb2O5 phase to NbO2 due to the interfacial oxide layer formation between the reactive top electrode and Nb2O5. Additionally, reliability tests for both resistive switching and threshold characteristics are also performed to confirm switching stabilities.


Tissue Pathogens and Cancers: A Review of Commonly Seen Manifestations in Histo- and Cytopathology.
Tissue pathogens are commonly encountered in histopathology and cytology practice, where they can present as either benign mimickers of malignancy or true malignancies. The aim of this review is to provide a timely synthesis of our understanding of these tissue pathogens, with an emphasis on pertinent diagnostic conundrums associated with the benign mimickers of malignancy that can be seen with viral infections and those which manifest as granulomas. The oncogenic pathogens, including viruses, bacteria, and parasites, are then discussed with relationship to their associated malignancies. Although not exhaustive, the epidemiology, clinical manifestations, pathogenesis, and histological findings are included, along with a short review of emerging therapies.


Plasmin potency estimates: influence of the substrate used in assay.
A urokinase-activated plasmin (UK-plasmin) preparation was assayed against the International Reference Preparation for Plasmin (IRP-plasmin) using caseinolytic, fibrinolytic, fibrinogenolytic and chromogenic assay methods. The relative potency (using multi-dose bioassays) was estimated by the fibrinolytic method to be about twice that obtained by the caseinolytic, fibrinolytic and chromogenic assay methods. It was found that the UK-plasmin binds to fibrin to a greater extent than does the IRP-plasmin and this is advanced as an explanation for the discrepancy between assay methods. This difference in the binding of the two plasmins to fibrin may mean that it will be difficult to compare the fibrinolytic activities of various plasmin preparations. It is also shown that, during thermal degradation, the IRP-plasmin loses fibrinolytic activity more rapidly than amidolytic activity.


The Effects of Providing Advance Notice and Stress-Coping Traits on Physiological Stress of Patients during Dental Treatment.
Patients tend to feel stress in association with dental treatment due to uneasiness and fear. We investigated the effects of providing advance notice and stress-coping traits on the physiological stress of patients during dental treatment. Sixty non-dental professionals (male, n = 26; female, n = 34; mean age, 49.9 years) were recruited for this study and informed consent was obtained. Subjects were given simulated dental treatment including three stimuli, air, percussion and running of an air turbine, with or without advance notice of the stimulation during dental treatment. Real-time sympathetic nerve activity (SN) and parasympathetic nerve activity (PN) during the treatment were measured using a biological information monitor. The stress-coping traits of each subject were examined using the Lazarus-Type Stress Coping Inventory (SCI). Correlations between the nerve-activity and scores of eight stress-coping strategies of SCI and the presence/absence of advance notice were analyzed. Age, types of stimuli and order of stimuli significantly affected SN, while age, types of stimuli, and the pattern of stimulation significantly affected PN. The interaction of the stress-coping trait and presence/absence of advance notice significantly affected PN. Providing advance notice may have different effects on physiological stress depending on how the patient copes with stress.


Designer recombinases: Tools to cut and paste genomic DNA sequences.
Extract: Gene therapy is still very much in its infancy. In future, we are likely to see more and more sophisticated intervention in order to repair or replace faulty genes, or to introduce beneficial new genes. Site-specific recombinases have the potential to be engineered into tools for "cut and paste" genome editing. Our paper in PNAS shows how this might be done. Site-specific recombinases rearrange DNA sequences, by catalyzing cleavage and rejoining of DNA strands at specific sites to which they bind. They can promote excision, integration, or orientation inversion of DNA segments bounded by the sites. They are now used extensively in mammals and other eukaryotes, for experimental research or projected gene therapy and biotechnology applications. The most commonly used enzymes are Cre and FLP, both "tyrosine recombinases" (so-called because the nucleophilic residue that breaks the DNA strands is a tyrosine), but members of another, unrelated "serine recombinase" family also function in eukaryotes.


The past, present, and future of continuing medical education. Achievements and opportunities, computers and recertification.
Changes in continuing medical education (CME) during the past three decades have been controversial and complex. A 1950s-style, small-scale, voluntary activity has evolved, in 1987, into broad-scale programs with accredited sponsors and with ties to relicensure. Within the next three decades, CME will be directed by methods chosen by specialty boards for recertification and by exploitation of computer and telecommunication technology. Written recertification examinations can waste physicians' time studying material that will not improve care of their patients. We hope improved methods of analysis of individual practices, on-the-spot access to pertinent medical information, and better communication among physicians can be incorporated into recertification procedures. Policies established now will shape CME for decades to come. We encourage a coordinated effort by medical specialty boards, medical societies, hospitals, medical schools, computer corporations, telecommunication firms, granting agencies, and the National Library of Medicine to ensure the most effective and efficient recertification and CME policies.


[Personal resources in inpatient psychotherapy - Relationships and development].
We examined how personal resources develop during inpatient psychodynamic psychotherapy, their relationship to symptom development, Operationalized Psychodynamic Diagnosis (OPD-2), and sociodemographic aspects. 483 patients were examined using self-assessment questionnaires (Brief Symptom Inventory (BSI), Hospital Anxiety and Depression Scale (HADS), Inventory of Interpersonal Problems (IIP), Questionnaire of Actual Resource Realization (RES)) as well as the expert ratings Global Assessment of Functioning Scale (GAF), Impairment Score (IS), OPD-2, and the Heidelberg Structural Change Scale (HSCS), both at the beginning and the end of treatment. There was a marked improvement in the realization of personal resources during inpatient psychotherapy, which showed significant correlations to the improvement of symptoms. The extent of improvement of resources correlated with the duration of psychotherapy and the assessment of the psychodynamic therapy foci on the HSCS. The results show that personal resources are activated when successfully working on the psychodynamic foci in psychodynamic inpatient treatment, and that this corresponds to an improvement of symptoms.


Co-occurrence of 12-month mood and anxiety disorders and personality disorders in the US: results from the national epidemiologic survey on alcohol and related conditions.
The objective of this study was to determine the prevalence and co-occurrence of DSM-IV personality disorders (PDs) among individuals with current DSM-IV mood and anxiety disorders in the US population and among individuals who sought treatment for such mood or anxiety disorders. Face-to-face interviews were conducted with 43,093 individuals, 18 years and older, in the National Institute on alcohol abuse and alcoholism's 2001-2002 National epidemiologic survey on alcohol and related conditions (NESARC). Odds ratios (ORs) were calculated to determine the prevalence and associations between current DSM-IV axis I and axis II disorders. Associations between mood, anxiety and PDs were all positive and statistically significant. Avoidant and dependent PDs were more strongly related to mood and anxiety disorders than other PDs. Associations between obsessive-compulsive PD and mood and anxiety disorders were significant, but much weaker. Paranoid and schizoid PDs were most strongly related to dysthymia, mania, panic disorder with agoraphobia, social phobia and generalized anxiety disorder, while histrionic and antisocial PDs were most strongly related to mania and panic disorder with agoraphobia. Results of this study highlight the need for further research on overlapping symptomatology, factors giving rise to the associations and the treatment implications of these disorders when comorbid.


[Epidemiological and clinical aspects of prurigo in HIV infected patients in Fousseyni N'Daou hospital of Kayes, Mali].
Prurigo is one of the most common dermatological conditions during HIV infection and AIDS. It appears as an immunosuppression marker associated with HIV infection. The study objective was to determine the prevalence of prurigo in people living with the human immunodeficiency virus (PLHIV) in Fousseyni N'Daou Hospital of Kayes, and to describe the socio-demographic aspects of patients and lesions associated with prurigo among PLHIV. It was a descriptive cross-sectional study included all cases of HIV infected patients with prurigo in the Dermatology-Venomology Department of Fousseyni N'DAOU Hospital from January 1, 2015 to August 31, 2015. We collected 121 cases of prurigo. The hospital prevalence was 14.5% among PLHIV with 65% Female and the average age was 34.8 years old (SD: 15-81 years). The elementary lesions associated with prurigo were seropapules (40.2%), vesiculo-crusts (13%), excoriated papules (33.3%), lichenified papules (10.8%), and cicatricial lesions (2.7%). The prurigo was generalized in 68.5% of cases and localized in 31.24%. More than half of our patients had weight loss, fever, diarrhea and oral candidiasis in their medical history. Patients were infected with HIV1 in 60.03% and HIV1+ 2 in 24.3%. More than the half of our patients had a CD4 count inferior to 250 cells/mm3 at the time of prurigo diagnosis. In our study, prurigo remains a common condition in PLHIV, particularly in patients with low CD4 counts. Early detection and rapid antietroviral therapy can reduce the frequency of prurigo in PLHIV.


The absence of a surface protease, OmpT, determines the intercellular spreading ability of Shigella: the relationship between the ompT and kcpA loci.
A large plasmid-encoded protein, VirG, on the bacterial surface is essential for the spreading of Shigella by eliciting polar deposition of filamentous actin in the cytoplasm of epithelial cells. VirG expression from the large plasmid is diminished greatly when it is introduced into Escherichia coli K-12 from Shigella. In an attempt to identify factors affecting VirG expression, we found that the absence of the ompT gene, encoding outer membrane protease OmpT, restored full production of VirG protein to E. coli K-12. Conversely, upon introduction of the ompT gene of E. coli K-12 into Shigella, spreading ability was completely abolished, probably because of the proteolytic degradation of VirG protein by OmpT. Analysis of the DNA sequence of the ompT region indicated that the absence of the ompT gene occurred in Shigella and enteroinvasive E. coli strains, and that the absent DNA segment corresponded to a remnant lambdoid phage structure found in E. coli K-12, which encompasses a 21 kb DNA segment spanning from argU through to the ompT genes. Since ompT is located near purE in E. coli K-12 and a virulence locus for provoking keratoconjunctivitis in the eyes of guinea-pigs, named kcpA, is located near purE in S. flexneri, and the two loci are involved in VirG expression, the KcpA- mutants of S. flexneri 2a constructed were examined for correlation between acquisition of ompT and VirG degradation. Our data suggest that the previous recognition of a kcpA locus in S. flexneri is the result of transfer of the ompT gene from E. coli K-12, giving rise to a KcpA- phenotype. These results indicate that the lack of OmpT protease confers upon Shigella the ability to spread into adjacent epithelial cells.


Beta-adrenoceptor-mediated inhibition of mediator release from human peripheral blood-derived mast cells.
1. Mast cells cultured from human peripheral blood have been used as a cell model for functional studies of human mast cells, particularly human lung mast cells. However, the beta-adrenoceptor subtype expressed by these cultured cells has not been identified. The aim of the present study was to characterize pharmacologically the beta-adrenoceptors involved in the suppression of IgE-mediated release of mediators, including histamine, prostaglandin (PG) D2 and leukotriene (LT) C4 from cultured mast cells. 2. Mast cells were cultured from mast cell progenitors isolated from peripheral blood in the presence of 200 ng/mL stem cell factor and 50 ng/mL interleukin-6. Mast cells were sensitized with human myeloma IgE, treated with beta-adrenoceptor agonists or antagonist and then challenged with anti-human IgE. The release of histamine, PGD2 and LTC4 from mast cells was determined. 3. Both isoprenaline and salbutamol inhibited anti-IgE-induced release of histamine, PGD2 and LTC4 from cultured mast cells in a dose-dependent manner. Isoprenaline was a more potent inhibitor than salbutamol. The pD2 values for the inhibition of the release of histamine, PGD2 and LTC4 were 7.37 +/- 0.12, 8.38 +/- 0.23, 8.85 +/- 0.23, respectively, for isoprenaline and 6.96 +/- 0.12, 7.65 +/- 0.36, 7.91 +/- 0.64, respectively, for salbutamol. The selective beta3-adrenoceptor agonist BRL-37344 failed to affect anti-IgE-induced histamine release from cultured mast cells. 4. The selective beta2-adrenoceptor antagonist ICI 118 551 (108 mol/L) strongly reversed the concentration-dependent suppression of histamine release by isoprenaline and salbutamol; however, the selective beta1-adrenoceptor antagonist atenolol (106 mol/L) did not have any effect. 5. These results indicate that both isoprenaline and salbutamol act at beta2-adrenoceptors to suppress IgE-mediated mediator release from cultured human mast cells.


Community-associated methicillin-resistant Staphylococcus aureus, Minnesota, 2000-2003.
We compared characteristics of community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) skin and soft tissue infections (SSTIs) and CA-MRSA invasive disease identified in Minnesota from 2000 through 2003. A total of 586 patients with SSTIs and 65 patients with invasive disease were identified. Patients with invasive disease were more likely to be smokers (p = 0.03), and report a history of immunosuppressive therapy (p = 0.03), emphysema (p = 0.011), or injection drug use (p = 0.020) than were SSTI patients. Invasive disease isolates were less likely to be susceptible to ciprofloxacin (p = 0.002) and clindamycin (p = 0.001) and more likely to have healthcare-associated pulsed-field gel electrophoresis subtypes than SSTI isolates (p<0.001). Patients with invasive disease may have had healthcare exposures that put them at risk of acquiring healthcare-associated MRSA and which were not exclusion criteria in the CA-MRSA case definition. Continued surveillance of MRSA is needed to better characterize CA-MRSA infections.


Chronic exposure to superantigen induces regulatory CD4(+) T cells with IL-10-mediated suppressive activity.
The repeated injection of bacterial superantigens (SAg), such as staphylococcus enterotoxin (SE) A or B, has been shown in mice to induce a state of unresponsiveness characterized by the lack of secretion of Th1 lymphokines, such as IL-2 and IFN-gamma, following subsequent SAg challenge. We made the observation, in vivo as well as in vitro, that unresponsiveness to SAg could be transferred from SEA- to SEB-reactive T cells (and reversibly from SEB- to SEA-specific T cells) in C57BL/6 mice but not in BALB/c mice. Since C57BL/6 mice, unlike BALB/c mice, possess TCR V(beta)3+ and V(beta)11+ T cells able to react with both SEA and SEB, we hypothesized that SAg-unresponsive V(beta)3(+) and V(beta)11+ T cells could mediate linked suppression of other SAg-reactive T cells. To analyze further this possibility, spleen cells from BALB/c mice made unresponsive to SEB were tested for their capacity to suppress the response of normal BALB/c cells to SEB. The production of both IFN-gamma and IL-2 following SEB stimulation was greatly impaired in co-cultures containing CD4(+) T cells, but not CD8(+) T cells, isolated from unresponsive animals. In vivo, the production of both IFN-gamma and IL-2 responses to SEB was dramatically reduced in animals adoptively transferred with unresponsive spleen cells. This suppression was abrogated in recipients injected with neutralizing anti-IL-10 antibodies. Moreover, in animals made unresponsive to SEB, SAg-reactive CD4(+) T cells were found to express high levels of CTLA-4, a molecule recently described to play an essential role in the suppressive function of regulatory T cells. Taken together these results demonstrate that the repetitive injection of SAg induces the differentiation of regulatory CD4(+) T cells capable of suppressing SAg-reactive naive T cells.


Availability of insecticidal molecules to control Aedes albopictus (Skuse).
Following the implementation of the Directive 98/8/CE a few changes in the availability of insecticidal molecules to control Ae. albopictus have been outlined. Available products for larvicidal treatments will predominantly be based upon two growth regulators (diflubenzuron and pyriproxyfen). For the control of the adult forms there will mostly be active ingredients belonging to the pyrethroid group. Importance of surveillance for the onset of tolerance or resistance phenomena.


Expression and function of miRNA in postoperative radiotherapy sensitive and resistant patients of non-small cell lung cancer.
To investigate the different miRNA expression profiles of postoperative radiotherapy sensitive and resistant patients of non-small cell lung cancer, explore their potential role and find some radio-sensitivity markers. Thirty non-small cell lung cancer patients who have been treated by postoperative radiotherapy were selected and were divided into radiotherapy sensitive group and resistant group according to overall survival and local or distant recurrence rate. Expression profile of miRNA in these two groups was detected by a microarray assay and the results were validated by quantitative RT-PCR and Northern blot. At the molecular level, the effect of one differently expressed miRNA (miR-126) on the growth and apoptosis of SK-MES-1 cells induced by irradiation was examined. Comparing with resistant patients, five miRNAs (miRNA-126, miRNA-let-7a, miRNA-495, miRNA-451 and miRNA-128b) were significantly upregulated and seven miRNAs (miRNA-130a, miRNA-106b, miRNA-19b, miRNA-22, miRNA-15b, miRNA-17-5p and miRNA-21) were greatly downregulated in radiotherapy sensitive group. Overexpression of miRNA-126 inhibited the growth of SK-MES-1 cells and promoted its apoptosis induced by irradiation. The expression level of p-Akt decreased in miRNA-126 overexpression group. After treating with phosphoinositidyl-3 kinase (PI3K) constitutively activator (IGF-1) and inhibitor (LY294002), miRNA-126 overexpression had no significant effects on the apoptosis of SK-MES-1 cells. We found 12 differently expressed miRNAs in the radiotherapy sensitive and resistant non-small cell lung cancer samples. Moreover, our results showed miRNA-126 promoted non-small cell lung cancer cells apoptosis induced by irradiation through the PI3K-Akt pathway.


[Gains in life expectancy by sex and age in Mexico].
"This work estimates the gains in life expectancy, or average years a person lives, at a national level [in Mexico] during the 1950-1980 period by sex and age of the population structure, referring as well to the changes undergone in such gains. An exhaustive presentation of the Pollard method is made in order to enable us to appreciate the advantages and limitations of the methodology used to quantify the gains in life expectancy. It also shows, graphically, the differences by sex and age from which the gain distributions are derived." (SUMMARY IN ENG)


In vivo function of posterior cruciate ligament before and after posterior cruciate ligament-retaining total knee arthroplasty.
The object of this study was to investigate the in vivo function of the posterior cruciate ligament (PCL) in patients before and after a PCL-retaining total knee arthroplasty (TKA). Eleven patients with advanced osteoarthritis (OA) of the knee were recruited. Magnetic resonance scans of each OA knee were obtained, and 3D computer models, including the femoral and tibial insertion areas of the anterolateral and posteromedial bundles of the PCL, were created. Before and after PCL-retaining TKA, dual fluoroscopic images of each knee were acquired during weight-bearing knee flexion. The images and computer models were used to reproduce the in vivo motion of the knee. The function of the PCL bundles was described in terms of elongation, elevation and deviation. Twenty-two healthy controls were also included as normal references. PCL bundles of the OA knees were overstretched during late knee flexion and orientated more medially throughout flexion compared with normal knees. After PCL-retaining TKA, PCL bundles were further overstretched during late flexion and changed from medially directed in normal and OA knees to almost sagittally directed, which may compromise function in controlling knee rotation. The current PCL-retaining TKA systems and surgical techniques may not adequately re-establish normal biomechanics of PCL bundles after PCL-retaining TKA.


A colorimetric assay for high-throughput screening of inhibitors of herpes simplex virus type 1 alkaline nuclease.
Herpes simplex virus type 1 (HSV-1) encodes a deoxyribonuclease that is frequently referred to as alkaline nuclease (AN) because of its elevated pH optimum. Studies with recombinant viruses which contain deletions in the HSV-1 gene encoding AN have indicated that this enzyme is required for efficient virus replication and therefore represents a potential target for novel antiviral therapies. A simple colorimetric assay for deoxyribonuclease activity employing a DNA-methyl green substrate was adapted for use in a high-throughput screen to identify small molecule inhibitors of this enzyme. This screen identified 1,2-benzoisothiazolin-3-one as a specific inhibitor of AN, since it exhibited activity against AN but was completely inactive against bovine pancreatic DNaseI. Subsequent studies revealed that this compound most likely inhibited AN by forming disulfide linkages with one or more exposed cysteine residues on the surface of the enzyme and that AN was sensitive to sulfhydryl-group-modifying reagents in general. These results demonstrated the utility of this DNA-methyl green substrate-based assay in both the rapid identification and the characterization of novel small molecule inhibitors of the AN encoded by HSV-1 and other herpesviruses.


Cultural significance of the flora of a tropical dry forest in the Doche vereda (Villavieja, Huila, Colombia).
In Colombia, ethnobotanical studies regarding plant cultural significance (CS) in tropical dry forests are scarce and mainly focused on the Caribbean region. Different authors have indicated that the plants with the most uses are those of greater cultural importance. Additionally, gender differences in knowledge and interest in natural resources has been widely recorded. This study evaluated the cultural significance of plants in the Doche community, in the Department of Huila. Furthermore, it evaluates the richness of plant knowledge among local inhabitants, looking for testing the hypothesis that the CS of plants positively correlates to the number of uses people inform about, and that there are significant differences on the richness of ethnobotanical knowledge between men and women in this community. The ethnobotanical categories: "food," "condiment," "economy," "fodder," "firewood," "timber", "medicine," and "others" were established to carry out semi-structured interviews, social cartography, and ethnobotanical walks. The frequency of mention was calculated as a measure of CS. The richness of knowledge of each collaborator was obtained. Non-parametric tests were performed to determine whether differences between the numbers of mentioned species existed between genders and ethnobotanical categories. Finally, Pearson correlation tests determined the relationship between CS and the number of ethnobotanical categories. A hundred useful species were registered in crops and forests. The most abundant categories were medicinal (45 species), firewood (30), and fodder (28). The most culturally significant species according to frequency of mention were Pseudosamanea guachapele, Guazuma ulmifolia, Manihot esculenta, and Musa balbisiana. The species with the most registered uses (five) were Guazuma ulmifolia and Gliricidia sepium. We found a correlation between CS and the number of uses per ethnobotanical category, but no significant difference between genders regarding ethnobotanical knowledge. Frequency of mention provides relevant information about the CS of species. Furthermore, it aids to establish sustainable use of tropical dry forests without loss of resources parting from strategies designed from within the Doche community and based on their ethnobotanical knowledge. We found that the number of uses of a plant is correlated with its degree of cultural importance. On the other hand, no significant differences were found between genders regarding ethnobotanical knowledge; that is, both men and women have similar roles in the community, which allows them to recognize the same uses per species.


Increased number of accessible sugar epitopes defined with monoclonal antibody AM-3 on colonic mucins is associated with malignant transformation of colonic mucosa.
Monoclonal antibody AM-3 detects a mucin sugar epitope (AM-3 epitope) the expression of which increases in the course of human colon carcinogenesis parallel to the gradual morphological alterations (so called adenoma-carcinoma sequence). In the present report the AM-3-positive mucin has been purified from human normal and carcinomatous colonic tissue. About 300-fold enrichment of the epitope per protein from both sources was achieved after ultracentrifugation, gel filtration on Sepharose CL-6B and isopyknic gradient centrifugation. Slot-blot and enzyme-linked immunosorbent assays of the purified preparation indicated not only different amounts of the mucins but also a consistent qualitative difference between the molecules from both sources. The qualitative difference could be obliterated by a partial removal of the AM-3 epitope from the tumor-derived mucin with neuraminidase. The visualization of the molecules by rotary shadowing indicated that the mucins from both sources have similar length distribution, 80% of the molecules being 100-600 nm long. The reaction with AM-3 antibody followed by rotary shadowing showed that in the purified preparations more than 95% of the tumor-derived molecules and 74% of the normal colon tissue-derived molecules carried the epitope. The tumor-derived mucins bound, on the average, 34 +/- 15 (SD) antibodies/1000 nm of the protein core while the mucin from normal colon tissue carried 12 +/- 11 antibodies/1000 nm of the protein core. These data indicate that the increased expression of AM-3 epitopes during malignant transformation of the human colon is due to accumulation of AM-3-positive mucin as well as a higher number of accessible AM-3 epitopes on this mucin.


Quercetin increases mitochondrial proteins (VDAC and SDH) and downmodulates AXL and PIM-1 tyrosine kinase receptors in NRAS melanoma cells.
Melanoma is a type of skin cancer with low survival rates after it has metastasized. In order to find molecular differences that could represent targets of quercetin in anti-melanoma activity, we have chosen SKMEL-103 and SKMEL-28 melanoma cells and human melanocytes as models. Firstly, we observed that quercetin was able in reducing SKMEL-103 cell viability, but not in SKMEL-28. Besides that, quercetin treatment caused inhibition of AXL in both cell lines, but upregulation of PIM-1 in SKMEL-28 and downregulation in SKMEL-103. Moreover, HIF-1 alpha expression decreased in both cell lines. Interestingly, quercetin was more effective against SKMEL-103 than kinases inhibitors, such as Imatinib, Temsirolimus, U0126, and Erlotinib. Interestingly, we observed that while the levels of succinate dehydrogenase and voltage-dependent anion channel increased in SKMEL-103, both proteins were downregulated in SKMEL-28 after quercetin's treatment. Furthermore, AKT, AXL, PIM-1, ABL kinases were much more active and chaperones HSP90, HSP70 and GAPDH were highly expressed in SKMEL-103 cells in comparison with melanocytes. Our findings indicate, for the first time, that the efficacy of quercetin to kill melanoma cells depends on its ability in inhibiting tyrosine kinase and upregulating mitochondrial proteins, at least when SKMEL-103 and SKMEL-28 cells response were compared.


Development of RNA aptamer that inhibits methyltransferase activity of dengue virus.
To develop an RNA aptamer specific for the methyltransferase (MTase) of dengue virus (DENV) which is essential for viral genome replication and translation acting directly on N-7 and 2'-O-methylation of the type-I cap structure of the viral RNA. We identified 2'-fluoro-modified RNA aptamers that can specifically bind DENV serotype 2 (DENV2) MTase using systematic evolution of ligands by exponential enrichment technology. We truncated the chosen aptamer into a 45-mer RNA sequence that can bind DENV2 MTase with K d ~ 28 nM and inhibit N-7 methylation activity of the protein. Moreover, the 45-mer truncated aptamer could not only bind with an K d ~ 15.6 nM but also inhibit methylation activity of DENV serotype 3 (DENV3) MTase. The 45-mer aptamer competitively impeded binding of both DENV2 and DENV3 genomic RNA to MTase of each serotype. The selected 45-mer truncated RNA aptamer specifically and avidly bound DENV MTase and competitively inhibited its methylation activity, and thus could be useful for the development of anti-DENV agents.


The complete mitochondrial genome sequence of Hemibagrus sp. (Siluriformes: Bagridae).
At present, the phylogenetic and taxonomic positions of Bagridae and Siluriformes have long been unclear. In this paper, we sequenced and annotated the complete mitogenome of Hemibagrus sp. The total length was 16,529 bp, and was typically consist of 37 genes, including 13 protein-coding genes, 2 rRNAs, 22 tRNA, a light-strand replication origin (OL) and a single large control region (CR). Except for eight tRNA and ND6 genes, all other mitochondrial genes were encoded on the heavy strand (H-strand). The gene content, organization and arrangement were identical to that of a typical bony fish. The overall base composition was 26.5%, 26.8%, 31.7% and 15.0% for T, C, A and G, respectively, with a slight bias on AT content (58.2%). This information could not only contribute to provide useful molecular data for the species identification, but also to further taxonomic and phylogenetic studies of Hemibagrus and Bagridae.


Subpectoral Implantation of Internal Pulse Generators for Deep Brain Stimulation: Technical Note for Improved Cosmetic Outcomes.
Deep brain stimulation is increasingly used to treat a variety of disorders. As the prevalence of this technology increases, greater demands are placed on neurosurgical practitioners to improve cosmetic results, maximize patient comfort, and minimize complication rates. We have increasingly employed subpectoral implantation of internal pulse generators (IPGs) to improve patient satisfaction. To determine the complication rates of subpectorally placed IPGs as compared to those placed in a subcutaneous location. We reviewed a series of 301 patients from a single institution. Complication rates including infection, hematoma, and lead fracture were recorded. Rates were compared for subcutaneously and subpectorally located devices. Of the records reviewed, we found 301 patients who underwent 308 procedures for initial IPG implantation. Of these, 275 were subpectoral IPG implantation, 19 were infraclavicular subcutaneous implantation, and 14 were subcutaneous implantation in the abdomen. A total of 6 IPG pocket infections occurred, 2 subpectoral and 4 infraclavicular subcutaneous. Of the IPG infections, 2 of the infraclavicular subcutaneous devices had associated erosions. Two patients had their devices relocated from a subpectoral pocket to a subcutaneous pocket in the abdomen due to discomfort. Two patients in the subpectoral group suffered from hematoma requiring evacuation. Two patients in the infraclavicular subcutaneous group had lead fracture occur. Subpectoral implantation of deep brain stimulation IPGs is a viable alternative with a low complication rate. This technique may offer a lower rate of infection and wound erosion.


Computational identification of transcription factor binding sites via a transcription-factor-centric clustering (TFCC) algorithm.
While microarray-based expression profiling has facilitated the use of computational methods to find potential cis-regulatory promoter elements, few current in silico approaches explicitly link regulatory motifs with the transcription factors that bind them. We have thus developed a TF-centric clustering (TFCC) algorithm that may provide such missing information through incorporation of biological knowledge about TFs. TFCC is a semi-supervised clustering algorithm which relies on the assumption that the expression profiles of some TFs may be related to those of the genes under their control. We examined this premise and found the vicinities of TFs in expression space are often enriched with the genes they regulate. So, instead of clustering genes based on the mutual similarity of their expression profiles to each other, we used TFs as seeds to group together genes whose expression patterns correlate with that of a particular TF. Then a Gibbs sampling algorithm was applied to search for shared cis-regulatory elements in promoters of clustered genes. Our working hypothesis was that if a TF-centric cluster indeed contains many targets of the seeding TF, at least one of the discovered motifs would be the site bound by the very same TF. We tested the TFCC approach on eight cell cycle and sporulation regulating TFs whose binding sites have been previously characterized in Saccharomyces cerevisiae, and correctly identified binding site motifs for half of them. In addition, we also made de novo predictions for some unknown TF binding sites.


A pilot study of temsirolimus and body composition.
Body weight and composition play a role in cancer etiology, prognosis, and treatment response. Therefore, we analyzed the weight, body composition changes, and outcome in patients treated with temsirolimus, an mTor inhibitor that has weight loss as one of its side effects. Sixteen patients with advanced solid tumors treated with temsirolimus were studied; body composition was evaluated utilizing computerized tomography images. Sarcopenia was defined as skeletal muscle index lower than 38.5 cm(2)/m(2) for women and 52.4 cm(2)/m(2) for men. Five of 16 patients (31 %) were men; median age, 60 years. Forty-four percent (7/16) of patients were sarcopenic. Fatigue, anemia, hyperglycemia, and hyperlipidemia were common. Baseline sarcopenia and body composition did not correlate with worse toxicity or treatment outcome. However, there was a trend for greater loss of adipose area (p = 0.07), fat mass (p = 0.09), and adipose index (p = 0.07) for patients with grade 3 or 4 toxicities versus those with grade 1 and 2 side effects. Patients with higher grade toxicities tended to lose more body fat, suggesting a possible end-organ metabolic effect of temsirolimus. These observations merit exploration in a larger cohort of patients.


Base mismatch-specific endonuclease activity in extracts from Saccharomyces cerevisiae.
An endonuclease activity (called MS-nicking) for all possible base mismatches has been detected in the extracts of yeast, Saccharomyces cerevisiae. DNAs with twelve possible base mismatches at one defined position are cleaved at different efficiencies. DNA fragments with A/G, G/A, T/G, G/T, G/G, or A/A mismatches are nicked with greater efficiencies than C/T, T/C, C/A, and C/C. DNA with an A/C or T/T mismatch is nicked with an intermediate efficiency. The MS-nicking is only on one particular DNA strand, and this strand disparity is not controlled by methylation, strand break, or nature of the mismatch. The nicks have been mapped at 2-3 places at second, third, and fourth phosphodiester bonds 5' to the mispaired base; from the time course study, the fourth phosphodiester bond probably is the primary incision site. This activity may be involved in mismatch repair during genetic recombination.


A novel route to the F-ring of halichondrin B. Diastereoselection in Pd(0)-mediated meso and C(2) biol desymmetrization.
Both sigma and C(2) symmetric diol diacetates were synthesized via two-directional chain elongation. A palladium-mediated, ligand-controlled C(2) diol desymmetrization provided the desired tetrahydrofuran with the correct relative and absolute stereochemistries. Simple functional group manipulation led to the desired F-ring of halichondrin B. Desymmetrization of the meso substrate enantioselectively provided the diastereomer, leading to a refinement of our understanding of the transition state model. [reaction: see text]


Dose escalation study with respiratory-gated carbon-ion scanning radiotherapy using a simultaneous integrated boost for pancreatic cancer: simulation with four-dimensional computed tomography.
Pancreatic cancer is a difficult to treat disease with a persistently high mortality rate. We evaluated dose distribution simulation with respiratory-gated carbon-ion pencil beam scanning (C-PBS) with a simultaneous integrated boost (SIB) to increase tumour dose, sparing organs at risk (OARs). Using four-dimensional CT data of 12 patients, we delineated gross tumour volume and two clinical target volumes (CTVs). To consider beam range intrafractional uncertainty, we calculated field-specific target volumes, from which two planning target volumes (PTVs) were generated. PTV1 would receive a planned dose of 55.2 Gy [relative biological effectiveness (RBE)-weighted absorbed dose] in 12 fractions, and PTV2 would receive an SIB dose up to 67.2 Gy (RBE). Dose assessments were conducted with regard to the targets and OARs. CTV2 dose covering 95% of the volume (D95%) increased from 50.3 ± 5.1 Gy (RBE) to 62.5 ± 3.5 Gy (RBE) for a planned dose from 55.2 Gy (RBE) to 67.2 Gy (RBE). For 4 of 12 patients with a distance of ≥5 mm between the tumour and the gastrointestinal tract, CTV2 D95% was ≥95% of planned dose at all dose levels. We quantified dose escalation with respiratory-gated C-PBS using SIB for pancreatic cancer and revealed that OAR dose was not affected to the same degree as the tumour dose. Advances in knowledge: A simulation study on respiratory-gated C-PBS with SIB for pancreatic cancer was performed. The results indicated the feasibility of dose escalation for pancreatic cancer, which should be confirmed in clinical trials.


Stress and burnout among health-care workers in the coronavirus disease 2019 intensive care unit.
Since late December 2019, the city of Wuhan in China has reported a novel pneumonia caused by coronavirus disease 2019 (COVID-19), which has now spread domestically and internationally. The COVID-19 pandemic has caused unthinkable consequences and has challenged and, in numerous cases, beaten the capacity of hospitals and intensive care units (ICUs) worldwide to handle it. Apart from the obvious burden of patient care, extended work timings, and fear of personal safety, health-care workers (HCWs) also suffer from occupational stress as a result of lack of skills, organizational factors, and low social support at work leading to distress, burnout, and psychosomatic problems. This leads to stress, direct deterioration in quality of life as well as the quality of service provided. In this article, the authors navigate on the root cause of stress that is peculiar to the HCWs deployed in the ICU and suggest recommendations to alleviate it. To aid in the research, we searched the Cochrane Central Register of Controlled Trials, MEDLINE, EMBASE, PsycINFO, CINAHL, NIOSHTIC-2, and Web of Science up to May 2021.


Cilia, Alström syndrome--molecular mechanisms and therapeutic perspectives.
Over the past ten years, several studies demonstrated the connections between cilia, basal bodies and human diseases with a wide phenotypic spectrum, including randomization of body symmetry, obesity, cystic kidney diseases and retinal degeneration. Alström syndrome (OMIM 203800) first described in 1959, is a rare autosomal recessive disorder caused by mutations in a novel gene of unknown function, ALMS1, located on the short arm of chromosome 2. Central features of Alström syndrome include obesity, insulin resistance, and type 2 diabetes. About 500 individuals with Alström syndrome are known worldwide. ALMS1 is widely expressed and localizes to centrosomes and to the base of cilia. We discuss the possible molecular mechanisms, clinical features, and future therapeutic options in a patient diagnosed with this rare disease. Monogenic defects causing human obesity actually disrupt hypothalamic pathways with a profound effect on satiety and food intake. A potential contributor to obesity- cilia with impaired function or abnormal structure, creates a new link to be studied in the future, between these organelles and the genetics of obesity.


Turning Waste into Value: Nanosized Natural Plant Materials of Solanum incanum L. and Pterocarpus erinaceus Poir with Promising Antimicrobial Activities.
Numerous plants are known to exhibit considerable biological activities in the fields of medicine and agriculture, yet access to their active ingredients is often complicated, cumbersome and expensive. As a consequence, many plants harbouring potential drugs or green phyto-protectants go largely unnoticed, especially in poorer countries which, at the same time, are in desperate need of antimicrobial agents. As in the case of plants such as the Jericho tomato, Solanum incanum, and the common African tree Pterocarpus erinaceus, nanosizing of original plant materials may provide an interesting alternative to extensive extraction and isolation procedures. Indeed, it is straightforward to obtain considerable amounts of such common, often weed-like plants, and to mill the dried material to more or less uniform particles of microscopic and nanoscopic size. These particles exhibit activity against Steinernema feltiae or Escherichia coli, which is comparable to the ones seen for processed extracts of the same, respective plants. As S. feltiae is used as a model nematode indicative of possible phyto-protective uses in the agricultural arena, these findings also showcase the potential of nanosizing of crude "waste" plant materials for specific practical applications, especially-but not exclusively-in developing countries lacking a more sophisticated industrial infrastructure.


Roles of Mineralocorticoid Receptors in Cardiovascular and Cardiorenal Diseases.
Mineralocorticoid receptor (MR) activation in the heart and vessels leads to pathological effects, such as excessive extracellular matrix accumulation, oxidative stress, and sustained inflammation. In these organs, the MR is expressed in cardiomyocytes, fibroblasts, endothelial cells, smooth muscle cells, and inflammatory cells. We review the accumulating experimental and clinical evidence that pharmacological MR antagonism has a positive impact on a battery of cardiac and vascular pathological states, including heart failure, myocardial infarction, arrhythmic diseases, atherosclerosis, vascular stiffness, and cardiac and vascular injury linked to metabolic comorbidities and chronic kidney disease. Moreover, we present perspectives on optimization of the use of MR antagonists in patients more likely to respond to such therapy and review the evidence suggesting that novel nonsteroidal MR antagonists offer an improved safety profile while retaining their cardiovascular protective effects. Finally, we highlight future therapeutic applications of MR antagonists in cardiovascular injury.


Human adenovirus type 26 basic biology and its usage as vaccine vector.
Due to their nature, adenoviruses have been recognised as promising candidates for vaccine vector development. Since they mimic natural infection, recombinant adenovirus vectors have been proven as ideal shuttles to deliver foreign transgenes aiming at inducing both humoral and cellular immune response. In addition, a potent adjuvant effect can be exerted due to the adenovirus inherent stimulation of various elements of innate and adaptive immunity. Due to its low seroprevalence in humans as well as induction of favourable immune response to inserted transgene, human adenovirus type 26 (HAdV-D26) has been recognised as a promising platform for vaccine vector development and is studied in number of completed or ongoing clinical studies. Very recently HAdV-D26 based Ebola and Covid-19 vaccines were approved for medical use. In this review, current state of the art regarding HAdV-D26 basic biology and its usage as vaccine vector will be discussed.


A novel selective androgen receptor modulator (SARM) MK-4541 exerts anti-androgenic activity in the prostate cancer xenograft R-3327G and anabolic activity on skeletal muscle mass & function in castrated mice.
The androgen receptor (AR) is a member of the nuclear hormone receptor super family of transcription factors. Androgens play an essential role in the development, growth, and maintenance of male sex organs, as well as the musculoskeletal and central nervous systems. Yet with advancing age, androgens can drive the onset of prostate cancer, the second leading cause of cancer death in males within the United States. Androgen deprivation therapy (ADT) by pharmacologic and/or surgical castration induces apoptosis of prostate cells and subsequent shrinkage of the prostate and prostate tumors. However, ADT is associated with significant musculoskeletal and behavioral adverse effects. The unique pharmacological activity of selective androgen receptor modulator (SARM) MK-4541 recently has been reported as an AR antagonist with 5α-reductase inhibitor function. The molecule inhibits proliferation and induces apoptosis in AR positive, androgen dependent prostate cancer cells. Importantly, MK-4541 inhibited androgen-dependent prostate growth in male rats yet maintained lean body mass and bone formation following ovariectomy in female rats. In the present study, we evaluated the effects of SARM MK-4541 in the androgen-dependent Dunning R3327-G prostate carcinoma xenograft mouse model as well as on skeletal muscle mass and function, and AR-regulated behavior in mice. MK-4541 significantly inhibited the growth of R3327-G prostate tumors, exhibited anti-androgen effects on the seminal vesicles, reduced plasma testosterone concentrations in intact males, and inhibited Ki67 expression. MK-4541 treated xenografts appeared similar to xenografts in castrated mice. Importantly, we demonstrate that MK-4541 exhibited anabolic activity in androgen deficient conditions, increasing lean body mass and muscle function in adult castrated mice. Moreover, MK-4541 treatment restored general activity levels in castrated mice. Thus, MK-4541 exhibits an optimum profile as an adjuvant therapy to ADT which may provide potent anti-androgenic activity at the prostate yet protective activity on skeletal muscle and behavior in patients.


The treatment of acetabular bone defects with an associated pelvic discontinuity.
Pelvic discontinuity is encountered frequently during acetabular revision in patients with severe acetabular bone loss. Prompt recognition of the discontinuity and appropriate intraoperative treatment are essential for a successful clinical outcome. The treatment of the discontinuity is dependent on the remaining host bone, the potential for healing of the discontinuity, and the potential for biologic ingrowth of acetabular components. If healing potential of the discontinuity exists, the discontinuity should be treated in compression with a posterior column plate and structural allograft or with the use of trabecular metal acting as an internal plate. If healing potential for the discontinuity does not exist, the discontinuity should be bridged and treated in distraction with an acetabular transplant supported with a cage, a trabecular metal component with trabecular metal augmentation, or with the use of a custom triflange implant. Therapeutic study, Level III-1 (case-control study). See the Guidelines for Authors for a complete description of levels of evidence.


Phase I, randomized, double-blind, placebo-controlled, multiple intravenous, dose-ascending study of sirukumab in cutaneous or systemic lupus erythematosus.
We undertook a 2-part, phase I, double-blind, placebo-controlled study to evaluate the safety and pharmacokinetics of multiple intravenous infusions of sirukumab, a human anti-interleukin-6 monoclonal antibody, in patients with cutaneous lupus erythematosus (CLE) or systemic lupus erythematosus (SLE). In part A, patients with histologically confirmed CLE were randomized to 4 infusions of placebo or 1, 4, or 10 mg/kg sirukumab every 2 weeks. In part B, SLE patients diagnosed according to American College of Rheumatology criteria with a score of 5-12 on the Safety of Estrogens in Lupus Erythematosus National Assessment version of the SLE Disease Activity Index were randomized to 4 infusions of placebo or 10 mg/kg sirukumab every 2 weeks. We treated 31 CLE patients (23 with sirukumab, 8 with placebo) and 15 SLE patients (10 with sirukumab, 5 with placebo). Adverse events (AEs) occurred more often with sirukumab than placebo in CLE patients (91% versus 63%) and in SLE patients (90% versus 80%). Sirukumab led to sustained, dose-independent decreases in white blood cell counts, absolute neutrophil counts (neutropenia), and platelet counts (thrombocytopenia) and to minor elevations in total cholesterol levels. The majority of infections were mild respiratory infections. which were reported similarly across CLE cohorts but more often in sirukumab-treated than in placebo-treated SLE patients. Two serious AEs of infection occurred (pneumonia in the 10 mg/kg-treated group and iatrogenic wound infection in the 4 mg/kg-treated group). Sirukumab showed linear pharmacokinetics in CLE patients. Systemic exposure and half-life were comparable between CLE and SLE patients. No patient developed antibodies to sirukumab through 22 weeks. C-reactive protein and serum amyloid A mean concentrations were suppressed with sirukumab from week 1 to week 14. Treatment with intravenous sirukumab infusions was generally well tolerated in both CLE and SLE patients with mild, stable, active disease. Sirukumab demonstrated linear pharmacokinetics over the dose range studied and comparable systemic exposure and half-life in CLE and SLE patients.


Discrepancies in Assessing Diastolic Function in Pre-Clinical Heart Failure Using Different Algorithms-A Primary Care Study.
Current guidelines on diastolic function (DF) by the American Society of Echocardiography and the European Association of Cardiovascular Imaging (ASE/EACVI) have been disputed and two alternative algorithms have been proposed by Johansen et al. and Oh et al. We sought (a) to assess the concordance of ASE/EACVI guidelines on DF using these proposed alternative approaches and (b) to evaluate the prevalence of indeterminate diastolic dysfunction (DD) by each method, exploring means for reducing their number. We retrospectively analyzed the echocardiographic reports of 1158 outpatients including subjects at risk of heart failure without (n = 644) or with (n = 241) structural heart disease, and 273 healthy individuals. Concordance was calculated using the k coefficient and overall proportion of DD reclassification rate. The effectiveness of pulmonary vein flow (PVF), Valsalva maneuver, and left atrial volume index/late diastolic a'-ratio (LAVi/a') over indeterminate grading was assessed. The DD reclassification rate was 30.1% (k = 0.35) for ASE/EACVI and OH, 36.5% (k = 0.27) for ASE/EACVI and JOHANSEN and 31.1% (k = 0.37) for OH and JOHANSEN (p < 0.0001 for all comparisons). DF could not be graded only by ASE/EACVI and OH in 9% and 11% patients, respectively. The majority of patients could be reclassified using PVF or Valsalva maneuver or LAVi/a', with the latter being the single most effective parameter. Inconsistencies between updated guidelines and independent approaches to assess and grade DF impede their interchangeable clinical use. The inconclusive diagnoses can be reconciled by conventional echocardiography in most patients, and LAVi/a' emerges as a simple and effective approach to this aim.


In vitro assessment of dialysis membrane as an endotoxin transfer barrier: geometry, morphology, and permeability.
High-flux dialysis membranes used with bicarbonate dialysis fluid increase the risk of back diffusion of bacterial endotoxin into the blood during hemodialysis. Endotoxin transfer of various synthetic fiber membranes was tested with bacterial culture filtrates using an in vitro system testing both diffusive and convective conditions. Membranes were tested in a simulated dialysis mode with endotoxin challenge material (approximately 420 EU/mL) added to the dialysis fluid, with saline used to model both blood and dialysis fluid. Samples were taken of both blood and dialysis fluid, and analyzed using a kinetic turbidimetric Limulus amoebocyte lysate assay. Endotoxin was found in all of the blood circuit samples, except for the Fresenius Optiflux F200NR(e) and thick-wall membranes. All membranes tested removed approximately 95% of the endotoxin from solution, with the residual approximately 5% recirculating within the dialysis fluid compartment. Endotoxin distribution through the fiber membrane was examined using a fluorescent-labeled endotoxin conjugate. Fluorescence images indicate that adsorption occurs throughout the membrane wall, with the greatest concentration of endotoxin located at the inner lumen. Contact angle analysis was able to show that all membranes exhibit a more hydrophilic lumen and a more hydrophobic outer surface except for the polyethersulfone membranes, which were of equal hydrophobicity. Resulting data indicate that fiber geometry plays an important role in the ability of the membrane to inhibit endotoxin transfer, and that both adsorption and filtration are methods by which endotoxin is retained and removed from the dialysis fluid circuit.


Distant metastases from laryngeal and hypopharyngeal cancer.
A retrospective tumor registry analysis of patients with squamous cell carcinoma (SCC) of the larynx and hypopharynx who were treated with curative intent in the Department of Otolaryngology--Head and Neck Surgery at Washington University School of Medicine and Barnes Hospital between January 1971 and December 1991. In 2,550 patients, the mean age, sex and tumor differentiation did not affect the incidence of distant metastases. The overall incidence of distant metastases was 8.5% (217/2,550 patients) with the following distribution: glottis 4.4%, supraglottis 3.6%, subglottis 14%, aryepiglottic fold 16%, pyriform sinus 17% and posterior hypopharynx 17.6%. The overall 5-year disease-specific survival for distant metastases was 6.4%. Distant metastases were related to advanced local disease (T3 + T4), lymph node metastases at presentation (N+), tumor location (hypopharynx) and locoregional tumor recurrence (p < or = 0.028). A meta-analysis of variables which predispose to a higher incidence of distant metastases indicate that tumor location (hypopharynx > larynx), advanced primary disease (T3 + T4), regional disease (N+), locoregional recurrences, and advanced regional metastases (N2 + N3) are statistically significant.


Toughening thin-film structures with ceramic-like amorphous silicon carbide films.
A significant improvement of adhesion in thin-film structures is demonstrated using embedded ceramic-like amorphous silicon carbide films (a-SiC:H films). a-SiC:H films exhibit plasticity at the nanoscale and outstanding chemical and thermal stability unlike most materials. The multi-functionality and the ease of processing of the films have potential to offer a new toughening strategy for reliability of nanoscale device structures.


The ROP vesicle release factor is required in adult Drosophila glia for normal circadian behavior.
We previously showed that endocytosis and/or vesicle recycling mechanisms are essential in adult Drosophila glial cells for the neuronal control of circadian locomotor activity. In this study, our goal was to identify specific glial vesicle trafficking, recycling, or release factors that are required for rhythmic behavior. From a glia-specific, RNAi-based genetic screen, we identified eight glial factors that are required for normally robust circadian rhythms in either a light-dark cycle or in constant dark conditions. In particular, we show that conditional knockdown of the ROP vesicle release factor in adult glial cells results in arrhythmic behavior. Immunostaining for ROP reveals reduced protein in glial cell processes and an accumulation of the Par Domain Protein 1ε (PDP1ε) clock output protein in the small lateral clock neurons. These results suggest that glia modulate rhythmic circadian behavior by secretion of factors that act on clock neurons to regulate a clock output factor.


Effects of Topically Applied Betulinic Acid and NVX-207 on Melanocytic Tumors in 18 Horses.
The naturally occurring betulinic acid (BA) and its derivative NVX-207 induce apoptosis in equine melanoma cells in vitro. After topical application, high concentrations of the substances can be reached in healthy equine skin. With the aim to investigate the effect and safety of topically applied BA and NVX-207 in horses with melanocytic tumors, the longitudinal, prospective, randomized, double-blind, placebo-controlled study protocol included eighteen Lipizzaner mares with early-stage cutaneous melanoma assigned to three groups. Melanocytic lesions were topically treated either with a placebo, 1% BA or 1% NVX-207 twice a day for 91 days. Caliper measurements, clinical examinations and blood tests were performed to assess the effects and safety of the medication. The topical treatment was convenient and safe. The volumes of tumors treated with BA were significantly reduced over time as compared to tumors treated with the placebo from day 80 of the study. Although treatment with NVX-207 seemed to decrease tumor volume, these results did not reach statistical significance. The findings must be regarded as preliminary due to the limited group size and need to be replicated in a larger cohort with modified pharmaceutical test formulations. Accordingly, the treatment protocol cannot yet be recommended in its current form.


ERK2 Mediates Metabolic Stress Response to Regulate Cell Fate.
Insufficient nutrients disrupt physiological homeostasis, resulting in diseases and even death. Considering the physiological and pathological consequences of this metabolic stress, the adaptive responses that cells utilize under this condition are of great interest. We show that under low-glucose conditions, cells initiate adaptation followed by apoptosis responses using PERK/Akt and MEK1/ERK2 signaling, respectively. For adaptation, cells engage the ER stress-induced unfolded protein response, which results in PERK/Akt activation and cell survival. Sustained and extreme energetic stress promotes a switch to isoform-specific MEK1/ERK2 signaling, induction of GCN2/eIF2α phosphorylation, and ATF4 expression, which overrides PERK/Akt-mediated adaptation and induces apoptosis through ATF4-dependent expression of pro-apoptotic factors including Bid and Trb3. ERK2 activation during metabolic stress contributes to changes in TCA cycle and amino acid metabolism, and cell death, which is suppressed by glutamate and α-ketoglutarate supplementation. Taken together, our results reveal promising targets to protect cells or tissues from metabolic stress.


Primary graft dysfunction in lung transplantation: the role of CD26/dipeptidylpeptidase IV and vasoactive intestinal peptide.
Enzymatic activity inhibition of CD26/dipeptidylpeptidase IV (CD26/DPP IV) attenuated short-term post-Tx (transplantation) ischemia-reperfusion injury after 18-hr-ischemia. Here, we investigated the effect of intragraft CD26/DPP IV catalytic inhibition on primary graft dysfunction during 7 day post-Tx, following extended ischemia. A syngeneic rat (LEW [Lewis abstract]) orthotopic lung Tx model was used, grafts exposed to 18 hr cold ischemia before Tx. Controls were flushed and preserved in Perfadex, and harvested after 1 day (CON1) or 7 day (CON7) post-Tx. Investigational groups IN1, IN3, and IN7 grafts were perfused with and stored in Perfadex + inhibitor (AB192) and harvested at 1, 3, and 7 days post-Tx, respectively. Blood gas analysis, peak airway pressure (PAwP), wet/dry weight ratio, myeloperoxidase thiobarbituric acid reactive substances (TBARS), and staining for vasoactive intestinal peptide (VIP) were analyzed. IN1 versus CON1 showed preserved histology, increased pO2 (P<0.01), lowered PAwP (P<0.01), less edema (P<0.05) and decreased TBARS (P<0.05). Survival was better for IN7 versus CON7 (P<0.01). The course of AB192-perfused grafts from 1 to 7 days displayed improved values for pO2 (P<0.01), PAwP (P<0.01), edema (P<0.05), TBARS (P<0.05), and myeloperoxidase (P<0.05). Compared with controls, VIP was preserved during 18 hr ischemia in alveolar macrophages (P=0.0001) and respiratory epithelial cells (P=0.001). Perfusion with an inhibitor of CD26/DPP IV enzymatic activity significantly reduced the incidence and severity of pulmonary primary graft dysfunction and enabled recovery after extended ischemia. This is the first report that CD26/DPPIV inhibitor treatment increases local pulmonary VIP levels, which correlate with preserved ventilatory function and pulmonary structural integrity.


Success and predictability of provox prosthesis voice rehabilitation.
To determine the success rate and relating clinical factors of voice prosthesis rehabilitation and to analyze the discrimination ability of the multidimensional Harrison-Robillard-Shultz Tracheoesophageal Puncture Rating Scale (HRS Rating Scale). Prospective clinical study. University Department of Otorhinolaryngology, Head and Neck Surgery, University Hospital Inselspital, Bern, Switzerland. From 1992 through 1998, 87 patients with advanced squamous cell carcinoma of the larynx and/or hypopharynx underwent primary tracheoesophageal puncture after total laryngectomy. Clinician otolaryngologists and speech/language pathologists independently used the HRS Rating Scale for success assessment of voice prosthesis rehabilitation. Age, sex, tumor localization, tumor stage, and radiation therapy had no influence on the success of voice prosthesis rehabilitation. Overall, voice rehabilitation success rates between 40% and 62% were achieved. Speech/language pathologists and clinician otolaryngologists evaluated the same patient group without significant statistical differences. The HRS Rating Scale analysis showed an equal distribution of the subscale parameter care in functional and nonfunctional speakers and a strong correlation between the subscale parameters quality and use. Because of its safety and simplicity, tracheoesophageal puncture has become a state of the art method for voice rehabilitation after total laryngectomy. The short-term superiority of voice prosthesis in voice rehabilitation over esophageal speech rehabilitation must be seen in light of comparable long-term success rates of the 2 methods.


Experiences and Needs of Parents Caring for Children with Cerebral Palsy: A Systematic Review.
Caregiver burden of parents of children with disabilities is negatively associated with parental quality of life and psychosocial well-being and has been identified as a public health concern. Reviews have consolidated the experiences of these parents, but little is understood about the unique challenges faced by parents of children with cerebral palsy (CP). We aim to conduct an in-depth synthesis of the experiences and needs of parents of children with CP to better support them. Six electronic databases were searched systematically for qualitative research, and a thematic synthesis was performed. Seventeen studies from 1988 to 2019 encapsulating the experiences of 226 parents were included in this review. Four themes and 11 subthemes were identified: (1) self: the caregiver-parent trajectory (coming to terms with the fact, physical and emotional demands, and a nice outlet), (2) family: maintaining the family unit (juggling multiple children and parenting as a co-occupation), (3) society: social repercussions (facing public discrimination, rejection by extended family, and loss of freedom), and (4) parents' wish list (open communication and caring attitudes, funding and health care accessibility, social integration, and inclusivity). Apart from emotional adaptation and physical childcare difficulty, parents indicated a need for more information on health care services, funding, better attitudes and empathy from health care providers, properly trained educators, disability-friendly fixtures, and more public awareness. The findings in this review provide insight into the true experiences and needs of parents caring for children with CP and will aid clinicians in the development of family-centered care pathways for these patients.


Serum copper, zinc and magnesium in maternal and cord blood at delivery.
Serum concentrations of copper, zinc and magnesium were determined in maternal and cord blood at delivery in term pregnancies among healthy Nigeria women. Maternal serum copper concentration was significantly higher than cord serum concentration. There were no significant differences in maternal and cord serum levels of zinc and magnesium.


Masking of foveal and parafoveal vision during eye fixations in reading.
A window or visual mask as moved across text in synchrony with the reader's eye movements. The size of the window or mask was varied so that either information in foveal or parafoveal vision was masked on each fixation. In another experiment, the onset of the mask was delayed for a certain amount of time following the end of the saccade. The results of the experiments point out the relative importance of foveal and parafoveal vision for reading and further indicate that most of the visual information necessary for reading can be acquired during the first 50 msec that information is available during an eye fixation.