Diaryl urea analogues of SB-334867 as orexin-1 receptor antagonists.
As a part of our program to develop OX1-CB1 bivalent ligands, we required a better understanding of the basic structure-activity relationships (SARs) of orexin antagonists. A series of SB-334867 analogues were synthesized and evaluated in calcium mobilization assays. SAR results suggest that the 2-methylbenzoxazole moiety may be replaced with a disubstituted 4-aminophenyl group without loss of activity and an electron-deficient system is generally preferred at the 1,5-naphthyridine moiety for OX1 antagonist activity. In particular, substitution of larger potential linkers such as n-hexyl provided compound 33 with equivalent activity at the OX1 receptor compared to the lead compound SB-334867. These compounds should be of value in the development of ligands targeting the orexin-1 receptor and its potential heterodimers.


Platelet binding sites for atrial natriuretic factor in humans. Characterization and effects of sodium intake.
Platelets bear receptors for vasoactive peptides such as angiotensin II and vasopressin. The presence of binding sites for another vasoactive peptide, atrial natriuretic factor, was therefore investigated in human platelets. 125I-labeled synthetic atrial natriuretic factor bound specifically to human platelets. Steady-state and kinetic experiments demonstrated the existence of one class of high-affinity low-capacity binding sites for atrial natriuretic factor in platelets with a dissociation constant of 30 pM. The order of potency of atrial natriuretic factor fragments showed that the structural requirements of the high-affinity binding site detected were similar to those of receptors for atrial natriuretic factor in rat blood vessels and adrenal zona glomerulosa. To study the regulation of these binding sites by sodium, normal young men were subjected successively in random order to a low-sodium (40 mmol per day) and high-sodium (300 mmol per day) diet for 4 days. Binding of atrial natriuretic factor to platelets was higher with the low-sodium diet (10.3 +/- 1.0 sites per cell) than with the high-sodium diet (7.1 +/- 0.7 sites per cell). In conclusion, human platelets bear binding sites for atrial natriuretic factor, the density of which may be modulated by sodium intake. The platelet is a useful model for investigating atrial natriuretic factor receptors in different physiopathological conditions in humans.


All-optical AND and NAND gates based on cascaded second-order nonlinear processes in a Ti-diffused periodically poled LiNbO(3) waveguide.
All-optical AND and NAND gates have been demonstrated in a Ti-diffused periodically poled LiNbO(3) channel waveguide which has two second-harmonic phase-matching peaks by cascaded sum-frequency-generation/difference-frequency-generation (cSFG/DFG) and sum-frequency-generation (SFG) processes. The conversion efficiency of signal to idler (AND gate signal) was approximately 0 dB in cSFG/DFG process. In the second SFG process, more than 15 dB extinction ratio between signal and dropped signal (NAND gate signal) has been observed.


Reduction in potassium concentration of stored red blood cell units using a resin filter.
Hyperkalemia is a serious complication of rapid and massive blood transfusion due to high plasma potassium (K) in stored red blood cell (RBC) units. A potassium adsorption filter (PAF) was developed in Japan to remove K by exchanging with sodium (Na). We performed an in vitro evaluation of its efficacy and feasibility of use. Three AS-3 RBC units were filtered by each PAF using gravity; 10 PAFs were tested. Blood group, age, flow rate, and irradiation status were recorded. Total volume, K, Na, Cl, Mg, total Ca (tCa), RBC count, hemoglobin (Hb), hematocrit (Hct), and plasma Hb were measured before and after filtering each unit. Ionized Ca (iCa), pH, and glucose were measured for some units. After filtration, the mean decrease in K was 97.5% in the first RBC unit, 91.2% in the second unit, and 64.4% in the third unit. The mean increases in Na, Mg, and tCa were 33.0, 151.4, and 116.1%, respectively. iCa and pH remained low; glucose was unchanged. RBC count, Hb, and Hct decreased slightly after filtration of first units; plasma Hb was unchanged. After filtration, there was no visual evidence of increased hemolysis or clot formation. The PAF decreased K concentration in stored AS-3 RBC units to minimal levels in the first and second RBC units. Optimally, one filter could be used for 2 RBC units. Although Na increased, the level may not be clinically significant. PAF may be useful for at-risk patients receiving older units or blood that has been stored after gamma irradiation.


Patient-reported, observer-reported and performance outcomes in qualification procedures at the European Medicines Agency 2013-2018.
To describe characteristics of applicant, tool, outcomes, regulatory responses and general learnings from European Medicines Agency (EMA) Qualification Procedures on patient-reported outcomes (PROs), observer-reported outcomes (ObsROs) and performance outcomes (PerfOs) finalized between January 2013 and December 2018. Descriptive analysis, and qualitative review of the regulatory outcomes of the study procedures. Seventeen qualification programmes for PROs, 6 for ObsRO tools and 11 for PerfO tools were submitted by consortia, large and small/medium companies. Gastroenterology and neurology were the most frequent therapeutic areas. There was a high level of regulators' partial agreement (above 70%) with applicant's approaches with constructive input; EMA published Letters of Support for PRO (6), ObsRO (2) and PerfO (4) tools, and Qualification Opinions on PROs (2) and PerfOs (1). General issues related to Qualification Procedures on PROs raised by EU regulatorsincluded: population, appropriate studies to demonstrate ability to detect change, tool validation in interventional trials, anchoring, identification of the minimally important difference, item selection, weighting, and multiple domains. In addition, specific issues for ObsROs and PerfO tools validation are identified. Regulators discussed principles and challenges of validation tailored to specific setting in tool development, providing constructive feedback. Regulatory outputs supportive of further development were published in over one-third of programs. We encourage applicants intending to use or develop novel PRO, ObsRO and PerfO tools that will generate evidence for regulatory submissions on medicines to consider Qualification procedures for novel methods to seek feedback on the development and validation of these tools.


Virulence characteristics and genetic affinities of multiple drug resistant uropathogenic Escherichia coli from a semi urban locality in India.
Extraintestinal pathogenic Escherichia coli (ExPEC) are of significant health concern. The emergence of drug resistant E. coli with high virulence potential is alarming. Lack of sufficient data on transmission dynamics, virulence spectrum and antimicrobial resistance of certain pathogens such as the uropathogenic E. coli (UPEC) from countries with high infection burden, such as India, hinders the infection control and management efforts. In this study, we extensively genotyped and phenotyped a collection of 150 UPEC obtained from patients belonging to a semi-urban, industrialized setting near Pune, India. The isolates representing different clinical categories were analyzed in comparison with 50 commensal E. coli isolates from India as well as 50 ExPEC strains from Germany. Virulent strains were identified based on hemolysis, haemagglutination, cell surface hydrophobicity, serum bactericidal activity as well as with the help of O serotyping. We generated antimicrobial resistance profiles for all the clinical isolates and carried out phylogenetic analysis based on repetitive extragenic palindromic (rep)-PCR. E. coli from urinary tract infection cases expressed higher percentages of type I (45%) and P fimbriae (40%) when compared to fecal isolates (25% and 8% respectively). Hemolytic group comprised of 60% of UPEC and only 2% of E. coli from feces. Additionally, we found that serum resistance and cell surface hydrophobicity were not significantly (p = 0.16/p = 0.51) associated with UPEC from clinical cases. Moreover, clinical isolates exhibited highest resistance against amoxicillin (67.3%) and least against nitrofurantoin (57.3%). We also observed that 31.3% of UPEC were extended-spectrum beta-lactamase (ESBL) producers belonging to serotype O25, of which four were also positive for O25b subgroup that is linked to B2-O25b-ST131-CTX-M-15 virulent/multiresistant type. Furthermore, isolates from India and Germany (as well as global sources) were found to be genetically distinct with no evidence to espouse expansion of E. coli from India to the west or vice-versa.


[Attitudes of dietitians in relation to obese individuals - an exploratory study].
The scope of this study was to assess attitudes of dietitians in relation to obesity; involving beliefs about the characteristics attributed to obese people, the reasons that lead to obesity and obesity itself. Dietitians (N = 344; 97.1% women) were contacted via their professional council and filled out the online survey. The survey questions were translated and adapted from international studies on this subject and the responses were analyzed for concordance rate. The results pointed to strong stigmatization of obesity and prejudice against the obese, attributing characteristics such as greed (67.4%), unattractiveness (52.0%), ungainliness (55.1%), lack of willpower (43.6%) and laziness (42.3%). The most important causal factors were considered to be emotional and mood changes, food addiction and low self-esteem. Research on this topic should be enhanced since these attitudes can affect the efficacy of treatment and also to foster broad discussion and training regarding the significance of obesity and to ensure more individualized and humanized treatment for obese patients.


Influence of ANG II on cytoplasmic sodium in cultured rabbit nonpigmented ciliary epithelium.
Angiotensin (ANG) II receptors have been reported in the nonpigmented ciliary epithelium (NPE) of the eye. In cultured NPE, we found ANG II caused a dose-dependent rise of cytoplasmic sodium. The sodium increase was inhibited by the AT(1)-AT(2) receptor antagonist saralasin (IC(50) = 3.7 nM) and the AT(1) antagonist losartan (IC(50) = 0.6 nM) but not by the AT(2) antagonist PD-123319. ANG II also caused a dose-dependent increase in the rate of ouabain-sensitive (86)Rb uptake. The ANG II-induced cell sodium increase and (86)Rb uptake increase were reduced by dimethylamiloride (DMA; 10 microM). On the basis of this finding, we propose that Na(+)/H(+) exchange is stimulated by ANG II. Simultaneously, ANG II appears to inhibit H(+)-ATPase-mediated proton export. Thus Ang II (10 nM) did not alter the baseline cytoplasmic pH (pH(i)) but reduced pH(i) in cells that were also exposed to 10 microM DMA. Consistent with the notion of H(+)-ATPase inhibition in ANG II-treated NPE, bafilomycin A(1) (100 nM) (BAF) and ANG II were both observed to suppress the pH(i) increase that occurs upon exposure to a mixture of epinephrine (1 microM) and acetylcholine (10 microM) and the pH(i) increase elicited by depolarization. In ATP hydrolysis measurements, H(+)-ATPase activity (bafilomycin A(1)-sensitive ATP hydrolysis) was reduced significantly in cells that had been pretreated 10 min with 10 nM ANG II. In summary, these studies suggest that ANG II causes H(+)-ATPase inhibition and an increase of cell sodium due to activation of Na(+)/H(+) exchange.


Population dynamics of some Pakistan mosquitoes: changes in adult relative abundance over time and space.
Time series and spatial changes in the relative abundance of 14 mosquito species were described from weekly or biweekly collections at nine localities in Punjab province, Pakistan, from January 1976 to June 1980. Comparisons between indoor aspirator catches and outdoor mechanical sweeper collections enabled mosquito resting habits to be classified as: (1) endophilic (Anopheles culicifacies, An. fluviatilis, An. stephensi, An. subpictus); (2) partially exophilic (An. annularis, An. pulcherrimus, An. nigerrimus, Culex bitaeniorhynchus, Cx, pseudovishnui, Cx, quinquefasciatus, Cx. tritaeniorhynchus), or exophilic (Cx. fuscocephala, Aedes caspius, Mansonia uniformis). Temporal abundance patterns were grouped by seasonality, overwintering strategies and the magnitude of fluctuation. Seasonal patterns were: (1) unimodal-spring (Cx. quinquefasciatus, Ae. capius); (2) unimodal-monsoon (An. nigerrimus, An. subpictus, Cx. bitaeniorhynchus, Cx. fuscocephala); (3) bimodal-spring dominant (An. annularis, An. culcifacies, An. stephensi), and (4) bimodal-monsoon/post-monsoon dominant (An. fluviatilis, An. pulcherrimus, Cx. pseudovishnui, Cx. tritaeniorhynchus, Ma. uniformis). Mosquito overwintering strategies included: (1) adults with slowed reproductive activity (An. annularis, An. culicifacies, An. fluviatilis, An. pulcherrimus, An. stephensi, Cx. quinquefasciatus); (2) females with interrupted reproductive activity (An. nigerrimus, Cx. fuscocephala, Cx. pseudovishnui, Cx. tritaeniorhynchus); (3) immature stages (Ae. caspius, Ma. uniformis) and (4) annual extinction and re-introduction (An. subpictus). The magnitude of seasonal change was classified by the number of standard deviations from the overall mean exhibited by the annual maxima or minima: (1) stationary, less than 1 standard deviation (An. culicifacies, An. fluviatilis, Cx. bitaeniorhynchus, Ae. caspius), (2) fluctuating moderately, one to two standard deviations (An. annularis, An. nigerrimus, An. pulcherrimus, An. stephensi, Cx. fuscocephala, Cx. quinquefasciatus, Ma. uniformis) and (3) markedly fluctuating greater than two standard deviations (An. subpictus, Cx. pseudovishnui, Cx. tritaeniorhynchus). The seasonal increases of An. culicifacies and An. stephensi abundance did not closely parallel the periods of maximal malaria transmission. Temporal patterns exhibited by several culicine species indicated that the period of greatest West Nile virus activity would probably occur during the post-monsoon season.


Expression profile of kalliklectin, a soluble-type mannose receptor, during embryogenesis and early larval development in fugu (Takifugu rubripes).
Kalliklectin is a unique fish-specific lectin, whose sequence is similar to the heavy chain of mammalian plasma kallikrein and coagulation factor XI. In this study, we aimed to evaluate dynamic expression profiles of the lectin gene, during early developmental stages, in fugu, Takifugu rubripes. Reverse transcription-polymerase chain reaction (RT-PCR) showed that the kalliklectin gene was not expressed until 14 h post-fertilization (hpf), while the mRNA was detected after 30 hpf. In real-time quantitative PCR (qPCR), the gene was first expressed at 10.5 hpf; then, the expression level increased with a peak at 30 hpf and then gradually decreased. On the other hand, western blotting with specific antibody detected the lectin protein at all tested stages, including the unfertilized egg, which suggests that the lectin detected in the early stages was a maternal factor. Immunohistochemistry demonstrated that kalliklectin was localized at the basement membranes of the newly hatched larvae, while the lectin was widely detected in epidermal cells in larva at 5 dph. A 40-kDa lectin was partially purified from unfertilized eggs using mannose-affinity chromatography, and the lectin was determined as kalliklectin by liquid chromatography with quadrupole time-of-flight tandem mass spectrometry (LC/Q-TOF-MS) analysis, which indicated that the lectin is functional in the eggs. The egg lectin can bind to Gram-positive bacterial pathogens of fish, such as Lactococcus garvieae and Streptococcus iniae. We conclude that fugu kalliklectin might be an important immunocomponent, transferred from mother to offspring.


Subspecies IIIa and IIIb Salmonellae are defective for colonization of murine models of salmonellosis compared to Salmonella enterica subsp. I serovar typhimurium.
Non-subspecies I salmonellae are commensals of cold-blooded vertebrates and cause sporadic disease in mammals. The reasons why non-subspecies I salmonellae do not circulate in populations of warm-blooded vertebrates, but instead only cause occasional disease in this niche, are unknown. We examined the ability of Salmonella enterica subsp. IIIa (subsp. arizonae) and subsp. IIIb (subsp. diarizonae) isolates to grow competitively with subspecies I (serovar Typhimurium) ATCC 14028 in vitro, to colonize Salmonella-sensitive BALB/c mice, and to persist in the intestine of Salmonella-resistant CBA/J mice in competitive infections. Subspecies IIIa had severely reduced intestinal colonization, intestinal persistence, and systemic spread in mice. Subspecies IIIa is nonmotile on swarming agar and thus may also have reduced motility under viscous conditions in vivo. Surprisingly, subspecies IIIb colonizes the intestinal tract of BALB/c mice normally yet does not spread systemically. Subspecies IIIb colonization of the intestine of CBA/J mice is reduced late in infection. In order to understand why these isolates do not colonize systemic sites, we determined that subspecies IIIa and IIIb are not internalized well and do not replicate in J774-A.1 murine macrophages, despite normal adherence to these cells. We further show that selected effectors of both type III secretion systems 1 and 2 are secreted by subspecies IIIa and IIIb in vitro but that each of these isolates secretes a different combination of effectors. We outline the phenotypic differences between these subspecies and subspecies I and provide a possible explanation for the inability of these strains to spread systemically in murine models.


Magnetic propulsion of colloidal microrollers controlled by electrically modulated friction.
Precise control over the motion of magnetically responsive particles in fluidic chambers is important for probing and manipulating tasks in prospective microrobotic and bio-analytical platforms. We have previously exploited such colloids as shuttles for the microscale manipulation of objects. Here, we study the rolling motion of magnetically driven Janus colloids on solid substrates under the influence of an orthogonal external electric field. Electrically induced attractive interactions were used to tune the load on the Janus colloid and thereby the friction with the underlying substrate, leading to control over the forward velocity of the particle. Our experimental data suggest that the frictional coupling required to achieve translation, transitions from a hydrodynamic regime to one of mixed contact coupling with increasing load force. Based on this insight, we show that our colloidal microrobots can probe the local friction coefficient of various solid surfaces, which makes them potentially useful as tribological microsensors. Lastly, we precisely manipulate porous cargos using our colloidal rollers, a feat that holds promise for bio-analytical applications.


Application of different genotyping methods for Pseudomonas aeruginosa in a setting of endemicity in an intensive care unit.
Colonization with Pseudomonas aeruginosa was studied by taking serial swab specimens from the oropharynges and anuses and tracheal and gastric aspirates from patients in an intensive care unit during a 10-month period in a setting of endemicity. Nineteen (10%) of the 192 patients included in the study were colonized on admission, while another 30 (16%) patients acquired P. aeruginosa while in the hospital. Typing of 353 isolates was performed by random amplified polymorphic DNA (RAPD) analysis, and 56 strains were selected for further typing by RAPD analysis, pulsed-field gel electrophoresis (PFGE), and amplified fragment length polymorphism (AFLP) analysis. By these methods, 42, 44, and 44 genotypes were found, respectively. Computer-aided cluster analysis indicated that similar groups of related isolates were obtained by each method. By taking admission periods into account, analysis of the typing results suggested cross-acquisition of P. aeruginosa for five patient pairs. The small number of transfers and the large number of genotypes found indicate that most P. aeruginosa strains were derived from the patients themselves. The numbers of observed typing patterns and band differences between related isolates were counted for each typing method. AFLP analysis with primers without a selective base proved to be the most discriminatory method, followed by PFGE, AFLP analysis (with one selective base), and RAPD analysis. On the basis of a comparison with established strain differentiation criteria for PFGE, the criteria for differentiation of P. aeruginosa by AFLP analysis are presented.


Dielectric Breakdown in Chemical Vapor Deposited Hexagonal Boron Nitride.
Insulating films are essential in multiple electronic devices because they can provide essential functionalities, such as capacitance effects and electrical fields. Two-dimensional (2D) layered materials have superb electronic, physical, chemical, thermal, and optical properties, and they can be effectively used to provide additional performances, such as flexibility and transparency. 2D layered insulators are called to be essential in future electronic devices, but their reliability, degradation kinetics, and dielectric breakdown (BD) process are still not understood. In this work, the dielectric breakdown process of multilayer hexagonal boron nitride (h-BN) is analyzed on the nanoscale and on the device level, and the experimental results are studied via theoretical models. It is found that under electrical stress, local charge accumulation and charge trapping/detrapping are the onset mechanisms for dielectric BD formation. By means of conductive atomic force microscopy, the BD event was triggered at several locations on the surface of different dielectrics (SiO2, HfO2, Al2O3, multilayer h-BN, and monolayer h-BN); BD-induced hillocks rapidly appeared on the surface of all of them when the BD was reached, except in monolayer h-BN. The high thermal conductivity of h-BN combined with the one-atom-thick nature are genuine factors contributing to heat dissipation at the BD spot, which avoids self-accelerated and thermally driven catastrophic BD. These results point to monolayer h-BN as a sublime dielectric in terms of reliability, which may have important implications in future digital electronic devices.


A new interpretation of spontaneous sway measures based on a simple model of human postural control.
This study shows that center-of-pressure (COP) traces that closely resemble physiologically measured COP functions can be produced by an appropriate selection of model parameters in a simple feedback model of the human postural control system. Variations in the values of stiffness, damping, time delay, and noise level determine the values of 15 sway measures commonly used to characterize spontaneous sway. Results from model simulations indicate that there is a high degree of correlation among these sway measures, and the measures cluster into three different groups. Only two principal components accounted for about 92% of the variation among the different sway measures analyzed. This model can be used to formulate hypotheses regarding the cause of postural control deficits reported in the literature. This is accomplished using a multidimensional optimization procedure to estimate model parameters from a diverse set of spontaneous sway measures. These model parameters describe physiologically meaningful features of the postural control system as opposed to conventional sway measures that provide only a parametric description of sway. To show the application of this method, we applied it to published data of spontaneous sway from elderly subjects and contrasted it to the data of young healthy subjects. We found that modest increases in stiffness and damping and a fairly large increase in noise level with aging could account for the variety of sway measures reported in the literature for elderly subjects.


Mutable Collagenous Tissue Isolated from Echinoderms Leads to the Production of a Dermal Template That Is Biocompatible and Effective for Wound Healing in Rats.
The mutable collagenous tissue (MCT) of echinoderms possesses biological peculiarities that facilitate native collagen extraction and employment for biomedical applications such as regenerative purposes for the treatment of skin wounds. Strategies for skin regeneration have been developed and dermal substitutes have been used to cover the lesion to facilitate cell proliferation, although very little is known about the application of novel matrix obtained from marine collagen. From food waste we isolated eco-friendly collagen, naturally enriched with glycosaminoglycans, to produce an innovative marine-derived biomaterial assembled as a novel bi-layered skin substitute (Marine Collagen Dermal Template or MCDT). The present work carried out a preliminary experimental in vivo comparative analysis between the MCDT and Integra, one of the most widely used dermal templates for wound management, in a rat model of full-thickness skin wounds. Clinical, histological, and molecular evaluations showed that the MCDT might be a valuable tool in promoting and supporting skin wound healing: it is biocompatible, as no adverse reactions were observed, along with stimulating angiogenesis and the deposition of mature collagen. Therefore, the two dermal templates used in this study displayed similar biocompatibility and outcome with focus on full-thickness skin wounds, although a peculiar cellular behavior involving the angiogenesis process was observed for the MCDT.


Metal Positions and Translocation Pathways of the Dodecameric Ferritin-like Protein Dps.
Iron storage in biology is carried out by cage-shaped proteins of the ferritin superfamily, one of which is the dodecameric protein Dps. In Dps, four distinct steps lead to the formation of metal nanoparticles: attraction of ion-aquo complexes to the protein matrix, passage of these complexes through translocation pores, oxidation of these complexes at ferroxidase centers, and, ultimately, nanoparticle formation. In this study, we investigated Dps from Listeria innocua to structurally characterize these steps for Co2+, Zn2+, and La3+ ions. The structures reveal that differences in their ion coordination chemistry determine alternative metal ion-binding sites on the areas of the surface surrounding the translocation pore that captures nine La3+, three Co2+, or three Zn2+ ions as aquo clusters and passes them on for translocation. Inside these pores, ion-selective conformational changes at key residues occur before a gating residue to actively move ions through the constriction zone. Ions upstream of the Asp130 gate residue are typically hydrated, while ions downstream directly interact with the protein matrix. Inside the cavity, ions move along negatively charged residues to the ferroxidase center, where seven main residues adapt to the three different ions by dynamically changing their conformations. In total, we observed more than 20 metal-binding sites per Dps monomer, which clearly highlights the metal-binding capacity of this protein family. Collectively, our results provide a detailed structural description of the preparative steps for amino acid-assisted biomineralization in Dps proteins, demonstrating unexpected protein matrix plasticity.


Identical cancers in husband and wife.
Eleven married couples having histologically identical malignant tumors have been encountered: two couples with melanoma; three with carcinoma of the breast; two with carcinoma of the kidney, and one couple each with fibrosarcoma; carcinoma of the floor of the mouth; carcinoma of the colon and rectum, and carcinoma of the nasopharynx. The interval until the spouse of the patient with cancer was diagnosed to have cancer was within five years in all but three instances. The clinical course of the disease tended to be similar in both husband and wife. Generally, the longer the couple had been married, the more successful was their treatment. Those tumors described, as well as those identical cancers of husband and wife previously reported, have been suggested either experimentally or indirectly to have viral relationships.


Penetratin improves tumor retention of single-chain antibodies: a novel step toward optimization of radioimmunotherapy of solid tumors.
Single-chain Fv (scFv) antibody fragments exhibit improved pharmacokinetics and biodistribution compared with intact IgG. The tumor uptake of scFvs is rapid, and the serum half-life is shorter than IgG. However, scFvs exhibit lower net dose deposition in the tumor due to a shorter residence time that limits their use in radioimmunotherapy. To improve the tumor uptake and retention of scFvs, we investigated the utility of cell-penetrating peptides, penetratin and transactivator of transcription (TAT). Biodistribution studies were done in LS174T tumor-bearing mice with divalent scFv derived from anti-tumor-associated glycoprotein 72 monoclonal antibody (mAb) CC49. Penetratin increased the tumor retention of scFvs without affecting the peak dose accumulation. The percentage of doses retained in tumors at 24 hours post-administration with a control (no peptide), penetratin, and TAT were 27.25%, 79.84%, and 48.55%, respectively, of that accumulated at 8 hours postinjection. The tumor-to-blood ratios at 24 hours postadministration were 7.14, 19.53, and 16.48 with control, penetratin, and TAT treatment, respectively, whereas the pharmacokinetics were unaltered. Coinjection with TAT, however, resulted in increased uptake of the radioconjugate by the lungs. Autoradiography of the excised tumors indicated a more homogenous distribution of the radiolabeled scFv with both penetratin and TAT in comparison with the control treatment. Real-time whole-body imaging of the live animals confirmed improved tumor localization with penetratin without any increase in the uptake by normal tissues. In conclusion, a significant improvement in the tumor retention of sc(Fv)2 was achieved by administration of penetratin. Therefore, the combination of penetratin and scFvs has the potential of improving the utility of mAb-based radiopharmaceuticals.


Human Chondrocytes, Metabolism of Articular Cartilage, and Strategies for Application to Tissue Engineering.
Hyaline cartilage, which is characterized by the absence of vascularization and innervation, has minimal self-repair potential in case of damage and defect formation in the chondral layer. Chondrocytes are specialized cells that ensure the synthesis of extracellular matrix components, namely type II collagen and aggregen. On their surface, they express integrins CD44, α1β1, α3β1, α5β1, α10β1, αVβ1, αVβ3, and αVβ5, which are also collagen-binding components of the extracellular matrix. This article aims to contribute to solving the problem of the possible repair of chondral defects through unique methods of tissue engineering, as well as the process of pathological events in articular cartilage. In vitro cell culture models used for hyaline cartilage repair could bring about advanced possibilities. Currently, there are several variants of the combination of natural and synthetic polymers and chondrocytes. In a three-dimensional environment, chondrocytes retain their production capacity. In the case of mesenchymal stromal cells, their favorable ability is to differentiate into a chondrogenic lineage in a three-dimensional culture.


Generalized discriminant analysis using a kernel approach.
We present a new method that we call generalized discriminant analysis (GDA) to deal with nonlinear discriminant analysis using kernel function operator. The underlying theory is close to the support vector machines (SVM) insofar as the GDA method provides a mapping of the input vectors into high-dimensional feature space. In the transformed space, linear properties make it easy to extend and generalize the classical linear discriminant analysis (LDA) to nonlinear discriminant analysis. The formulation is expressed as an eigenvalue problem resolution. Using a different kernel, one can cover a wide class of nonlinearities. For both simulated data and alternate kernels, we give classification results, as well as the shape of the decision function. The results are confirmed using real data to perform seed classification.


Childhood onset in familial prion disease with a novel mutation in the PRNP gene.
Up to 15% of cases of prion diseases are due to the autosomal dominant inheritance of coding PRNP mutations. To describe the unique clinical and genetic findings in a family of East Indian origin with autosomal dominant inheritance of a novel PRNP mutation. Detailed neurological examination and sequencing analysis of the MAPT and PRNP genes. Toronto Western Hospital, Toronto, Ontario. Five available members of a family of East Indian origin with a rapidly progressive neurodegenerative disorder characterized by dementia, motor decline, and ataxia. We identified a novel Pro105Thr mutation in the PRNP gene in all of the 3 clinically affected family members but not in their unaffected relatives or normal controls. Although 5 of 6 affected family members had a relatively homogeneous phenotype and age at onset (range, 33-41 years), 1 of the 6 patients developed the disease at age 13 years. A novel mutation in the PRNP gene was identified in all of the available, clinically affected members of this family with a rapidly progressive neurodegenerative disease. To our knowledge, the propositus represents the youngest individual with inherited prion disease described to date.


Brainstem glucose metabolism predicts reward dependence scores in treatment-resistant major depression.
It has been suggested that individual differences in temperament could be involved in the (non-)response to antidepressant (AD) treatment. However, how neurobiological processes such as brain glucose metabolism may relate to personality features in the treatment-resistant depressed (TRD) state remains largely unclear. To examine how brainstem metabolism in the TRD state may predict Cloninger's temperament dimensions Harm Avoidance (HA), Novelty Seeking (NS), and Reward Dependence (RD), we collected 18fluorodeoxyglucose positron emission tomography (18FDG PET) scans in 40 AD-free TRD patients. All participants were assessed with the Temperament and Character Inventory (TCI). We applied a multiple kernel learning (MKL) regression to predict the HA, NS, and RD from brainstem metabolic activity, the origin of respectively serotonergic, dopaminergic, and noradrenergic neurotransmitter (NT) systems. The MKL model was able to significantly predict RD but not HA and NS from the brainstem metabolic activity. The MKL pattern regression model identified increased metabolic activity in the pontine nuclei and locus coeruleus, the medial reticular formation, the dorsal/median raphe, and the ventral tegmental area that contributed to the predictions of RD. The MKL algorithm identified a likely metabolic marker in the brainstem for RD in major depression. Although 18FDG PET does not investigate specific NT systems, the predictive value of brainstem glucose metabolism on RD scores however indicates that this temperament dimension in the TRD state could be mediated by different monoaminergic systems, all involved in higher order reward-related behavior.


The systematic relationships and biogeographic history of ornithischian dinosaurs.
The systematic relationships of taxa traditionally referred to as 'basal ornithopods' or 'hypsilophodontids' remain poorly resolved since it was discovered that these taxa are not a monophyletic group, but rather a paraphyletic set of neornithischian taxa. Thus, even as the known diversity of these taxa has dramatically increased over the past two decades, our knowledge of their placement relative to each other and the major ornithischian subclades remained incomplete. This study employs the largest phylogenetic dataset yet compiled to assess basal ornithischian relationships (255 characters for 65 species level terminal taxa). The resulting strict consensus tree is the most well-resolved, stratigraphically consistent hypothesis of basal ornithischian relationships yet hypothesized. The only non-iguanodontian ornithopod (=basal ornithopod) recovered in this analysis is Hypsilophodon foxii. The majority of former 'hypsilophodontid' taxa are recovered within a single clade (Parksosauridae) that is situated as the sister-taxon to Cerapoda. The Parksosauridae is divided between two subclades, the Orodrominae and the Thescelosaurinae. This study does not recover a clade consisting of the Asian taxa Changchunsaurus, Haya, and Jeholosaurus (=Jeholosauridae). Rather, the former two taxa are recovered as basal members of Thescelosaurinae, while the latter taxon is recovered in a clade with Yueosaurus near the base of Neornithischia.The endemic South American clade Elasmaria is recovered within the Thescelosaurinae as the sister taxon to Thescelosaurus. This study supports the origination of Dinosauria and the early diversification of Ornithischia within Gondwana. Neornithischia first arose in Africa by the Early Jurassic before dispersing to Asia before the late Middle Jurassic, where much of the diversification among non-cerapodan neornithischians occurred. Under the simplest scenario the Parksosauridae originated in North America, with at least two later dispersals to Asia and one to South America. However, when ghost lineages are considered, an alternate dispersal hypothesis has thescelosaurines dispersing from Asia into South America (via North America) during the Early Cretaceous, then back into North America in the latest Cretaceous. The latter hypothesis may explain the dominance of orodromine taxa prior to the Maastrichtian in North America and the sudden appearance and wide distribution of thescelosaurines in North America beginning in the early Maastrichtian. While the diversity of parksosaurids has greatly increased over the last fifteen years, a ghost lineage of over 40 myr is present between the base of Parksosauridae and Cerapoda, indicating that much of the early history and diversity of this clade is yet to be discovered. This new phylogenetic hypothesis provides a comprehensive framework for testing further hypotheses regarding evolutionary patterns and processes within Ornithischia.


Multidrug resistance-associated protein: a protein distinct from P-glycoprotein involved in cytotoxic drug expulsion.
1. Multidrug resistance (MDR) is a phenomenon originally seen in cultured tumor cells that, following selection for resistance to a single anticancer agent, become resistant to a range of chemically diverse anticancer agents. These MDR cells show a decrease in intracellular drug accumulation due to active efflux by transporter proteins. The transporter best characterized is P-glycoprotein (Pgp). This protein has been identified in many cancers and has been the target for agents able to inhibit its action, thereby reversing resistance. 2. More recently, another transporter, multidrug resistance-associated protein (MRP) has been identified in a number of MDR human tumor cell lines that do not apparently express Pgp. The presence of MRP at the cell surface of these cells is associated with alterations in drug accumulation and distribution. 3. The gene-encoding MRP has been cloned and sequenced and shown by transfection studies to be able to confer resistance and changes in drug accumulation in sensitive tumor cells. The profile of anticancer drugs expelled in the presence of MRP is similar, but not identical, to that of Pgp. 4. MRP has been identified in a number of different types of cancers, but it is not yet clear to what extent it is involved with clinical resistance. Furthermore, resistance modulators useful against Pgp are less effective in reversing MRP-mediated resistance. 5. It is not fully understood how MRP brings about drug efflux, but it is clear that the underlying mechanisms are different from those responsible for Pgp-mediated drug efflux. In particular, glutathione (GSH) is required for the effective expulsion of the anticancer agents. 6. Unlike Pgp, MRP is able to transport metallic oxyanions and glutathione and other conjugates, including peptidyl leukotrienes. Agents that inhibit organic anion transport, such as probenecid, can block MRP activity. 7. Like Pgp, MRP is expressed not only in resistant tumor cells, but also in normal human tissues. These include the epithelial cells lining the airways and the gastrointestinal tract. In cells in normal tissues, MRP appears to be located within the cytoplasm, which may mean that it functions here in a manner slightly different to that in malignant cells. It is now also recognized in cells and tissues from other species, such as the rat and mouse.


Health-related behaviors of women physicians vs other women in the United States.
To examine the health-related behaviors of women physicians compared with those of other women of high and not high socioeconomic status and with national goals. We examined the results of a questionnaire-based survey of a stratified random sample, the Women Physicians' Health Study, and a US telephone survey (Behavioral Risk Factor Surveillance System of the Centers for Disease Control and Prevention, Atlanta, Ga). We analyzed 3 samples of women aged 30 to 70 years: (1) respondents from the Women Physicians' Health Study (n = 4501); (2) respondents from the Behavioral Risk Factor Surveillance System (n = 1316) of the highest socioeconomic status; and (3) all other respondents from the Behavioral Risk Factor Surveillance System (n = 35,361). Women physicians were more likely than other women of high socioeconomic status and even more likely than other women not to smoke. The few physicians (3.7%) who smoked reported consuming fewer cigarettes per day, and physicians who had stopped smoking reported quitting at a younger age than women in the general population. Women physicians were less likely to report abstaining from alcohol, but those who drank reported consuming less alcohol per episode than other women and were less likely to report binging on alcohol than women in the general population. Unlike women in the general population and even other women of high socioeconomic status, women physicians' reported behaviors exceeded national goals for the year 2000 in all examined behaviors and screening habits. Women physicians report having generally good health habits even when compared with other socioeconomically advantaged women and report exceeding all examined national goals for personal screening practices and other personal health behaviors. Women physicians' behaviors may provide useful standards for other women in the United States.


Intraosseous Regional Prophylactic Antibiotics Decrease the Risk of Prosthetic Joint Infection in Primary TKA: A Multicenter Study.
Recent studies have demonstrated that the administration of regional prophylactic antibiotics by intraosseous (IO) injection achieves tissue concentrations around the knee that are 10- to 15-fold higher than intravenous (IV) delivery of prophylactic antibiotics. It is currently unknown whether the use of regional prophylactic antibiotics for primary TKA would result in a lower risk of prosthetic joint infection (PJI). (1) Is IO injection of prophylactic antibiotics associated with a decreased risk of early (< 12 months) deep PJI compared with traditional IV prophylactic antibiotics? (2) What other patient factors are associated with an increased risk of early PJI after TKA, and do regional prophylactic antibiotics influence these risk factors? (3) Can IO antibiotics be administered to all patients, and what complications occurred from the delivery of IO prophylactic antibiotics? A retrospective comparative study of all primary TKAs (1909 TKAs) over a 5-year period (January 2013 to December 2017) was performed to determine the risk of early PJI. Three primary TKAs did not meet the study inclusion criteria and were excluded from the study, leaving a total of 1906 TKAs (725 IO, 1181 IV) for analysis at a minimum of 12 months after index procedure. Both cohorts exhibited similar ages, BMI, and American Society of Anesthesiologists (ASA) grades; however, a greater proportion of patients in the IO cohort were smokers (p = 0.01), while a greater proportion of patients were diabetic in the IV cohort (p = 0.006). The PJI risk between IO and IV delivery techniques was compared while adjusting for patient demographics and medical comorbidities. Complications related to IO delivery-inability to administer via IO technique, compartment syndrome, fat embolism, and red man syndrome with vancomycin use-were recorded. The delivery of regional prophylactic antibiotics by the IO technique resulted in a lower PJI risk than IV prophylactic antibiotics (0.1% [1 of 725] compared with 1.4% [16 of 1181]; relative risk 0.10 [95% CI 0.01 to 0.77]; p = 0.03). BMI (β = -0.17; standard error = 0.08; p = 0.02), diabetes (β = -1.80; standard error = 0.75; p = 0.02), and renal failure (β = -2.37; standard error = 0.84; p = 0.01) were factors associated with of PJI, while smoking, sex, and ASA score were not contributing factors (p > 0.05). Although BMI, diabetes, and renal failure were identified as infection risk factors, the use of IO antibiotics in these patients did not result in a lower PJI risk compared with IV antibiotics (p > 0.05). IO antibiotics were able to be successfully administered to all patients in this cohort, and there were no complications related to the delivery of IO antibiotics. Surgeons should consider administering regional prophylactic antibiotics in primary TKA to reduce the risk of early PJI. Future randomized prospective clinical trials are needed to validate the efficacy of regional prophylactic antibiotics in reducing the PJI risk in primary TKA. Level III, therapeutic study.


Improved performance of progeny of broiler parent chickens vaccinated with infectious bursal disease oil-emulsion vaccine.
Field trials were conducted on nine broiler chicken flocks, which were the progeny of parents vaccinated with inactivated infectious bursal disease (IBD) oil-emulsion vaccine (OEV). The average increase in overall weight gains was 7.97% of weight gains of similar birds from live-vaccinated parents. Food-conversion ratios (FCR) were calculated for five of the trials. The average improvement in FCR over that of the control chickens was 2.89%. Every broiler flock contracted subclinical IBD, but there was no significant difference in overall mortality rates, except in one trial. In that trial, inclusion body hepatitis and Escherichia coli septicemia in the progeny of the parents that received only live vaccines caused increased mortality. Weekly monitoring of IBD antibodies in two of the trials showed that, in one trial, maternally derived antibody (MDA) persisted in the test chicks until at least 22 days of age and in the controls until at least day 15. In the other trial, MDA persisted until at least day 15 and day 8, respectively, in the test and control flocks. Field IBD challenge occurred at about day 30 and day 37 in the control and test chicks, respectively, in one trial and on day 17 and day 30 in the other. Overall weight gains and FCRs were not related to stocking densities.


The aging lacrimal gland: changes in structure and function.
The afferent nerves of the cornea and conjunctiva, efferent nerves of the lacrimal gland, and the lacrimal gland are a functional unit that works cooperatively to produce the aqueous component of tears. A decrease in the lacrimal gland secretory function can lead to dry eye disease. Because aging is a risk factor for dry eye disease, study of the changes in the function of the lacrimal gland functional unit with age is important for developing treatments to prevent dry eye disease. No one mechanism is known to induce the changes that occur with aging, although multiple different mechanisms have been associated with aging. These fall into two theoretical categories: programmed theories of aging (immunological, genetic, apoptotic, and neuroendocrine) and error theories of aging (protein alteration, somatic mutation, etc). Lacrimal glands undergo structural and functional alteration with increasing age. In mouse models of aging, it has been shown that neural stimulation of protein secretion is an early target of aging, accompanied by an increase in mast cells and lipofuscin accumulation. Hyperglycemia and increased lymphocytic infiltration can contribute to this loss of function at older ages. These findings suggest that an increase in oxidative stress may play a role in the loss of lacrimal gland function with age. For the afferent and efferent neural components of the lacrimal gland functional unit, immune or inflammatory mediated decrease in nerve function could contribute to loss of lacrimal gland secretion with age. More research in this area is critically needed.


How Networks of Informal Trails Cause Landscape Level Damage to Vegetation.
When visitors are not constrained to remain on formal trails, informal trail networks can develop and damage plant communities in protected areas. These networks can form in areas with low growing vegetation, where formal trails are limited, where there is limited regulation and where vegetation is slow to recover once disturbed. To demonstrate the extent of impacts from unregulated recreational use, we assessed damage to alpine vegetation by hikers and pack animals in the highest protected area in the southern Hemisphere: Aconcagua Park, in the Andes. Within the 237 ha area surveyed in the Horcones Valley, over 19 km of trails were found, nearly all of which (94%) were informal. This network of trails resulted in the direct loss of 11.5 ha of vegetation and extensive fragmentation of alpine meadows (21 fragments) and steppe vegetation (68 fragments). When levels of disturbance off these trails were quantified using rapid visual assessments, 81% of 102 randomly located plots showed evidence of disturbance, with the severity of disturbance greatest close to trails. As a result, vegetation in 90% of the Valley has been damaged by visitor use, nearly all of it from unregulated use. These results highlight the extent to which informal trails and trampling off-trail can cause landscape damage to areas of high conservation value, and hence the importance of better regulation of visitor use. The methodology used for off-trail impact assessment can be easily applied or adapted for other popular protected areas where trampling off-trail is also an issue.


[Regeneration of the gastric mucosa under conditions of massive blood loss].
The autoradiographic method was used for a comparative study of regeneration of the gastric mucosa after wounding the rat with a preliminary loss of blood and without it. The H3-thymidin label index was determined in the portions of the mucosa of the wound edge and far from it. A histological analysis of healing the wounds failed to reveal any difference in terms of this process in rats with a loss of blood and without it. A momentary massive loss of blood did not influence the strain and intracellular regeneration processes in the healing wound of the stomach: the amount of cells synthesizing DNA at the edges of the gastrotonic wound proved to be the same in experiments both with a loss of blood and without it. The authors believe that it may serve as an indirect evidence of a somewhat exaggerated danger of surgery at the height of gastroduodenal bleeding.


Tracking the Re-organization of Motor Functions After Disconnective Surgery: A Longitudinal fMRI and DTI Study.
Objective: Mechanisms of motor plasticity are critical to maintain motor functions after cerebral damage. This study explores the mechanisms of motor reorganization occurring before and after surgery in four patients with drug-refractory epilepsy candidate to disconnective surgery. Methods: We studied four patients with early damage, who underwent tailored hemispheric surgery in adulthood, removing the cortical motor areas and disconnecting the corticospinal tract (CST) from the affected hemisphere. Motor functions were assessed clinically, with functional MRI (fMRI) tasks of arm and leg movement and Diffusion Tensor Imaging (DTI) before and after surgery with assessments of up to 3 years. Quantifications of fMRI motor activations and DTI fractional anisotropy (FA) color maps were performed to assess the lateralization of motor network. We hypothesized that lateralization of motor circuits assessed preoperatively with fMRI and DTI was useful to evaluate the motor outcome in these patients. Results: In two cases preoperative DTI-tractography did not reconstruct the CST, and FA-maps were strongly asymmetric. In the other two cases, the affected CST appeared reduced compared to the contralateral one, with modest asymmetry in the FA-maps. fMRI showed different degrees of lateralization of the motor network and the SMA of the intact hemisphere was mostly engaged in all cases. After surgery, patients with a strongly lateralized motor network showed a stable performance. By contrast, a patient with a more bilateral pattern showed worsening of the upper limb function. For all cases, fMRI activations shifted to the intact hemisphere. Structural alterations of motor circuits, observed with FA values, continued beyond 1 year after surgery. Conclusion: In our case series fMRI and DTI could track the longitudinal reorganization of motor functions. In these four patients the more the paretic limbs recruited the intact hemisphere in primary motor and associative areas, the greater the chances were of maintaining elementary motor functions after adult surgery. In particular, DTI-tractography and quantification of FA-maps were useful to assess the lateralization of motor network. In these cases reorganization of motor connectivity continued for long time periods after surgery.


From how small is too small to how much is too much. Ethical issues at the limits of neonatal viability.
In recent years, survival of the extremely low birthweight infant has dramatically improved. Morbidity, both short- and long-term, however, remains very high Costs in the currency of pain and suffering for the newborn, stress and financial burdens for the family, and dollars for society are extensive. The controversial questions that are discussed include: How small is too small? How much is too much? How far should we go? How much can we afford? Who should decide? What strategies should be followed to ensure such decisions are just?


Beta-hydroxydecanoyl thio ester dehydrase does not catalyze a rate-limiting step in Escherichia coli unsaturated fatty acid synthesis.
The intracellular level of beta-hydroxydecanoyl thio ester dehydrase, the product of the fabA gene of Escherichia coli, was increased by isolation of a putative promotor mutant (termed fabAup) or by molecular cloning of the wild-type fabA gene into plasmid pBR322. The fabAup and plasmid-carrying strains overproduced dehydrase by about 15- and 10-fold, respectively. The phospholipids of all strains that overproduced the dehydrase contained significantly higher levels of saturated fatty acids than isogenic strains producing a normal level of dehydrase. No increased levels of unsaturated fatty acids were observed. This result indicates that, although the dehydrase is required for unsaturated fatty acid synthesis, the level of dehydrase activity in wild-type cells does not limit the rate of unsaturated fatty acid synthesis. The introduction of a plasmid carrying the structural gene for beta-ketoacyl acyl carrier protein synthase I into a fabAup strain overcame the effect of dehydrase overproduction on fatty acid composition.


Determining priorities of the Georgia Student Society for Medical Technology: results of a survey.
The opinions of student laboratory personnel regarding the objectives of their student organization were obtained and analyzed. By documenting their perceptions in this manner, the use of unspoken assumptions accompanying formulation and change in organizational direction has been abolished. Additionally, this has allowed them to acquire an understanding of the current perceptions held by their peers, and of the goals cited for making their professional organization more responsive to current and future needs. As a result, it is anticipated that motivation will be heightened and that professional unity, identity, growth, and development will improve.


Quantitative models reveal the organization of diverse cognitive functions in the brain.
Our daily life is realized by the complex orchestrations of diverse brain functions, including perception, decision-making, and action. The essential goal of cognitive neuroscience is to reveal the complete representations underlying these functions. Recent studies have characterised perceptual experiences using encoding models. However, few attempts have been made to build a quantitative model describing the cortical organization of multiple active, cognitive processes. Here, we measure brain activity using fMRI, while subjects perform 103 cognitive tasks, and examine cortical representations with two voxel-wise encoding models. A sparse task-type model reveals a hierarchical organization of cognitive tasks, together with their representation in cognitive space and cortical mapping. A cognitive factor model utilizing continuous, metadata-based intermediate features predicts brain activity and decodes tasks, even under novel conditions. Collectively, our results show the usability of quantitative models of cognitive processes, thus providing a framework for the comprehensive cortical organization of human cognition.


Studies of selected phenyltropanes at monoamine transporters.
3-Phenyltropane analogues of cocaine are useful neurobiologic tools for examining mechanisms of neurotransmitter transporters and psychostimulant drugs. They are also potential substitute medications for psychostimulant abuse. In this study, 18 3-phenyltropane analogues were characterized in uptake and binding studies at dopamine (DAT), norepinephrine (NET) and serotonin (SERT) transporters from the rat, and in binding at DAT in rat, rhesus monkey, and human brain tissue. In rat brain tissue, potency in inhibiting uptake generally correlated with the potency in inhibiting binding at all three transporters suggesting that none of these compounds have antagonist properties. At the DAT, there was a significant correlation of inhibitory potencies between the rat and monkey, the monkey and human, and the rat and human transporters although some compounds showed some species difference. These findings suggest that with regard to the 3-phenyltropane series, there is generally little pharmacologic difference between DATs from the three species examined, although binding data from rat may not be a perfect predictor of uptake inhibition in human.


Left-right locomotor circuitry depends on RhoA-driven organization of the neuroepithelium in the developing spinal cord.
RhoA is a key regulator of cytoskeletal dynamics with a variety of effects on cellular processes. Loss of RhoA in neural progenitor cells disrupts adherens junctions and causes disorganization of the neuroepithelium in the developing nervous system. However, it remains essentially unknown how the loss of RhoA physiologically affects neural circuit formation. Here we show that proper neuroepithelial organization maintained by RhoA GTPase in both the ventral and dorsal spinal cord is critical for left-right locomotor behavior. We examined the roles of RhoA in the ventral and dorsal spinal cord by deleting the gene in neural progenitors using Olig2-Cre and Wnt1-Cre mice, respectively. RhoA-deleted neural progenitors in both mutants exhibit defects in the formation of apical adherens junctions and disorganization of the neuroepithelium. Consequently, the ventricular zone and lumen of the dysplastic region are lost, causing the left and right sides of the gray matter to be directly connected. Furthermore, the dysplastic region lacks ephrinB3 expression at the midline that is required for preventing EphA4-expressing corticospinal neurons and spinal interneurons from crossing the midline. As a result, aberrant neuronal projections are observed in that region. Finally, both RhoA mutants develop a rabbit-like hopping gait. These results demonstrate that RhoA functions to maintain neuroepithelial structures in the developing spinal cord and that proper organization of the neuroepithelium is required for appropriate left-right motor behavior.


Differential shrinkage of mesophyll cells in transpiring cotton leaves: implications for static and dynamic pools of water, and for water transport pathways.
Shrinkage of palisade cells during transpiration, previously measured for sclerophyllous leaves of Eucalyptus where cells shrank equally, was compared with shrinkage in thin mesophytic leaves of cotton (Gossypium hirsutum L.). Selected vapour pressure differences (Δe) from 0.6 to 2.7kPa were imposed during steady-state photosynthesis and transpiration. Leaves were then cryo-fixed and cryo-planed paradermally, and images obtained with a cryo-scanning electron microscope (CSEM). Diameters of palisade 'cavity cells' within sub-stomatal cavities, and surrounding palisade 'matrix cells' were measured on CSEM images. Cavity and spongy mesophyll cells shrank progressively down to Δe=2.7kPa, while matrix cells remained at the same diameter at all Δe. Diameters were also measured of cavity and matrix cells quasi-equilibrated with relative humidities (RHs) from 100% to 86%. In leaves quasi-equilibrated with 95% RH, the cavity cells shrank so much as to be almost unmeasurable, while matrix cells shrank by only 6%. These data suggest that there are two distinct pools of water in cotton leaves: cavity plus spongy mesophyll cells (two-thirds of leaf volume) which easily lose water; and matrix cells (one-third of leaf volume), which retain turgor down to relative water loss=0.4, providing structural rigidity to prevent wilting. This phenomenon is probably widespread among mesophytic leaves.


Health plan responses to Medicaid managed care policy in New York City.
Nearly all states in the United States have instituted managed care programs to serve Medicaid recipients and are developing policies to increase program participation. State practices regarding managed care contracting, premiums, and enrollment have implications for whether managed care plans will respond in a manner consistent with overall state policy objectives for the Medicaid managed care program. The experience of expanding the Medicaid managed care program in New York City, where the number of Medicaid beneficiaries exceeds the number in all but three states, has provided an interesting opportunity to look at the relationship between Medicaid managed care policy and plan enrollment. This paper analyzes trends in Medicaid managed care enrollment in New York City from January 1991 to September 1998, a period of critical changes in Medicaid managed care policy in New York.


A randomized, placebo-controlled trial of topiramate in amyotrophic lateral sclerosis.
To determine if long-term topiramate therapy is safe and slows disease progression in patients with ALS. A double-blind, placebo-controlled, multicenter randomized clinical trial was conducted. Participants with ALS (n = 296) were randomized (2:1) to receive topiramate (maximum tolerated dose up to 800 mg/day) or placebo for 12 months. The primary outcome measure was the rate of change in upper extremity motor function as measured by the maximum voluntary isometric contraction (MVIC) strength of eight arm muscle groups. Secondary endpoints included safety and the rate of decline of forced vital capacity (FVC), grip strength, ALS functional rating scale (ALSFRS), and survival. Patients treated with topiramate showed a faster decrease in arm strength (33.3%) during 12 months (0.0997 vs 0.0748 unit decline/month, p = 0.012). Topiramate did not significantly alter the decline in FVC and ALSFRS or affect survival. Topiramate was associated with an increased frequency of anorexia, depression, diarrhea, ecchymosis, nausea, kidney calculus, paresthesia, taste perversion, thinking abnormalities, weight loss, and abnormal blood clotting (pulmonary embolism and deep venous thrombosis). At the dose studied, topiramate did not have a beneficial effect for patients with ALS. High-dose topiramate treatment was associated with a faster rate of decline in muscle strength as measured by MVIC and with an increased risk for several adverse events in patients with ALS. Given the lack of efficacy and large number of adverse effects, further studies of topiramate at a dose of 800 mg or maximum tolerated dose up to 800 mg/day are not warranted.


Application of computers in clinics in Singapore: status and doctors' perceptions.
A survey of 67 private clinics in Singapore indicates a positive trend towards use of computer-based information systems for administrative and medical applications. The computerised clinics are generally large in terms of number of staff and number of patients, and have been in operation for 3 to 15 years. Specialist clinics have a greater tendency to use computers. The widely used applications in clinics with computers are word processing, accounting, database, medical billing, drugs inventory, laboratory billing and reporting, medical records tracking, and prescription processing. Clinics with computers find: (1) software are generally suitable, (2) objectives of computerisation have been generally achieved, (3) a slight increase in operating costs, (4) a slight increase in staff productivity, and (5) satisfactory vendor support. The responding doctors rate their knowledge of computers from average to poor. Doctors with computers in their clinics hold the perception that: (1) computers can adequately support clinic operations, (2) computer systems are reliable, (3) a genuine need exists for computers in clinics, (4) computers are useful as research tools, and (5) legal and ethical constraints may inhibit use of computers in medical applications. Further analysis of the data has identified some interesting correlations among clinic variables and among doctors' perception variables.


Nightly biting cycles of malaria vectors in a heterogeneous transmission area of eastern Amazonian Brazil.
The biting cycle of anopheline mosquitoes is an important component in the transmission of malaria. Inter- and intraspecific biting patterns of anophelines have been investigated using the number of mosquitoes caught over time to compare general tendencies in host-seeking activity and cumulative catch. In this study, all-night biting catch data from 32 consecutive months of collections in three riverine villages were used to compare biting cycles of the five most abundant vector species using common statistics to quantify variability and deviations of nightly catches from a normal distribution. Three communities were selected for study. All-night human landing catches of mosquitoes were made each month in the peridomestic environment of four houses (sites) for nine consecutive days from April 2003 to November 2005. Host-seeking activities of the five most abundant species that were previously captured infected with Plasmodium falciparum, Plasmodium malariae or Plasmodium vivax, were analysed and compared by measuring the amount of variation in numbers biting per unit time (co-efficient of variation, V), the degree to which the numbers of individuals per unit time were asymmetrical (skewness = g1) and the relative peakedness or flatness of the distribution (kurtosis = g2). To analyse variation in V, g1, and g2 within species and villages, we used mixed model nested ANOVAs (PROC GLM in SAS) with independent variables (sources of variation): year, month (year), night (year X month) and collection site (year X month). The biting cycles of the most abundant species, Anopheles darlingi, had the least pronounced biting peaks, the lowest mean V values, and typically non-significant departures from normality in g1 and g2. By contrast, the species with the most sharply defined crepuscular biting peaks, Anopheles marajoara, Anopheles nuneztovari and Anopheles triannulatus, showed high to moderate mean V values and, most commonly, significantly positive skewness (g1) and kurtosis (g2) moments. Anopheles intermedius was usually, but not always, crepuscular in host seeking, and showed moderate mean V values and typically positive skewness and kurtosis. Among sites within villages, significant differences in frequencies of departures from normality (g1 and g2) were detected for An. marajoara and An. darlingi, suggesting that local environments, such as host availability, may affect the shape of biting pattern curves of these two species. Analyses of co-efficients of variation, skewness and kurtosis facilitated quantitative comparisons of host-seeking activity patterns that differ among species, sites, villages, and dates. The variable and heterogeneous nightly host-seeking behaviours of the five exophilic vector species contribute to the maintenance of stable malaria transmission in these Amazonian villages. The abundances of An. darlingi and An. marajoara, their propensities to seek hosts throughout the night, and their ability to adapt host-seeking behaviour to local environments, contribute to their impact as the most important of these vector species.


Prospective Associations of Dietary and Nutrient Patterns with Fracture Risk: A 20-Year Follow-Up Study.
Studies on long-term exposure to foods/nutrients and its associations with fracture risk are scarce. Using data from the China Health and Nutrition Survey (CHNS), we determined the prospective association of dietary and nutrient patterns with fractures. Data from 15,572 adults aged ≥18 years were analyzed. Fracture occurrence was self-reported and dietary intake data were collected using a 24-h recall method for three consecutive days, for each individual across nine waves (1989-2011). We used cumulative and overall mean, recent and baseline dietary and nutrient exposures. Hazard ratios (HR) were used to determine the associations. Two dietary (traditional and modern) and two nutrient (plant- and animal-sourced) patterns were identified. After adjusting for potential confounders, study participants in the third tertiles (highest intake) of the modern dietary and animal-sourced nutrient patterns' cumulative scores had a 34% (HR = 1.34; 95% CI: 1.06-1.71) and 37% (HR = 1.37; 95% CI: 1.08-1.72) increase in fracture risks compared to those in the first tertiles, respectively. While the overall mean factor scores of dietary and nutrient patterns had a similar (or stronger) pattern of association as the cumulative scores, no association between recent and baseline scores and fracture was found. Greater adherence to a modern dietary and/or an animal-sourced nutrient pattern is associated with a higher risk of total fractures. This suggests that a modern animal based diet is related to bone fragility. A repeated three-day 24-h recall dietary assessment provides a stronger association with fracture compared to a recent or baseline exposure.


[Study progress on chemical compounds, pharmacological action and clinical application of Naoxintong capsule].
Naoxintong capsule (NXTC) is an oral Chinese preparation produced by modern technology, derived from the classic preparation of Buyang Huanwu decoction which was recorded by WANG Qing-ren (Qing dynasty) in Yilingaicuo Juanxia Tanweilun. NXTC is composed of 16 herbs including insect herbs and some blood circulation herbs, with the effects of supplementing Qi and activating blood circulation, dispersing blood stasis and dredging collaterals. In clinical application, it is mainly used for stroke, cerebral infarction, vascular dementia, ischemic cerebrovascular disease, transient ischemic attack, coronary heart disease, angina pectoris, ischemic cardiomyopathy, diabetic cardiomyopathy, myocardial infarction, chronic heart failure, chronic complications of diabetes, essential hypertension, hyperlipidemia and other cardiovascular and cerebrovascular diseases, and has achieved good therapeutic effect on above diseases or their concurrent diseases. Its clinical efficacy is mainly achieved through the improvement in related links of brain protection, neuroprotection, cardioprotection, hemorheology, et al. Nearly 200 chemical constituents identified in NXTC are important pharmacological basis for its functions. At present, however, most of its pharmacological basic researches are focused on plant herbs, and the three kinds of insect herbs remain to be further studied. The researches on clinical effectiveness are more comprehensive; the safety study of long-term application in real world is ongoing by our team, and its results are yet to be published after finishing the study. Through the systematic and comprehensive combing and elaboration of the research progresses on the chemical compositions, pharmacological action and clinical application of NXTC, it can provide a reference for the in-depth study of this preparation, with a great significance for the quality control, secondary development and internationalization promotion of NXTC.


Simultaneous Encapsulation of Nano-Si in Redox Assembled rGO Film as Binder-Free Anode for Flexible/Bendable Lithium-Ion Batteries.
The emerging ubiquitous flexible/wearable electronics are in high demand for compatible flexible/high-energy rechargeable batteries, which set a collaborative goal to promote the electrochemical performance and the mechanical strength of the fundamental flexible electrodes involved. Herein, freestanding flexible electrode of Si/graphene films is proposed, which is fabricated through a scalable, zinc-driven redox layer-by-layer assembly process. In the hybrid films, silicon nanoparticles are intimately encapsulated and confined in multilayered reduced graphene oxide (rGO) nanosheet films. The designed monolithic rGO/Si film possesses several structural benefits such as high mechanical integrity and three-dimensional conductive framework for accessible charge transport and Li+ diffusion upon cycling. When adopted as binder-free electrode in half-cells, the optimized hybrid rGO/Si film delivers high gravimetric capacity (981 mA h g-1 at 200 mA g-1 with respect to the total weight of the electrode) and exceptional cycling stability (0.057% decay per cycle over 1000 cycles at 1000 mA g-1). Besides, the binder-free rGO/Si film anode is further combined with a commercial LiCoO2 foil cathode for completely flexible full cell/battery, which exhibits excellent cycling performance and a high capacity retention of over 95% after 30 cycles under continuous bending. This solution-processable, elaborately engineered, and robust Si/graphene films will further harness the potential of silicon-carbon composites for advanced flexible/wearable energy storage.


The surgical management of childhood bronchiectasis. A review of 96 consecutive pulmonary resections in children with nontuberculous bronchiectasis.
In 195 children with nontuberculous bronchiectasis, periodic bronchography and clinical examinations were conducted over a period of 16 years (average 9.4 years). This was provided a critical assessment of surgical accomplishments in 96 consecutive resections and a parallel observation of 111 cases not submitted to resection. The final clinical assessment of the surgical cases shows 75% to be well or much improved, 22% to be improved, and 4% unchanged, while patients not submitted to resection have remained largely unchanged (69%) or have become worse (23%). The isolated superior segment can be preserved in children with good results, provided there is clear bronchographic evidence that the segment is entirely free of disease. When partially diseased segments are retained and required to fill a large volume, there is a tendency for even slightly altered bronchi to deteriorate postoperatively. Serial bronchography has proved helpful in determining when the disease has reached a mature, stable state and in planning the extent of resection.


GeoExplainer: A Visual Analytics Framework for Spatial Modeling Contextualization and Report Generation.
Geographic regression models of various descriptions are often applied to identify patterns and anomalies in the determinants of spatially distributed observations. These types of analyses focus on answering why questions about underlying spatial phenomena, e.g., why is crime higher in this locale, why do children in one school district outperform those in another, etc.? Answers to these questions require explanations of the model structure, the choice of parameters, and contextualization of the findings with respect to their geographic context. This is particularly true for local forms of regression models which are focused on the role of locational context in determining human behavior. In this paper, we present GeoExplainer, a visual analytics framework designed to support analysts in creating explanative documentation that summarizes and contextualizes their spatial analyses. As analysts create their spatial models, our framework flags potential issues with model parameter selections, utilizes template-based text generation to summarize model outputs, and links with external knowledge repositories to provide annotations that help to explain the model results. As analysts explore the model results, all visualizations and annotations can be captured in an interactive report generation widget. We demonstrate our framework using a case study modeling the determinants of voting in the 2016 US Presidential Election.


Comparison of clinical characteristics of pediatric autoimmune neuropsychiatric disorders associated with streptococcal infections and childhood obsessive-compulsive disorder.
The objectives of this study were to identify unique clinical characteristics of children with pediatric autoimmune neuropsychiatric disorders associated with streptococcal infections (PANDAS) compared with a control group of children with non-PANDAS obsessive-compulsive disorder (OCD) with respect to ancillary symptoms, types of obsessions and compulsions, symptom severity, and co-morbid DSM-IV diagnoses. Classification of PANDAS was based on review of pediatric and psychiatric records using the criteria developed by Swedo and colleagues. Children aged 6-14 with PANDAS (n = 21) and non-PANDAS OCD (n = 18) were assessed by blind independent evaluators using the PANDAS Questionnaire, Children's Yale-Brown Obsessive Compulsive Scale, Yale Global Tic Severity Scale, and Anxiety Disorders Interview Schedule for DSM-IV. PANDAS children were significantly more likely to present with separation anxiety, urinary urgency, hyperactivity, impulsivity, deterioration in handwriting, and decline in school performance during their initial episode of neuropsychiatric illness compared with children with OCD. Total tics and vocal tics were more severe in PANDAS children. Separation anxiety disorder and social phobia were more prevalent in non-PANDAS OCD children. Children with non-PANDAS OCD were significantly more likely to include others in their rituals. There were no significant differences between groups on demographics or severity of OCD. Distinguishing clinical characteristics in PANDAS, which included urinary urgency, hyperactivity, impulsivity, and deterioration in handwriting, are linked to basal ganglia functions. These clinical characteristics will aid in the differentiation of PANDAS children for research and clinical purposes and ultimately advance our understanding and treatment of this disorder.


Hydrogels in spinal cord injury repair strategies.
Nowadays there are at present no efficient therapies for spinal cord injury (SCI), and new approaches have to be proposed. Recently, a new regenerative medicine strategy has been suggested using smart biomaterials able to carry and deliver cells and/or drugs in the damaged spinal cord. Among the wide field of emerging materials, research has been focused on hydrogels, three-dimensional polymeric networks able to swell and absorb a large amount of water. The present paper intends to give an overview of a wide range of natural, synthetic, and composite hydrogels with particular efforts for the ones studied in the last five years. Here, different hydrogel applications are underlined, together with their different nature, in order to have a clearer view of what is happening in one of the most sparkling fields of regenerative medicine.


Differences in the central serotonergic effects of methylenedioxymethamphetamine (MDMA) in mice and rats.
The effects of subcutaneous injection of 3,4-methylenedioxymethamphetamine (MDMA), a psychoactive amphetamine congener, on mouse central monoaminergic systems were assessed and compared to effects in rats. Whereas neostriatal concentrations of 5-hydroxytryptamine and 5-hydroxyindoleacetic acid in mouse were transiently decreased after a single moderately high dose of MDMA (15 mg/kg), mouse neostriatal or hippocampal tryptophan hydroxylase activity was not significantly affected, even after a dose of 60 mg/kg. These results are in contrast to effects in rats, in which a single 10 mg/kg dose of MDMA induced immediate and prolonged decreases in both central tryptophan hydroxylase activity and 5-hydroxyindole concentrations. Decreases in mouse central tryptophan hydroxylase activity, and prolonged decreases in mouse 5-hydroxyindole concentrations, were observed only after multiple doses of MDMA, suggesting that the duration of exposure may be an important determinant of toxic effects. These results show mice to be less susceptible than rats to MDMA-induced neurotoxicity, and are discussed in terms of possible interspecies differences in MDMA pharmacokinetics.


The effect of intravenous maropitant on blood pressure in healthy awake and anesthetized dogs.
To evaluate the effects of intravenous maropitant on arterial blood pressure in healthy dogs while awake and under general anesthesia. Experimental crossover study. Eight healthy adult Beagle dogs. All dogs received maropitant (1 mg kg-1) intravenously under the following conditions: 1) awake with non-invasive blood pressure monitoring (AwNIBP), 2) awake with invasive blood pressure monitoring (AwIBP), 3) premedication with acepromazine (0.005 mg kg-1) and butorphanol (0.2 mg kg-1) intramuscularly followed by propofol induction and isoflurane anesthesia (GaAB), and 4) premedication with dexmedetomidine (0.005 mg kg-1) and butorphanol (0.2 mg kg-1) intramuscularly followed by propofol induction and isoflurane anesthesia (GaDB). Heart rate (HR), systolic (SAP), diastolic (DAP), and mean blood pressures (MAP) were recorded before injection of maropitant (baseline), during the first 60 seconds of injection, during the second 60 seconds of injection, at the completion of injection and every 2 minutes post injection for 18 minutes. The data were compared over time using a Generalized Linear Model with mixed effects and then with simple effect comparison with Bonferroni adjustments (p <0.05). There were significant decreases from baseline in SAP in the GaAB group (p < 0.01) and in MAP and DAP in the AwIBP and GaAB (p < 0.001) groups during injection. A significant decrease in SAP (p < 0.05), DAP (p < 0.05), and MAP (p < 0.05) occurred at 16 minutes post injection in GaDB group. There was also a significant increase in HR in the AwIBP group (p < 0.01) during injection. Clinically significant hypotension occurred in the GaAB group with a mean MAP at 54 ± 6 mmHg during injection. Intravenous maropitant administration significantly decreases arterial blood pressure during inhalant anesthesia. Patients premedicated with acepromazine prior to isoflurane anesthesia may develop clinically significant hypotension.


[Discovering the authentic Hildegard. Comments on the only in Florance 'Physica' handwritten texts].
The recently discovered ms. of Hildegard's, Physica' (Florence, B. Med.-Laurenz. Ashburnh. 1323) contains much more text than the Migne edition, based on Paris, BN.lat. 6952, does. Did the Florence ms. add or the Paris ms. omit? A comparison of all extant mss. and the edition princeps of 1533 reveals that the corpus of items and their order remain essentially the same, whereas the number of recipes and applications is in permanent decrease. We also observe an increasing reduction of explanatory remarks unique to Hildegard's medical doctrine. The more copious text of the Florence ms. must, therefore, be regarded as authentic, and it should be made the base of a future edition of the 'Physica'.


Making sense of the cause of Crohn's - a new look at an old disease.
The cause of Crohn's disease (CD) has posed a conundrum for at least a century. A large body of work coupled with recent technological advances in genome research have at last started to provide some of the answers. Initially this review seeks to explain and to differentiate between bowel inflammation in the primary immunodeficiencies that generally lead to very early onset diffuse bowel inflammation in humans and in animal models, and the real syndrome of CD. In the latter, a trigger, almost certainly enteric infection by one of a multitude of organisms, allows the faeces access to the tissues, at which stage the response of individuals predisposed to CD is abnormal. Direct investigation of patients' inflammatory response together with genome-wide association studies (GWAS) and DNA sequencing indicate that in CD the failure of acute inflammation and the clearance of bacteria from the tissues, and from within cells, is defective. The retained faecal products result in the characteristic chronic granulomatous inflammation and adaptive immune response. In this review I will examine the contemporary evidence that has led to this understanding, and look for explanations for the recent dramatic increase in the incidence of this disease.


Supervisory neglect and adolescent alcohol use disorders: effects on AUD onset and treatment outcome.
While parental supervision has been demonstrated to influence adolescent alcohol involvement, lack of a threshold defining inadequate supervision and little study of DSM defined alcohol use disorders (AUDs) have limited clinical applications. Utilizing a four-item supervision scale, this study identified adolescents with inadequate supervision, and examined effects on AUD development and treatment outcome. Two groups of adolescent subjects (ages 14-17) were recruited: (1) a representative community sample of adolescents (n = 170) and (2) adolescents with AUDs from drug and alcohol or psychiatric treatment programs (n = 194). Supervision was determined by the adolescent's responses to items inquiring whether parents typically had knowledge of the adolescent's activities away from home, including with whom, where, when, and communication items. Adolescents with unusually low parental supervision (i.e., Supervisory Neglect) were identified by scale distributions. Follow-up assessments were conducted 1, 3, and 5 years after the initial assessment. Among community subjects, adolescents with inadequate supervision were significantly more likely to drink alcohol in a variety of situations, were more likely to have AUDs at the initial assessment and, among those without AUDs, were more likely to develop AUDs in the follow-up period. Among adolescents receiving treatment for AUDs, those with inadequate supervision were less likely to be free of AUD symptoms over a 1-year follow-up period. These results indicate that the described method was a straightforward and potentially clinically applicable approach to identifying adolescents with inadequate parental supervision.


Safety of distilled water as an irrigating fluid for transurethral resection of the prostate.
Irrigating fluid is necessary for transurethral resection of the prostate (TURP). Initially, distilled water was used as irrigating fluid and some advantages and complications were noted. Although other solutions were used to substitute for distilled water, we collected data for changes in electrolytes and hematocrit (Hct) during surgery to evaluate the safety of distilled water as irrigating fluid for TURP with suprapubic trocar cystostomy. Fifty-three patients who underwent TURP using distilled water as irrigating fluid were investigated. TURP was performed with suprapubic trocar cystostomy routinely. Preoperative and postoperative blood samples were collected and analyzed for electrolytes and Hct to study any changes during surgery. The change in plasma sodium concentration was not statistically significant, but the decreases in blood Hct and plasma potassium concentration were statistically significant. There was no major operative complication and no clinical-evident related postoperative renal impairment. Distilled water is a relatively safe solution as an irrigating fluid for TURP under drainage with suprapubic trocar cystostomy, but attention must be paid to the inevitable blood loss during surgery.


A critical analysis of the resource-based relative value scale.
BACKGROUND--There is a general perception that procedural medical services are reimbursed at an inappropriately greater rate than cognitive services. By congressional mandate, the Health Care Financing Administration (HCFA) has been directed to establish a Medicare fee schedule to shift funding under a budget-neutral assumption from procedural to cognitive services. To provide a rational basis for this change, Hsiao et al (Harvard-Hsiao) developed a resource-based relative value scale (RBRVS) that equates the value of a service to the resources necessary to generate the service. METHODS--Instead of focusing on relative values and fee schedules ("price-per-unit-service"), the present study employs the standard commercial/industrial method of determining reimbursement rate (income divided by hours of labor) for 15 medical and surgical specialties. Data from independent sources are used to determine income and hours of professional effort for each of the specialties studied. Harvard-Hsiao and HCFA predicted the percent change in income for each of the specialties under the initial RBRVS and the HCFA fee schedule. The predicted income was then employed in this study to recompute reimbursement rates under the newly proposed payment systems. CURRENT PAYMENT SYSTEM--Average annual incomes for medical and surgical specialties are $124,500 and $176,600, respectively, a 42% difference (P = .03). Average weekly work hours (nominal hours, as adjusted for overtime) for medical and surgical specialties are 70.6 and 87.8, respectively (P = .005). Average hourly reimbursement rates for medical ($33.90) and surgical ($38.80) specialties are not substantially different (P, not significant). The difference in annual income is explained by the 17.2 hours per week of additional work hours by surgeons. The erroneous perception that procedurists are reimbursed at a higher rate than cognitive practitioners likely arises from differences in billing methods by which surgeons shift charges for cognitive work hours to the 18% of their time spent in the operating room. PROPOSED RBRVS AND HCFA PAYMENT SYSTEM--The income of all specialties is equalized about a mean of $132,500 (+/- $21,400 [1 SD]) by varying reimbursement rates in such a way that the effect of working hours is fully discounted. Reimbursement rates under the proposed payment system make no recognition of the hours of professional effort, postgraduate specialty training, or putative differences in the nature of the physician's work. --The RBRVS, and the HCFA fee schedule to the extent that it is based on that scale, are inappropriate bases for the reform of the physician reimbursement system.


Genetic identification of the site of DNA contact in the yeast heat shock transcription factor.
The heat shock transcription factor (HSF), a trimeric transcription factor, activates the expression of heat shock genes in eukaryotes. We have isolated mutations in the HSF1 gene from Saccharomyces cerevisiae that severely compromise the ability of HSF to bind to its normal binding site, repeats of the module nGAAn. One of these mutations, Q229R, shows a "new specificity" phenotype, in which the protein prefers the mutant sequence nGACn. These results identify the region of HSF that contacts DNA, in complete agreement with the crystal structure of HSF of Kluyveromyces lactis and the nuclear magnetic resonance data from HSF of Drosophila melanogaster. To determine the orientation of the DNA-binding domain on the nGAAn motif, we performed site-specific cross-linking between cysteine residues of single-cysteine substitutions. Cysteines placed at the N terminus of the DNA contact helix formed cross-links readily, while cysteines placed at the C terminus of the helix did not.


Heatwaves do not limit recovery following defoliation but alter leaf drought tolerance traits.
As heatwave frequency increases, they are more likely to coincide with other disturbances like insect defoliation. But it is unclear if high temperatures after defoliation impact canopy recovery or leaf traits which may affect response to further stressors like drought. To examine these stressor interactions, we subjected defoliated (DEF) and undefoliated (UNDEF) oak saplings to a simulated spring heatwave of +10°C for 25 days. We measured gas exchange, leaf area recovery, carbohydrate storage, turgor loss point (ΨTLP ), and minimum leaf conductance (gmin ). During the heatwave, stem respiration exhibited stronger thermal acclimation in DEF than UNDEF saplings, while stomatal conductance and net photosynthesis increased. The heatwave did not affect leaf area recovery or carbohydrate storage of DEF saplings, but reflush leaves had higher gmin than UNDEF leaves, and this was amplified by the heatwave. Across all treatments, higher gmin was associated with higher daytime stomatal conductance and a lower ΨTLP . The results suggest defoliation stress may not be exacerbated by higher temperatures. However, reflush leaves are less conservative in their water use, limiting their ability to minimise water loss. While lower ΨTLP could help DEF trees maintain gas exchange under mild drought, they may be more vulnerable to dehydration under severe drought.


Preclinical Evaluation of the Hsp70 Peptide Tracer TPP-PEG24-DFO[89Zr] for Tumor-Specific PET/CT Imaging.
High precision in vivo PET/CT imaging of solid tumors improves diagnostic credibility and clinical outcome of patients. An epitope of the oligomerization domain of Hsp70 is exclusively exposed on the membrane of a large variety of tumor types, but not on normal cells, and thus provides a universal tumor-specific target. Here we developed a novel PET tracer TPP-PEG24-DFO[89Zr] based on the tumor cell-penetrating peptide probe TPP, which specifically recognizes membrane Hsp70 (mHsp70) on tumor cells. The implemented PEG24 moiety supported tracer stability and improved biodistribution characteristics in vivo The K d of the tracer ranged in the low nanomolar range (18.9 ± 11.3 nmol/L). Fluorescein isothiocyanate (FITC)-labeled derivatives TPP-[FITC] and TPP-PEG24-[FITC] revealed comparable and specific binding to mHsp70-positive 4T1, 4T1+, a derivative of the 4T1 cell line sorted for high Hsp70 expression, and CT26 tumor cells, but not to mHsp70-negative normal fibroblasts. The rapid internalization kinetics of mHsp70 into the cytosol and the favorable biodistribution of the peptide-based tracer TPP-PEG24-DFO[89Zr] in vivo enabled a tumor-specific accumulation with a high tumor-to-background contrast and renal body clearance. The tumor-specific enrichment of the tracer in 4T1+ (6.2 ± 1.1%ID/g), 4T1 (4.3 ± 0.7%ID/g), and CT26 (2.6 ± 0.6%ID/g) mouse tumors with very high, high, and intermediate mHsp70 densities, respectively, reflected mHsp70 expression profiles of the different tumor types, whereas benign mHsp70-negative fibroblastic hyperplasia showed no tracer accumulation (0.2 ± 0.03%ID/g). The ability of our chemically optimized peptide-based tracer TPP-PEG24-DFO[89Zr] to detect mHsp70 in vivo suggests its broad applicability in targeting and imaging with high specificity for any tumor type that exhibits surface expression of Hsp70.Significance: A novel peptide-based PET tracer against the oligomerization domain of Hsp70 has potential for universal tumor-specific imaging in vivo across many tumor type. Cancer Res; 78(21); 6268-81. ©2018 AACR.


Excitations and stripe phase formation in a two-dimensional dipolar Bose gas with tilted polarization.
We present calculations of the ground state and excitations of an anisotropic dipolar Bose gas in two dimensions, realized by a nonperpendicular polarization with respect to the system plane. For sufficiently high density, an increase of the polarization angle leads to a density instability of the gas phase in the direction where the anisotropic interaction is strongest. Using a dynamic many-body theory, we calculate the dynamic structure function in the gas phase which shows the anisotropic dispersion of the excitations. We find that the energy of roton excitations in the strongly interacting direction decreases with increasing polarization angle and almost vanishes close to the instability. Exact path integral ground state Monte Carlo simulations show that this instability is indeed a quantum phase transition to a stripe phase, characterized by long-range order in the strongly interacting direction.


The cell cycle of parasitic protozoa: potential for chemotherapeutic exploitation.
There is an urgent need to develop new drugs against eukaryotic parasitic protozoa such as Plasmodium, Trypanosoma and Leishmania, which cause the diseases malaria, trypanosomiasis and the leishmaniases respectively. The biology of these organisms has many unusual facets that might be exploited for drug design, and the recent availability of parasite genome sequence data has facilitated the search for novel drug targets. Here we review current understanding of the cell cycle in these parasites and show that important structural and functional differences exist between parasite and mammalian cell cycle control machineries and signal transduction pathways, which might be utilised for rational drug design. Potential targets include protein kinases from the cyclin-dependent kinase, cAMP-dependent kinase and mitogen activated protein kinase families.