Evaluation of the Haemoglobin Colour Scale and comparison with the HemoCue haemoglobin assay.
To evaluate the Haemoglobin Colour Scale developed by WHO for estimating haemoglobin concentration and to compare the results obtained using it and the HemoCue assay with those determined using a reference method, the Technicon H3 analyser. The Colour Scale and HemoCue assay were used to test 408 blood samples. Subsequently, Bland-Altman plots were determined and the proximity of the test results to those obtained using the reference method was determined. The mean difference between the Haemoglobin Colour Scale and the reference method was 0.19 g/dl (95% confidence interval: 3.50 g/dl below to 3.11 g/dl above); the corresponding value for the HemoCue assay was 0.50 g/dl (1.16 g/dl below to 0.16 g/dl above). Only 46.08% of the results obtained by means of the Colour Scale were within 1.0 g/dl of the reference method, whereas 95.34% of the HemoCue results fell within this limit; 22.79% of the Colour Scale results but none of the HemoCue results lay more than 2.0 g/dl from the reference method. The Haemoglobin Colour Scale test is too inaccurate for general use, particularly if devices such as the HemoCue are available.


Third-Order Muscle Models: The Role of Oscillatory Behavior In Force Control.
This paper presents the analysis of a third-order linear differential equation representing a muscle-tendon system, including the identification of critical damping conditions. We analytically verified that this model is required for a faithful representation of muscle-skeletal muscles and provided numerical examples using the biomechanical properties of muscles and tendon reported in the literature. We proved the existence of a theoretical threshold for the ratio between tendon and muscle stiffness above which critical damping can never be achieved, thus resulting in an oscillatory free response of the system, independently of the value of the damping. Oscillation of the limb can be compensated only by active control, which requires creating an internal model of the limb mechanics. We demonstrated that, when admissible, over-damping of the muscle-tendon system occurs for damping values included within a finite interval between two separate critical limits. The same interval is a semi-infinite region in second-order models. Moreover, an increase in damping beyond the second critical point rapidly brings the system to mechanical instability.


Familial temporal lobe epilepsy with aphasic seizures and linkage to chromosome 10q22-q24.
To describe the phenotypic expression of a new family with familial lateral temporal lobe epilepsy with aphasic seizures, and to compare the findings with the clinical features of previously reported families linked to chromosome 10q22-q24. Medical records were collected from 12 living affected members. The patients underwent a personal interview and a clinical neurologic examination. Results from interictal scalp EEGs and neuroimaging examinations were obtained. The cardinal ictal symptom was a brief sensory aphasia in eight of the patients. In four, this was accompanied by auditory symptoms, usually in the form of monotonous unformed sounds. Simple partial seizures with psychic or somatosensory seizures also were present. Visual ictal symptoms and complex partial seizures were absent. All patients had generalized tonic-clonic seizures. Magnetic resonance imaging (MRI) or computed tomography (CT) did not reveal morphologic correlates. Improvement with age seemed to occur in many patients. Significant linkage to chromosome 10q22-q24 was established by testing 17 polymorphic microsatellite markers. The epilepsy of this family appears to represent a variety of autosomal dominant lateral temporal lobe epilepsy. Aphasic seizures and a peculiar seizure-precipitating effect of the activation of speech (initiation or perception) may serve as markers for identifying further families with this phenotype.


Opportunistic infections caused by protozoan parasites.
Parasitic protozoa have played a major role in the discovery of the HIV epidemic. It was the occurrence of clustered cases of Pneumocystis pneumonia which caught the attention of the epidemiologists indicating that something unusual was going on. Very recently remarkable progress was made in understanding the epidemiology. Knowledge is evolving that Pneumocystis is host-specific to an extent which seems to justify species rank for the cause of death in AIDS patients for many years. Toxoplasma gondii infections were also recognized very early in AIDS patients. Cerebral toxoplasmosis has now replaced Pneumocystis pneumonia as a fatal infection in many cases. AIDS patients with CD4 cells below 200/microl have a high risk to develop this complication. Cryptosporidium parvum was recognized as the cause of diarrhea in AIDS patients and also in immunocompetent humans where it only leads to a diarrheal episode lasting a few days. In AIDS patients the persisting diarrhea leading to loss of fluid and electrolytes can be life-threatening. Another group of parasites previously unknown to infect humans are the microsporidia, which are wide-spread parasites in nature. Increasing numbers of such infections are observed but many questions regarding pathogenesis and epidemiology are still unclear.


Phytochemical analysis of traditional Chinese medicine using liquid chromatography coupled with mass spectrometry.
Traditional Chinese medicine (TCM) is commonly considered to operate due to the synergistic effects of all the major and minor components in the medicines. Hence sensitive and comprehensive analytical techniques are needed to acquire a better understanding of the pharmacological basis of the herb and to enhance the product quality control. The present review mainly focuses on the phytochemical analysis of TCMs using high-performance liquid chromatography coupled with mass spectrometry (HPLC-MS). Atmospheric pressure chemical ionization (APCI) and electrospray ionization (ESI) are the two commonly used ion sources. Triple quadrupole, ion trap (IT), Fourier transform ion cyclotron resonance (FTICR) and time-of-flight (TOF) mass spectrometers are used as on-line analyzer. The relationship between structural features and fragmentation patterns should be investigated as thoroughly as possible and hence be applied in the on-line analysis to deduce the structures of detected peaks. Characteristic fragmentation behaviors of the reference standards, as well as information regarding polarity obtained from retention time data, on-line UV spectra, data from the literature and bio-sources of the compounds allowed the identification of the phytochemical constituents in the crude extracts. Although a mass spectrometer is not a universal detector, high-performance liquid chromatography coupled with multistage mass spectrometry (HPLC-MS(n)) technique was still proved to be a rapid and sensitive method to analyze the majority of the many constituents in herbal medicines, particularly for the detection of those present in minor or trace amounts. The methods established using HPLC-MS techniques facilitate the convenient and rapid quality control of traditional medicines and their pharmaceutical preparations. However, the quantitative analysis is not the topic of this review.


[A role of radionuclide studies for gastrointestinal and hepatobiliary diseases].
Radionuclide studies for gastrointestinal and hepatobiliary diseases were presented. Some are routinely done, but others are rarely done due to complicated technique. For the evaluation of functional abnormalities in gastrointestinal and hepatobiliary diseases, an important role of the radionuclide studies was discussed.


Mice deficient in heparan sulfate N-deacetylase/N-sulfotransferase 1.
Ndsts (N-deacetylase/N-sulfotransferases) are enzymes responsible for N-sulfation during heparan sulfate (HS) and heparin biosynthesis. In this review, basic features of the Ndst1 enzyme are covered and a brief description of HS biosynthesis and its regulation is presented. Effects of Ndst1 deficiency on embryonic development are described. These include immature lungs, craniofacial dysplasia and eye developmental defects, branching defect during lacrimal gland development, delayed mineralization of the skeleton, and reduced pericyte recruitment during vascular development. A brief account of the effects of Ndst1 deficiency in selective cell types in adult mice is also given.


Extension distribution for DNA confined in a nanochannel near the Odijk regime.
DNA confinement in a nanochannel typically is understood via mapping to the confinement of an equivalent neutral polymer by hard walls. This model has proven to be effective for confinement in relatively large channels where hairpin formation is frequent. An analysis of existing experimental data for Escherichia coli DNA extension in channels smaller than the persistence length, combined with an additional dataset for λ-DNA confined in a 34 nm wide channel, reveals a breakdown in this approach as the channel size approaches the Odijk regime of strong confinement. In particular, the predicted extension distribution obtained from the asymptotic solution to the weakly correlated telegraph model for a confined wormlike chain deviates significantly from the experimental distribution obtained for DNA confinement in the 34 nm channel, and the discrepancy cannot be resolved by treating the alignment fluctuations or the effective channel size as fitting parameters. We posit that the DNA-wall electrostatic interactions, which are sensible throughout a significant fraction of the channel cross section in the Odijk regime, are the source of the disagreement between theory and experiment. Dimensional analysis of the wormlike chain propagator in channel confinement reveals the importance of a dimensionless parameter, reflecting the magnitude of the DNA-wall electrostatic interactions relative to thermal energy, which has not been considered explicitly in the prevailing theories for DNA confinement in a nanochannel.


The role of galactolipids in spinach chloroplast lamellar membranes: I. Partial purification of a bean leaf galactolipid lipase and its action on subchloroplast particles.
A galactolipid lipase has been isolated and partially purified from the chloroplast fraction of the primary leaves of Phaseolus vulgaris var. Kentucky Wonder. The lipase hydrolyzed monogalactosyl diglyceride rapidly and phosphatidyl choline relatively slowly. Triolein and p-nitrophenyl stearate were not hydrolyzed.Spinach subchloroplast particles were excellent substrates for the lipase. Initial rates of fatty acid release from subchloroplast particles at 30 C by the lipase as high as 60 microequivalents per minute per milligram protein were observed. At completion of the reaction, about 2.7 microequivalents of fatty acid were liberated per milligram of chlorophyll in the subchloroplast particles, indicating that major amounts of lipid in the particles were rapidly attacked by the lipase.The treatment of subchloroplast particles with the lipase resulted in a rapid inhibition of light-dependent electron flow. This inhibition was largely prevented when the incubation was carried out in the presence of high concentrations of defatted bovine serum albumin. These results suggest that when precautions are taken to prevent the binding of fatty acids to the subchloroplast particles, large amounts of lipid may be removed without a marked effect on electron flow.


Selective inhibition of Colorado potato beetle cathepsin H by oryzacystatins I and II.
The use of oryzacystatins I and II, two cysteine proteinase inhibitors naturally produced in rice grains, represents an attractive way for the control of Coleoptera insect pests. The present study was done to analyze the inhibitory effect of recombinant oryzacystatins produced in Escherichia coli as fusion proteins against digestive proteinases of the major pest Colorado potato beetle (Leptinotarsa decemlineata Say). Both inhibitors had a significant effect on total proteolytic activity, but maximal inhibitions ranged from 20 to 80% for pHs varying from 5.0 to 7.0, respectively. This pH-dependent efficiency of plant cystatins was due to the selective inactivation of potato beetle cathepsin H, as demonstrated by the use of inhibitors with different specificities against cathepsins B and H. These results demonstrate the importance of having an adequate knowledge of insect proteinases specifically recognized by the inhibitors to be used in pest control strategies.


[Research progress on the expression and carcinogenic mechanisms of SETD2 in malignant tumors].
The human epigenetic gene of SET domain containing 2 (SETD2) is located at the cytogenetic band p21.31 of chromosome 3, which encodes the histone3 lysine36 trimethyltransferase SETD2, the major enzyme that catalyzes the trimethylation of lysine 36 on histone 3 (H3K36me3) of human. SETD2 involves in many pathologic and physiological processes such as transcriptional elongation, DNA damage repair, alternative splicing, epigenetic modifications of gene expression, viral immunology, embryonic development, and angiogenesis. A growing list of tumor types including renal cell carcinoma and mesothelioma develops with mutation or inactivation of SETD2. SETD2 loss-of-function promotes the occurrences and development of cancers by inhibiting process of transcription extension, damage repair, cell cycles, apoptosis and cell metabolism. The under expression and mutation of SETD2 is synthetically lethal with the inhibition of G2M checkpoint and PI3K-AKT pathway and SETD2 is considered as a potential epigenetic therapy targets. Furthermore, a loss of SETD2 indicates worse pathological features. There are huge prospects in the diagnosis and treatment of related cancers.


Recombinant human thrombopoietin alleviates infection-associated thrombocytopenia: a retrospective study in senile patients.
To examine the efficacy and adverse events of recombinant human thrombopoietin (rhTPO) in the treatment of infection-associated thrombocytopenia in senile patients. The current study is a retrospective analysis of the patients receiving rhTPO for infection-associated thrombocytopenia in our hospital. Forty-nine cases were included in the analysis as rhTPO group. The absolute platelet count after treatment, increase in platelet count, and the overall response rate were considerably higher in the rhTPO group than that in the control group. Improvement in bleeding score was higher in the rhTPO treatment group than that in the control group (2.1 ± 5.4 vs 0.4 ± 1.7). Bleeding event was stopped in 68.2% of the patients after rhTPO treatment and in 35% of the patients in the control group (P = .032). A stratified analysis indicated that the therapeutic efficacy is much better in patients without organ failure. Recombinant human TPO is effective in alleviating infection-associated thrombocytopenia and hemorrhage in senile patients, particularly if given prior to the emergence of organ failure.


Chemosensitizing effect of nordihydroguaiaretic acid and its tetra-acetylated derivative on parental and multiresistant TA3 mouse mammary adenocarcinoma cells.
Multidrug resistance (MDR) continues being the major obstacle for successful anticancer chemotherapy. The action of nordihydroguaiaretic acid (NDGA) and its tetra-acetylated derivative (NDGATA) on TA3 mouse mammary adenocarcinoma cells and their ability to restore doxorubicin (DOX), cisplatin (CPT) and methotrexate (MTX) sensitivity of the multiresistant variant TA3-MTX-R was examined. Both NDGA and NDGATA synergistically enhanced the cytotoxicity of DOX, CPT and MTX, with a more evident effect in the TA3-MTX-R than in the TA3 cells. NDGATA was more effective than NDGA, as analyzed by the isobologram method. The combination of NDGATA and DOX also reduced the tumor growth rate in mice. Although it did not prolong the median survival time, 30% of mice showed no vestiges of tumor 200 days after implantation with either TA3 or TA3-MTX-R cells. Moreover, NDGA and NDGATA increased the accumulation of DOX and rhodamine (RHO) 123 in both cell lines. NDGA and NDGATA are able to chemosensitize tumor cells and combination therapy with NDGATA and DOX is effective at inhibiting tumor growth in mice.


Timeliness of breast cancer diagnosis and initiation of treatment in the National Breast and Cervical Cancer Early Detection Program, 1996-2005.
To determine the effects of program policy changes, we examined service delivery benchmarks for breast cancer screening in the National Breast and Cervical Cancer Early Detection Program (NBCCEDP). We analyzed NBCCEDP data for women with abnormal mammogram or clinical breast examination (n=382 416) from which 23 701 cancers were diagnosed. We examined time to diagnosis and treatment for 2 time periods: 1996 to 2000 and 2001 to 2005. We compared median time for diagnostic, treatment initiation, and total intervals with the Kruskal-Wallis test. We calculated adjusted proportions (predicted marginals) with logistic regression to examine diagnosis and treatment within program benchmarks (<or=60 days) and time from screening to treatment (<or=120 days). Median diagnostic intervals decreased by 2 days (25 vs 23; P<.001). Median treatment initiation intervals increased by 2 days (12 vs 14; P<.001). Total intervals decreased by 3 days (43 vs 40; P<.001). Women meeting the 60-day benchmark for diagnosis improved the most for women with normal mammograms and abnormal clinical breast examinations from 77% to 82%. Women screened by the NBCCEDP received diagnostic follow-up and initiated treatment within preestablished program guidelines.


Quality of life in type 2 diabetic patients and related effective factors.
Diabetes type 2, as one of the most important chronic diseases in the world, reduces quality of life in patients. Our research--as a descriptive analytical cross-sectional study--was performed on 120 type 2 diabetics in Tehran 503 hospital in 2008. Diverse domain scores of patients' quality of life were evaluated by SF-36 standard questionnaire. Gained data analyzed by SPSS 16.0 software through statistical tests including Pearson's correlation coefficient, Chi-square test, Independent T-test, Kolmogrov-Smirnov test, Mann-Whitney test, Kruskal Wallis test, ANOVA, and Fisher exact test. Age had significant reverse relationship with physical functioning, vitality, mental health, and mental component score. Education level was just directly correlated to bodily pain. Marital status was correlated with social functioning and general health domains. Family income level was directly related to mental health and mental component score domains. No relation was found between years of disease duration, smoking status, workout status, diet follow-up, lack/presence of hypertension, and diverse domains of these patients' quality of life. Hyperlipidemia had meaningful reverse relationship to physical functioning, bodily pain, social functioning, and physical component score domains. Heart diseases had meaningful reverse relationship to physical functioning and mental health domains. Patients' physical component score had direct relationship to mental component score. Observed scores were meaningfully low in comparison with both healthy people and similar surveys.


Two-stage testing using selection schemes.
In this paper, we consider pooling schemes in which samples are to be tested in two-stages. We show that when batch size is limited as well as pool size, selection schemes tend to be more efficient and flexible. Formulae for the efficiencies of square arrays in all dimensions and for all selection schemes are given in the Appendix. A Fortran program has been written to find whether the most efficient scheme under constraints of batch and pool size be 'square' or 'selection'.


Efficience quantique et efficience quantique de détection des photomultiplicateurs et des matériaux photographiques.
The cathode quantum efficiency of several photomultipliers is evaluated at He-Ne laser wavelength from manufacturer's data. The quantum efficiency of the photographic process is defined as the ratio of developed grains (fog grains deduced) in an area A over the number of incident quanta. Five photographic materials were tested, and the quantum efficiency lies between 4 x 10(-4) and 1.1 x 10(-2); the quantum efficiency of photographic materials may be comparable to that of the cathode of photomultipliers. The detective quantum efficiency (EQD) of any detector is defined as the mean square of the SNR over the square of the same ratio obtained with a noiseless detector. The EQD of the photomultiplier 1P21 is equal to 1.4 x 10(-4). Following Shaw, the EQD of five photographic materials is evaluated at zero spatial frequency. The range of the EQD values is 3 x 10(-6) to 1.6 x 10(-4). The EQD values of photomul ipliers and photographic materials may thus be comparable.


Cx26 affects the in vitro reconstruction of human epidermis.
To study the function of connexins in human keratinocytes, we have used a three-dimensional culture system, in which a tissue is reconstructed using cells from the outer root sheet of hair follicles. This tissue reproduces in vitro the histological organisation of human epidermis in situ and the normal distribution of several keratinocyte markers. Furthermore, it shows characteristics of a differentiating epidermis, including the expression of connexin26. Connexin26 protein expression is increased under physiological and pathological conditions resulting in increased keratinocyte turnover. Loss of this protein in keratinocytes, obtained from patients carrying a stop mutation, resulted in a reduced stratification of the in vitro reconstructed tissue, probably due to a lower proliferation and migration capacity of the keratinocytes, although dye coupling and persistence of other gap junctions is maintained. No changes were seen in tissues reconstructed with keratinocytes from patients carrying a non stop mutation of connexin30. The data indicate that, at least in vitro, connexin26 affects the function of human keratinocytes, independently of obvious changes in coupling.


Upregulation of endothelin-1 binding in tissues of salt-loaded stroke-prone spontaneously hypertensive rats.
Upon maintained on a 1% NaCl drinking solution beginning at 7 weeks of age, the stroke-prone spontaneously hypertensive rat (SHRsp) developed severe hypertension and stroke; most died by 16 weeks. The mechanism by which these diseases evolve remains unclear. Endothelin-1 (ET-1) is a potent, peptidic vasoconstrictor and is implicated in the pathogenesis of various cardiovascular, renal, and central nervous system diseases. The purpose of the present study was to compare the binding of [125I]ET-1 to the brain, heart, kidney, liver, and spleen membrane preparations of 16-week-old SHRsp and age-matched normotensive Wistar-Kyoto rats (WKY). The KD values for [125I]ET-1 binding to the corresponding tissues of the two strains were not significantly different, except in the brain (SHRsp: 17 +/- 1 pM; WKY: 24 +/- 1 pM). In contrast, the Bmax values measured in the brain, heart, kidney, and liver of SHRsp were 1.5- to 2.1-fold greater than those of their WKY counterparts. Competition of [125I]ET-1 binding to the membrane preparations by the specific ETA receptor antagonist BQ-123 or the specific ETB receptor agonist sarafotoxin S6c revealed a similar proportion of ETA and ETB receptor subtypes in the corresponding tissues of the two rat strains. These results indicate that ET-1 binding is upregulated in SHRsp and suggest that ET-1 may play a pathophysiological role in this animal model of genetic hypertension.


Flow disturbance due to venous valves: a cause of graft failure.
Late (greater than 1 year) postoperative angiographic studies in five patients revealed evidence of flow disturbance at the distal valve, when two valves were present in the body of a saphenous vein graft (SVG). This was associated with delayed clearance of radiopaque dye due to reduced velocity of flow. On follow-up, occlusion, thrombus formation, or development of atherosclerosis seemed to have occurred in anatomic relation to these valves. All five patients had one additional graft in a good state of preservation; three internal mammary artery (IMA) grafts, and 2 SVGs. The latter had only one venous valve, and the velocity of flow was good. It appears that good velocity of flow is essential to the integrity of coronary bypass grafts. The presence of more than one venous valve in a graft appears to cause flow disturbance and sluggish velocity, factors adversely affecting their integrity.


Implant-supported mandibular overdentures in very old adults: a randomized controlled trial.
The purpose of this study was (1) to investigate denture satisfaction following the conversion of existing mandibular complete dentures to implant overdentures (IOD) in very old edentulous patients who depend on help for activities of daily living and (2) to evaluate secondary end points, such as functional, structural, nutritional, and patient-centered aspects. For this randomized clinical trial, 2 interforaminal short implants were placed in the intervention group (n = 16, 85.0 ± 6.19 yrs) to retain mandibular IODs; the control group (n = 18, 84.1 ± 5.55 yrs) received conventional relines. During the first year, no implant was lost; however, 2 patients died. IODs proved more stable, and participants in the intervention group demonstrated significantly higher denture satisfaction as well as an increased oral health-related quality of life compared to the control group. Maximum voluntary bite force improved significantly with IODs, yet the chewing efficiency was not different between groups. Masseter muscle thickness increased with IODs, mainly on the preferred chewing side. Body mass index decreased in both groups, but the decline tended to be smaller in the intervention group; blood markers and the Mini Nutritional Assessment did not confirm this tendency. These results indicate that edentulous patients who depend on help for activities of daily living may benefit from IODs even late in life.


Molecular Evolution of Alternative Oxidase Proteins: A Phylogenetic and Structure Modeling Approach.
Alternative oxidases (AOXs) are mitochondrial cyanide-resistant membrane-bound metallo-proteins catalyzing the oxidation of ubiquinol and the reduction of oxygen to water bypassing two sites of proton pumping, thus dissipating a major part of redox energy into heat. Here, the structure of Arabidopsis thaliana AOX 1A has been modeled using the crystal structure of Trypanosoma brucei AOX as a template. Analysis of this model and multiple sequence alignment of members of the AOX family from all kingdoms of Life indicate that AOXs display a high degree of conservation of the catalytic core, which is formed by a four-α-helix bundle, hosting the di-iron catalytic site, and is flanked by two additional α-helices anchoring the protein to the membrane. Plant AOXs display a peculiar covalent dimerization mode due to the conservation in the N-terminal region of a Cys residue forming the inter-monomer disulfide bond. The multiple sequence alignment has also been used to infer a phylogenetic tree of AOXs whose analysis shows a polyphyletic origin for the AOXs found in Fungi and a monophyletic origin of the AOXs of Eubacteria, Mycetozoa, Euglenozoa, Metazoa, and Land Plants. This suggests that AOXs evolved from a common ancestral protein in each of these kingdoms. Within the Plant AOX clade, the AOXs of monocotyledon plants form two distinct clades which have unresolved relationships relative to the monophyletic clade of the AOXs of dicotyledonous plants. This reflects the sequence divergence of the N-terminal region, probably due to a low selective pressure for sequence conservation linked to the covalent homo-dimerization mode.


Quantum Spin-Quantum Anomalous Hall Insulators and Topological Transitions in Functionalized Sb(111) Monolayers.
Electronic and topological behaviors of Sb(111) monolayers decorated with H and certain magnetic atoms are investigated by using ab initio methods. The drastic exchange field induced by the magnetic atoms, together with strong spin-orbit coupling (SOC) of Sb atoms, generates one new category of valley polarized topological insulators, called quantum spin-quantum anomalous Hall (QSQAH) insulators in the monolayer, with a band gap up to 53 meV. The strong SOC is closely related to Sb px and py orbitals, instead of pz orbitals in usual two-dimensional (2D) materials. Topological transitions from quantum anomalous Hall states to QSQAH states and then to time-reversal-symmetry-broken quantum spin Hall states are achieved by tuning the SOC strength. The behind mechanism is revealed. Our work is helpful for future valleytronic and spintronic applications in 2D materials.


De novo expression of pp125FAK in human macrophages regulates CSK distribution and MAP kinase activation but does not affect focal contact structure.
The protein tyrosine kinase pp125FAK (focal adhesion kinase, or FAK) is expressed by a variety of cell types and has been implicated in integrin-mediated signaling events. We explored the potential functions of FAK by expressing it de novo in a cell type lacking FAK. We showed previously that cultured human macrophages lack FAK yet still have well-formed focal contacts. Adenovirus-mediated expression of FAK results in the appearance of FAK protein, which localizes to focal contacts and becomes tyrosine-phosphorylated without perturbing overall cell morphology or focal contacts. FAK associates with CSK 48 h after infection and recruits it to focal contacts. Tyrosine phosphorylation of p130cas but not of paxillin is stimulated after FAK expression. The phosphorylation of p130cas is lost at 48 h in parallel with CSK accumulation in focal contacts. The ERK2 form of MAP kinase is similarly activated at 12-24 h, but it also returns to low levels at 48 h. These findings demonstrate that FAK can be reconstituted to focal contacts in cells that lack it without affecting cell morphology or focal contact structure. FAK can regulate the distribution and activities of elements of the MAP kinase signaling pathway.


Impacts of early life exposure to estrogen on subsequent breeding behavior and reproductive success in zebrafish.
Impacts of exposure to environmental estrogens on reproductive development are well documented, but recently wider concern has been raised due to evidence that such exposures can disrupt normal patterns of reproductive behavior, dominance, and parentage, with potential population level implications. It is fundamental therefore to understand any such effects for effective risk assessment. This study investigated the impact of a transient exposure to ethinylestradiol (EE(2)) during early life (from 20-60 days post fertilization), including at a dosing level within the environmental range, on the subsequent reproductive behavior and success in both male and female zebrafish (Danio rerio) in competitive breeding scenarios. There were no obvious effects of the early life EE(2) exposures on the subsequent gonadal phenotypes in either mature males or females. In fact, reproductive success in males exposed to 2.76 ng EE(2)/L was increased in competitive spawning scenarios. In contrast, exposure of females to EE(2) (9.86 ng/L) during early life reduced their subsequent reproductive success in competitive spawning scenarios. Mate choice experiments suggested this was a consequence of the females' diminished courting behavior toward males, rather than any male preference for unexposed females. Reproductive capability of females is generally considered a key determinant in population demographics and dynamics, and therefore the effect of exposure to EE(2) on female reproductive success may have significant implications for exposed fish populations.


Preparation of Core-Shell Silver Nanoparticles@Mesoporous Silica Nanospheres with Catalytic Activities.
Core-shell silver nanoparticles@mesoporous silica spherical nanoparticles (Ag NPs@MSNs) were prepared by a two-step method. First, Ag NPs were synthesized by chemical reduction using silver nitrate (AgNO₃) as the precursor, cetyl trimethyl ammonium bromide (CTAB) as the stabilizer, and sodium borohydride (NaBH₄) as the reductant. Then, MSNs were obtained by employing CTABstabilized Ag NPs as the template and hydrolyzing tetraethoxysilane (TEOS) precursor in the presence of the alkaline precipitant, triethanolamine (TEOA). The effects of different preparation routes (core-first vs. shell-first), type of reductants as well as extraction methods and agents were studied. The obtained core-shell NPs were characterized by infrared spectroscopy (IR) and transmission electron microscopy (TEM). Our results showed that the core-first route was viable to produce uniform Ag NPs@MSNs with ordered mesostructures. Afterwards, those NPs were used as the catalyst to catalyze the reduction of rhodamine, a model dye compound representing organic pollutants in waste water, in the presence of NaBH₄. It was found that Ag NPs@MSNs not only were efficient catalysts but also participated as coreductants in the reaction. Moreover, they exhibited almost no loss of catalytic efficacy after several reduction cycles, which indicated their promising future use as efficient recyclable catalysts for organic pollutant treatments.


When to base clinical policies on observational versus randomized trial data.
Physicians must decide when the evidence is sufficient to adopt a new clinical policy. Analysis of large clinical and administrative databases is becoming an important source of evidence for changing clinical policies. Because such analysis cannot control for the effects of all potential confounding variables, physicians risk drawing the wrong conclusion about the cause-and-effect relation between a change in clinical policy and outcomes. Randomized studies offer protection against drawing a conclusion that would lead to adoption of an inferior policy. However, a randomized study may be difficult to justify because of the extra costs of collecting data for a randomized study and concerns that a study will not directly benefit the patients enrolled in the study. This article reviews the advantages and disadvantages of basing clinical policy on analysis of large databases compared with conducting a randomized study. A technique is described and illustrated for accessing the potential costs and benefits of conducting such a study. This type of analysis formed the basis for a physician-managed health care organization deciding to sponsor a randomized study among patients with end-stage renal disease as part of a quality-improvement initiative.


Trolox inhibits peroxynitrite-mediated oxidative stress and apoptosis in rat thymocytes.
Peroxynitrite is a strong oxidant that reacts with a variety of biomolecules in vivo and in vitro. When rat thymocytes in phosphate buffer are exposed to 25 microM peroxynitrite for 10 min, DNA single strand breaks (SSB) can be detected. These SSB are repaired if the cells are incubated in fresh media at 37 degrees C for 120 min. In addition, DNA protein cross-links and apoptosis are observed 1 and 6 h, respectively, after peroxynitrite exposure. Peroxynitrite mediates the formation of thiobarbituric acid-reactive substances (TBARS) that may be responsible for the DNA-protein crosslinks (DPXL). Both TBARS and DPXL formation are lowered by posttreating the cells immediately after the 10-min exposure to peroxynitrite with Trolox, a water-soluble vitamin E analog. These results suggest that oxidative stress mediated by peroxynitrite can trigger a critical sequence of events ending in programmed cell death and that intracellular oxidation is a component of the apoptosis of thymocytes, since both oxidative processes and apoptosis can be prevented by Trolox. In addition to Trolox, we obtained partial data on three other phenolic antioxidants (3-tert-butyl-4-hydroxyanisole, butylated hydroxytoluene, and 2,6-diisopropylphenol). We find that Trolox and these three phenols similarly protect rat thymocytes from apoptosis mediated by peroxynitrite.


Carbon Nano-Fiber/PDMS Composite Used as Corrosion-Resistant Coating for Copper Anodes in Microbial Fuel Cells.
The development of high-performance anode materials is one of the greatest challenges for the practical implementation of Microbial Fuel Cell (MFC) technology. Copper (Cu) has a much higher electrical conductivity than carbon-based materials usually used as anodes in MFCs. However, it is an unsuitable anode material, in raw state, for MFC application due to its corrosion and its toxicity to microorganisms. In this paper, we report the development of a Cu anode material coated with a corrosion-resistant composite made of Polydimethylsiloxane (PDMS) doped with carbon nanofiber (CNF). The surface modification method was optimized for improving the interfacial electron transfer of Cu anodes for use in MFCs. Characterization of CNF-PDMS composites doped at different weight ratios demonstrated that the best electrical conductivity and electrochemical properties are obtained at 8% weight ratio of CNF/PDMS mixture. Electrochemical characterization showed that the corrosion rate of Cu electrode in acidified solution decreased from (17 ± 6) × 103 μm y-1 to 93 ± 23 μm y-1 after CNF-PDMS coating. The performance of Cu anodes coated with different layer thicknesses of CNF-PDMS (250 µm, 500 µm, and 1000 µm), was evaluated in MFC. The highest power density of 70 ± 8 mW m-2 obtained with 500 µm CNF-PDMS was about 8-times higher and more stable than that obtained through galvanic corrosion of unmodified Cu. Consequently, the followed process improves the performance of Cu anode for MFC applications.


Tracheostomy care: a clinician's guide.
Tracheostomies have become a typical component of the management of patients with prolonged respiratory failure. There are, however, relatively few studies from which to establish an accepted standard of care with regard to the specific features, daily care, and removal of tracheostomy tubes. Consequently, these decisions are sometimes guided by myth and misconception. In this article, we review the different types of tracheostomy tubes with their respective advantages and disadvantages, basic principles of care, recognition of complications, speech with a tracheostomy tube, and the process by which they may sometimes be removed.


Dexamethasone restores hormonal inducibility of ornithine decarboxylase in primary cultures of rat hepatocytes.
Induction of ornithine decarboxylase by various hormones was studied in quiescent primary cultures of adult rat hepatocytes maintained in a chemically defined medium. The following results were obtained: Enzyme activity rose transiently during the first day of cultivation in hormone-untreated cells. During this phase, insulin increased ornithine decarboxylase activity. Inducibility by insulin was maintained for more than 40 h only after pretreatment with 0.1 microM dexamethasone. Enzyme activity could be induced by 1 nM insulin and peaked after 7 h. Inducibility by glucagon and growth hormone required pretreatment with the glucocorticoid hormone. Ornithine decarboxylase activity was maximal 5 h after glucagon addition. Concentrations down to 0.1 nM were effective. Pretreatment with dexamethasone was most effective, when the hormone was present during the first 20 h of cultivation. The effect of the glucocorticoid during the pretreatment phase was diminished by colchicine and to a lesser extent by cytochalasine B. We suggest that part of the permissive effect of dexamethasone could be mediated by changes in the cytoskeleton and the function of hormone receptors. The fact that induction of ornithine decarboxylase was exerted by several hormones despite the absence of cell proliferation and DNA synthesis may indicate that polyamine biosynthesis has an important role in the quiescent hepatocyte.


Electrocochleographic evaluation of the guinea pig model of endolymphatic hydrops.
Electrocochleography (ECochG) was used to evaluate cochlear function in guinea pigs with experimentally induced endolymphatic hydrops (ELH) before and after osmotic dehydration with either glycerol or urea. We surgically induced ELH in the right ears of 9 guinea pigs, while the right ears of 6 guinea pigs received a sham operation. The left ears of the 15 animals constituted the normal group. Eight weeks after surgery, summating potential (SP) and action potential (AP) amplitudes were measured prior to and following the administration of glycerol or urea. The SPs and SP/AP ratios were reduced in all groups, with no significant differences among groups or between dehydrating agents. Some of the hydropic ears, however, did show an increased AP threshold and a recruitment effect. In measurements from 6 additional animals, serum osmolarity increased more with urea than with glycerol. The guinea pig model remains valuable for investigation of ELH, even though it differs in significant respects from ELH in humans.


Promoting Effects of Titanium Implants Coated with Dipterocarpus tuberculatus Extract on Osseointegration.
Titanium (Ti) is the most commonly used biomaterial for dental implants. When inserting Ti implants into jawbones, the main issue is the lack of strong bonding between the Ti implant and the host bone (osseointegration). Inspired by the outstanding adhesion performance of natural phenolic compounds on metal substrates and promoting effect for cell adhesion, we coated a natural plant extract, Dipterocarpus tuberculatus (MED), on Ti implants. We tested three groups of Ti plates and screw-shaped fixtures: nontreated Ti as commercially produced, ozone-treated Ti as commonly used surface modification for dental implants, and MED-coated Ti. Interestingly, the MED coating on the Ti plate preserved the surface wetting property for 20 days, whereas the hydrophilic wetting of ozone-treated Ti was readily transformed to hydrophobic within a day. Computerized tomography and histopathological analysis revealed that MED coating enhanced new bone tissue formation and regeneration. The gene expression level of integrin as a bone cell adhesion receptor and its downstream key regulators was significantly increased than that of ozone-treated Ti. Therefore, we suggest considering MED-mediated cell signaling pathways in screening natural products for cell adhesion and osseointegration, and MED as a suitable coating agent for improving Ti implantation.


Factors related to the use of antenatal care services in Ethiopia: Application of the zero-inflated negative binomial model.
The major causes for poor health in developing countries are inadequate access and under-use of modern health care services. The objective of this study was to identify and examine factors related to the use of antenatal care services using the 2011 Ethiopia Demographic and Health Survey data. The number of antenatal care visits during the last pregnancy by mothers aged 15 to 49 years (n = 7,737) was analyzed. More than 55% of the mothers did not use antenatal care (ANC) services, while more than 22% of the women used antenatal care services less than four times. More than half of the women (52%) who had access to health services had at least four antenatal care visits. The zero-inflated negative binomial model was found to be more appropriate for analyzing the data. Place of residence, age of mothers, woman's educational level, employment status, mass media exposure, religion, and access to health services were significantly associated with the use of antenatal care services. Accordingly, there should be progress toward a health-education program that enables more women to utilize ANC services, with the program targeting women in rural areas, uneducated women, and mothers with higher birth orders through appropriate media.


[Effects of high glucose on in vitro invasiveness of human breast cancer cell line MDA-MB-435].
To explore the effects and underlying mechanisms of high glucose on in vitro invasiveness of human breast cancer cell line MDA-MB-435. The invasiveness of MDA-MB-435 was determined by Matrigel-coated transwell chambers. Reverse transcription-polymerase chain reaction (RT-PCR) and Western blot were employed to analyze the cellular expression of matrix metalloproteinase-9/matrix metalloproteinase-2/E-cadherin (MMP-9/MMP-2/E-cadherin) gene/protein. The invasive breast cancer cell numbers of each group (Glu 5.5, 11 and 25 mmol/L) were 50 ± 5, 65 ± 6 and 77 ± 3 respectively. Cellular invasion was dramatically enhanced in the Glu 11 and 25 mmol/L group compared with the 5.5 mmol/L group. The MMP-9/MMP-2 protein expression increased significantly in the Glu 11 and 25 mmol/L groups compared with 5.5 mmol/L group while high glucose (Glu 11 and 25 mmol/L group) down-regulated significantly the E-cadherin mRNA/protein expression. High glucose can promote the in vitro invasiveness of human breast cancer cells through the altered expression of MMP-9/MMP-2/E-cadherin.


Host-Guest Chemistry for Simultaneous Imaging of Endogenous Alkali Metals and Metabolites with Mass Spectrometry.
Sodium and potassium are biological alkali metal ions that are essential for the physiological processes of cells and organisms. In combination with small-molecule metabolite information, disturbances in sodium and potassium tissue distributions can provide a further understanding of the biological processes in diseases. However, methods using mass spectrometry are generally tailored toward either elemental or molecular detection, which limits simultaneous quantitative mass spectrometry imaging of alkali metal ions and molecular ions. Here, we provide a new method by including crown ether molecules in the solvent for nanospray desorption electrospray ionization mass spectrometry imaging (nano-DESI MSI) that combines host-guest chemistry targeting sodium and potassium ions and quantitative imaging of endogenous lipids and metabolites. After evaluation and optimization, the method was applied to an ischemic stroke model, which has highly dynamic tissue sodium and potassium concentrations, and we report 2 times relative increase in the detected sodium concentration in the ischemic region compared to healthy tissue. Further, in the same experiment, we showed the accumulation and depletion of lipids, neurotransmitters, and amino acids using relative quantitation with internal standards spiked in the nano-DESI solvent. Overall, we demonstrate a new method that with a simple modification in liquid extraction MSI techniques using host-guest chemistry provides the added dimension of alkali metal ion imaging to provide unique insights into biological processes.


Alteration of X and Y chromosomes in human esophageal squamous cell carcinoma.
The incidence of human esophageal squamous cell carcinoma in males is well-known to be higher than in females and its biological action in male patients is generally much more aggressive than that of the female. Recently, aberrations and/or other abnormalities of the sex chromosomes, especially the Y chromosome, have been postulated to be involved in some of the differences in the incidence and/or biological action of human malignancies between male and female patients. Therefore, in this study, we examined abnormalities of the sex chromosomes in cell smears obtained from 30 male patients diagnosed with esophageal squamous cell carcinoma. In addition, TE series cell lines, derived from esophageal squamous cell carcinomas, were studied for sex chromosome abnormalities by utilizing a simultaneous double color fluorescent in situ hybridization (FISH) and these findings were correlated with various clinicopathological parameters in order to examine its likely biological significance. In esophageal squamous cell carcinoma, Y chromosome loss was detected in all cases studied (1.6-86.9%, mean 22.98 +/- 22.04%), but the loss of the X chromosome was encountered in only 6 of the cases (7.1-40.6%, mean 15.90 +/- 12.46%). There was no significant association between the rate of Y chromosome loss in carcinoma cells and any of the clinicopathological parameters examined including age and stage of the cancer. Loss of the Y chromosome was observed in only two cases of adjacent non-pathological esophageal squamous cell epithelium. Among the TE series examined, the cell lines derived from male patients demonstrated loss of the Y chromosome in all cell lines (1.4-92.9%, mean 44.92 +/- 42.55%), but the great majority of cell lines derived from female patients were associated with the karyotype of XX. These results indicated that the loss of the Y chromosome is associated with the malignant phenotype in human esophageal squamous epithelium, but possibly not with biological behavior. These results also suggested that at least one X chromosome is indispensable for the survival of esophageal squamous cell carcinoma.


Do patients want to be informed? A study of consent for cardiac catheterisation.
One hundred and four patients were asked to recall what they had been told before they gave consent for cardiac catheterisation to be performed. Of these, 14% had received no explanation, 24% had not been warned to expect specific symptoms or side effects, 40% had been told that complications were possible, and 26% had been warned that there was a risk of death. Most patients did not know about these risks, and two thirds would not have wanted to be told about them. Doctors should neither insist on gaining fully informed consent from anxious patients nor deny detailed information to inquiring ones.


Investigation of the optothermal properties of a new polymeric blend: polymethyl-methacrylate-poly (ethylene-co-vinylacetate).
A new polymeric blend, polymethyl methacrylate-poly (ethylene-co-vinylacetate) (PMMA-EVA) has been produced recently in our laboratories. The optothermal characterization of PMMA-EVA was carried out by measuring the ultraviolet photoacoustic spectra of PMMA, EVA, and PMMA-EVA (25 degrees C, 220-350 nm), of a PMMA-EVA blend (250-750 nm) at two temperatures (19 degrees C and 65 degrees C), and by measuring the scattering coefficient (at 514.5 nm) of PMMA-EVA versus the temperature in the range 38-86 degrees C. Our measurements show that, in the explored thermal range, the blend undergoes a phase transition when its optical properties change drastically. The peculiar optical and thermal properties observed appear attractive for possible applications as temperature-controlled optical devices.


Intestinal intussusception.
Intussusception is defined as the invagination of one portion of the bowel into an immediately adjacent portion. Etiology, symptoms, diagnosis, and treatment are different in the pediatric and adult populations. In the pediatric population, most cases are idiopathic and result in the common scenario of ileocolic intussusception. Factors involved in causation include anatomic features of the developing gastrointestinal tract and infectious influences. In adults, the intussusceptum is typically the result of a mucosal, intramural, or extrinsic lead point that acts as a focal area of traction pulling the proximal portion of bowel into the peristalsing distal portion. The diagnosis and management in the pediatric population is relatively standardized with nonoperative reduction via air or contrast enemas attempted first. In the adult population, intussusception presents a preoperative diagnostic challenge; although surgical intervention is mandatory, intraoperative management remains controversial.


Unidirectional transport of glucose and lactate into brain of fetal sheep and guinea-pig.
The first-passage multiple-indicator dilution method was used to measure blood to brain transport of D- and L-glucose, D- and L-lactate and sucrose relative to 22Na, an impermeable reference tracer, in fetal sheep. Fractional extraction for D-glucose was 0.315 +/- 0.051 (S.E.M.) at normal glucose levels and fell to 0.198 +/- 0.041 at 5.2 +/- 0.4 mM-glucose. Fractional extractions for L-glucose, D- and L-lactate and sucrose were not different from zero. No specific blood-brain transport system was detected for L-lactate in fetal sheep in vivo (fractional extraction = -0.024 +/- 0.019). Uptake of L-lactate into isolated microvessels from fetal sheep cerebrum in vitro showed a slightly higher rate (32.2 +/- 8.9 pmol min-1 (mg protein)-1) than that for D-lactate (22.6 +/- 5.6). In fetal guinea-pigs, the carotid arterial injection method with tritiated water as the permeable reference was used to measure the brain uptake index (BUI). BUI was determined for D-glucose (0.304 +/- 0.065) sucrose (0.008 +/- 0.001), L-lactate (0.418 +/- 0.112) and D-lactate (0.071 +/- 0.024). Unidirectional influx calculated from these measurements and estimates of cerebral blood flow showed that transport would be rate-limiting for cerebral glucose utilization at arterial glucose levels below 0.5 mM in fetal sheep and 1.7 mM in fetal guinea-pig. In fetal sheep, but not in fetal guinea-pigs, lactate efflux may be limited by brain-blood transport.


Noncatalytic bromination of benzene: A combined computational and experimental study.
The noncatalytic bromination of benzene is shown experimentally to require high 5-14 M concentrations of bromine to proceed at ambient temperatures to form predominantly bromobenzene, along with detectable (<2%) amounts of addition products such as tetra and hexabromocyclohexanes. The kinetic order in bromine at these high concentrations is 4.8 ± 0.06 at 298 K and 5.6 ± 0.11 at 273 K with a small measured inverse deuterium isotope effect using D6 -benzene of 0.97 ± 0.03 at 298 K. These results are rationalized using computed transition states models at the B3LYP+D3/6-311++G(2d,2p) level with an essential continuum solvent field for benzene applied. The model with the lowest predicted activation free energies agrees with the high experimental kinetic order in bromine and involves formation of an ionic, concerted, and asynchronous transition state with a Br8 cluster resembling the structure of the known Br9 (-). This cluster plays three roles; as a Br(+) donor, as a proton base, and as a stabilizing arm forming weak interactions with two adjacent benzene CH hydrogens, these aspects together combining to overcome the lack of reactivity of benzene induced by its aromaticity. The computed inverse kinetic isotope effect of 0.95 agrees with experiment, and arises because C-Br bond formation is essentially complete, whereas C-H cleavage has not yet commenced. The computed free energy barriers for the reaction with 4Br2 and 5Br2 for a standard state of 14.3 M in bromine are reasonable for an ambient temperature reaction, unlike previously reported theoretical models involving only one or two bromines.


How do we decide whom our friends are? Defining levels of friendship in Poland and the United States.
Participants were 57 U.S. college students and 56 Polish university students and copper mine workers who judged the intimacy of 9 hypothetical relationships and also rated the intensity of their relationships with a best friend, a friend, and an acquaintance on the Friendship Intensity Measurement Scale (FIMS; T. S. Arunkumar & B. Dharmangadan, 2001). The present results confirmed that people perceive (a) relationships with best friends as more intense and intimate than other friendships and (b) other friendships as more intense and intimate than acquaintanceships. The results also indicated that Americans perceive all of their relationships, ranging from mere acquaintanceships to intimate friendships, as more intense and intimate than do Poles. It was somewhat surprising that there were no sex differences in either country in the perception of relationships. The authors discussed the research in the context of the difficulty of defining what friendship is and how an individual's cultural background might interact with person variables such as age and sex.


The effect of sustained phonation at high and low pitch on vocal jitter and shimmer.
Looking for possible signs of vocal fatigue, acoustic waveform perturbation was measured in normal female subjects during sustained phonation at various fundamental frequencies (Fo). At none of the pitch levels a rise of the jitter or the shimmer was found after 25 min of vocalization. On the other hand, an effect of Fo was seen from the start: above the habitual speaking Fo of our subjects' voices there was a tendency for the jitter to be higher and for the shimmer to be lower.


Effect of prosthetic sugar groups on the pharmacokinetics of glucose-oxidase.
The administration of enzymes is of potential therapeutic value in many disease states, e.g. lysosomal storage diseases, provided problems in the metabolism and targeting of large proteins can be overcome. We have addressed ourselves to these problems by studying the pharmacokinetics and distribution of glucose-oxidase (GO) and some of its derivatives in mice. A saturable mechanism was responsible for GO uptake by mononuclear phagocytes. After construction of a pharmacokinetic model, the Kuptake (850 nmol/l) and the number of capturing cells were determined; uptake was half the initial plasma concentration in about 10 min. Deglycosylated GO's had half-lives of about 100 min and were taken up by the same organs that took up native GO. Galactosylated GO had a half-life of 4 min and a different distribution; it was taken up preferentially by the liver in hepatocytes. Our results illustrate the role sugars might play in the targeting of foreign proteins to different cell types, and the feasibility of determining in vivo microscopic constants such as the affinity between molecules and certain cells.


Education and training of mandated reporters: innovative models, overcoming challenges, and lessons learned.
Several Archstone Foundation funded projects developed and implemented training curricula on elder abuse for mandated reporters such as dentists, adult protective services workers, paramedics, and coroner investigators. Common education and training issues emerged, including the need to provide basic content on normal aging and the need for creating standardized trainings. Strategies include integrating elder abuse and neglect content into existing courses, building relationships with stakeholders, and customizing content and delivery to student needs and preferences. Projects developed relevant, practice-based content, decided on curriculum delivery methods, engaged learners, and provided feedback to them. A main outcome is the permanent institution of elder abuse content in training curricula.


Human immunodeficiency virus infection: musculoskeletal manifestations.
The authors retrospectively reviewed the charts, radiographs, and other accompanying imaging studies of 45 patients with musculoskeletal abnormalities associated with human immunodeficiency virus (HIV) infection. These included 19 patients with osseous infection, including eight with osteomyelitis, seven with bacillary angiomatosis (six of whom were described in a previous report), and four with septic arthritis; 10 with bacterial myositis (six of whom were described in a previous report); seven with non-Hodgkin lymphoma; five with hypointense marrow signal intensity at magnetic resonance imaging; two with Kaposi sarcoma; one with polymyositis; and one with psoriasis. The musculoskeletal system can be affected by a variety of abnormalities in association with HIV infection. Knowledge of their existence and characteristic appearance is valuable to radiologists for diagnosis and to clinicians for detection and appropriate treatment.


Heavy-atom labeling of RNA by PLOR for de novo crystallographic phasing.
Due to the paucity of known RNA structures, experimental phasing is crucial for obtaining three-dimensional structures of RNAs by X-ray crystallography. Covalent attachment of heavy atoms to RNAs is one of the most useful strategies to facilitate phase determination. However, this approach is limited by the inefficiency or inability to synthesize large RNAs (>60 nucleotides) site-specifically labeled with heavy atoms using traditional methods. Here, we applied our recently reported method, PLOR (position-selective labeling of RNA) to incorporate 5-iodouridine at specific positions in the adenine riboswitch RNA aptamer domain, which was then used for crystallization and subsequent de novo SAD phasing. PLOR is a powerful tool to improve the efficiency of obtaining RNA structures de novo by X-ray crystallography.


High BrdU Sensitivity of Passeriformes Chromosomes: Conservation of BrdU-Sensitive Fragile Sites on Their Z Chromosomes during Evolution.
Amongst 15 bird species, representative of 7 orders, recurrent breakages evocating the presence of fragile sites were detected in the chromosomes of the 5 species belonging to Passeriformes. These breaks appeared when 5-bromodeoxyuridine (BrdU) was added to the cell culture medium at a dose inefficient for inducing chromosome structure alterations in other birds and mammals. They involved, similarly in male and female, 3 loci on the Z chromosome of 3 Turdus species (Turdidae). Labeling by BrdU antibody confirmed the correlation between BrdU incorporation into DNA and breakage, especially around and in the sites of breakage. Thus, 3 BrdU-sensitive fragile sites were present in the Z chromosomes of these birds. Three fragile sites were also detected at different locations in the Z chromosomes of the European robin (Erithacus rubecula, Muscicapidae), suggesting that a structural rearrangement occurred during the evolution of Turdidae and Muscicapidae. Chromosome banding confirmed this interpretation. Finally, in the more distantly related species Parus major (Paridae), the almost acrocentric Z chromosome displayed a single BrdU-sensitive fragile site in its short arm, and the W appeared to be pulverized by BrdU incorporation. Although it cannot be excluded that the BrdU-sensitive fragile sites may be involved in rearrangements, their conservation in many species, and possibly all Passeriformes, provides evidence that they do not constitute a pejorative character during evolution.


Population-based screening versus case detection.
India has a large burden of blindness and population-based screening is a strategy commonly employed to detect disease and prevent morbidity. However, not all diseases are amenable to screening. This communication examines the issue of "population-based screening" versus "case detection" in the Indian scenario. Using the example of glaucoma, it demonstrates that given the poor infrastructure, for a "rare" disease, case detection is more effective than population-based screening.


Thermal response of rat fibroblasts stably transfected with the human 70-kDa heat shock protein-encoding gene.
The major heat shock protein hsp70 is synthesized by cells of a wide variety of organisms in response to heat shock or other environmental stresses and is assumed to play an important role in protecting cells from thermal stress. We have tested this hypothesis directly by transfecting a constitutively expressed recombinant human hsp70-encoding gene into rat fibroblasts and examining the relationship between the levels of human hsp70 expressed and thermal resistance of the stably transfected rat cells. Successful transfection and expression of the gene for human hsp70 were characterized by RNA hybridization analysis, two-dimensional gel electrophoresis, and immunoblot analysis. When individual cloned cell lines were exposed to 45 degrees C and their thermal survivals were determined by colony-formation assay, we found that the expression of human hsp70 conferred heat resistance to the rat cells. These results reinforce the hypothesis that hsp70 has a protective function against thermal stress.


Rhizobacteria-mediated induced systemic resistance (ISR) in Arabidopsis is not associated with a direct effect on expression of known defense-related genes but stimulates the expression of the jasmonate-inducible gene Atvsp upon challenge.
Selected strains of nonpathogenic rhizobacteria from the genus Pseudomonas are capable of eliciting broad-spectrum induced systemic resistance (ISR) in plants that is phenotypically similar to pathogen-induced systemic acquired resistance (SAR). In Arabidopsis, the ISR pathway functions independently of salicylic acid (SA) but requires responsiveness to jasmonate and ethylene. Here, we demonstrate that known defense-related genes, i.e. the SA-responsive genes PR-1, PR-2, and PR-5, the ethylene-inducible gene Hel, the ethylene- and jasmonate-responsive genes ChiB and Pdf1.2, and the jasmonate-inducible genes Atvsp, Lox1, Lox2, Pall, and Pin2, are neither induced locally in the roots nor systemically in the leaves upon induction of ISR by Pseudomonas fluorescens WCS417r. In contrast, plants infected with the virulent leaf pathogen Pseudomonas syringae pv. tomato (Pst) or expressing SAR induced by preinfecting lower leaves with the avirulent pathogen Pst(avrRpt2) exhibit elevated expression levels of most of the defense-related genes studied. Upon challenge inoculation with Pst, PR gene transcripts accumulated to a higher level in SAR-expressing plants than in control-treated and ISR-expressing plants, indicating that SAR involves potentiation of SA-responsive PR gene expression. In contrast, pathogen challenge of ISR-expressing plants led to an enhanced level of Atvsp transcript accumulation. The otherjasmonate-responsive defense-related genes studied were not potentiated during ISR, indicating that ISR is associated with the potentiation of specific jasmonate-responsive genes.


The 3D-structure-mediated growth of zero-dimensional Cs4SnX6 nanocrystals.
Innovations in approaches to synthesize high-quality lead perovskite nanocrystals have enabled the prosperous development of nanocrystal-based optoelectronic devices in recent years. However, the transfer of these approaches to tin perovskite nanocrystals, which are the most promising lead-free perovskite candidates, remains unsuccessful. Herein, based on a three-dimensional (3D)-structure-mediated approach, monodispersed and highly luminescent inorganic zero-dimensional (0D) tin perovskite nanocrystals (NCs) are synthesized. The crystal growth kinetics are revealed via tracking the intermediate structures and using theoretical simulations. The luminescence quantum yield of Cs4SnBr6 NCs is as high as 52%, which is the highest value for inorganic tin perovskite NCs. Cs4SnI6 NCs with a luminescence quantum yield of 27% are synthesized, which is 35 times higher than previous results. Based on the Cs4SnBr6 NCs, an ultraviolet-light-pumped white-light-emitting device (WLED) with an excellent color-rendering index of 92 is fabricated.


Extent and distribution of calcification of both the aortic annulus and the left ventricular outflow tract predict aortic regurgitation after transcatheter aortic valve replacement.
We sought to analyse local distribution of aortic annulus and left ventricular outflow tract (LVOT) calcification in patients undergoing transcatheter aortic valve replacement (TAVR) and its impact on aortic regurgitation (AR) immediately after device placement. A group of 177 patients with severe aortic stenosis undergoing multislice computed tomography of the aortic root followed by TAVR were enrolled in this single-centre study. Annular and LVOT calcifications were assessed per cusp using a semi-quantitative grading system (0: none; 1 [mild]: small, non-protruding calcifications; 2 [moderate]: protruding [>1 mm] or extensive [>50% of cusp sector] calcifications; 3 [severe]: protruding and extensive calcifications). Any calcification of the annulus or LVOT was present in 107 (61%) and 63 (36%) patients, respectively. Prevalence of annulus/LVOT calcifications in the left coronary cusp was 42% and 25%, respectively, in the non-coronary cusp 28% and 13%, in the right coronary cusp 13% and 5%. AR grade 2 to 4 assessed by the method of Sellers immediately after TAVR device implantation was observed in 55 patients (31%). Multivariate regression analysis revealed that the overall annulus calcification (OR [95% CI] 1.48 [1.10-2.00]; p=0.0106), the overall LVOT calcification (1.93 [1.26-2.96]; p=0.0026), any moderate or severe LVOT calcification (5.37 [1.52-18.99]; p=0.0092), and asymmetric LVOT calcification were independent predictors of AR. Calcifications of the aortic annulus and LVOT are frequent in patients undergoing TAVR, and both the distribution and the severity of calcifications appear to be independent predictors of aortic regurgitation after device implantation.


3-hydroxy-3-methylglutaryl-CoA synthase intermediate complex observed in "real-time".
The formation of carbon-carbon bonds via an acyl-enzyme intermediate plays a central role in fatty acid, polyketide, and isoprenoid biosynthesis. Uniquely among condensing enzymes, 3-hydroxy-3-methylglutaryl (HMG)-CoA synthase (HMGS) catalyzes the formation of a carbon-carbon bond by activating the methyl group of an acetylated cysteine. This reaction is essential in Gram-positive bacteria, and represents the first committed step in human cholesterol biosynthesis. Reaction kinetics, isotope exchange, and mass spectroscopy suggest surprisingly that HMGS is able to catalyze the "backwards" reaction in solution, where HMG-CoA is cleaved to form acetoacetyl-CoA (AcAc-CoA) and acetate. Here, we trap a complex of acetylated HMGS from Staphylococcus aureus and bound acetoacetyl-CoA by cryo-cooling enzyme crystals at three different times during the course of its back-reaction with its physiological product (HMG-CoA). This nonphysiological "backwards" reaction is used to understand the details of the physiological reaction with regards to individual residues involved in catalysis and substrate/product binding. The structures suggest that an active-site glutamic acid (Glu-79) acts as a general base both in the condensation between acetoacetyl-CoA and the acetylated enzyme, and the hydrolytic release of HMG-CoA from the enzyme. The ability to trap this enzyme-intermediate complex may suggest a role for protein dynamics and the interplay between protomers during the normal course of catalysis.


Involvement of tyrosine phosphorylation in HMG-CoA reductase inhibitor-induced cell death in L6 myoblasts.
Our previous studies have shown that the HMG-CoA reductase (HCR) inhibitor (HCRI), simvastatin, causes myopathy in rabbits and kills L6 myoblasts. The present study was designed to elucidate the molecular mechanism of HCRI-induced cell death. We have demonstrated that simvastatin induces the tyrosine phosphorylation of several cellular proteins within 10 min. These phosphorylations were followed by apoptosis, as evidenced by the occurrence of internucleosomal DNA fragmentation and by morphological changes detected with Nomarski optics. Simvastatin-induced cell death was prevented by tyrosine kinase inhibitors. The MTT assay revealed that the addition of mevalonic acid into the culture medium partially inhibited simvastatin-induced cell death. Thus, these results suggested that protein tyrosine phosphorylation might play an important role in the intracellular signal transduction pathway mediating the HCRI-induced death of myoblasts.


Induction of activating transcription factor 1 by nickel and its role as a negative regulator of thrombospondin I gene expression.
Thrombospondin I (TSP I) is an extracellular matrix glycoprotein that influences cell adhesion, motility, and growth. On the basis of its effects on endothelial cell proliferation, TSP I has attracted interest as a potential regulator of solid tumor growth through modulation of tumor blood supply. The regulation of TSP I expression is of critical importance for designing new approaches in tumor therapy. Recently, we have shown that TSP I expression is lost in nickel-transformed cells. In this paper, we identified an activating transcription factor (ATF)/cAMP-responsive element-binding protein binding site as a negative regulatory site in the 5'-flanking sequence of mouse TSP I promoter. We identified ATF-1 as a major component of the ATF/cAMP-responsive element-binding protein binding complex. This Mr 35,000 nuclear ATF-1 protein was shown to be present in higher amounts in nickel-transformed 3T3 cells that do not express TSP 1. Acute treatment of 3T3 cells with NiCl2 resulted in the induction of this transcription factor, and this induction was correlated temporally with the suppression of TSP I expression in the same cells. These results show that nickel exposure causes accumulation of the ATF-1 transcription factor, which is responsible for the down-regulation of transcription of TSP I, and possibly other tumor suppressor genes during nickel-induced cellular transformation.


The Einstein effect provides global evidence for scientific source credibility effects and the influence of religiosity.
People tend to evaluate information from reliable sources more favourably, but it is unclear exactly how perceivers' worldviews interact with this source credibility effect. In a large and diverse cross-cultural sample (N = 10,195 from 24 countries), we presented participants with obscure, meaningless statements attributed to either a spiritual guru or a scientist. We found a robust global source credibility effect for scientific authorities, which we dub 'the Einstein effect': across all 24 countries and all levels of religiosity, scientists held greater authority than spiritual gurus. In addition, individual religiosity predicted a weaker relative preference for the statement from the scientist compared with the spiritual guru, and was more strongly associated with credibility judgements for the guru than the scientist. Independent data on explicit trust ratings across 143 countries mirrored our experimental findings. These findings suggest that irrespective of one's religious worldview, across cultures science is a powerful and universal heuristic that signals the reliability of information.


Propyl Gallate Exerts an Antimigration Effect on Temozolomide-Treated Malignant Glioma Cells through Inhibition of ROS and the NF-κB Pathway.
In this study, we demonstrated that temozolomide (TMZ) and propyl gallate (PG) combination enhanced the inhibition of migration in human U87MG glioma cells. PG inhibited the TMZ-induced reactive oxygen species (ROS) generation. The mitochondrial complex III and NADPH oxidase are two critical sites that can be considered to regulate antimigration in TMZ-treated U87MG cells. PG can enhance the antimigration effect of TMZ through suppression of metalloproteinase-2 and metalloproteinase-9 activities, ROS generation, and the NF-κB pathway and possibly provide a novel prospective strategy for treating malignant glioma.


Cervical metastasis in papillary carcinoma of the thyroid: a histopathological study.
The tendency of well differentiated thyroid carcinoma to remain localised, and the slow progression of these tumours, have supported the belief that this disease behaves as a low grade malignancy. However, with differentiated carcinomas of the thyroid the incidence of nodal metastasis is highest in the papillary subgroup, and the importance of cervical metastases is being increasingly recognised. We have performed and prospectively analysed 17 neck dissections in 13 patients with clinically N+ve necks and papillary carcinoma of the thyroid. In all 17 neck dissections, metastatic deposits of papillary carcinoma of the thyroid were seen, with metastases found in all levels. Our findings of positive lymph nodes in all levels of the neck, with no predictable pattern of spread, and skip metastases, support an aggressive surgical approach to the treatment of cervical metastatic papillary carcinoma of the thyroid.


Where is the sodium in self-assembled monolayers of single-stranded DNA?
Monolayers of single-stranded DNA (ssDNA) immobilized on surfaces form the basis of a number of important biotechnology applications, including DNA microarrays and biosensors. The organization of ssDNA as layer on a solid substrate allows one to investigate various properties of the DNA in a controlled manner and to use DNA for analytical applications as well as for exploring futuristic schemes for molecular electronics. It is commonly assumed that the adsorbed DNA layer contains some structural water and the cations. Here we show, based on XPS studies, that when monolayers of ssDNA are formed from sodium phosphate buffer and washed thoroughly, no Na+ signal is detected. A finite concentration of ions is observed when the DNA is made from a solution of Mg2+ ions, but it is still only a fifth of what it would be if all the phosphate ions were fully neutralized by the metal cations.


Effect of fluoxetine on intracellular Ca2+ levels in bladder female transitional carcinoma (BFTC) cells.
The effect of the antidepressant fluoxetine on Ca2+ signaling in cultured cells was largely unknown. The effect of various concentrations of fluoxetine on [Ca 2+] i in populations of bladder female transitional cancer (BFTC) cells was evaluated by using fura-2 as a Ca2+ probe. Fluoxetine increased [Ca 2+] i concentration dependently (20-100 microM) with an EC50 value of 30 microM. The response was inhibited by 50-60% on extracellular Ca2+ removal. In Ca2+ -free medium, pretreatment with 1 microM thapsigargin (an inhibitor of the endoplasmic reticulum Ca2+ pump) abolished 50 microM fluoxetine-induced Ca2+ release; whereas pretreatment with fluoxetine did not alter the thapsigargin-induced Ca2+ response. Addition of 3 mM Ca2+ increased [Ca 2+] i after pretreatment with 50 microM fluoxetine in Ca2+ -free medium, suggestive of fluoxetine-induced capacitative Ca2+ entry. Suppression of inositol 1,4,5-trisphosphate formation by 2 microM U73122 (a phospholipase C inhibitor) did not affect 50 microM fluoxetine-induced Ca2+ release. Collectively, this study shows that fluoxetine increased [Ca 2+] i in bladder cancer cells in a concentration-dependent fashion, by releasing Ca2+ from thapsigargin-sensitive Ca2+ stores in an IP3-independent manner, and by inducing Ca2+ influx from extracellular medium.


Dieckol, a SARS-CoV 3CL(pro) inhibitor, isolated from the edible brown algae Ecklonia cava.
SARS-CoV 3CL(pro) plays an important role in viral replication. In this study, we performed a biological evaluation on nine phlorotannins isolated from the edible brown algae Ecklonia cava. The nine isolated phlorotannins (1-9), except phloroglucinol (1), possessed SARS-CoV 3CL(pro) inhibitory activities in a dose-dependently and competitive manner. Of these phlorotannins (1-9), two eckol groups with a diphenyl ether linked dieckol (8) showed the most potent SARS-CoV 3CL(pro) trans/cis-cleavage inhibitory effects (IC(50)s = 2.7 and 68.1 μM, respectively). This is the first report of a (8) phlorotannin chemotype significantly blocking the cleavage of SARS-CoV 3CL(pro) in a cell-based assay with no toxicity. Furthermore, dieckol (8) exhibited a high association rate in the SPR sensorgram and formed extremely strong hydrogen bonds to the catalytic dyad (Cys145 and His41) of the SARS-CoV 3CL(pro).


Ethical considerations in conducting pediatric research.
The critical need for pediatric research on drugs and biological products underscores the responsibility to ensure that children are enrolled in clinical research that is both scientifically necessary and ethically sound. In this chapter, we review key ethical considerations concerning the participation of children in clinical research. We propose a basic ethical framework to guide pediatric research, and suggest how this framework might be operationalized in linking science and ethics. Topics examined include: the status of children as a vulnerable population; the appropriate balance of risk and potential benefit in research; ethical considerations underlying study design, including clinical equipoise, placebo controls, and non-inferiority designs; the use of data monitoring committees; compensation; and parental permission and child assent to participate in research. We incorporate selected national (USA) and international guidelines, as well as regulatory approaches to pediatric studies that have been adopted in the USA, Canada, and Europe.


Distribution and biodegradation of polycyclic aromatic hydrocarbons in contaminated sites of Hisar (India).
Fifty-two soil samples were collected from various location of the Hisar city. These samples were analysed for six polycyclic aromatic hydrocarbons (naphthalene, acenaphthene, phenanthrene, anthracene, fluoranthene, and pyrene). Total mean concentration of six PAHs varied from 51.79 to 148.82 mg kg(-1) dry weight of the soil, PAH concentration was higher in soil samples from local auto market while lower concentration was recorded in agricultural soil. Effect ofpH (5.0 to 9.0), temperature (20 degrees to 40 degrees C), and concentration of PAHs (5 to 20 mg kg(-1)) on biodegradation were optimized. Biodegradation of phenanthrene (3-ring) and pyrene (4-ring) was evaluated using two acclimatized microbial strains Pseudomonas putida and Pseudomonas paucimobilis. Biodegradation was maximum in sterilized artificial spiked soil with phenanthrene (5 mg kg(-1)) and pyrene (5 mg kg(-1)) at pH 7.0 and at 30 degrees C (optimized conditions) than the native unsterilized contaminated soil (without optimized conditions) in 42 days of incubation period with Pseudomonas putida and Pseudomonas paucimobilis. Phenanthrene was completely disappeared after 28 days with P. putida and after 35 days with P. paucimobilis. Whereas, pyrene was disappeared up to 97.40% with P. putida and 95.5% with P. paucimobilis after 42 days incubation period at optimum conditions. Under unoptimized conditions, disappearance of phenanthrene was 65.89% with P. putida and 57.81% with P. paucimobilis after 42 days, whereas the % disappearance of pyrene was 59.80% with P. putida and 52.07% with P. paucimobilis.


Effect of pH on the activity of ice-binding protein from Marinomonas primoryensis.
The ability of an ice-binding protein (IBP) from Marinomonas primoryensis (MpIBP) to influence ice crystal growth and structure in nonphysiological pH environments was investigated in this work. The ability for MpIBP to retain ice interactivity under stressed environmental conditions was determined via (1) a modified splat assay to determine ice recrystallization inhibition (IRI) of polycrystalline ice and (2) nanoliter osmometry to evaluate the ability of MpIBP to dynamically shape the morphology of a single ice crystal. Circular dichroism (CD) was used to relate the IRI and DIS activity of MpIBP to secondary structure. The results illustrate that MpIBP secondary structure was stable between pH 6 and pH 10. It was found that MpIBP did not interact with ice at pH ≤ 4 or pH ≥ 13. At 6 ≤ pH ≥ 12 MpIBP exhibited a reduction in grain size of ice crystals as compared to control solutions and demonstrated dynamic ice shaping at 6 ≤ pH ≥ 10. The results substantiate that MpIBP retains some secondary structure and function in non-neutral pH environments; thereby, enabling its potential utility in nonphysiological materials science and engineering applications.


Hepatic xanthine levels as viability predictor of livers procured from non-heart-beating donor pigs.
The aim of the present study was to evaluate hepatic content of adenine nucleotides and their degradation products in non-heart-beating donor (NHBD) pigs and its relationship with recipient survival. Thirty animals were transplanted with an allograft from NHBDs. After warm ischemia (WI) time (20, 30, or 40 min), cardiopulmonary bypass and normothermic recirculation (NR) were run for 30 min. Afterward, the animals were cooled to 15 degrees C and liver procurement was performed. Survival rate was 100% in the 20WI, 70% in the 30WI, and 50% in the 40WI. Livers from non-surviving animals had higher levels of xanthine after NR than livers from surviving animals. Logistic regression analysis revealed that xanthine at the end of NR was the only variable able to predict survival with a calculated sensitivity of 80% and a specificity of 60%. Prolongation of warm ischemic period leaded to a greater xanthine accumulation as well as increased plasma alpha-glutathione S-transferase levels at reperfusion. Xanthine at NR and alpha-glutathione S-transferase at reperfusion significantly correlated, indicating that donor xanthine contributes to some extent to the severity of the lesion by ischemia-reperfusion. It is suggested that xanthine content in the donor is able to predict survival after transplantation. Xanthine is significantly involved in the hepatic lesion elicited by warm ischemia and subsequent ischemia-reperfusion associated to liver transplantation from a NHBD.


Paraxial properties of three-element zoom system for laser beam expanders based on tunable-focus lenses.
The paper is focused on the problem of a theoretical analysis of paraxial imaging properties and initial optical design of the three-element zoom optical system for laser beam expanders using lenses with a tunable focal length. Equations which allow calculation of required optical powers of individual elements of the three-element zoom optical system for laser beam expander depending on the value of the axial position of the beam waist of the input Gaussian beam and the required magnification of the system are derived.


Ectoine production with indigenous Marinococcus sp. MAR2 isolated from the marine environment.
Ectoine has fostered the development of products for skin care and cosmetics. In this study, we employed the marine bacterial strain Marinococcus sp. MAR2 to increase ectoine production by optimizing medium constituents using Response Surface Methodology (RSM) and a fed-batch strategy. The results from the steepest ascent and central composite design indicated that 54 g/L of yeast extract, 14.0 g/L of ammonium acetate, 74.4 g/L of sodium glutamate, and 6.2 g/L of sodium citrate constituted the optimal medium with maximum ectoine production (3.5 g/L). In addition, we performed fed-batch culture in the bioreactor, combining pH and dissolved oxygen to produce ectoine by Marinococcus sp. MAR2. The ectoine production, content, and productivity of 5.6 g/L, 10%, and 3.9 g/L/day were further reached by a fed-batch culture. Thus, the ectoine production by Marinococcus sp. MAR2 using RSM and fed-batch strategy shows its potential for industrial production.


Extracellular NAD+ shapes the Foxp3+ regulatory T cell compartment through the ART2-P2X7 pathway.
CD4(+)CD25(+)FoxP3(+) regulatory T cells (T reg cells) play a major role in the control of immune responses but the factors controlling their homeostasis and function remain poorly characterized. Nicotinamide adenine dinucleotide (NAD(+)) released during cell damage or inflammation results in ART2.2-mediated ADP-ribosylation of the cytolytic P2X7 receptor on T cells. We show that T reg cells express the ART2.2 enzyme and high levels of P2X7 and that T reg cells can be depleted by intravenous injection of NAD(+). Moreover, lower T reg cell numbers are found in mice deficient for the NAD-hydrolase CD38 than in wild-type, P2X7-deficient, or ART2-deficient mice, indicating a role for extracellular NAD(+) in T reg cell homeostasis. Even routine cell preparation leads to release of NAD(+) in sufficient quantities to profoundly affect T reg cell viability, phenotype, and function. We demonstrate that T reg cells can be protected from the deleterious effects of NAD(+) by an inhibitory ART2.2-specific single domain antibody. Furthermore, selective depletion of T reg cells by systemic administration of NAD(+) can be used to promote an antitumor response in several mouse tumor models. Collectively, our data demonstrate that NAD(+) influences survival, phenotype, and function of T reg cells and provide proof of principle that acting on the ART2-P2X7 pathway represents a new strategy to manipulate T reg cells in vivo.


Sensitivity to changing contingencies in an impulsivity task.
Using a video-game-based escalating interest task, participants repeatedly encountered a reward that gradually increased in value over a 10-second interval. Responding early in the interval netted less immediate reward than responding later in the interval. Each participant experienced four different reward contingencies for waiting. These contingencies were changed three times as the experiment proceeded. Behavior tracked these changing contingencies, but wait times reflected long-term carryover from the previously assigned contingencies. Both the tendency to respond slowly and the optimality of behavior were affected by the order of contingencies experienced. Demographic variables only weakly predicted behavior, and delay discounting rate in a hypothetical money choice task predicted choice only when the contingencies in the game were weaker.


Nerve action potential recording method and equipment.
Nerve action potential (NAP) recording is a proven and useful tool in the surgical management of nerve injury. Understanding its materials and methods as well as its pitfalls and limitations is necessary for successful and routine use of this tool. The basic electrophysiology of nerve and its implications in the setting of nerve trauma and regeneration are reviewed. Electrode design and electrode placement are discussed. The use of simple electromyographic/nerve conduction machines for intraoperative recording of NAPs is developed, along with some of the technical problems that result from electrophysiological recording in an electrically hostile environment. NAP recording is a good indicator of useful recovery in nerve trauma. The presence and demonstration of an NAP is associated with a 90% chance of recovery to a useful motor state. NAP recording provides useful information regarding the regenerative potential of a damaged nerve long before that potential is clinically evident. This information is critical in the optimal management of nerve injuries. The techniques of NAP recording are straightforward, but attention to detail is important, and full understanding of the method is required.


Specific shRNA targeting of FAK influenced collagen metabolism in rat hepatic stellate cells.
To investigate the effects and mechanism of disruption of focal adhesion kinase (FAK) expression on collagen metabolism in rat hepatic stellate cells (HSC). The plasmids expressing FAK short hairpin RNA (shRNA) were transfected into HSC-T6 cells, and the level of FAK expression was determined by both real-time quantitative polymerase chain reaction (Q-PCR) and Western blotting analysis. The production of type I collagen and type III collagen in FAK-disrupted cells was analyzed by real-time Q-PCR. The level of collagen metabolism proteins, including matrix metalloproteinases-13 (MMP-13) and tissue inhibitors of metalloproteinases-1 (TIMP-1) was also determined by both real-time Q-PCR and Western blotting analysis. The transfection of FAK shRNA plasmids into HSC resulted in disrupted FAK expression. Compared with the HK group, the levels of type I collagen and type III collagen mRNA transcripts in FAK shRNA plasmid group were significantly decreased (0.69 +/- 0.03 vs 1.96 +/- 0.15, P = 0.000; 0.59 +/- 0.07 vs 1.62 +/- 0.12, P = 0.020). The production of TIMP-1 in this cell type was also significantly reduced at both mRNA and protein levels (0.49 +/- 0.02 vs 1.72 +/- 0.10, P = 0.005; 0.76 +/- 0.08 vs 2.31 +/- 0.24, P = 0.000). However, the expression of MMP-13 mRNA could be significantly up-regulated by the transfection of FAK shRNA plasmids into HSC (1.74 +/- 0.20 vs 1.09 +/- 0.09, P = 0.000). These data support the hypothesis that shRNA-mediated disruption of FAK expression could attenuate extracellular matrix (ECM) synthesis and promote ECM degradation, making FAK a potential target for novel anti-fibrosis therapies.


Biodiversity assessment of Phlebotomine (Diptera: Psychodidae) in an environmental impacted area in Rio de Janeiro State, Brazil.
From October 1994 to September 1996, in the municipality of Mangaratiba, Rio de Janeiro State, phlebotomines were captured on the walls of the houses by means of a manual suction tube and light traps, in the household, the peridomicile and in the forest. As it is an area undergoing a real expansion and an increment in the local tourism, with the purpose of assessing changes in the phlebotomine fauna caused by environmental changes in the region, new captures were made in the same location from October 2015 to September 2016, this time using only light traps. In the two phases of the study, a total of 6,681 phlebotomines were captured. The results indicated that Ny. intermedia and Mg. migonei are fully adapted to this anthropic environment, while Pi. fischeri was more abundant and eclectic, and was probably attracted to exercise hematophagy. Nyssomyia intermedia can be suggested as the main vector of the etiological agent of the American Tegumentary Leishmaniasis in the studied areas. Pintomyia fischeri can also transmit Leishmania braziliensis, both in the environment altered by human action, and in the wild environment. Phlebotomines were captured in greater numbers between 6 and 8 pm in the peridomicile and between 9 and 11 pm in the household. The highest densities of phlebotomines were recorded in December, January and February. Despite almost 20 years between the two studies in the municipality of Mangaratiba, there was no change in the profile of the phlebotomine fauna in general; however, there was a greater density of species of medical importance in areas that suffered environmental impacts.


Using Artificial Intelligence to Obtain More Evidence? Prediction of Length of Hospitalization in Pediatric Burn Patients.
Background: It is not only important for counseling purposes and for healthcare management. This study investigates the prediction accuracy of an artificial intelligence (AI)-based approach and a linear model. The heuristic expecting 1 day of stay per percentage of total body surface area (TBSA) serves as the performance benchmark. Methods: The study is based on pediatric burn patient's data sets from an international burn registry (N = 8,542). Mean absolute error and standard error are calculated for each prediction model (rule of thumb, linear regression, and random forest). Factors contributing to a prolonged stay and the relationship between TBSA and the residual error are analyzed. Results: The random forest-based approach and the linear model are statistically superior to the rule of thumb (p < 0.001, resp. p = 0.009). The residual error rises as TBSA increases for all methods. Factors associated with a prolonged LOS are particularly TBSA, depth of burn, and inhalation trauma. Conclusion: Applying AI-based algorithms to data from large international registries constitutes a promising tool for the purpose of prediction in medicine in the future; however, certain prerequisites concerning the underlying data sets and certain shortcomings must be considered.


Anesthesia-related periodic involuntary movement in an obstetrical patient for cesarean section under epidural anesthesia: a case report.
Anesthesia-related periodic involuntary movements in a patient with regional anesthesia or etomidate anesthesia can be alarming. This case report describes anesthesia-related periodic involuntary movement in a patient undergoing cesarean section with epidural anesthesia. The anesthesia-related periodic involuntary movement occurred postoperatively in the recovery room. The discussion includes a brief review of spinal cord anatomy and physiology, as well as a review of the literature describing similar events.


Evidence that a cell cycle regulator, E2F1, down-regulates transcriptional activity of the human immunodeficiency virus type 1 promoter.
Proliferation of eukaryotic cells is orchestrated by a series of cellular proteins which participate in various stages of the cell cycle to guide the cell through mitosis. Some of these proteins, including E2F1, play a critical role in G1 and S phases by coordinately regulating expression of several important cell cycle-associated genes. On the basis of recent observations indicating a block in human immunodeficiency virus type 1 (HIV-1) replication in cells arrested in G1/S phase of the cell cycle, we sought to evaluate the regulatory action of E2F1 on transcription from the HIV-1 long terminal repeat (LTR). Results from transient transfection of cells with an E2F1 expression plasmid indicated that E2F1 has the ability to suppress basal transcriptional activity of the LTR and to diminish the extent of the Tat-induced activation of the viral promoter. Deletion analysis of the HIV-1 LTR in transfection studies revealed the presence of two major elements responsive to E2F1 repression located distally (-454 to -381) and proximally (-117 to -80) with respect to the +1 transcription start site. E2F1-mediated suppression of LTR activity was observed in a wide range of human cell lines. Expression of E2F1 by a transgene showed an inhibitory effect on the levels of reverse transcriptase activity obtained upon introduction of the proviral genome into cells. The data presented in this study suggest that cellular regulatory proteins involved in the progression of cells through the mitotic cycle could play crucial roles in determining the efficiency of HIV-1 replication during the various stages of infection. The possible roles of these factors in viral latency and activation are discussed.


Muscarinic receptor subtype in isolated dog and monkey facial vein.
Using the cannula inserting method, we investigated vascular responses to ACh and McN-A-343 (a M1-agonist) in isolated and perfused canine and simian facial veins in non-preconstricted conditions. ACh usually induced only a vasoconstriction, but McN-A-343 did not induce any significant vasoconstriction. It is concluded that canine and simian facial veins contain very few receptors of the muscarinic M1-subtype; and according to previous studies, these vessels have abundant M3-receptors.