Topical Application of 0.5% Timolol Maleate Hydrogel for the Treatment of Superficial Infantile Hemangioma.
The therapeutic options for infantile hemangiomas (IHs) have been greatly altered since the introduction of oral propranolol for successful treatments of IHs. Recently, there is an increase in the application of topical timolol maleate for treating superficial IHs. In the present study, we developed a new formulation of timolol maleate 0.5% hydrogel and treated 321 patients with superficial IHs to evaluate its efficacy and safety in the treatment of superficial IHs. This new timolol hydrogel was applied three times daily with a mean duration of 7.1 months. Response to treatment was assessed according to cosmetic improvement by using visual analog scale (VAS). The average VAS improvement after treatment was 76.4, with 126 patients (39.3%) achieving excellent responses, 159 patients (49.5%) achieving good responses, 33 patients (10.3%) achieving fair responses, and three patients (0.9%) achieving poor responses. Age at treatment initiation (P = 0.0349) and lesion thickness (P = 0.0147) were significantly associated with therapeutic efficacy. No severe side effects were observed in all patients. In conclusion, this new topical timolol maleate 0.5% hydrogel appears to be a proper candidate for treating superficial IHs, and our study provides supportive evidence and experience of topical timolol maleate in treating superficial IHs.


Rectangular multilayer dielectric gratings with broadband high diffraction efficiency and enhanced laser damage resistance.
Broadband multilayer dielectric gratings (MDGs) with rectangular HfO2 grating profile were realized for the first time using a novel fabrication process that combines laser interference lithography, nanoimprint, atomic layer deposition and reactive ion-beam etching. The laser-induced damage initiating at the grating ridge was mitigated for two reasons. First, the rectangular grating profile exhibits the minimum electric-field intensity (EFI) enhancement inside the grating pillar compared to other trapezoidal profiles. Second, our etching process did not create nano-absorbing defects at the edge of the HfO2 grating where the peak EFI locates, which is unavoidable in traditional fabrication process. The fabricated MDGs showed a high laser induced damage threshold of 0.59J/cm2 for a Ti-sapphire laser with pulse width of 40 fs and an excellent broadband diffraction spectrum with 95% efficiency over 150 nm in TE polarization.


Thai elderly who do not coreside with their children.
In this paper, data from a nationally representative survey of elderly Thais living in private households are analyzed. The analysis focuses on situations of the 756 elderly who do not coreside with an adult child. Only a minority of those elderly who do not coreside with an adult child were childless. The majority have at least one noncoresident child with whom they could potentially live. Daily contact with children for elderly who live alone was not significantly different from that of elderly who live with their children, suggesting that households that are classified as being separate may in fact function as single households or that at least one non-coresident child may live in very close proximity to the elderly person. Differences between urban and rural elderly in terms of type of support received from non-coresident children as well as likelihood of living near a non-coresident child are discussed.


Malignant female adnexal tumors of probable Wolffian origin.
Female adnexal tumors of probable Wolffian origin are rare. Although the histological features of the benign tumors are recognized, features of malignancy are not well defined in published work because these tumors are particularly rare. In this study, we report an 81-year-old woman presenting with a 6-month history of postmenopausal bleeding. A pelvic transvaginal ultrasound scan demonstrated a midline, highly vascular, predominantly solid pelvic mass, approximately 14 cm, posterior to the uterus. CA-125 was moderately raised. She underwent total abdominal hysterectomy with bilateral salpingo-oophorectomy and infracolic omentectomy. Immunohistochemical staining and electron microscopy supported the diagnosis of female adnexal tumor of probable Wolffian origin. There was significant and widespread nuclear pleomorphism with increased mitotic activity endorsing the diagnosis of a malignant tumor. Seven months after initial surgery, follow-up magnetic resonance imaging showed a mass with a maximum diameter of 7 cm in the region of the right adnexa consistent with a recurrence. The patient declined further treatment.


Increased expression of delta-catenin/neural plakophilin-related armadillo protein is associated with the down-regulation and redistribution of E-cadherin and p120ctn in human prostate cancer.
delta-Catenin, or neural plakophilin-related armadillo protein, is a unique armadillo domain-containing protein in that it is neural-specific and primarily expressed in the brain. However, our recent analysis of the human genome revealed a consistent association of delta-catenin messenger RNA sequences with malignant cells, although the significance of these findings was unclear. In this study, we report that a number of delta-catenin epitopes were expressed in human prostate cancer cells. Western blot and tissue microarray revealed a close association between increased delta-catenin expression and human primary prostatic adenocarcinomas. The analyses of 90 human prostate cancer and 90 benign prostate tissue samples demonstrated that an estimated 85% of prostatic adenocarcinomas showed enhanced delta-catenin immunoreactivity. delta-Catenin expression increased with prognostically significant increased Gleason scores. By analyzing the same tumor cell clusters using consecutive sections, we showed that an increased delta-catenin immunoreactivity was accompanied by the down-regulation and redistribution of E-cadherin and p120ctn, major cell junction proteins whose inactivation is frequently associated with cancer progression. Furthermore, overexpression of delta-catenin in tumorigenic CWR-R1 cells that are derived from human prostate cancer xenograft resulted in reduced immunoreactivity for E-cadherin and p120ctn at the cell-cell junction. This is the first study comparing overexpression of delta-catenin with the E-cadherin/catenin system in cancer and shows that delta-catenin may be intimately involved in regulating E-cadherin/p120ctn cell-cell adhesion in prostate cancer progression.


Disrupting cortical astrocyte Ca2+ signaling in developing brain induces social deficits and depressive-like behaviors.
Astrocytes are present throughout the central nervous system and display complex intracellular Ca2+ signals. However, it is largely unknown regarding how astrocytic Ca2+ signals regulate neural microcircuits in developing brain and mammalian behavior in vivo. In this study, we specifically overexpressed the plasma membrane calcium-transporting ATPase2 (PMCA2) of cortical astrocytes and used immunohistochemistry, Ca2+ imaging, electrophysiology, and behavioral tests to investigate the effects of genetically reducing cortical astrocyte Ca2+ signaling during a critical developmental period in vivo. We found that reducing cortical astrocyte Ca2+ signaling during development led to social interaction deficits, depressive-like behaviors, and abnormal synaptic structure and transmission. In addition, restoring cortical astrocyte Ca2+ signaling using chemogenetic activation of Gq-coupled designer receptors exclusively activated by designer drugs rescued these synaptic and behavioral deficits. Together, our data demonstrate that the integrity of cortical astrocyte Ca2+ signaling in developing mice is critical for neural circuit development and may be involved in the pathogenesis of developmental neuropsychiatric diseases, such as autism spectrum disorders and depression.


Nosocomial outbreak of Legionnaires' disease: molecular epidemiology and disease control measures.
Molecular laboratory techniques were used to study the epidemiology of an outbreak of nosocomial Legionnaires' disease. All patient isolates were Legionella pneumophila serogroup 1 and showed identical plasmid profiles and reactions with serogroup-specific monoclonal antibodies. L pneumophila was also cultured from four of five cooling tower water samples; however, the isolate from only one tower was serogroup 1 of the same subtype as patient isolates. Since the cases were temporally clustered and epidemiologically associated with exposure to cooling tower aerosols, the single cooling tower implicated by molecular analysis was the most likely source of the outbreak. Chlorination of cooling tower ponds has eradicated the epidemic strain. Since potable water also harbored the infecting organism and was the probable source for cooling tower contamination, decontamination of the hospital water system was also undertaken. Superchlorination of hot water holding tanks to 17 ppm on a weekly basis has effectively eradicated L pneumophila from the potable water system and appears to be a reasonable, simple, and relatively inexpensive alternative to previously described methods of control.


The MGuard coronary stent: safety, efficacy, and clinical utility.
Atheromatous and thrombotic embolization during percutaneous coronary revascularization is a feared complication that may cause impaired myocardial reperfusion even with a patent epicardial vessel. The MGuard stent is a cobalt chromium bare metal stent with a porous net attached to its outer surface that has been designed to prevent thrombus fragmentation and distal embolization during stent implantation. This review summarizes the available evidence supporting the use of the MGuard stent in different scenarios such as lesions with high thrombus burden, saphenous vein graft interventions, coronary perforations, or carotid lesions.


Caspase-3 mediated release of SAC domain containing fragment from Par-4 is necessary for the sphingosine-induced apoptosis in Jurkat cells.
Prostate apoptosis response-4 (Par-4) is a tumor-suppressor protein that selectively activates and induces apoptosis in cancer cells, but not in normal cells. The cancer specific pro-apoptotic function of Par-4 is encoded in its centrally located SAC (Selective for Apoptosis induction in Cancer cells) domain (amino acids 137-195). The SAC domain itself is capable of nuclear entry, caspase activation, inhibition of NF-κB activity, and induction of apoptosis in cancer cells. However, the precise mechanism(s) of how the SAC domain is released from Par-4, in response to apoptotic stimulation, is not well explored. In this study, we demonstrate for the first time that sphingosine (SPH), a member of the sphingolipid family, induces caspase-dependant cleavage of Par-4, leading to the release of SAC domain containing fragment from it. Par-4 is cleaved at the EEPD131G site on incubation with caspase-3 in vitro, and by treating cells with several anti-cancer agents. The caspase-3 mediated cleavage of Par-4 is blocked by addition of the pan-caspase inhibitor z-VAD-fmk, caspase-3 specific inhibitor Ac-DEVD-CHO, and by introduction of alanine substitution for D131 residue. Moreover, suppression of SPH-induced Akt dephosphorylation also abrogated the caspase dependant cleavage of Par-4. Evidence provided here shows that Par-4 is cleaved by caspase-3 during SPH-induced apoptosis. Cleavage of Par-4 leads to the generation of SAC domain containing fragment which may possibly be essential and sufficient to induce or augment apoptosis in cancer cells.


Making Time: Conservation of Biological Clocks from Fungi to Animals.
The capacity for biological timekeeping arose at least three times through evolution, in prokaryotic cyanobacteria, in cells that evolved into higher plants, and within the group of organisms that eventually became the fungi and the animals. Neurospora is a tractable model system for understanding the molecular bases of circadian rhythms in the last of these groups, and is perhaps the most intensively studied circadian cell type. Rhythmic processes described in fungi include growth rate, stress responses, developmental capacity, and sporulation, as well as much of metabolism; fungi use clocks to anticipate daily environmental changes. A negative feedback loop comprises the core of the circadian system in fungi and animals. In Neurospora, the best studied fungal model, it is driven by two transcription factors, WC-1 and WC-2, that form the White Collar Complex (WCC). WCC elicits expression of the frq gene. FRQ complexes with other proteins, physically interacts with the WCC, and reduces its activity; the kinetics of these processes is strongly influenced by progressive phosphorylation of FRQ. When FRQ becomes sufficiently phosphorylated that it loses the ability to influence WCC activity, the circadian cycle starts again. Environmental cycles of light and temperature influence frq and FRQ expression and thereby reset the internal circadian clocks. The molecular basis of circadian output is also becoming understood. Taken together, molecular explanations are emerging for all the canonical circadian properties, providing a molecular and regulatory framework that may be extended to many members of the fungal and animal kingdoms, including humans.


Network algorithmics and the emergence of the cortical synaptic-weight distribution.
When a neuron fires and the resulting action potential travels down its axon toward other neurons' dendrites, the effect on each of those neurons is mediated by the strength of the synapse that separates it from the firing neuron. This strength, in turn, is affected by the postsynaptic neuron's response through a mechanism that is thought to underlie important processes such as learning and memory. Although of difficult quantification, cortical synaptic strengths have been found to obey a long-tailed unimodal distribution peaking near the lowest values (approximately lognormal), thus confirming some of the predictive models built previously. Most of these models are causally local, in the sense that they refer to the situation in which a number of neurons all fire directly at the same postsynaptic neuron. Consequently, they necessarily embody assumptions regarding the generation of action potentials by the presynaptic neurons that have little biological interpretability. We introduce a network model of large groups of interconnected neurons and demonstrate, making none of the assumptions that characterize the causally local models, that its long-term behavior gives rise to a distribution of synaptic weights (the mathematical surrogates of synaptic strengths) with the same properties that were experimentally observed. In our model, the action potentials that create a neuron's input are, ultimately, the product of network-wide causal chains relating what happens at a neuron to the firings of others. Our model is then of a causally global nature and predicates the emergence of the synaptic-weight distribution on network structure and function. As such, it has the potential to become instrumental also in the study of other emergent cortical phenomena.


Solubilization and regeneration of Vitreoscilla hemoglobin isolated from protein inclusion bodies.
Vitreoscilla hemoglobin (VHb), a homodimeric protein containing two heme groups in its native state, was used as a model to investigate inclusion body approtein solubilization, prosthetic group incorporation, and reactivation. High-level expression in recombinant Escherichia coli results in accumulation of a substantial portion of heme-free VHb in inclusion bodies. VHb can be solubilized from these inclusion bodies by relatively low concentrations of urea with the dissolution midpoint at approximately 3.2M urea. Dissolution in the presence of stoichiometric heme shifts the dissolution midpoint to approximately 4.5M urea without influencing the dissolution properties of contaminant proteins, suggesting the effect is specific for VHb. Denaturation of apoVHb and holoVHb obtained from purified native VHb has midpoints of 2.9M and 5.1M urea, respectively. VHb solubilized from inclusion bodies with urea at concentrations from 0 to 3.5M urea can be regenerated by heme addition without dilution of urea to yield active holoVHb. The fraction of solubilized VHb reconstituted upon heme addition is maximum at around 30% when solubilization and reconstitution is conducted in less than 1M urea. At these low urea concentrations, approximately 5% of inclusion body VHb is solubilized. These results show the utility of prosthetic group addition to reconstitute holoVHb in the presence of urea. Also, these findings suggest that some inclusion body protein has partially folded conformation and that a fractional dissolution and refolding process may be advantageous.


Denatured mammalian protein mixtures exhibit unusually high solubility in nucleic acid-free pure water.
Preventing protein aggregation is a major goal of biotechnology. Since protein aggregates are mainly comprised of unfolded proteins, protecting against denaturation is likely to assist solubility in an aqueous medium. Contrary to this concept, we found denatured total cellular protein mixture from mammalian cell kept high solubility in pure water when the mixture was nucleic acids free. The lysates were prepared from total cellular protein pellet extracted by using guanidinium thiocyanate-phenol-chloroform mixture of TRIzol, denatured and reduced total protein mixtures remained soluble after extensive dialysis against pure water. The total cell protein lysates contained fully disordered proteins that readily formed large aggregates upon contact with nucleic acids or salts. These findings suggested that the highly flexible mixtures of disordered proteins, which have fully ionized side chains, are protected against aggregation. Interestingly, this unusual solubility is characteristic of protein mixtures from higher eukaryotes, whereas most prokaryotic protein mixtures were aggregated under identical conditions. This unusual solubility of unfolded protein mixtures could have implications for the study of intrinsically disordered proteins in a variety of cells.


Effect of Calcium and Fluoride Addition to Hydrogen Peroxide Bleaching Gel On Tooth Diffusion, Color, and Microhardness.
The aim of this study was to evaluate the effect of calcium and fluoride addition to a 35% hydrogen peroxide (HP) bleaching gel with regard to its diffusion through the tooth structure, enamel microhardness, and bleaching efficacy. Eighty specimens (6 mm in diameter and 2 mm in height; 1 mm/enamel and 1 mm/dentin) were obtained from bovine incisors that were polished and divided into four groups (n=20) according to the remineralizing agent added to the gel: Ca = 0.5% calcium gluconate; F = 0.2% sodium fluoride; Ca+F = 0.5% calcium gluconate and 0.2% sodium fluoride; and control = no agent. Initial microhardness and color were assessed. The samples were positioned over simulated pulpal chambers filled with acetate buffer solution to capture the HP. Gels were applied over enamel for 30 minutes, and HP diffusion was assessed by spectrophotometry two hours after bleaching. Microhardness was measured immediately after bleaching and then the specimens were immersed into artificial saliva for seven days for final color assessment. Data were analyzed by one-way analysis of variance followed by Tukey test. Bleaching reduced microhardness for all groups (p=0.0001), but the Ca+F and F groups showed lower reductions after bleaching. The addition of Ca, F, and Ca+F decreased the peroxide penetration through the tooth structure (p=0.0001), but there were no differences in color change for ΔL (p=0.357), Δa (p=0.061), Δb (p=0.823), and ΔE (p=0.581). The addition of calcium and fluoride in the gel did not affect bleaching efficacy, but it was able to reduce both the peroxide diffusion and the bleached enamel microhardness loss.


Electron beam therapy with coil-generated magnetic fields.
This paper presents an initial study on the issues involved in the practical implementation of the use of transverse magnetic fields in electron beam therapy. By using such magnetic fields the dose delivered to the tumor region can increase significantly relative to that deposited to the healthy tissue. Initially we calculated the magnetic fields produced by the Helmholtz coil and modified Helmholtz coil configurations. These configurations, which can readily be used to generate high intensity magnetic fields, approximate the idealized magnetic fields studied in our previous publications. It was therefore of interest to perform a detailed study of the fields produced by these configurations. Electron beam dose distributions for 15 MeV electrons were calculated using the ACCEPTM code for a 3T transverse magnetic field produced by the modified Helmholtz configuration. The dose distribution was compared to those obtained with no magnetic field. The results were similar to those obtained in our previous work, where an idealized step function magnetic field was used and a 3T field was shown to be the optimal field strength. A simpler configuration was also studied in which a single external coil was used to generate the field. Electron dose distributions are also presented for a given geometry and given magnetic field strength using this configuration. The results indicate that this method is more difficult to apply to radiotherapy due to its lack of symmetry and its irregularity. For the various configurations dealt with here, a major problem is the need to shield the magnetic field in the beam propagation volume, a topic that must be studied in detail.


Calculation of Hamilton energy and control of dynamical systems with different types of attractors.
Strange attractors can be observed in chaotic and hyperchaotic systems. Most of the dynamical systems hold a finite number of attractors, while some chaotic systems can be controlled to present an infinite number of attractors by generating infinite equilibria. Chaos can also be triggered in some dynamical systems that can present hidden attractors, and the attractors in these dynamical systems find no equilibria and the basin of attraction is not connected with any equilibrium (the equilibria position meets certain restriction function). In this paper, Hamilton energy is calculated on the chaotic systems with different types of attractors, and energy modulation is used to control the chaos in these systems. The potential mechanism could be that negative feedback in energy can suppress the phase space and oscillating behaviors, and thus, the chaotic, periodical oscillators can be controlled. It could be effective to control other chaotic, hyperchaotic and even periodical oscillating systems as well.


Supporting in grief and burnout of the nursing team from pediatric units in Chilean hospitals.
Objective To know the levels of Burnout Syndrome and perception of grief support in nursing teams of oncology and pediatric intensive care in public hospitals in Chile. Method A study of descriptive cross-sectional design. The Maslach Burnout Inventory and the Grief Support Health Care Scale were applied to university-level and technical nursing professionals between March and November 2015. An analysis to compare the means was performed with use of the Student's t-test, and the level of significance was set at 5%. Results The study included 153 professionals. Results show 4% of professionals have Burnout Syndrome (BS) and 89% are at risk of having it. Oncology professionals are at higher risk as they present higher levels of emotional exhaustion and lower levels of personal fulfillment. Half of professionals have a high level of perception of grief support, which is greater in the oncology team. Conclusion Professionals working in pediatric oncology units are at increased risk of Burnout Syndrome and grief support may be a mediating factor in this process.


Inhibition of NK cell activity through TGF-beta 1 by down-regulation of NKG2D in a murine model of head and neck cancer.
In an orthotopic murine model of head and neck squamous cell carcinoma (SCC VII/SF) we studied NK cell-mediated immunity following vaccination with a recombinant vaccinia virus expressing IL-2 (rvv-IL-2). SCC VII/SF tumor cells were injected into the oral cavity of C3H/HeJ mice on day 0. Mice were vaccinated on days 7, 10, and 14 with rvv-IL-2 and control vaccines. Phenotypes, numbers, and biological activities of NK cells were determined following vaccination. Levels of expression of NK-activating receptor NKG2D and CD16 on NK cell surface were assayed in the vaccinated mice. Expression of NKG2D ligands, Rae1, and H60 on SCC VII/SF cells was also examined. Vaccination with rvv-IL-2 resulted in expansion of NK cells. NK cells isolated from rvv-IL-2-vaccinated mice had significantly higher biological activities compared with mice treated with control vaccines. NK cells from tumor-bearing mice expressed significantly lower levels of NKG2D and CD16 compared with rvv-IL-2 vaccinated mice. SCC VII/SF tumors expressed NKG2D ligand Rae 1, although H60 was not present. SCC VII/SF tumors expressed high levels of TGF-beta1, which were down-modulated by vaccination with rvv-IL-2. Incubation of NK cells with tumor homogenate or cultured supernatant of SCC VII/SF cells reduced the expression of NKG2D and CD16. This inhibition appeared to be mediated by TGF-beta1. SCC VII/SF tumors in the oral cavity of the mice secrete high quantities of TGF-beta1, which reduce the expression of NK cell receptor NKG2D as well as CD16 and inhibits biological functions of NK cells.


Application of laser optical tweezers in immunology and molecular genetics.
Optical tweezers, based on a compact diode pumped Nd:YAG laser providing 350 mW at 1,064 nm coupled into a Zeiss IM 35 microscope, were used to sort CD4+ T cells into a capillary for further mechanical handling and to establish contact between single human natural killer (NK) cells and human erythroleukemia cells (K562) as targets. After contact and a lag phase of a few tens of seconds, the target cell starts to change its morphology and membrane blebbing occurs. The kinetics of the attack of the NK cell on K562 cells is not straightforward but governed by temporal oscillations in the shape of the target cell (zeosis). In a second application, the optical tweezers are combined with a UV laser microbeam based on a pulsed UV laser and with flow cytometry and sorting. With the pulsed laser, segments of sorted chromosome 1 of the chinese hamster karyotype (CHV 79) can be easily micro-dissected and subsequently collected using the optical tweezers. This allows preparation of a few hundred chromosome segments per day without mechanical contact and in an absolutely sterile way and thus may provide an interesting basic technique in any type of genome sequencing project.


Domesticating biosurveillance: 'containment' and the politics of bioinformation.
Although the term biosurveillance is employed with increasing frequency there remain variances in way in which the concept is both understood and practiced in the US and the UK, respectively. In this paper I begin by exploring the different epistemological and geographical approaches to biosurveillance that are employed in each locality, paying particular attention to the scales at which they, respectively, operate. I also consider how the subjects of these systems (a State's citizenry) are monitored in each jurisdiction and with what effects. I contend in this paper, and illustrate through a study of the techniques of surveillance employed during the recent H1N1 (swine flu) pandemic, that these different 'registers' of biosurveillance are now being bought into the same frame of reference to create new, ever more robust and finely calibrated systems of biological surveillance. In thinking through the political implications of the emergent collision, I outline here, employing work from Cooper, Katz, and Lyon how biosurveillance is becoming progressively domesticated and reflect on the potential this has for creating new, expansive, and very pervasive, forms of biological 'governmentality'.


Posttransplant lymphoproliferative disorder in liver allograft biopsies: a comparison of three methods for the demonstration of Epstein-Barr virus.
Posttransplant lymphoproliferative disorder (PTLD) is associated with Epstein-Barr virus (EBV), and may clinically resemble acute allograft rejection. Three methods to show EBV in tissue were evaluated in 15 liver allograft biopsies from 12 patients including four with PTLD: (1) semiquantitative polymerase chain reaction (PCR) for EBV DNA; (2) in situ hybridization for EBV RNA (EBER); and (3) immunoperoxidase for EBV latent membrane protein (LMP). Index cases had a PCR dot blot result of "positive" or "weak positive." Findings were correlated with histology, clinical data, therapy, and outcome. All four PTLD patients had a clinical diagnosis of acute rejection. All four showed EBV: PCR 4, EBER 4, LMP 3, Liver function tests were elevated in three, but EBV viral capsid antigen (VCA) IgM was not increased in three, but EBV viral capsid antigen (VCA) IgM was not increased in three. Immunosuppression was withdrawn and all four patients underwent a second transplantation. One died 4 days posttransplant with disseminated PTLD, two died of sepsis at 1.5 and 14 months, and one is well at 3 years without PTLD. Eleven biopsies without PTLD showed: acute rejection 7, acute rejection and hepatitis 1, hepatitis B 1, and non-inflammatory changes 2. In this group, EBV results included: PCR weak positive in 10 and 1+ in one, EBER negative in ten and rare positive cells in one, LMP negative in 11. Liver function tests were elevated in 10, whereas VCA IgM was not increased in three and increased in one. Patients with acute rejection were treated with increased immunosuppression: none developed PTLD, with follow-up of at least 6 months in nine cases. Two patients died within 4 months of biopsy. One patient with PTLD in tonsils had a liver biopsy showing both acute rejection and EBV (PCR 1+, rare EBER + small cells). Histological studies combined with special EBV detection methods, can be useful to evaluate atypical lymphoid infiltrates in liver allograft biopsies and confirmation of a diagnosis of PTLD. All three methods are useful; EBER and PCR are the most sensitive. EBER and LMP can use paraffin sections.


An inhibitory role of NEK6 in TGFβ/Smad signaling pathway.
The NEK6 (NIMA-related kinases 6) is reported to play potential roles in tumorigenesis. Although it is suggested to function in several cellular pathways, the underlying mechanism in tumorigenesis is still largely unknown. In the present study, we discovered interaction of NEK6 with Smad4, a key member of transforming growth factor beta (TGFβ) pathway. Over-expression of NEK6 in hepatocellular carcinoma (HCC) cell lines suppresses TGFβ-mediated transcription activity in a kinase activity-dependent manner. In addition, NEK6 suppresses the cell growth arrest induced by TGFβ. Mechanically, NEK6 blocks nuclear translocation of Smad4, which is essential for TGFβ function. Moreover, we identified that NEK6 could be regulated by TGFβ and hypoxia. Our study sheds new light on the roles of NEK6 in canonical TGFβ/Smad pathway and tumorigenesis.


Improved sensitivity for quantification of proteins using triply charged cleavable isotope-coded affinity tag peptides.
Isotope-coded affinity tag (ICAT) methods, in conjunction with capillary liquid chromatography/tandem mass spectrometry (LC/MS/MS), represent a promising approach for accurate protein quantification. However, sensitivity remains a challenge for the quantification of low-copy proteins in complex biological matrices. Here we investigated the electrospray ionization (ESI) and collision-activated dissociation (CAD) behavior of peptides derivatized with the cleavable ICAT (cICAT) reagent. For cICAT-peptides that were either synthesized or obtained by digestion of model proteins, the cICAT moiety showed a tendency toward protonation under positive ESI, producing relatively intense triply charged cICAT-peptide ions ([IP+3H]3+). [IP+3H]3+ exhibited significantly higher CAD reactivity than did the doubly charged cICAT-peptide ([IP+2H]2+), and produced a greater abundance of fragments at lower collision energies. Fragmentation spectra of [IP+3H]3+ showed variable intensities of doubly charged y and b ions, and the amount of sequence information obtained was dependent on the position of the cICAT-labeled cysteine residue in the peptide sequence. However, the absolute abundances of major fragments of [IP+3H]3+ were much higher than for [IP+2H]2+. Although the efficiency of identification of cICAT-peptides was compromised by their charge distribution toward the triply charged state and by the unique CAD behavior of the [IP+3H]3+ ions, it was found that the triply charged ions provided higher sensitivity than [IP+2H]2+ for quantification using multiple reaction monitoring (MRM). ESI and CAD conditions for MRM of [IP+3H]3+ were optimized, and, for all cICAT-peptides studied, MRM using [IP+3H]3+ as precursors showed 2- to 8-fold higher sensitivity than obtained using [IP+2H]2+, without compromising quantitative accuracy. Using this approach, the time course of tyrosine aminotransferase induction by methylprednisolone was monitored in rat livers. A remarkably better signal-to-noise ratio was observed by using [IP+3H]3+ for quantification compared to [IP+2H]2+.


Is the Natural Instinct to Oviposit in Mated Female Oriental Fruit Fly, Bactrocera dorsalis More of a Brain-Independent Act?
What physiological and neuro-molecular changes control the female oviposition behavior post-mating in insects? The molecular changes that occur in a gravid female insect are difficult to dissect out considering the distinct behavioral patterns displayed by different insect groups. To understand the role of the brain center in Oriental fruit fly, Bactrocera dorsalis oviposition, egg-laying behavior was analyzed in γ-octalactone exposed, decapitated mated B. dorsalis females. Interestingly, the females displayed a possible urge to oviposit, which suggests a natural instinct to pass on the gene pool. Expression analysis of certain genes involved in oviposition behavior was also carried out in these insects to explore the molecular aspects of such behavior. This study tries to assess the involvement of brain center in egg-laying and also explore the role of certain neurotransmitter-related receptors in decapitated B. dorsalis oviposition behavior. Our results indicate that B. dorsalis oviposition behavior could potentially have a bypass route of neuronal control devoid of the brain. The study reported here establishes that decapitation in gravid females fails to abolish their ability to sense ovipositional cues and also to oviposit.


Toward an understanding of the sequence and structural basis of allosteric proteins.
Allostery is the most efficient means of regulating protein functions, ranging from the control of metabolic mechanisms to signal transduction pathways. Although allosteric regulation has been recognized for half a century, our knowledge is limited to the characteristics of allosteric proteins and the structural coupling of allosteric sites and modulators. In this paper, we present a comprehensive analysis of allosteric proteins that provides insight into the foundation of allosteric interactions by revealing a series of common features in the allosteric proteins. Allosteric proteins mainly appear in transferases, and phosphorylation is the most common type of modification found in allosteric proteins. Disorders related to allosteric proteins primarily comprise metabolic diseases and cancers. In general, allosteric proteins prefer to exist as monomers or even-numbered multimers. Greater stability and hydrophobicity are observed in allosteric proteins than in general proteins. Further analysis of the allosteric sites reveals a series of buried and compact pockets composed of significantly greater hydrophobic surface area than the corresponding orthosteric sites. The hydrophobicity of the allosteric sites plays a dominant role in the binding of allosteric modulators as observed in the analysis of 106 diverse allosteric protein-modulator pairs. These results may be of great significance in predicting which proteins are allosteric and in designing novel triggers to inhibit or activate proteins of interest.


Hyperhomocysteinemia and Risk of First Venous Thrombosis: The Influence of (Unmeasured) Confounding Factors.
Meta-analyses have reported a 2- to 3-fold increased risk of venous thrombosis (VT) in individuals with hyperhomocysteinemia. However, confounding factors were generally not considered. In contrast, randomized trials of homocysteine-lowering therapy and VT risk have been negative. We investigated whether hyperhomocysteinemia was associated with VT in the Multiple Environmental and Genetic Assessment of Risk Factors for Venous Thrombosis (MEGA) case-control study (1999-2004) from the Netherlands (1,689 cases and 1,726 controls), taking into account measured and unmeasured confounders. We compared patients with population controls to estimate odds ratios using unconditional logistic regression and adjusted for various potential confounders. We matched patients to their partners to additionally adjust for unmeasured confounders (e.g., lifestyle factors) using conditional logistic regression. We found that elevated homocysteine concentrations were not associated with an increased risk for VT when comparing patients with population controls, either as a continuous variable (odds ratio = 1.00, 95% confidence interval: 0.99, 1.01), in terms of 0.7-mg/L increase (odds ratio = 0.99, 95% confidence interval: 0.93, 1.05), or within different homocysteine categories. We obtained similar results when patients were compared with their partners. Stratification by sex, deep vein thrombosis, pulmonary embolism, provoked VT, and unprovoked VT also provided no evidence of an association. In conclusion, after extensive adjustments for confounding, hyperhomocysteinemia was not associated with an increased risk of venous thrombosis in this study.


Tooth loss and tooth retention in a multitribal group of Bantu-speaking South African blacks: a study on 500 dry mandibles.
Tooth loss and tooth retention in adult South African Bantu-speaking Negroes were studied from data derived from 500 dry mandibles. The tribe, sex, and stated age of the specimens were available. Mandibles were equally distributed over the third to seventh decade of life. The presence of dental units was recorded and analysed within and between decade age groups. The mandibles carried a total of 5,459 dental units. Between the third and fourth decades of life and between the sixth and seventh decades, the reduction of the frequency of dental units was mild and statistically not significant. It was, however, severe and significant between the fourth and fifth, and the fifth and sixth decades. In the third-decade group, most teeth were found in the anterior region with a tendency to diminish in number posteriorly. After the third decade there was a definite persistence of first premolars and canines with significant tendency toward loss of teeth anterior and posterior to them. Only the incisors and third molars showed significant reduction in number from the third to the fourth decade. The reduction of all teeth was statistically significant later, until the sixth decade of life. Although there was loss of teeth from the sixth to the seventh decade, it was not significant for any of the homologous dental units. There is a possibility that some dietary differences existed between the older subjects (50-70 years old) and the younger ones (20-49 years old) which may have had some influence on the results.


[Corticotherapy in the otorhinolaryngology service at Brazzaville university hospital].
Corticoids are one of the great therapeutic advances of modern medicine. A retrospective study of corticotherapy conducted in the otorhinolaryngology and cervicofacial surgery service at Brazzaville University Hospital (Centre hospitalier et universitaire de Brazzaville) examined 952 cases treated from 1996 through 2000 and enabled us to determine that: corticoids account for 14% of the treatments prescribed to ENT patients; the patients' average age was 27 years; the most common indications were infections and/or inflammatory disorders (98.2%); local administration was most often prescribed (78.7%, compared with systemic, 21.3%); dexamethasone (60.1%) and betamethasone (19.5%) were prescribed most often; the average duration of treatment was 8 days; adverse reactions were rare (2 cases); treatment was effective in 78% of the cases.


A rational use of immune enhancing diets: when should we use dietary arginine supplementation?
Controversies in any arena of human activity often result in the polarization of the individuals involved into 2 opposing camps. Controversy about the use of immune-enhancing diets (IEDs) is no exception. On one hand, some groups are proposing indiscriminate use of IEDs, whereas others have created guidelines advocating that their use should be banned for the critically ill. At stake is an emerging paradigm: that dietary manipulation of the immune system is possible and may become an important adjunct to other therapies, thus helping prevent or treat multiple diseases for millions worldwide. Under these circumstances, extremist claims of miraculous benefits or inappropriate assertions of evil can only delay the emergence of a nascent science. This paper is therefore a plea for moderation from both camps, lest we cause irreparable damage to our clinical practices and potential injury to individual patients. IEDs all contain arginine. However, they also contain other substances such as omega-3 fatty acids, and nucleotides. The use of all these nutrients together into commercial IEDs without adequate evaluation of their individual effects has prevented the development of mechanistic hypotheses of action. Despite this, IEDs have been tested extensively, allowing the development of guidelines for their use. IEDs should be used for surgical patients, especially those undergoing elective surgery. IEDs show no benefit and indeed can potentially harm patients with sepsis, especially in the nonsurgical group, and should not be used outside of research protocols. Advances in basic research have helped us understand mechanisms of how arginine contained in IEDs may help surgical patients but may be deleterious in patients with sepsis. A review of the basic mechanisms of action of arginine on the immune system is enclosed in this paper and should serve as a basis for the development of scientific principles that guide clinical use of IEDs.


Metastatic Cutaneous Melanoma of the Gallbladder.
Metastatic melanoma is an aggressive disease that can spread to many organs of the body. In rare cases, it can spread to the gallbladder causing secondary lesions, yet presenting with little to no symptoms. Therefore, most cases of metastatic melanoma lesions to the gallbladder go undiagnosed. Here, we present the case of a 41-year-old male with a four-month history of melanoma of the face, with a postresection status, who presented with right upper quadrant abdominal pain. Doppler ultrasound and computed tomography confirmed the presence of a mass on the gallbladder. Laparoscopic excision along with liver wedge resection was performed. Pathology staining revealed the presence of a malignant metastatic melanoma lesion of the gallbladder.


Role of neutrophils in the deposition of platelets during acute inflammation.
Platelets can release a variety of inflammatory mediators. These formed elements have been shown to accumulate early in acute inflammation, often prior to fibrin and thrombus formation. Polymorphonuclear leukocyte (PMNL) infiltration is also an early event and is linked to protein exudation and hyperemia. The relationship between PMNL and platelet accumulation was studied. Inflammation was induced in rabbits by intradermal injection of stimuli, and 51Cr-labeled leukocytes and 111In-labeled platelets were used to quantitate the rates of leukocyte and platelet accumulation in the dermal reactions. A temporal association between the rates of leukocyte infiltration (greater than 95% PMNL) and of platelet deposition at skin sites was observed when killed Escherichia coli, E. coli-derived chemotactic factors, zymosan-activated plasma, f-met-leu-phe, or endotoxin were injected intradermally. A linear correlation (r = 0.96; p less than 0.001) between these parameters was observed. 59Fe-labeled red cells did not accumulate in these lesions. Platelet deposition in response to inflammatory stimuli did not occur in PMNL-depleted (nitrogen mustard treatment) but platelet-sufficient rabbits, in spite of normal platelet deposition in thrombin-injected sites. Platelet responses to inflammatory stimuli were normal when neutropenia, after nitrogen mustard treatment, was prevented. In contrast, in situ PMNL reconstitution of the skin sites in neutropenic rabbits did not cause local platelet accumulation. Intravenous infusion of the synthetic chemotactic factor, f-met-leu-phe, induced transient neutropenia and thrombocytopenia (30% of control) with platelet sequestration primarily in the lung. This is also the known site of PMNL sequestration during f-met-leu-phe infusion. It is concluded that platelets selectively deposit in acutely inflamed tissues primarily during PMNL margination in, and emigration across, the microvasculature. Platelets and PMNLs likely coassociate on the vessel wall, and this interaction may influence the course of inflammation.


Consequences of genetic manipulations of gonadotrophins and gonadotrophin receptors in mice.
We have produced over the years several genetically modified mouse models (transgenic [TG], knockout [KO] and knockin [KI]) for the study of normal and aberrant functions of gonadotrophins and their receptors. We summarise in the present review some of our recent findings on these animal models. One is the cascade of extragonadal phenotypes triggered by ovarian hyperstimulation in TG mice overexpressing the human choriongonadotrophin (hCG) beta-subunit and presenting with elevated levels of serum luteinising hormone (LH)/hCG bioactivity. Massively elevated levels of serum progesterone, rather than oestrogens, are responsible for the induction of pituitary prolactinomas and the subsequently elevated prolactin (PRL) levels. Along with normal oestradiol and elevated progesterone levels, the increased concentration of PRL induces lobuloalveolar development of the mammary gland, with ultimate formation of oestrogen and progesterone receptor-negative malignant tumours. Another TG mouse model expressing a constitutively activating mutant form of the follicle-stimulating hormone receptor (FSHR) presents with a strong ovarian phenotype inducing advanced follicular development and depletion, haemorrhagic follicles, teratomas and infertility. A third TG mouse model, coexpressing binding- and signalling-deficient mutants of LHCGR in the KO background for the same receptor (R) gene provided convincing evidence that functional complementation through homo-di/oligomerisation is a physiologically relevant mode of activation of class A G protein-coupled receptors (GPCR). Taken together, genetically modified mouse models provide powerful tools for the elucidation of normal and pathological functions of gonadotrophins and their R.


Temperature effects on pacemaker generation, membrane potential, and critical firing threshold in Aplysia neurons.
Temperature increases cause a regular and reproducible increase in the frequency of generation of pacemaker potentials in most Aplysia neurons specialized for this type of activity which can only be explained as a direct stimulating effect of temperature upon the ionic mechanisms responsible for pacemaker potentials. At the same time all cells in the visceral ganglion undergo a membrane potential hyperpolarization of approximately 1-2 mv/ degrees C warmed. In spite of the marked variation in resting membrane potential the critical firing threshold remains at a constant membrane potential level at all temperatures in the absence of accommodative changes. The temperature-frequency curves of all types of cells are interpreted as a result of the interaction between the effects of temperature on the pacemaker-generating mechanism and resting membrane potential. Previous observations on the effects of temperature on excitability of mammalian neurons suggest that other types of neurons may undergo similar marked shifts in resting membrane potential with temperature variation.


Menstrual migraine. Methods of prevention and control.
In many women, migraine headaches are clearly linked to estrogen levels: the incidence rises at the menarche; attacks may be precipitated by falling estrogen levels before menses; and symptoms usually improve during pregnancy when there are noncyclic high levels of estrogen. Decreased estrogen production in the perimenopausal phase may trigger an exacerbation of migraine. However, after menopause when estrogen levels are noncyclic and low, there may be an improvement in migraine. The falling estradiol level rather than the absolute level provides the trigger for menstruation-associated migraine. Treatment involves both prophylactic and acute measures. Therapy for an acute attack is similar to that for nonmenstrual migraine. Sumatriptan is equally effective for both nonmenstrual and menstrual migraine.


Oral vinorelbine as first line chemotherapy in unfit elderly patients with hormone-refractory prostate cancer.
Hormone-refractory prostate cancer (HRPC) is a rapidly progressive disease which produces considerable morbidity and involves mostly men over 70, often comorbid and with poor tolerance to chemotherapy. Low-toxicity chemotherapy is a reasonable option in this setting. Vinorelbine and a corticosteroid show activity and clinical benefit responses in HRPC. An oral regimen is preferable for elderly patients. This study aimed to evaluate safety, prostate-specific antigen (PSA) response, clinical benefit and progression-free survival in chemonaive elderly HRPC patients. 33 men, median age 78.2, were treated with oral vinorelbine 60 mg/m2 days 1 and 8 every 3 weeks, escalable to 80 mg/m2 after the first cycle, and prednisone 5 mg b.i.d. The main toxicity was hematopoietic (mild at 60 mg/m2 and moderate at 80 mg/m2). Of 27 evaluable patients, 9 (33%) had PSA responses and 9 had clinical benefit, PSA-correlated in 5 cases (56%). Median progression-free survival was 13.4 weeks, median overall survival 45 weeks. Oral vinorelbine plus prednisone is safe and has moderate activity, with biochemical and clinical responses in about one-third of patients and could be an option in unfit elderly HRPC patients.


How do researchers determine the difference to be detected in superiority trials? Results of a survey from a panel of researchers.
There is currently no guidance for selecting a specific difference to be detected in a superiority trial. We explored 3 factors that in our opinion should influence the difference to be detected (type of outcome, patient age group, and presence of treatment side-effects), and 3 that should not (baseline level of risk, logistical difficulties, and cost of treatment). We conducted an experimental survey using a factorial design among 380 corresponding authors of randomized controlled trials indexed in Medline. Two hypothetical vignettes were submitted to participants: one described a trial of a new analgesic in mild trauma injuries, the other described a trial of a new chemotherapy among cancer patients. The first vignette tested the baseline level of risk, patient age-group, patient recruitment difficulties, and treatment side-effects. The second tested the baseline level of risk, patient age-group, type of outcome, and cost of treatment. The respondents were asked to select the smallest gain of effectiveness that should be detected by the trial. In vignette 1, respondents selected a median difference to be detected corresponding to an improvement of 7.0 % in pain control with the new treatment. In vignette 2, they selected a median difference to be detected corresponding to a reduction of 5.0 % in mortality or cancer recurrence with the new chemotherapy. In both vignettes, the difference to be detected decreased significantly with the baseline risk. The other factor influencing difference to be detected was the age group, but the impact of this factor was smaller. Cost, side-effects, outcome severity, or mention of logistical difficulties did not significantly impact the difference to be detected selected by participants. Three of the anticipated effects conformed to our expectations (the effect of patient age, and absence of effect of the cost of treatment and of patient recruitment difficulties) and the other three did not. These findings can guide future research in determining differences to be detected in trials that can translate to meaningful clinical decision-making.


OncodriveCLUST: exploiting the positional clustering of somatic mutations to identify cancer genes.
Gain-of-function mutations often cluster in specific protein regions, a signal that those mutations provide an adaptive advantage to cancer cells and consequently are positively selected during clonal evolution of tumours. We sought to determine the overall extent of this feature in cancer and the possibility to use this feature to identify drivers. We have developed OncodriveCLUST, a method to identify genes with a significant bias towards mutation clustering within the protein sequence. This method constructs the background model by assessing coding-silent mutations, which are assumed not to be under positive selection and thus may reflect the baseline tendency of somatic mutations to be clustered. OncodriveCLUST analysis of the Catalogue of Somatic Mutations in Cancer retrieved a list of genes enriched by the Cancer Gene Census, prioritizing those with dominant phenotypes but also highlighting some recessive cancer genes, which showed wider but still delimited mutation clusters. Assessment of datasets from The Cancer Genome Atlas demonstrated that OncodriveCLUST selected cancer genes that were nevertheless missed by methods based on frequency and functional impact criteria. This stressed the benefit of combining approaches based on complementary principles to identify driver mutations. We propose OncodriveCLUST as an effective tool for that purpose. OncodriveCLUST has been implemented as a Python script and is freely available from http://bg.upf.edu/oncodriveclust firstname.lastname@example.com or firstname.lastname@example.com Supplementary data are available at Bioinformatics online.


Definition of the immunogenic forms of modified human LDL recognized by human autoantibodies and by rabbit hyperimmune antibodies.
Humans and laboratory animals recognize human modified LDL as immunogenic. Immune complexes (ICs) isolated from human sera contain malondialdehyde-modified LDL (MDA-LDL) and N (epsilon)(carboxymethyl)lysine-modified LDL (CML-LDL) as well as antibodies reacting with MDA-LDL, copper-oxidized LDL (OxLDL), CML-LDL, and advanced glycosylation end product (AGE)-modified LDL. OxLDL and AGE-LDL antibodies isolated from human sera recognize the same LDL modifications and do not react with modified non-LDL proteins. Rabbit antibodies have different reactivity patterns: MDA-LDL antibodies react strongly with MDA-LDL and MDA-BSA but weakly with OxLDL; OxLDL antibodies react strongly with OxLDL and weakly with MDA-LDL; CML-LDL antibodies react with CML-LDL > CML-BSA > AGE-LDL > OxLDL; AGE-LDL antibodies react strongly with AGE-LDL, react weakly with OxLDL, and do not react with CML-LDL. Thus, human and rabbit antibodies seem to recognize different epitopes. Capture assays carried out with all rabbit antibodies showed binding of apolipoprotein B-rich lipoproteins isolated from ICs, suggesting that laboratory-generated epitopes are expressed by in vivo-modified LDL, although they are not necessarily recognized by the human immune system. Thus, the definition of immunogenic forms of modified LDL eliciting human autoimmune responses requires the isolation and characterization of autoantibodies and modified LDL from human samples, whereas rabbit antibodies can be used to detect in vivo-modified human LDL.


Human Somatosensory Processing and Artificial Somatosensation.
In the past few years, we have gained a better understanding of the information processing mechanism in the human brain, which has led to advances in artificial intelligence and humanoid robots. However, among the various sensory systems, studying the somatosensory system presents the greatest challenge. Here, we provide a comprehensive review of the human somatosensory system and its corresponding applications in artificial systems. Due to the uniqueness of the human hand in integrating receptor and actuator functions, we focused on the role of the somatosensory system in object recognition and action guidance. First, the low-threshold mechanoreceptors in the human skin and somatotopic organization principles along the ascending pathway, which are fundamental to artificial skin, were summarized. Second, we discuss high-level brain areas, which interacted with each other in the haptic object recognition. Based on this close-loop route, we used prosthetic upper limbs as an example to highlight the importance of somatosensory information. Finally, we present prospective research directions for human haptic perception, which could guide the development of artificial somatosensory systems.


The Multiple Roles of Peptidyl Prolyl Isomerases in Brain Cancer.
Peptidyl prolyl isomerases (PPIases) are broadly expressed enzymes that accelerate the cis-trans isomerization of proline peptide bonds. The most extensively studied PPIase family member is protein interacting with never in mitosis A1 (PIN1), which isomerizes phosphorylated serine/threonine⁻proline bonds. By catalyzing this specific cis-trans isomerization, PIN1 can alter the structure of its target proteins and modulate their activities in a number of different ways. Many proteins are targets of proline-directed phosphorylation and thus PIN1-mediated isomerization of proline bonds represents an important step in the regulation of a variety of cellular mechanisms. Numerous other proteins in addition to PIN1 are endowed with PPIase activity. These include other members of the parvulin family to which PIN1 belongs, such as PIN4, as well as several cyclophilins and FK506-binding proteins. Unlike PIN1, however, these other PPIases do not isomerize phosphorylated serine/threonine⁻proline bonds and have different substrate specificities. PIN1 and other PPIases are overexpressed in many types of cancer and have been implicated in various oncogenic processes. This review will discuss studies providing evidence for multiple roles of PIN1 and other PPIases in glioblastoma and medulloblastoma, the most frequent adult and pediatric primary brain tumors.


Genomic DNA analysis of genes encoding (hemi-)cellulolytic enzymes of the anaerobic fungus Piromyces sp. E2.
Anaerobic fungi contain more than one copy of genes encoding (hemi-)cellulases in their genome. The arrangement of these genes on the chromosomes was not known. A genomic DNA (gDNA) library of Piromyces sp. E2 was screened with different probes specific for (hemi-)cellulolytic enzymes. This screening resulted in three gDNA clones with genes encoding glycoside hydrolase enzymes of families 1 (beta-glucosidase), 6 (exoglucanase) and 26 (mannanase). Each clone contained two or more genes of the same family. Comparison of the gene copies on a clone revealed that they were highly homologous, and in addition, 54-75% of the substitutions was synonymous. One of the mannanase genes contained an intron. PCR with selected primers resulted in a gDNA clone with a new representative (cel9B) of glycoside hydrolase family 9 (endoglucanase). Comparison with cel9A revealed that cel9B had 67% homology on the nucleotide level. Furthermore, three introns were present. All results of this paper taken together provided evidence for duplications of (hemi-)cellulolytic genes, which resulted in clusters of almost identical genes arranged head-to-tail on the genome. In contrast to other eukaryotes, this phenomenon appears frequently in anaerobic fungi.


R&D during public health emergencies: the value(s) of trust, governance and collaboration.
In January 2021, Dr Tedros Adhanom Ghebreyesus, director-general of the WHO, warned that the world was 'on the brink of a catastrophic moral failure [that] will be paid with lives and livelihoods in the world's poorest countries'. We are now past the brink. Many high-income countries have vaccinated their populations (which, in some cases, includes third and even fourth doses) and are loosening public health and social measures, while low-income and middle-income countries are struggling to secure enough supply of vaccines to administer first doses. While injustices abound in the deployment and allocation of COVID-19 vaccines, therapies and diagnostics, an area that has hitherto received inadequate ethical scrutiny concerns the upstream structures and mechanisms that govern and facilitate the research and development (R&D) associated with these novel therapies, vaccines and diagnostics. Much can be learnt by looking to past experiences with the rapid deployment of R&D in the context of public health emergencies. Yet, much of the 'learning' from past epidemics and outbreaks has largely focused on technical or technological innovations and overlooked the essential role of important normative developments; namely, the importance of fostering multiple levels of trust, strong and fair governance, and broad research collaborations. In this paper, we argue that normative lessons pertaining to the conduct of R&D during the 2014-2016 Ebola epidemic in West Africa provide important insights for how R&D ought to proceed to combat the current COVID-19 pandemic and future infectious disease threats.


Retinopathy is related to the angiographically detected severity and extent of coronary artery disease in patients with type 2 diabetes mellitus.
Diabetic retinopathy (DR) is an early and frequent marker of other vascular complications of diabetes and its relation with coronary ischemia is known. The aim of the present study was to evaluate the association between DR and indices of coronary artery disease (CAD) severity and extent determined by coronary angiography. Sixty-nine diabetic patients undergoing coronary angiography for suspected CAD were evaluated. The severity and extent of CAD were scored from coronary angiograms by using 3 scores. Retinopathy was graded by fundus examination. There were differences in the severity score, extent score, number of vessels with disease, duration of diabetes, diabetes therapy, history of previous myocardial infarction (MI), and serum creatinine level among patients with and without DR. CAD severity was associated with the presence of DR (r: 0.53, P < 0.001), grade of DR (r: 0.52, P < 0.001), duration of diabetes (r: 0.28, P: 0.019), history of previous MI (r: 0.36, P: 0.002); and serum creatinine level (r: 0.24, P: 0.049) where the presence of DR was the only independent factor related to the severity score in multivariate analysis (r: 0.48, P < 0.001). The parameters related to the extent score were the presence of DR (r: 0.50, P < 0.001); grade of DR (r: 0.48, P < 0.001); previous MI (r: 0.37, P: 0.002) and age of the patient (r: 0.26, P: 0.033). Factors independently related to the extent score in multivariate analysis were the presence of DR (r: 0.37, P: 0.001), previous MI (r: 0.30, P: 0.006), and age of the patient (r: 0.22, P: 0.003). Among diabetics who are suspected of having CAD, those with retinopathy have more diffuse and severe coronary atherosclerosis, compared with diabetics without retinopathy. This cannot be explained by a longer duration or inferior control of the disease.


ATAC Sequencing Protocol For Cryopreserved Mammalian Cells.
ATAC-seq (assay for transposase-accessible chromatin with high-throughput sequencing) is a powerful method to evaluate chromatin accessibility and nucleosome positioning at a genome-wide scale. This assay uses a hyperactive Tn5 transposase, to simultaneously cut open chromatin and insert adapter sequences. After sequencing, the reads generated through this technique are generally indicative of transcriptional regulatory elements that are located in accessible chromatin. This method was originally developed by Buenrostro et al. (2013), and since then it has been improved by the same authors several times, until their last update called OMNI ATAC-seq ( Corces et al., 2017 ). Here, we describe an ATAC-seq protocol based on the OMNI-ATAC method, with a special focus on the initial steps of thawing cryopreserved cells, and the final steps of library purification using magnetic beads. This protocol can be of interest for laboratories working in a fast-paced environment. Graphic abstract: Flowchart of the protocol.


Episodic conjunctival inflammation after Stevens-Johnson syndrome.
The authors studied the histopathologic, ultrastructural, and immunopathologic characteristics of conjunctiva from patients with Stevens-Johnson syndrome (SJS). A small subset of SJS patients with recurrent conjunctival inflammation unassociated with external factors such as lid margin keratinization, sicca syndrome, trichiasis, or entropion was identified. The ultrastructural and immunopathologic characteristics of the conjunctiva from these patients were distinctly different from those of the conjunctiva from SJS patients without recurrent conjunctivitis, and suggested an active, immunologically mediated inflammation. Vasculitis or perivasculitis, immunoreactant deposition in vessel walls, vascular basement membrane disruption, thickening, and reduplication, and a preponderance of helper T-lymphocytes, macrophages, and Langerhans' cells were the notable distinguishing features in those patients with recurrent conjunctival inflammation. This rare clinical syndrome may represent the ocular counterpart to recurrent dermal or oral mucosal erythema multiforme.


SHARON process evaluated for improved wastewater treatment plant nitrogen effluent quality.
New stricter nitrogen effluent standards and increasing influent loads require existing wastewater treatment plans (WWTPs) to extend or optimize. At WWTPs with limited aeration capacity, limited denitrification capacity or shortage of aerobic sludge age, implementation of SHARON to improve nitrogen effluent quality can be a solution. SHARON is a compact, sustainable and cost-effective biological process for treatment of nitrogen-rich rejection waters. At WWTP Rotterdam-Dokhaven and WWTP Utrecht a SHARON has been in operation for several years. For both WWTPs the effect of SHARON on the nitrogen effluent quality has been evaluated. WWTP Rotterdam-Dokhaven has limited aeration capacity. By implementation of SHARON, the ammonia load of the effluent was reduced by 50%. WWTP Utrecht had limited denitrification capacity. The implementation of SHARON improved the effluent nitrate load by 40%. The overall TN removal efficiency increased from 65% to over 75% and strict nitrogen effluents standards (TN = 10 mg N/l) could be reached. Through modelling and supported by full scale practice it has been shown that by implementation of SHARON in combination with enhanced influent pre-treatment, the aerobic sludge age can be extended to maintain total nitrogen removal at lower temperatures.


5-methylcytosine modification of an Epstein-Barr virus noncoding RNA decreases its stability.
Many cellular noncoding RNAs contain chemically modified nucleotides that are essential for their function. The Epstein-Barr virus expresses two highly abundant noncoding RNAs called EBV-encoded RNA 1 (EBER1) and EBER2. To examine whether these viral RNAs contain modified nucleotides, we purified native EBERs from EBV-infected cells and performed mass spectrometry analysis. While EBER2 contains no modified nucleotides at stoichiometric amounts, EBER1 was found to carry 5-methylcytosine (m5C) modification. Bisulfite sequencing indicated that a single cytosine of EBER1 is methylated in ∼95% of molecules, and the RNA methyltransferase NSUN2 was identified as the EBER1-specific writer. Intriguingly, ablation of NSUN2 and thus loss of m5C modification resulted in an increase in EBER1 levels. We further found that EBER1 is a substrate for the RNase Angiogenin and cleavage in vivo is dependent on the presence of m5C, providing an explanation as to why loss of m5C increases EBER1 levels. Taken together, our observations indicate that m5C, a modification previously shown for tRNAs to oppose Angiogenin-mediated degradation, can also adversely affect RNA stability.


Rapid Determination of Bucinnazine in Blood by UPLC-MS/MS.
To establish a rapid method for the analysis of bucinnazine in blood by UPLC-MS/MS and to apply the method to the practical case. After the internal standard was added to blood, the protein was precipitated with 900 μL mixed solution (Vacetonitrile∶Vwater=8∶2). After vortex and centrifugation, the protein was measured through 0.22 μm filter membrane. The separation was performed on C18 chromatography column, with acetonitrile and 5 mmol/L ammonium acetate containing 0.1% formic acid aqueous as mobile phase gradient elution at the flow rate of 0.4 mL/min. Multiple reaction monitoring scan was performed in electrospray positive ion mode, quantitative measurement was performed by internal standard method, and methodological verification was carried out. The linear relationship of bucinnazine in blood was good in the range of 0.5-200 μg/L, the correlation coefficient (r) was 0.999 7, the limit of detection was 0.1 μg/L, the limit of quantitation was 0.5 μg/L, and the recovery was 78.3%-83.8% at 1, 10 and 100 μg/L mass concentration levels. The matrix effect was 69.4%-73.8%, the intra-day precision was 1.9%-2.8%, and the inter-day precision was 2.8%-3.2%, the accuracy was 3.1%-3.5%. The stability test results of 1 and 100 μg/L mass concentrations at -25 ℃ showed that the accuracy (bias) of 10 d was less than 4.5%. This method has the advantages of simple pre-treatment process, fast sample processing speed, high sensitivity of instrument analysis, good stability of content determination and reliable identification results, and can meet the needs of case identification.


Vaccines for women to prevent neonatal tetanus.
Tetanus is an acute, often fatal, disease caused by an exotoxin produced by Clostridium tetani. It occurs in newborn infants born to mothers who do not have sufficient circulating antibodies to protect the infant passively, by transplacental transfer. Prevention may be possible by the vaccination of pregnant and/or non-pregnant women with tetanus toxoid, and the provision of clean delivery services. Tetanus toxoid consists of a formaldehyde-treated toxin which stimulates the production of antitoxin. To assess the effectiveness of tetanus toxoid, administered to women of childbearing age or pregnant women, to prevent cases of, and deaths from, neonatal tetanus. We searched the Cochrane Pregnancy and Childbirth Group Trials Register (December 2004) , The Cochrane Library (Issue 1, 2005), MEDLINE (1966 to December 2004), EMBASE (1974 to December 2004). We also used the results from handsearching and consultations with manufacturers and authors. Randomised or quasi-randomised trials evaluating the effects of tetanus toxoid in pregnant women or women of childbearing age on numbers of neonatal tetanus cases and deaths. Three review authors independently assessed trials for inclusion, data extraction and trial quality. Two trials (10,560 infants) were included. One study (1919 infants) assessed the effectiveness of tetanus toxoid in preventing neonatal tetanus deaths. After a single dose, the relative risk (RR) was 0.57 (95% confidence interval (CI) 0.26 to 1.24), and the vaccine effectiveness was 43%. With a two or three dose course, the RR was 0.02 (95% CI 0.00 to 0.30); vaccine effectiveness was 98%. No effect was detected on causes of death other than tetanus. The RR of cases of neonatal tetanus after at least one dose of tetanus toxoid was 0.20 (95% CI 0.10 to 0.40); vaccine effectiveness was 80%. Another study, involving 8641 children, assessed the effectiveness of tetanus-diptheria toxoid in preventing neonatal mortality after one or two doses. The RR was 0.68 (95% CI 0.56 to 0.82); vaccine effectiveness was 32%. In preventing deaths at 4 to 14 days, the RR was 0.38 (95% CI 0.27 to 0.55), and vaccine effectiveness 62% (95% CI 45% to 73%). Available evidence supports the implementation of immunisation practices on women of childbearing age or pregnant women in communities with similar, or higher, levels of risk of neonatal tetanus, to the two study sites. More information is needed on possible interference of vaccination by malaria chemoprophylaxis on the roles of malnutrition and vitamin A deficiency, and on the quality of tetanus toxoid production and storage.


Generation and validation of structurally defined antibody-siRNA conjugates.
Gene silencing by RNA interference (RNAi) has emerged as a powerful treatment strategy across a potentially broad range of diseases. Tailoring siRNAs to silence genes vital for cancer cell growth and function could be an effective treatment, but there are several challenges which must be overcome to enable their use as a therapeutic modality, among which efficient and selective delivery to cancer cells remains paramount. Attempts to use antibodies for siRNA delivery have been reported but these strategies use either nonspecific conjugation resulting in mixtures, or site-specific methods that require multiple steps, introduction of mutations, or use of enzymes. Here, we report a method to generate antibody-siRNA (1:2) conjugates (ARCs) that are structurally defined and easy to assemble. This ARC platform is based on engineered dual variable domain (DVD) antibodies containing a natural uniquely reactive lysine residue for site-specific conjugation to β-lactam linker-functionalized siRNA. The conjugation is efficient, does not compromise the affinity of the parental antibody, and utilizes chemically stabilized siRNA. For proof-of-concept, we generated DVD-ARCs targeting various cell surface antigens on multiple myeloma cells for the selective delivery of siRNA targeting β-catenin (CTNNB1). A set of BCMA-targeting DVD-ARCs at concentrations as low as 10 nM revealed significant CTNNB1 mRNA and protein knockdown.


The Role of Duplex Ultrasound in Microsurgical Reconstruction: Review and Technical Considerations.
With the technological advances, microsurgery has evolved to the era of supermicrosurgery since its inception. With the popularization of these flaps, proficiency in using tools such as color Doppler ultrasound (CDU) has become crucial. Despite the increased interest, studies regarding the role of ultrasound in microsurgical reconstruction are scarce. Therefore, in this study, we discussed currently available imaging modalities for reconstructive surgeons, types of ultrasound, and the role of ultrasound in microsurgical reconstruction, and made recommendations for ultrasound use in reconstructive surgery. We performed a computerized search of ultrasound in reconstructive surgery using the MEDLINE database. Data regarding ultrasound indications, usage, and outcomes were collected. Of the 115 articles identified, the majority was written on nonreconstructive ultrasound uses (51.4%). For the reconstructive uses, mapping of perforators was the most common usage (39.1%), followed by flap monitoring and volume measurement. In addition, we found that there are a limited number of literature on the role of CDU in reconstructive surgery, especially on its intraoperative and postoperative use. CDU is a valuable and powerful tool for any reconstructive surgeons who are interested in performing microsurgery or supermicrosurgery. As the demand of customized flaps using superthin flaps, perforator-to-perforator anastomosis, and supermicrosurgery increases, understanding and becoming versatile in CDU will be critical. We hope our experience with using ultrasound and refinements that we made are helpful for those who would like to include it as a part of their armamentarium.


Measurement of quality of life in clinical trials of therapy.
This article discusses some of the issues involved in the use of quality of life methods to evaluate treatment, primarily in trials of cardiovascular disease. The choice of aspects to measure is influenced by the severity and nature of the disease, the expected benefits and adverse effects of treatment and pragmatic considerations such as the length of the trial, and availability of the methods. The choice of specific instruments should take account of good measurement criteria such as validity and reliability, and, most importantly, sensitivity to the effects of treatment. The advantages and disadvantages of interviewer and self-completed questionnaires are discussed. The choice of respondent is primarily the patient. In some circumstances it may be appropriate to also include family members, either for their views on the patient, or for an assessment of their own quality of life as this may also be disrupted. Withdrawals are a major problem in the analysis of all trial results. The inclusion of a Health Index is one method of obtaining quality of life scores from patients who withdraw or die. The interpretation of quality of life results from trials is not simple, for example, the benefit of trial inclusion needs to be distinguished from the effect of treatment. Moreover, changes in quality of life scores should correspond to changes in the patients' own experiences of their daily lives.


Nonintubated Transareolar Endoscopic Thoracic Sympathectomy with a Flexible Endoscope: Experience of 58 Cases.
Natural orifice transluminal endoscopic surgery (NOTES) has recently become a hot spot in the field of minimally invasive surgery. But, most of the procedures are still in the early stages of development and limited to animal experiments. Transareolar endoscopic surgery could work as a viable intermediate step before thoracic NOTES. Under intravenous anesthesia without endotracheal intubation, transareolar endoscopic thoracic sympathectomy (ETS) with a flexible endoscope has rarely been attempted. The objective of this study is to evaluate the feasibility and safety of this novel minimally invasive technique in managing primary palmar hyperhidrosis (PPH). From June 2012 to July 2014, a total of 58 male patients with severe PPH underwent transareolar ETS by use of a flexible endoscope. Under intravenous anesthesia without endotracheal intubation, a flexible endoscope was introduced through the incision on the edge of the areola into the thoracic cavity. The thoracic sympathetic chain was ablated at the level of the fourth rib. All procedures were successfully performed with a mean operating time of 33.6 ± 8.3 min. All patients regained consciousness rapidly and none of them complained about sore throat after surgery. There were no operative mortality and conversion to open procedure. The symptoms of all patients disappeared as soon as the sympathetic chain was cut off. Fifty six patients (96.6%) were discharged from the hospital on the first postoperative day. The postoperative complications were minor, and no patients developed Horner's syndrome. At 3 months postoperatively, there was no obvious surgical scar on the chest wall, and none of the patients complained about postoperative pain. Compensatory hyperhidrosis (CH) appeared in 19 patients. No recurrent symptoms were observed in our study. One year follow-up revealed an excellent cosmetic result and degree of satisfaction. Nonintubated transareolar ETS with a flexible endoscope is a safe, effective and minimally invasive therapeutic procedure, which has the possible advantages of thoracic NOTES and can be performed in routine clinical practice for male PPH patients.


Influence of sympathetic nervous system on dexamethasone-stimulated renal tubular transport of p-aminohippurate in young rats.
After simultaneous administration of dexamethasone together with 6-hydroxydopamine or adrenergic neuronal blocking agents (alpha-methyldopa, reserpine, guanethidine) the dexamethasone-stimulated increase in renal excretion of p-aminohippurate (PAH) is distinctly diminished. The same is true for dopamine. This depressive effect could be proved by measurement of transport rates of PAH in renal cortical slices in vitro. The increase in kidney weight and in the protein content of kidney tissue in dexamethasone-treated rats is lower in rats simultaneously treated with adrenergic neuronal blocking agents of 6-hydroxydopamine. Adrenoceptor agonists and antagonists (pholedrine, orciprenaline, phentolamine, propranolol) are without influence on the dexamethasone-stimulated increase in PAH transport.


Impact of platelet inhibition level on subsequent no-reflow in patients undergoing primary percutaneous coronary intervention for ST-segment elevation myocardial infarction.
High P2Y12 platelet reactivity (PR) level after primary percutaneous coronary intervention (PPCI) for ST-segment elevation myocardial infarction (STEMI) affects prognosis and may induce the no-reflow phenomenon. To investigate the role of PR in the genesis of microvascular obstruction. Patients with STEMI undergoing PPCI within 12hours of symptoms onset were included prospectively. All patients received a 600mg clopidogrel-loading dose before PPCI and 250mg aspirin. PR was measured thereafter during PPCI while wiring the culprit lesion and before coronary dilatation, using the P2Y12 VerifyNow® assay. No-reflow was defined as ST-segment regression<50% observed 90minutes after PPCI. Between January 2014 and November 2015, 140 STEMI patients were included, and divided into two groups: a low PR group (LPR) defined as PR<209P2Y12 reaction units (PRU); and a high PR group (HPR) defined as PR≥209PRU. There were no differences in baseline characteristics between LPR and HPR groups, including age (57.8±11.9 vs. 59.4±13.2 years, respectively; P=0.44) and weight (76.1±15.1 vs. 74.8±10.9kg, respectively; P=0.55). Delay to revascularization was 270.1±175.5 vs. 295.6±206.2minutes (P=0.49) and time between clopidogrel-loading and PR measurement was 53±37 vs 65±54minutes (P=0.29) in the LPR and HPR groups, respectively. No-reflow was more frequent in the HPR group (44 [47.3%] vs. 9 [19.1%]; P=0.0012). Mean PR was higher in patients with no-reflow: 268.3±53 vs. 223.8±50.1 PRU (P=0.002). In multivariable analysis, HPR was an independent predictor of no-reflow. Area under the receiver operating characteristic curve was 0.745 (0.654, 0.835); the cut-off value predicting no-reflow was 254PRU. High PR level measured at PPCI is independently associated with no-reflow.


Using phylogenomics to understand the link between biogeographic origins and regional diversification in ratsnakes.
Globally distributed groups may show regionally distinct rates of diversification, where speciation is elevated given timing and sources of ecological opportunity. However, for most organisms, nearly complete sampling at genomic-data scales to reduce topological error in all regions is unattainable, thus hampering conclusions related to biogeographic origins and rates of diversification. We explore processes leading to the diversity of global ratsnakes and test several important hypotheses related to areas of origin and enhanced diversification upon colonizing new continents. We estimate species trees inferred from phylogenomic scale data (304 loci) while exploring several strategies that consider topological error from each individual gene tree. With a dated species tree, we examine taxonomy and test previous hypotheses that suggest the ratsnakes originated in the Old World (OW) and dispersed to New World (NW). Furthermore, we determine if dispersal to the NW represented a source of ecological opportunity, which should show elevated rates of species diversification. We show that ratsnakes originated in the OW during the mid-Oligocene and subsequently dispersed to the NW by the mid-Miocene; diversification was also elevated in a subclade of NW taxa. Finally, the optimal biogeographic region-dependent speciation model shows that the uptick in ratsnake diversification was associated with colonization of the NW. We consider several alternative explanations that account for regionally distinct diversification rates.


Microemboli in aortic valve replacement.
Microembolic signals (MES) can be detected in many recipients of mechanical aortic valve prostheses by transcranial Doppler ultrasound. The nature and etiology of these MES have remained unclear for a long time. The solid and gaseous nature of MES are discussed, as well as whether or not MES may reflect artifacts. Recently, the gaseous nature of these MES has been widely established. To understand the physics of bubble formation related to mechanical heart valve prostheses, it is necessary to discuss the different types of cavitation occurring at the prostheses and the conditions leading to the degassing of blood. We describe the history of transcranial Doppler ultrasound-techniques and the current techniques in the measurement of these signals. Furthermore, the possible clinical impact of MES, as well as strategies for the design of new prostheses and surgical alternatives to diminish their load are discussed.


Protection of human influenza vaccines against a reassortant swine influenza virus of pandemic H1N1 origin using a pig model.
Since the pandemic H1N1 emergence in 2009 (pdmH1N1), many reassortant pdmH1N1 viruses emerged and found circulating in the pig population worldwide. Currently, commercial human subunit vaccines are used commonly to prevent the influenza symptom based on the WHO recommendation. In case of current reassortant swine influenza viruses transmitting from pigs to humans, the efficacy of current human influenza vaccines is of interest. In this study, influenza A negative pigs were vaccinated with selected commercial human subunit vaccines and challenged with rH3N2. All sera were tested with both HI and SN assays using four representative viruses from the surveillance data in 2012 (enH1N1, pdmH1N1, rH1N2 and rH3N2). The results showed no significant differences in clinical signs and macroscopic and microscopic findings among groups. However, all pig sera from vaccinated groups had protective HI titers to the enH1N1, pdmH1N1 and rH1N2 at 21DPV onward and had protective SN titers only to pdmH1N1and rH1N2 at 21DPV onward. SN test results appeared more specific than those of HI tests. All tested sera had no cross-reactivity against the rH3N2. Both studied human subunit vaccines failed to protect and to stop viral shedding with no evidence of serological reaction against rH3N2. SIV surveillance is essential for monitoring a novel SIV emergence potentially for zoonosis.


Applicative Value of Serum CA19-9, CEA, CA125 and CA242 in Diagnosis and Prognosis for Patients with Pancreatic Cancer Treated by Concurrent Chemoradiotherapy.
To evaluate the application value of serum CA19-9, CEA, CA125 and CA242 in diagnosis and prognosis of pancreatic cancer cases treated with concurrent chemotherapy. 52 patients with pancreatic cancer, 40 with benign pancreatic diseases and 40 healthy people were selected. The electrochemiluminescence immunoassay method was used for detecting levels of CA19-9, CEA and CA125, and a CanAg CA242 enzyme linked immunoassay kit for assessing the level of CA242. The Kaplan-Meier method was used for analyzing the prognostic factors of patients with pancreatic cancer. The Cox proportional hazard model was applied for analyzing the hazard ratio (HR) and 95% confidential interval (CI) for survival time of patients with pancreatic cancer. The levels of serum CA19-9, CEA, CA125 and CA242 in patients with pancreatic cancer were significantly higher than those in patients with benign pancreatic diseases and healthy people (P<0.001). The sensitivity of CA19-9 was the highest among these, followed by CA242, CA125 and CEA. The specificity of CA242 is the highest, followed by CA125, CEA and CA19-9. The sensitivity and specificity of joint detection of serum CA19-9, CEA, CA125and CA242 were 90.4% and 93.8%, obviously higher than single detection of those markers in diagnosis of pancreatic cancer. The median survival time of 52 patients with pancreatic cancer was 10 months (95% CI7.389~12.611).. Patients with the increasing level of serum CA19-9, CEA, CA125, CA242 had shorter survival times (P=0.047. 0.043, 0.0041, 0.029). COX regression analysis showed that CA19-9 was an independent prognostic factor for patients with pancreatic cancer (P=0.001, 95%CI 2.591~38.243). The detection of serum tumor markers (CA19.9, CEA, CA125 and CA242) is conducive to the early diagnosis of pancreatic cancer and joint detection of tumor markers helps improve the diagnostic efficiency. Moreover, CA19-9 is an independent prognostic factor for patients with pancreatic cancer.


Changes in patterns of mental illness over different hospital admissions.
Different diagnostic groups were compared in order to find out how constant individual patterns of illness remained over different hospital admissions. First, each of the syndrome scales in the Inpatient Multidimensional Psychiatric Scale and the Clinical Self-Rating Scales was examined to see how stable the patients' scores remained over two different admissions. Low correlations between scores on the two admissions were seen in the scales for mania and depression, reflecting changes in the clinical symptom patterns of patients with affective disorders. Many other scales, however, were fairly stable and were more closely connected with the given diagnosis. Secondly, each patient's pattern of illness on the first admission was compared with his or her pattern on the second admission. It was observed that patients with diagnoses of schizophrenia or schizo-affective psychosis tended to have less stable patterns of illness.


Executive functions, Personality traits and ADHD symptoms in adolescents: A mediation analysis.
Certain personality traits and cognitive domains of executive functions (EF) are differentially related to attention deficit hyperactivity disorder (ADHD) symptoms in adolescents. This study aimed to analyze the five-factor model (FFM) personality characteristics in adolescents with ADHD, and to examine whether EF mediate the relationships between FFM personality traits and ADHD symptoms. A comprehensive diagnostic assessment, including ADHD clinical interviews, ADHD rating scales, neuropsychological EF testing (i.e., working memory, flexibility and inhibition) and a personality assessment was carried out in a sample of 118 adolescents (75 ADHD and 43 control participants, 68% males), aged 12 to 16 years, and their parents and teachers. Adolescents with ADHD had lower scores than control participants on Conscientiousness and Agreeableness, and higher scores on Neuroticism. Structural equation models (SEM) showed that Conscientiousness directly influenced inattentive and hyperactive-impulsive symptoms, while Neuroticism, Agreeableness, and Extraversion directly affected hyperactive-impulsive symptoms. Only Conscientiousness exerted indirect effects on inattention, but not on hyperactivity-impulsivity symptoms, via EF; higher scores on Conscientiousness were related to higher scores on EF, which in turn were related to lower scores on inattentive symptoms. These findings corroborate the relationships between ADHD symptoms, FFM personality traits and EF and indicate the mediating effect of EF on the relationship between Conscientiousness and inattention.


Cell fractionation of anterior pituitary glands from beef and pig.
Fresh anterior pituitary glands from beef and pig were separated by differential centrifugation into subcellular fractions. Nuclei and debris were obtained at 700 g for 15 minutes, secretory granules at 7000 g for 20 minutes, mitochondria at 34,000 g for 15 minutes, and microsomes at 78,000 g for 3 hours. Electron micrographs were taken of the individual fractions. Each fraction was analyzed for nitrogen, pentosenucleic acid (PNA), and phospholipide. Beef and pig anterior lobes were quite similar in their intracellular composition as seen in the subcellular fractions. Succinic dehydrogenase was localized in mitochondria, while alkaline phosphatase was concentrated in the microsomes. A proteinase with pH optimum at 8.2 was exclusively localized. in microsomal and supernatant fractions. Acid phosphatase, acid ribonuclease, and acid proteinase were distributed among the subcellular fractions in another pattern, indicating the presence of a particle type distinct from mitochondria and microsomes. The distribution of cytoplasmic PNA paralleled that of alkaline phosphatase.


Preliminary evaluation of a series of amphiphilic timolol prodrugs: possible evidence for transscleral absorption.
A series of amphiphilic esters of timolol malonate (octanoyl, decanoyl, dodecanoyl, myristoyl and palmitoyl timolol) were tested in rabbits for their capacity to antagonise the isoproterenol-induced ocular hypotension, using timolol maleate as reference standard. The most active prodrug, palmitoyl timolol malonate (PTM) was also evaluated for its capacity: (a) to decrease IOP in a model of bethamethasone-induced ocular hypertension, and (b) to permeate "in vitro" through rabbit corneal tissues. PTM, the prodrug with the longest aliphatic chain and therefore the greatest amphiphilic/lipophilic character, showed "in vivo" significant activity differences with respect to timolol maleate: the beta-antagonism was more important at earlier and later experimental times, and the IOP decrease was more marked at longer times. The prodrug, however, showed "in vitro" an inferior corneal permeability when compared with timolol maleate. The significant differences observed for the beta-antagonism of PTM at earlier times of the test might be attributed to transscleral absorption, due to the physicochemical characteristics of the prodrug, while the prolonged action (also observed in the IOP-depression test) might be due to sustained release, resulting from accumulation of the prodrug in the corneal epithelium. The present preliminary results are indicative of the potentiality of amphiphilic properties in a prodrug molecule.


Long-term effects of nutrient intervention on markers of bone remodeling and calciotropic hormones in late-postmenopausal women.
Adequate intakes of calcium and vitamin D reduce bone loss and fracture risk in the elderly. Other nutrients also affect bone health, and adequate intakes may influence bone turnover and balance. We compared the long-term effects on bone turnover markers and calciotropic hormones of a multinutrient supplement, a calcium and vitamin D supplement, and dietary instruction aimed at increasing calcium intake through foods. Ninety-nine healthy postmenopausal women participated in a 3-y, randomized trial, receiving either 1) supplemental calcium (1450 mg/d) and vitamin D [10 microg (400 IU)/d], 2) calcium, vitamin D, and other nutrients (multinutrient supplement), or 3) dietary instruction (dietary control group). Data are from 83 subjects who completed the trial. Increases over baseline in calcium intakes and serum 25-hydroxyvitamin D concentrations were sustained over 3 y in all treatment groups. Circulating parathyroid hormone concentrations were reduced at year 1 in all treatment groups but trended toward baseline thereafter. Bone turnover markers followed a similar pattern, and none of the changes in biochemical concentrations differed significantly between groups. All 3 interventions offer long-term feasibility for increasing calcium intake and serum 25-hydroxyvitamin D concentrations. The dietary addition of micronutrients implicated in skeletal physiology confers no obvious bone-sparing effect in healthy postmenopausal women beyond that of calcium and vitamin D alone. The attenuation over time in suppression of parathyroid hormone and bone turnover might help explain why nutrient intervention tends to have less of a bone-sparing effect than do skeletally active medications such as estrogen or bisphosphonates.


Biomarkers, erectile dysfunction, and cardiovascular risk prediction: the latest of an evolving concept.
A number of circulating and imaging biomarkers are robustly associated with cardiovascular (CV) risk. The overall expectation from a biomarker in the erectile dysfunction (ED) setting is to enhance the optimal management of a man with this disorder but no clinical atherosclerosis. Evidence demonstrating that these biomarkers enhance risk prediction for individuals with ED is at this stage still limited for most of them. A better identification of the subsets of the ED population that require further risk stratification, as well as the initiation of randomized trials that will formally test the ability of biomarkers to predict CV risk, could make biomarker-guided prevention an attainable goal.


Acute scrotum in setting of acute pancreatitis.
Acute scrotum is a urologic emergency with many aetiologies. Acute scrotum in setting of acute pancreatitis is a rare occurrence and results from an effusion of pancreatic juices into the inguinal canal along a retroperitoneal tract. Knowledge regarding the existence of this obscure condition is essential for its diagnosis. It is thus important for medical professionals, particularly internists, surgical trainees and emergency physicians, to be aware of the condition and the options for its management.