Encapsulation of AGE-Breaker Alagebrium by Cucurbit[7]uril Improved the Stability of Both Its Carbonyl α-Hydrogen and Thiazolium C2-Hydrogen.
As determined by both 1 H NMR and UV/Vis spectroscopic titration, ESI-MS, isothermal titration calorimetry, and DFT molecular modeling, advanced glycation end products (AGE) breaker alagebrium (ALA) formed 1:1 guest-host inclusion complexes with cucurbit[7]uril (CB[7]), with a binding affinity, Ka , in the order of magnitude of 105 m-1 , thermodynamically driven by both enthalpy (ΔH=-6.79 kcal mol-1 ) and entropy (TΔS=1.21 kcal mol-1 ). For the first time, a dramatic inhibition of keto-enol tautomerism of the carbonyl α-hydrogen of ALA has been observed, as evidenced by over an order of magnitude decrease of both the first step rate constant, k1 , and the second step rate constant, k2 , during hydrogen/deuterium exchange in D2 O. Meanwhile, as expected, the reactivity of C2-hydrogen was also inhibited significantly, with an upshift of 2.09 pKa units. This discovery will not only provide an emerging host molecule to modulate keto-enol tautomerism, but also potentially lead to a novel supramolecular formulation of AGE-breaker ALA for improved stability and therapeutic efficacy.


Apolipoprotein A-IV: A Multifunctional Protein Involved in Protection against Atherosclerosis and Diabetes.
Apolipoprotein A-IV (apoA-IV) is a lipid-binding protein, which is primarily synthesized in the small intestine, packaged into chylomicrons, and secreted into intestinal lymph during fat absorption. In the circulation, apoA-IV is present on chylomicron remnants, high-density lipoproteins, and also in lipid-free form. ApoA-IV is involved in a myriad of physiological processes such as lipid absorption and metabolism, anti-atherosclerosis, platelet aggregation and thrombosis, glucose homeostasis, and food intake. ApoA-IV deficiency is associated with atherosclerosis and diabetes, which renders it as a potential therapeutic target for treatment of these diseases. While much has been learned about the physiological functions of apoA-IV using rodent models, the action of apoA-IV at the cellular and molecular levels is less understood, let alone apoA-IV-interacting partners. In this review, we will summarize the findings on the molecular function of apoA-IV and apoA-IV-interacting proteins. The information will shed light on the discovery of apoA-IV receptors and the understanding of the molecular mechanism underlying its mode of action.


T1/ST2 signaling establishes it as a member of an expanding interleukin-1 receptor family.
Through data base searches, we have discovered new proteins that share homology with the signaling domain of the type I interleukin-1 receptor (IL-1RI): human "randomly sequenced cDNA 786" (rsc786), murine MyD88, and two partial Drosophila open reading frames, MstProx and STSDm2245. Comparisons between these new proteins and known IL-1RI homologous proteins such as Toll, 18-Wheeler, and T1/ST2 revealed six clusters of amino acid similarity. We tested the hypothesis that sequence similarity between the signaling domain of IL-1RI and the three mammalian family members might indicate functional similarity. Chimeric IL-1RI receptors expressing the putative signaling domains of T1/ST2, MyD88, and rsc786 were assayed by three separate IL-1 responsive assays, NF-kappaB, phosphorylation of an epidermal growth factor receptor peptide, and an interleukin 8 promoter-controlled reporter construct, for their ability to transduce an IL-1-stimulated signal. All three assays were positive in response to the T1/ST2 chimera, while the MyD88 and rsc786 chimeras failed to respond. These data indicate that the sequence homology between IL-1RI and T1/ST2 indicates a functional homology as well.


Glycine betaine fluxes in Lactobacillus plantarum during osmostasis and hyper- and hypo-osmotic shock.
Bacteria respond to changes in medium osmolarity by varying the concentrations of specific solutes in order to maintain constant turgor. The primary response of Lactobacillus plantarum to an osmotic upshock involves the accumulation of compatible solutes such as glycine betaine, proline, and glutamate. We have studied the osmotic regulation of glycine betaine transport in L. plantarum by measuring the overall and unidirectional rates of glycine betaine uptake and exit at osmostasis, and under conditions of osmotic upshock and downshock. At steady state conditions, a basal flux of glycine betaine (but no net uptake or efflux) is observed that amounts to about 20% of the rate of "activated"' uptake (uptake at high osmolarity). No direct exchange of 14C-labeled glycine betaine in the medium for unlabeled glycine betaine in the cytoplasm was observed in glucose metabolizing and resting cells, indicating that a separate glycine betaine efflux system is responsible for the exit of glycine betaine. Upon osmotic upshock, the uptake system for glycine betaine is rapidly activated (within seconds), whereas the basal efflux is inhibited. These two responses account for a rapid accumulation of glycine betaine until osmostasis is reached. Upon osmotic downshock, glycine betaine is rapidly released by the cells in a process that has two kinetic components, i.e. one with a half-life of less than 2 s which is unaffected by the metabolic status of the cells, the other with a half-life of 4-5 min in glucose-metabolizing cells which is dependent on internal pH or a related parameter. We speculate that the former activity corresponds to a stretch-activated channel, whereas the latter may be facilitated by a carrier protein. Glycine betaine uptake is strongly inhibited immediately after an osmotic downshock, but slowly recovers in time. These studies demonstrate that in L. plantarum osmostasis is maintained through positive and negative regulation of both glycine betaine uptake and efflux, of which activation of uptake upon osmotic upshock and activation of a "channel-like" activity upon osmotic downshock are quantitatively most important.


Roles of histidine-194, aspartate-163, and a glycine-rich sequence of NAD(P)H:quinone oxidoreductase in the interaction with nicotinamide coenzymes.
NAD(P)H:(quinone-acceptor) oxidoreductase (NQOR, EC 220.127.116.11), an enzyme catalyzing the obligatory two electron reduction of quinones, can utilize both NADH and NADPH as electron donors at similar efficiencies. Based on site-directed mutagenesis studies, we previously suggested that the glycine-rich region of rat liver NQOR is important for the binding of NAD(P)H (Ma et al., J. Biol. Chem. 267, 22298-22304, 1992). However, the mode of interactions between the active site and NADH or NADPH is not clearly known. In this study, we conducted site-directed mutagenesis experiments and identified H194 and D163 of NQOR as key residues affecting the Km of NADPH. Steady-state kinetic analysis for the reduction of dichloroindophenol (DCIP) showed that Km(NADPH) values of purified mutant proteins H194D, H194A, and D163V were 288-, 14-, and 96-fold higher, respectively, than that of NQOR; but the Km(NADH) values were only slightly higher. The kcat(NADPH) values were almost the same as that of NQOR in the reduction of DCIP at the respective pH optima which were affected by the mutations. The kcat(NADH) values of these mutant enzymes were 30 to 60% that of NQOR. In the reduction of menadione, the mutations also caused much larger increases in km(NADPH) than Km(NADH). The results suggest that H194 and D163 are important for the interaction with the 2'-phosphate group of NADPH. NAD(P)H analogues, N-methyldihydronicotinamide and dihydronicotinamide mononucleotide, can also serve as electron donors for NQOR, but the Km values were 4.5- and 495-fold higher, respectively, than that with NADH. Mutations at H194 and D163 and at the glycine-rich region of NQOR, which increased Km(NADH) and Km(NADPH), did not substantially affect the Km values of these two analogues. This result is consistent with the suggested roles of these amino acid residues in the interaction with nicotinamide coenzymes. Based on these results, a model of the NAD(P)H binding site is proposed showing the interaction of the pyrophosphate group with the glycine-rich region and the interaction of 2'-phosphate group with H194 and D163.


New training model using chickens intestine for pediatric intestinal anastomosis.
To develop a new low-cost, easy-to-make and available training model using chickens' intestine for infant intestinal anastomosis. Segments of chicken intestine were used to create an intestinal anastomosis simulator. We tried to perform an end-to-end, end-to-side and side-to-side anastomosis. Handsewn sutured anastomosis were performed in single layered with interrupted prolene 5-0 suture. The parameters analyzed were cost, intestine's diameter and length, anastomosis patency and flow-through and leakage amount. In all cases it was possible to make the anastomosis in double layered without difficulties, different from the usual ones. There was a positive patency at all anastomoses after the end of the procedure, with no need for reinterventions. The new training model using chickens' intestine for infant intestinal anastomosis is low-cost, easy-to-make and easy available.


A neutravidin-based assay for reverse transcriptase suitable for high throughput screening of retroviral activity.
A non-isotopic neutravidin-based reverse transcriptase (RT) assay adapted for high throughput screening of HIV activity is described. Using a 96-well microtitre plate, HIV particles are lysed and the RT enzyme released into a reaction mixture containing poly(A) RNA, biotinylated oligo d(T) and fluorescein-labelled dUTP (FI-dUTP). With poly(A) as a template and oligo d(T) as primer, the viron RT incorporates FI-dUTP into an elongating DNA strand. The resulting product is captured on a neutravidin-coated 96-well plate and the unincorporated nucleotides removed by a series of washing steps. A simple ELISA is subsequently performed using a monoclonal antifluorescein antibody conjugated to alkaline phosphatase. Quantification of RT activity is facilitated by a colorimetric readout. The assay was validated in the context of a diagnostic HIV-1 phenotyping assay. Using supernatants from HIV-1 infected lymphocyte cultures the assay was shown to be as sensitive as a radioactive assay and the RT activity correlated well with levels of cell-associated HIVp24. Importantly, even minor reductions of RT activity by virus variants with reduced fitness could be distinguished.


Which visual pathways cause fixation-related inhibition?
Visual stimuli can both inhibit and activate motor mechanisms. In one well-known example, the latency of saccadic eye movements is prolonged in the presence of a fixation stimulus, relative to the case in which the fixation stimulus disappears before the target appears. This automatic sensory-motor effect, known as the gap effect or fixation-offset effect, has been associated with inhibitory connections within the superior colliculus (SC). Visual information is provided to the SC and other oculomotor areas, such as the frontal eye fields (FEF), mainly by the magnocellular geniculostriate pathway, and also by the retinotectal pathway. We tested whether signals in these pathways are necessary to create fixation-related inhibition, by using stimuli invisible to them. We found that such stimuli, visible only to short-wave-sensitive cones (S cones), do produce fixation-related inhibition (including when warning effects were equated). We also demonstrate that this fixation-related inhibition cannot be explained by residual activation of luminance pathways and must be caused by a route separate from that of luminance fixation signals. Thus there are at least two routes that cause fixation-related inhibition, and direct sensory input to the SC or FEF by the magnocellular or retinotectal pathways is not required. We discuss the implications that there may be both cortical and collicular mechanisms.


Identification of autophosphorylation sites in eukaryotic elongation factor-2 kinase.
eEF2K [eEF2 (eukaryotic elongation factor 2) kinase] phosphorylates and inactivates the translation elongation factor eEF2. eEF2K is not a member of the main eukaryotic protein kinase superfamily, but instead belongs to a small group of so-called α-kinases. The activity of eEF2K is normally dependent upon Ca(2+) and calmodulin. eEF2K has previously been shown to undergo autophosphorylation, the stoichiometry of which suggested the existence of multiple sites. In the present study we have identified several autophosphorylation sites, including Thr(348), Thr(353), Ser(366) and Ser(445), all of which are highly conserved among vertebrate eEF2Ks. We also identified a number of other sites, including Ser(78), a known site of phosphorylation, and others, some of which are less well conserved. None of the sites lies in the catalytic domain, but three affect eEF2K activity. Mutation of Ser(78), Thr(348) and Ser(366) to a non-phosphorylatable alanine residue decreased eEF2K activity. Phosphorylation of Thr(348) was detected by immunoblotting after transfecting wild-type eEF2K into HEK (human embryonic kidney)-293 cells, but not after transfection with a kinase-inactive construct, confirming that this is indeed a site of autophosphorylation. Thr(348) appears to be constitutively autophosphorylated in vitro. Interestingly, other recent data suggest that the corresponding residue in other α-kinases is also autophosphorylated and contributes to the activation of these enzymes [Crawley, Gharaei, Ye, Yang, Raveh, London, Schueler-Furman, Jia and Cote (2011) J. Biol. Chem. 286, 2607-2616]. Ser(366) phosphorylation was also detected in intact cells, but was still observed in the kinase-inactive construct, demonstrating that this site is phosphorylated not only autocatalytically but also in trans by other kinases.


[Limb perfusion in soft tissue sarcomas: twenty years of experience].
To evaluate the long-term results of 20 years of experience with isolated limb perfusion (ILP) with tumour necrosis factor α (TNF-α) and melphalan, followed by surgical resection and adjuvant radiotherapy, for the treatment of advanced soft tissue sarcomas of the extremities. Retrospective cohort study. From 1991 to 2011, 113 patients with primary irresectable soft tissue sarcomas underwent 117 ILPs at the University Medical Centre Groningen. 96 ILPs (82%) were performed in the lower limb, and 21 (18%) in the upper limb. The dosages used were 1-4 mg TNF-α and 10-13 mg/l melphalan. After a median follow up of 8 (range 2-15) weeks after ILP, 107 tumours were resected: 81 (76%) of the resection margins were tumor-free. After the resection, 69 patients (61%) received adjuvant radiotherapy. In total, 85 ILPs resulted in a tumoural response; 16 patients (14%) developed a local recurrence and after 46 treatments (39%), distant metastases had developed. After a median follow-up of 51 months, the limb had been spared in 88 patients (78%). The 10- year disease-specific survival was 53.8%. There was a median follow-up period of 76 months (range: 7-234); still alive at the end of this period were 56 patients (50%). A total of 83 perfusion- or resection-related complications occurred from 58 ILPs (50%): 55 (66%) early and 28 (34%) late treatment-related complications. None of the patients died as a result of the treatment. ILP is a safe and effective procedure in the treatment of advanced primary irresectable soft tissue sarcoma that can prevent amputation in many cases. It is however associated with significant morbidity and is burdensome for the patient.


[Synthesis of 2-{3-[4-(4-fluorophenyl)-1-piperazinyl]-2-hydroxy-propoxy}-phenylcarbamic acid alkylesters and in vitro evaluation of their beta-antiadrenergic and vasodilatative activities].
Synthesis of 2-{3-[4-(4-fluorophenyl)-1-piperazinyl]-2-hydroxy-propoxy}-phenylcarbamic acid alkylesters and in vitro evaluation of their beta-antiadrenergic and vasodilatative activities In effort to obtain effective compounds able to favourably influence pathologically changed cardiovascular functions, such as hypertension and ischemic cardiac disease, a new series of aryloxyaminopropanols were synthesized. Four of the compounds, which differ in the alkyl substitution of phenylcarbamate (methyl, ethyl, propyl, butyl), were chosen for basic in vitro pharmacological analyses. In experiments on the isolated spontaneously beating guinea pig atria all compounds at conc. of 1.0.10(-6) mol.l(-1) decreased the basic heart rate (7.6-13.6%) and inhibited the positive chronotropic effect of isoprenaline (pA2 = 6.28-6.81). The compounds manifest only a slight relaxation effect on KCl pre-contracted aortal strips of rats (not until conc. of 1.0.10(-5) mol.l(-1)). The compounds with propyl and butyl substitution appear more effective than the methyl and ethyl derivatives.


Long-term outcome of trabeculectomy in terms of intraocular pressure.
To evaluate the outcome, long-term results and contributing prognostic factors of trabeculectomy in terms of intraocular pressure (IOP) and to compare the results in primary open-angle glaucoma (POAG) and exfoliation glaucoma (EG). This study involved retrospective evaluation of 138 consecutive patients (138 eyes) with either POAG or EG. All patients had undergone primary trabeculectomy performed by the same surgeon between November 1994 and August 1996. Only one eye per patient was included. Operations performed with the use of antimetabolites were excluded. All patients were aged over 40 years and were white. Follow-up for all subjects lasted at least 2 years. Successful control of IOP was defined as achieving IOP < or = 21 mmHg without medication (complete success) or with a single topical medication (qualified success). Success rates were determined using the Kaplan-Meier survival curve, and risk factors were analysed with proportional hazards regression. According to the Kaplan-Meier survival curve, success rates (complete or qualified) were 82% at 1 year, 70% at 2 years, 64% at 3 years and 52% at 4 years. A total of 63% were complete successes at 1 year, 54% at 2 years, 45% at 3 years and 40% at 4 years. Complete success rates were significantly better in the POAG group than in the EG group. Proportional hazards regression analysis showed that the presence of EG and early postoperative IOP > 30 mmHg decreased the possibility of complete success, while a cataract operation performed during follow-up improved it. The IOP-reducing effect of trabeculectomy decreases gradually. After 4 years, 52% of operated eyes had IOP < or = 21 mmHg with or without a single topical medication. Diagnosis of EG implied a worse long-term outcome for trabeculectomy in terms of IOP.


A staining protocol for identifying secondary compounds in Myrtaceae.
Here we propose a staining protocol using toluidine blue (TBO) and ruthenium red to reliably identify secondary compounds in the leaves of some species of Myrtaceae. • Leaves of 10 species representing 10 different genera of Myrtaceae were processed and stained using five different combinations of ruthenium red and TBO. Optimal staining conditions were determined as 1 min of ruthenium red (0.05% aqueous) and 45 s of TBO (0.1% aqueous). Secondary compounds clearly identified under this treatment include mucilage in the mesophyll, polyphenols in the cuticle, lignin in fibers and xylem, tannins and carboxylated polysaccharides in the epidermis, and pectic substances in the primary cell walls. • Potential applications of this protocol include systematic, phytochemical, and ecological investigations in Myrtaceae. It might be applicable to other plant families rich in secondary compounds and could be used as a preliminary screening method for extraction of these elements.


Effects of Intensive Alfalfa Feeding on Meat Quality and Fatty Acid Profile of Korean Native Black Goats.
The aim of this study was to determine meat quality characteristics and fatty acid composition of Korean native black goats (KNBG) finished on intensive feeding of alfalfa (ALF) and conventional feeding of commercial concentrate pellets (CCP) with low-energy common grasses. Ten KNBG (12 months old) were divided into two groups and subjected to either ALF or CCP treatments. The goats were slaughtered after 6 months of feeding with experimental diets to investigate meat quality characteristics and fatty acid compositions of longissimus lumborum muscle. There were no significant differences in proximate chemical composition, collagen, or myoglobin content between ALF and CCP groups of goats. Meat color, water-holding capacity, or tenderness was not significantly different between the two groups either. However, proportions of monounsaturated fatty acids and polyunsaturated fatty acids were significantly (p<0.05) different between the two groups. The proportion of oleic acid was significantly (p<0.05) higher in ALF goat whereas proportion of linoleic and arachidonic acids were significantly (p<0.05) higher in CCP goat. Results suggest that KNBG finished with intensive feeding of alfalfa could produce goat meat with desirable fatty acids for human diets.


Current provision of myelopathy education in medical schools in the UK: protocol for a national medical student survey.
Degenerative cervical myelopathy (DCM) is a common, disabling and progressive neurological condition triggered by chronic compression of the cervical spinal cord by surrounding degenerative changes. Early diagnosis and specialist management are essential to reduce disability, yet time to diagnosis is typically prolonged. Lack of sufficient representation of DCM in undergraduate and postgraduate medical curricula may contribute to the poor recognition of DCM by non-specialist doctors in clinical practice.In this study, our objective, therefore, is to assess DCM teaching provision in medical schools throughout the UK and to assess the impact of teaching on the DCM knowledge of UK medical students. A 19-item questionnaire capturing data on medical student demographics, myelopathy teaching and myelopathy knowledge was designed. Ethical approval was granted by the Psychology Research Ethics Committee, University of Cambridge. An online survey was hosted on Myelopathy.org, an international myelopathy charity. Students studying at a UK medical school are eligible for inclusion. The survey is advertised nationally through university social media pages, university email bulletins and the national student network of Myelopathy.org. Advertisements are scheduled monthly over a 12-month recruitment period. Participation is incentivised by entering consenting participants of completed surveys to an Amazon voucher prize draw. Responses are anonymised using participant-chosen unique identifier codes. A participant information sheet followed by an explicit survey question captures participant informed consent. Regular updates on the progress of the study will be published on Myelopathy.org. Ethical approval for the study was granted by the Psychology Research Ethics Committee, University of Cambridge (PRE.2018.099). The findings of the study described in this protocol, and all other related work, will be submitted for publication in a peer-reviewed journal and will be presented at scientific conferences.


Rapid integrated rheo-optical and polarized Fourier-transform infrared spectrometry measurement system for polymer films undergoing chemo-mechanical changes.
The design and performance of a multisensory instrument to track physical and chemical changes of thin polymer films (typically 5 μm < thickness < 100 μm) subjected to thermal and mechanical treatments are described in this paper. For the first time, real-time measurements of spectral birefringence, true stress, true strain, and temperature are integrated together with ultra-rapid-scan polarized FT-IR spectrometer (URS-FT-IR) to investigate the relationships between true mechanical measures and structural features at different length scales. The rheo-optical properties (birefringence-true stress-true strain) are collected at a rate of 10 data points∕s and URS-FT-IR data are collected at a rate of 300 complete spectra∕s. The IR dichroism measurement is performed by exposing the sample to non-polarized IR beam in transmission mode with two mutually perpendicular polarizations, parallel and perpendicular to the stretching direction, received by detector unit. This design allows to analyze both polarizations simultaneously wavenumbers in the range of 500 cm(-1)-4000 cm(-1). Controlled processing parameters include air speed, air temperature, stretching rate, stretching ratio, stretch cycling, and holding times; while simultaneously measuring optical retardation, sample width, temperature, load cell, and both parallel and perpendicular IR spectra. Calibration and performance of this instrument is demonstrated with several film samples. These are: A polystyrene standard, an atactic polystyrene (homo-polymer), a polyurethane (consists of hard and soft segments) for physical changes during uniaxial deformation, and a polyamic acid during imidization reaction. This measurement system is particularly useful in unraveling molecular level details of complex physical and chemical events that take place during very fast deformation schemes (uniaxial stretching, retraction, relaxation, annealing, etc.) relevant to industrial processes. These include specific orientation behavior of each phase, block or filler, crystallization, relaxation and orientation state. It is also suited to track reaction rates and products in polymers undergoing thermal or photo curing.


Coverage-dependent morphology of PEGylated lysozyme layers adsorbed on silica.
Neutron reflection was used to characterize the adsorbed layer structure for lysozyme conjugated at the N-terminus with a single perdeuterated methoxy poly(ethylene glycol) (PEG) chain at the silica/water interface. Adsorbed layers were produced with two different surface concentrations corresponding to opposite sides of a pronounced transition in the adsorption isotherm for mono-PEGylated lysozyme. The transition was previously ascribed, on the basis of less direct characterization by normal force measurements, to a change in the distribution of the conjugated PEG chain segments in the interfacial region in response to lateral repulsions (S.M. Daly, T.M. Przybycien, R.D. Tilton, Langmuir 21 (2005) 1328-1337). Neutron reflectivity was measured for both surface concentrations using three different sets of neutron scattering length density contrast conditions, and models for the distribution of PEG and lysozyme content in the layers were obtained by simultaneous regression of all contrast condition data sets. This analysis indicated that the surface proximal volume is occupied almost equally by PEG and lysozyme at the low surface concentration, while at the higher surface concentration PEG is preferentially shifted away from the surface, with all lysozyme remaining in the surface proximal region. The distal regions of the high surface concentration layer contain only PEG, consistent with the previous interpretation of normal force measurements.


Dual regulation of protein maturation in viral infected rat HTC hepatoma cells by glucocorticoids and progesterone.
Dexamethasone, a synthetic glucocorticoid, is required for full posttranslational maturation of mouse mammary tumor virus (MMTV) phosphoproteins and glycoproteins in M1.54 cells, a viral infected rat hepatoma (HTC) cell line. Pulse-chase radiolabeling with [35S]methionine revealed that steroids with known glucocorticoid activity (such as dexamethasone and hydrocortisone) regulated the maturation of both MMTV polyproteins in a manner proportional to their occupancy for glucocorticoid receptors and their biological potency. In contrast, progesterone selectively induced the proteolytic processing of MMTV phosphoproteins but simultaneously antagonized the dexamethasone-regulated maturation of MMTV glycoproteins and all other tested glucocorticoid responses. Exposure to suboptimal concentrations of both progesterone and dexamethasone fully stimulated the processing of MMTV phosphoproteins, suggesting that steroid receptors occupied with combinations of either steroid functionally interact at the putative maturation gene. Moreover, treatment with either actinomycin D, a potent inhibitor of de novo RNA synthesis, or RU38486, a synthetic antagonist of glucocorticoid and progesterone action, prevented both the dexamethasone and progesterone-regulated induction of MMTV phosphoprotein maturation. Sedimentation velocity and saturation binding analysis revealed that the sizes and concentrations of hepatoma cell progesterone and dexamethasone binding activities are similar while specific binding of the active progestin R5020 was not detected in either M1.54 cells or the glucocorticoid receptor deficient HTC cell line MSN6.10.2. Taken together, our results demonstrate that two distinct classes of steroid hormones can uniquely alter the posttranslational maturation of a specific subset of phosphoprotein substrates by a common glucocorticoid receptor-dependent process.


Recent Advances in Conservation-Dissipation Formalism for Irreversible Processes.
The main purpose of this review is to summarize the recent advances of the Conservation-Dissipation Formalism (CDF), a new way for constructing both thermodynamically compatible and mathematically stable and well-posed models for irreversible processes. The contents include but are not restricted to the CDF's physical motivations, mathematical foundations, formulations of several classical models in mathematical physics from master equations and Fokker-Planck equations to Boltzmann equations and quasi-linear Maxwell equations, as well as novel applications in the fields of non-Fourier heat conduction, non-Newtonian viscoelastic fluids, wave propagation/transportation in geophysics and neural science, soft matter physics, etc. Connections with other popular theories in the field of non-equilibrium thermodynamics are examined too.


Patients' responses to incidentally discovered silent brain infarcts - a qualitative study.
Incidentally discovered silent brain infarcts (id-SBIs) are an understudied condition with probable clinical significance, but it is not known how patients respond to or prioritize this condition. We sought to assess reporting of id-SBIs and how patients approach their diagnosis. Patients with id-SBIs were identified from sequential scans between 12/2015-5/2016, were referred by treating clinicians, or self-referred for the study. This study used qualitative semi-structured interviews. Purposeful sampling was used to achieve diversity in acuity, setting, and recruitment strategy. Interviews were audio-recorded and transcribed. A constant comparative method was used to develop a coding schema, find consensus, and iteratively explore emergent themes until thematic saturation was achieved. Only 10 of 102 patients prospectively identified by neuroimaging were informed of the imaging findings. Twelve participants in total were interviewed. Among the study participants, the primary themes were cognitive, emotional, and behavioral responses to diagnostic, prognostic, and therapeutic uncertainty regarding id-SBIs. Clinicians described id-SBIs to participants as an ambiguous condition. Participants feared potential consequences of id-SBIs, including symptomatic stroke, dementia, and disability. Participants attempted to reduce uncertainty with strategies including equating id-SBIs with symptomatic stroke, self-education about stroke, and seeking second opinions. Participants considered id-SBIs to be a serious medical condition. Ambiguous counseling by clinicians on id-SBIs provoked or failed to attenuate fear, leading to participants adopting strategies aimed at reducing uncertainty.


Characterization of the integrity of three-dimensional trabecular bone microstructure by connectivity and shape analysis using high-resolution magnetic resonance imaging in vivo.
Bone mineral density and bone structure are the main determinants of bone strength in osteoporosis. In this study we used high-resolution magnetic resonance imaging to visualize the bone microstructure in the finger phalanges in vivo and to assess the topological three-dimensional connectivity of the trabecular network and the shape of the trabeculae as measures of bone quality. We visualized the phalanges of young and elderly healthy volunteers in vivo with a spatial resolution of 152 microm x 152 microm x 280 microm. Image processing software to quantify three measures of connectedness was developed and tested: connectivity, global connectivity density, and local connectivity density. Global three-dimensional connectivity ranged from 904 to 1,607 connections. Global connectivity density ranged from 2.9 to 4.7 connections per mm with large intersubject differences. We found a decrease of local connectivity density with growing distance from the joint ranging from 5.1 to 0.2 connections per mm. These preliminary results represent a quantitative description of the well-known rarefication of the trabecular network when moving from epiphysis to the diaphysis. Three-dimensional visualization showed a dense network consisting mostly of rod-like trabeculae at the epiphysis changing to a less dense network of a few plate-like structures near the medullary canal. An algorithm for the quantitative classification of trabecular architecture with regard to plate or rod-like shape was tested for feasibility. We conclude that in vivo assessment of three-dimensional properties of the trabecular network is possible in human phalanges. Determination of connectivity and shape will allow quantification of structural aspects of osteoporotic changes and may improve assessment of fracture risk.


Role of the bed nucleus of the stria terminalis (BST) in the expression of conditioned fear.
Male Sprague-Dawley rats were trained and tested in the fear-potentiated startle (FPS) paradigm to examine the involvement of the bed nucleus of the stria terminalis (BST) in the expression of conditioned fear. Studies were designed to (a) detect physiological changes in the BST that might correlate with different levels of FPS expression and (b) determine if chemical inactivation of the BST with muscimol (1 ng) had any effect on FPS expression. The data suggest that the BST plays a role in the expression of conditioned fear and that GABA-mediated inhibition at this level may influence the level of this expression.


In situ controlled synthesis of thermosensitive poly(N-isopropylacrylamide)/Au nanocomposite hydrogels by gamma radiation for catalytic application.
Thermosensitive poly(N-isopropylacrylamide) (PNIPAM)/Au nanoparticle (NP) nanocomposite hydrogels are synthesized by in situ γ-radiation-assisted polymerization of N-isopropylacrylamide monomer aqueous solution in the presence of HAuCl₄·4H₂O. In this reaction, the PNIPAM hydrogels and the Au NPs are formed simultaneously, thus demonstrating an easy and straightforward synthetic strategy for the preparation of a uniform nanocomposite. The results suggest that increasing the monomer content during the preparation of nanocomposite materials can increase the sizes of Au NPs. The effects of irradiation dose and concentration of HAuCl₄·4H₂O on the optical and thermal properties of the hydrogel are also investigated. The PNIPAM/Au nanocomposite hydrogels act as an excellent catalyst for the conversion of o-nitroaniline to 1,2-benzenediamine, and the catalytic activity of the composite hydrogel can be tuned by the volume transition of PNIPAM. The in situ polymerization of monomer and reduction of metal ions initiated by a "clean" and "green" γ-radiation technique can be extended to the efficient synthesis of other nanocomposite materials.


Factors influencing recovery from headache after common whiplash.
To assess the relation between pretraumatic and trauma related headache in patients suffering from whiplash. Follow up study of patients examined a mean (SD) of 7.4 (4.2) days after trauma and again at three and six months. Patients referred from primary care. 117 patients (mean age 30.8 (9.5) years. Prevalence of trauma related headache and the predictive relation by multiple logistic regression between different somatic and psychological variables and trauma related headache at each examination. Prevalence of trauma related headache decreased from 57% to 27%. History of pretraumatic headache proved a significant risk factor for presenting with trauma related headache. A significant relation between trauma related headache and the following variables was found: at seven days the initial wellbeing score, early onset of neck pain, depression scale from the personality inventory, and the initial intensity of neck pain; at three months, intensity of neck pain, and history of pretraumatic headache; and at six months neck pain, pain intensity, and history of pretraumatic headache. History of pretraumatic headache significantly increases the likelihood of presenting with trauma related headache but only in combination with findings indicative of clinically important injury to the cervical spine.


[Colorectal neoplasms in patients with ulcerative rectocolitis. Their surgical treatment and follow-up].
Patients with ulcerative colitis (UC), especially those with a long-term standing or severe disease, show a higher incidence of colorectal neoplasms. Dysplasia represents the most important risk-factor for malignancy in patients with UC. From 1989 to 1999, 71 patients with UC, were surgically treated at our Institution. Indications for surgery were: non responding disease or refractory bleeding in 61; pathologic evidence of severe dysplasia during endoscopic surveillance in 5 patients, evidence of a neoplasm in the remaining 5 patients. Operative mortality was 1.4%. Infections of the surgical wound has been the most common postoperative complication. In 4 cases pouchitis has been reported. In 7 patients because of progression of the proctitis a second surgical treatment has been made. When surgical treatment is indicated in the course of ulcerative colitis, several operations may be performed. In the large majority of patients, total colectomy, mucosectomy and ileal pouch-anal anastomosis is the procedure of choice.


Early clinical and radiographic outcomes of anatomic total shoulder arthroplasty with a biconvex posterior augmented glenoid for patients with posterior glenoid erosion: minimum 2-year follow-up.
Glenoid bone loss in anatomic total shoulder arthroplasty (aTSA) remains a controversial and challenging clinical problem. Previous studies have shown high rates of glenoid loosening for aTSA in shoulders with retroversion, posterior bone loss, and posterior humeral head subluxation. This study is the first to present minimum 2-year follow-up data of an all-polyethylene, biconvex augmented anatomic glenoid component for correction of glenoid retroversion and posterior humeral head subluxation. This study is a multicenter, retrospective review of prospectively collected data on consecutive patients from 7 global clinical sites. All patients underwent aTSA using the biconvex posterior augmented glenoid (PAG). Inclusion criteria were preoperative computed tomographic (CT) scan, minimum 2 years since surgery, preoperative and minimum 2-year postoperative range of motion examination, and patient-reported outcome measures (PROMs). Glenoid classification, glenoid retroversion, and posterior humeral head subluxation were measured from preoperative CT and radiography and postoperative radiography. Statistical comparisons between pre- and postoperative values were performed with a paired t test. Eighty-six of 110 consecutive patients during the study period (78% follow-up) met the inclusion criteria and were included in our analysis. Mean follow-up was 35 ± 10 months, with a mean age of 68 ± 8 years (range 48-85). Range of motion statistically improved in all planes from pre- to postoperation. Mean visual analog scale score improved from 5.2 preoperation to 0.7 postoperation, Single Assessment Numeric Evaluation score from 43.2 to 89.5, Constant score from 41.8 to 76.9, and American Shoulder and Elbow Surgeons Standardized Shoulder Assessment Form score from 49.8 to 86.7 (all P < .0001). Mean glenoid retroversion improved from 19.3° to 7.4° (P < .0001). Posterior subluxation improved from 69.1% to 53.5% and posterior decentering improved from 5.8% to -3.0% (P < .0001). There was 1 patient with both a prosthetic joint infection and radiographic glenoid loosening that required revision. Seventy-nine of 86 patients had a Lazarus score of 0 (no radiolucency seen about peg or keel) at final follow-up. This study shows that at minimum 2-year follow-up, a posterior-augmented all-polyethylene glenoid can correct glenoid retroversion and posterior humeral head subluxation. Clinically, there was significant improvement in both range of motion and PROMs.


Functional magnetic resonance imaging-controlled neuronavigator-guided brain surgery: a case report.
The effectiveness of functional magnetic resonance imaging (f-MRI)-controlled and navigator-guided brain surgery for a patient with a recurrent astrocytoma is demonstrated. Preoperative f-MRI was performed in order to identify the motor area and ensure that the tumour was in the left prefrontal area. A more aggressive operation was planned for the recurrent tumour. The f-MRI data were input to the MKM navigation system and during the operation the contours of the tumour and motor area were visualised b y the microscope of the navigation system. The tumour and surrounding gliotic brain tissue were removed completely. The diagnosis was a grade III astrocytoma. The combination of the navigation system and f-MRI was useful for preoperative design of the surgical strategy, and tumour orientation during the operation, enabling aggressive surgery to be performed without functional deficits ensuing.


Assessment of anthropometric indexes of children and adolescents with Down syndrome.
To compare the agreement in the classification of the weight-for-age (W/A) and height-for-age (HA) indexes for children and adolescents with Down syndrome (DS) according to selected international reference distributions. A cross-sectional study was carried out in children (2 to 9.9 years old) and adolescents (10 to 17.9 years old) with DS from cities in the state of Rio de Janeiro, Brazil, in 2005. The W/A and H/A indexes were classified according to the percentiles of two curves developed for individuals with DS and one distribution developed for healthy subjects. The cut-off limits applied for categorization were: below the 5th percentile (< P5) and above the 95th percentile (> 95). The weighted Kappa index was estimated to assess agreement between the classifications (p < 0.05). Information was obtained on 98 children and 40 adolescents. From 1.0 to 18.4% of the children were < P5 for W/A, and the agreement for this index was considered weak (Kappa = 0.16; 95%CI -0.03-0.34; p < 0.01); no agreement was observed between the H/A classifications. For adolescents, W/A < P5 varied from 2.5 to 5.0%; once more there was no agreement for this classification (Kappa = 0.16; 95%CI -0.15-0.48; p > 0.05). There was good agreement for the H/A index (Kappa = 1.00; 95%CI 0.23-1.00; p < 0.01). There was weak agreement between classifications of anthropometric indexes according to three different distributions. The data indicated that the construction of specific curves for individuals with DS would facilitate the identification of overweight, which is often observed among these patients.


Intrasplenic hepatocellular transplantation corrects hepatic encephalopathy in portacaval-shunted rats.
The aim of this work was to evaluate the effect of intrasplenic hepatocellular transplantation on hepatic encephalopathy in an experimental model of chronic liver failure induced by end-to-side portacaval shunt in the rat. Inbred male Wistar Furth rats were divided into three groups: rats subjected to portacaval shunt (n = 10), rats subjected to portacaval shunt and intrasplenic hepatocellular transplantation of 10(7) hepatocytes isolated from livers of syngeneic rats (n = 10) and sham-operated rats (n = 10). Behavior tests were performed in a blind fashion at 3 wk, at 2 mo and at 3 mo after surgery. Spontaneous activity and nose-poke exploration by individual rats were studied in automated open field boxes equipped with infrared cells. Each cell beam interruption was automatically recorded on a microcomputer and transformed into a score index (counts/hour). Plasma levels of amino acids, ammonia and total biliary acids were measured. Portacaval shunt rats showed reduced spontaneous activity and nose-poke exploration scores. Intrasplenic hepatocellular transplantation significantly increased spontaneous activity after 2 mo and improved nose-poke exploration after 3 wk. At 3 mo, spontaneous activity and nose-poke exploration in portacaval shunt/intrasplenic hepatocellular transplantation rats were not significantly different from those of sham rats. Increases in plasma ammonia levels after portacaval shunt were not corrected. Amino acid imbalance and bile acid concentration in plasma were partially corrected by intrasplenic hepatocellular transplantation. These data show that intrasplenic hepatocellular transplantation can correct the neurological symptoms of hepatic encephalopathy in an experimental model of chronic liver failure and suggest that intrasplenic hepatocellular transplantation might be of therapeutic interest in chronic liver failure.


Managing the risk of bacterial meningitis among healthcare workers.
Meningococcal disease is generally caused by A, B, C, W and Y subgroups of Neisseria meningitidis. In 2015, the Italian mass media focused on this disease due to the death of two nurses. This generated alarm in the general population, especially in healthcare workers (HCWs). The Occupational Medicine department of the Foundation PTV Polyclinic Tor Vergata (PTV) offered free MenACWY vaccine to HCWs and to students of the University of Rome Tor Vergata. To analyse the prevalence of N. meningitidis in PTV, a large teaching hospital, and to evaluate preventive measures offered to individuals at risk (i.e. HCWs and students). The Hospital Informatic Service provided data about patients admitted to PTV from January 2012 to December 2016. Hospital Discharge Registers were analysed and all cases of meningitis were selected. MenACWY vaccine administered to HCWs and students of PTV in 2017 was analysed. There was just one case of meningococcal disease in 2016. Of the 117 MenACWY vaccines administered, 42% were given to students, 32% to doctors, 8% to nurses, 2% to clerks and 16% to other healthcare professionals. International guidelines recommend MenACWY vaccine for microbiologists exposed to N. meningitidis isolates. The Italian Legislative Decree 81/2008, however, states that occupational physicians can adopt special protective measures such as vaccination programmes for HCWs. The response rate highlights how perceived risk may influence behaviour even in populations with a higher background knowledge than the general population.


Female zebra finches prefer symmetrically banded males, but only during interactive mate choice tests.
We used a four-arm choice chamber to investigate whether the symmetry of artificial traits (leg bands) influenced how attractive male zebra finches, Taeniopygia guttata, were to females, and whether a female's preference was influenced by the opportunity to interact with males during choice tests. Females (N=10) were tested separately and moved freely between the chamber's arms, each of which contained a male wearing bands (two per leg) in symmetric, asymmetric or cross-asymmetric configurations. In three of the six tests experienced by each female, one-way glass (mounted diagonally within each arm) prevented males from seeing the female or their own reflection. During the other three tests, transparent glass was used, so males could see and react to the female. A male's band combination had a significant influence on the amount of time that females spent with him, but only during transparent-glass tests, where symmetric males were preferred. The results of our transparent-glass tests were similar to those reported by Swaddle & Cuthill (1994, Nature, 367, 165-166), in a study where females selected between males that were interacting with their own mirror images (reflected by one-way glass). We discuss why females display the same apparent preference when tested with males engaged in intra- (Swaddle & Cuthill's study) versus intersexual activities (our study). Since females in our study displayed a preference only when males could interact with them, caution may be required when extrapolating the results of static choice tests (e.g. involving models, one-way glass or video images) to situations where the participants can interact. Copyright 1999 The Association for the Study of Animal Behaviour.


Genetic and biochemical studies of partially active tryptophan synthetase mutants of Saccharomyces cerevisiae.
Approximately 20% of the tryptophan synthetase mutants (tr(5)) of Saccharomyces cerevisiae retain activity in one of the half reactions catalyzed by this enzyme and have been identified as indole-accumulating or indole-utilizing tr(5) mutants by complementation tests. Ten indole-accumulating and six indole-utilizing mutants have been studied. For the half reactions they catalyze, these partially active mutants have from about one-half to twice the specific activities of the wild-type enzyme. Indole-accumulating mutant enzymes showed varying responses to pyridoxal phosphate and serine in the assay mixture. The partially active mutants were further characterized by their patterns of allelic complementation and their distribution on the fine-structure map of the locus. It was concluded that these mutants define two distinct functional regions of the tr(5) locus, corresponding to the two half reactions.


Multifaceted Interactions Between Endophytes and Plant: Developments and Prospects.
Microbial endophytes are present in all known plant species. The ability to enter and thrive in the plant tissues makes endophytes unique, showing multidimensional interactions within the host plant. Several vital activities of the host plant are known to be influenced by the presence of endophytes. They can promote plant growth, elicit defense response against pathogen attack, and can act as remediators of abiotic stresses. To date, most of the research has been done assuming that the interaction of endophytes with the host plant is similar to the plant growth-promoting (PGP) microbes present in the rhizosphere. However, a new appreciation of the difference of the rhizosphere environment from that of internal plant tissues is gaining attention. It would be interesting to explore the impact of endosymbionts on the host's gene expression, metabolism, and other physiological aspects essential in conferring resistance against biotic and abiotic stresses. A more intriguing and inexplicable issue with many endophytes that has to be critically evaluated is their ability to produce host metabolites, which can be harnessed on a large scale for potential use in diverse areas. In this review, we discuss the concept of endophytism, looking into the latest insights related to the multifarious interactions beneficial for the host plant and exploring the importance of these associations in agriculture and the environment and in other vital aspects such as human health.


Using engineered 2-O-sulfotransferase to determine the activity of heparan sulfate C5-epimerase and its mutants.
Heparan sulfate (HS) is involved in essential physiological and pathophysiological functions. HS is a highly sulfated polysaccharide consisting of glucuronic acid (or iduronic acid) linked to glucosamine carrying various sulfo groups. Biosynthesis of HS involves sulfotransferases and an epimerase. The HS C(5)-epimerase converts glucuronic acid to iduronic acid. The method for determining the activity has been cumbersome due to the use of a site-specifically (3)H-labeled polysaccharide substrate. Here, we report a two-enzyme coupling assay to determine the activity of C(5)-epimerase. HS 2-O-sulfotransferase (2OST) transfers the sulfo group to the 2-OH-position of glucuronic or iduronic acid. Unlike the wild type protein, 2-O-sulfotransferase mutant (2OST Y94I) transfers sulfate to the iduronic acid but not to the glucuronic acid. Thus, 2OST Y94I cannot sulfate N-sulfated heparosan, a polysaccharide containing glucuronic acid. Incubating N-sulfated heparosan with C(5)-epimerase converts some of the glucuronic acid to iduronic acid, thus becoming a substrate for 2OST Y94I. The susceptibility of the C(5)-epimerase-treated N-sulfated heparosan to 2OST Y94I modification directly correlates to the amount of the activity of C(5)-epimerase, proving that this two-enzyme coupling system can be used to assay for C(5)-epimerase. The method was further used to determine the activities of various C(5)-epimerase mutants. Our approach will significantly reduce the complexity for assaying the activity of C(5)-epimerase and facilitate the structural and functional analysis of C(5)-epimerase.


Treating epilepsy with options other than antiepileptic medications.
Epilepsy is a common health burden worldwide. Epilepsy is linked to variety of factors, including infectious, vascular, immune, structural, genetic, and metabolic etiologies. Despite the existence of multiple antiepileptic drugs (AEDs), many patients are diagnosed with intractable epilepsy. Many nonpharmacological options are available for epilepsy. Some types of epilepsy respond to cofactors. Other patients may be candidates for a ketogenic diet. Inflammatory mediators, such as intravenous immunoglobulins (IVIgs) and steroids, are other options for epilepsy. Recently, cannabinoids have been approved for epilepsy treatment. Refractory epilepsy can be treated with surgical interventions. Focal resections, hemispherectomies, and corpus callosotomies are some common epilepsy surgery approaches. Neuromodulation techniques are another option. Thermal ablation is a minimally invasive approach for epilepsy treatment. Epilepsy outcomes are improving, and treatment modalities are expanding. Trials of nonpharmacological options for epilepsy patients are recommended. This article summarizes available nonpharmacological options other than AEDs for the treatment of epilepsy.


The Effect of Exercise on a Novel Dual-Task Assessment for Sport Concussion.
This study aimed to determine the effect of exercise on cognitive and motor performance and the subsequent test-retest reliability of a novel dual-task (DT) assessment in nonconcussed college students. Sixty nonconcussed college students (53.3% female) with an average age (±SD) of 20.5 ± 1.34 yr, height of 171.7 ± 9.33 cm, and mass of 69.3 ± 12.23 kg were included in the study. Participants were assigned to an exercise ( n = 30) or rest ( n = 30) intervention group and completed two study visits that were separated by a 2-wk test-retest interval. At each visit, participants completed a novel DT assessment that consisted of the concurrent administration of the Standardized Assessment of Concussion (SAC) and tandem gait (TG) before the exercise or rest intervention. After the DT assessment at the first visit, participants in the exercise group performed moderate-intensity exercise, whereas the rest group sat quietly for 30 min. After the intervention, both groups were readministered the DT assessment. At the second visit, the same procedures were followed, except that each group was administered the opposite intervention (e.g., the exercise group completed the rest intervention). A composite TG (cTG) score was calculated by summing the average time to complete the TG pattern during each SAC domain (immediate memory, digits backwards, months in reverse order, delayed recall). An ANCOVA was conducted to assess postintervention differences while controlling for preintervention performance. Test-retest reliability was assessed using intraclass correlation coefficients (ICC 3,2 ) with 95% confidence intervals, with all analyses performed with α = 0.05. SAC and cTG performance was similar ( P values > 0.05) from preintervention to postintervention for the rest or exercise protocols. Good (rest: ICC = 0.77 (0.62-0.87); exercise: ICC = 0.84 (0.73-0.90)) and excellent (rest: ICC = 0.97 (0.94-0.98); exercise: ICC = 0.93 (0.88-0.96)) test-retest reliabilities were observed for the SAC composite score and cTG score, respectively. Our DT assessment was robust to the influence of moderate-intensity exercise and demonstrated good-to-excellent test-retest reliability in a healthy collegiate sample.


In vitro and in vivo antifungal activities of DU-6859a, a fluoroquinolone, in combination with amphotericin B and fluconazole against pathogenic fungi.
DU-6859a is an investigational fluoroquinolone agent with potent bactericidal activity, but by itself it has no antifungal activity. When combined with amphotericin B (AmB), however, DU-6859a clearly enhanced the in vitro antifungal activity of AmB against Candida albicans, Candida tropicalis, Candida krusei, Candida glabrata, and Cryptococcus neoformans in microdilution checkerboard studies. Positive interactions of DU-6859a with AmB against Aspergillus fumigatus were dependent on the medium used; yeast nitrogen base supplemented with amino acids, ammonium sulfate, and 1% glucose was better for demonstrating synergism, while in RPMI 1640 medium, unexpected antagonism between the drugs occurred against three of the strains tested. In combination with fluconazole (Flu), DU-6859a increased the activity of Flu against C. albicans both in synthetic amino acid medium fungal and in supplemented yeast nitrogen base. An in vitro time-kill study revealed that DU-6859a combined with AmB significantly suppressed the regrowth of C. albicans compared with the suppression brought about by AmB used alone in a concentration-dependent fashion. Furthermore, in a model of C. albicans infection in mice, the fungal load in infected kidneys was significantly less in mice given the combination treatment of DU-6859a plus either AmB or Flu, and thus, the combination treatment resulted in prolonged survival of infected mice compared with treatment with either antifungal alone. The prolonged survival in mice given the combined treatment was also observed in mice with A. fumigatus infection, indicating that DU-6859a potentiated the actions of the antifungal agents in vivo as well as in vitro.


Sarcoidosis presenting as papulonecrotic tuberculid-like lesions: report of a case.
A 50-year-old Thai woman presented with papulonecrotic tuberculid-like eruptions on her back and inframammary area with fever, nonproductive cough and weight loss. Chest radiograph showed diffuse bilateral reticulonodular opacities in both lungs with bilateral hilar lymph node enlargement. High resolution computed tomography (HRCT) of the lungs showed peribronchovascular interstitial thickening with multiple lymph nodes enlargement. Sputum for AFB was negative. Monotest (PPD) was negative. Skin biopsy revealed multiple naked granuloma compatible with sarcoidosis. She was treated with isoniazid, 300 mg/d, rifamipicin, 600 mg/d, ethambutal, 800, mg/d and pyrazinamide, 1000 mg/d for 2 months without improvement of skin and lung lesions. Prednisolone 45 mg/d was then administered adjunctive with isoniazid and rifampicin. After two weeks of treatment with prednisolone, the cutaneous and pulmonary lesions markedly improved Prednisolone was tapered in 6 months. Skin lesions, fever; dry cough disappeared and chest radiograph, HRCT of the chest were markedly improved.


Gut microbiome and pancreatic cancer cachexia: An evolving relationship.
Nearly 80% of patients with pancreatic ductal adenocarcinoma (PDAC) develop cachexia along their disease course. Cachexia is characterized by progressive weight loss, muscle wasting, and systemic inflammation and has been linked to poorer outcomes and impairments in quality of life. Management of PDAC cachexia has historically involved a multidisciplinary effort comprised of nutritional support, pancreatic enzyme replacement therapy, and/or pharmacologic interventions. Despite current interventions to mitigate PDAC cachexia, a significant proportion of patients continue to die from complications associated with cachexia underscoring the need for novel insights and treatments for this syndrome. We highlight the feasibility and effectiveness of a recent enteral feeding prospective trial at our institution to improve cachexia outcomes in patients with advanced PDAC. Additionally, we were among the first to characterize the stool microbiome composition in patients with advanced PDAC receiving enteral feeding for the treatment of cachexia. Novel insights into the relationship between enteral nutritional support, cachexia, and the gut microbiome are presented. These promising results are discussed in the context of a potential ability to modulate the stool microbiome as a new interventional strategy to mitigate PDAC cachexia.


Corpus Callosotomy in 3 Cavalier King Charles Spaniel Dogs with Drug-Resistant Epilepsy.
Corpus callosotomy (CC) is an established palliative surgery for human patients with drug-resistant epilepsy (DRE), especially those with generalized seizures and multiple or unknown epileptogenic focus. However, there are no reports to describe CC in canine patients with epilepsy. Three client-owned Cavalier King Charles Spaniels with DRE are included in this case series. In presurgical evaluations, an apparent epileptogenic zone was not detected in each dog and CC was conducted. Total CC was performed in one dog, whereas the other two received partial CC. One dog recovered from surgery without any complications, but died suddenly by an unknown cause at 10 h after surgery. For the other two dogs, postoperative evaluations including seizure outcomes, complications, and quality of life of the dogs and owners were assessed for at least 12 months. Both dogs showed a remarkable decrease in seizure frequency (averaged 80.3% reduction) and severity after surgery. The antiseizure medications were maintained, and not only the mentation and activity of the dogs, but also the quality of life of dogs and owners were improved postoperatively. Although technical improvement and more large-scale studies are needed, CC is a treatment option for dogs with DRE in veterinary medicine.


Regulation of porcupine-dependent Wnt signaling is essential for uterine development and function.
Six members of the Wnt family are expressed in the female reproductive tract. Their collective function ensures proper development of the uterus, preparing it for pregnancy during adulthood. Here, we take advantage of the fact that Porcn, a prerequisite for all Wnt secretion, is located on the X chromosome, to generate females that were mosaic for Porcn throughout the reproductive tract. Porcnflox/+ females were mated with progesterone receptor (Pgr)-Cre males (PgrCre/+ ) to generate females that were heterozygous for Porcupine in all tissues of the female reproductive tract, resulting in mosaicism due to random X-inactivation. We demonstrated that Porcn mosaic females are extremely subfertile and exhibit a large spectrum of phenotypes ranging from morphologically normal uteri to uteri with extremely enlarged cystic glands. Decreased fertility in Porcupine mosaic females was not associated with phenotype severity and was observed regardless of whether or not cystic glands were enlarged. By crossing-in a GFP reporter on the wild-type X chromosome, we were able to correlate endometrial gland hyperplasia with a mostly Porcupine mutant stroma, demonstrating the role of stromal Wnts in the regulation of endometrial gland proliferation. Finally, we demonstrated that fertility issues within mosaic females were due to a reduced response to estrogen and to abnormal Tcf/Lef signaling across the mesometrial-anti-mesometrial axis during the window of implantation.


Results of MIS TKA at mean nine year follow-up.
The first 103 consecutive minimally invasive total knee arthroplasties performed by a single surgeon were assessed to determine the clinical and radiographic outcomes at a mean of 9-year follow-up (range, 8 to 10 years). Patients who died before final follow-up were 10 (13 knees), leaving 90 knees in 69 patients for final review. Outcome was evaluated using Knee Society pain and functional scores. Survival with revision as an end point was 97.1%. Knee Society scores and range of motion improved significantly to a mean of 96 and 92 points and a mean of 115 degrees, respectively. There were five patients who required surgical intervention for unexplained pain with findings of retained cement and adhesions. Radiographic analysis did not reveal any progressive radiolucencies. These results suggest that excellent long-term outcomes can be achieved with minimally invasive total knee arthroplasty.


The central nervous system in systemic lupus erythematosus. Part 2. Pathogenetic mechanisms of clinical syndromes: a literature investigation.
To identify the pathogenetic mechanisms of central nervous system (CNS) syndromes of systemic lupus erythematosus (SLE) as described in the literature. Using PUBMED, we performed a systematic search of publications from 1980 onwards. Studies were eligible if they had been performed on patients or material from patients with CNS manifestations and definite SLE and when the CNS manifestations were not secondary. Criteria were formulated for the identification of pathogenetic mechanisms. The single most important cause of the CNS syndromes of SLE is ischaemia due to narrowing or occlusion of small vessels, arteries and veins. Antiphospholipid antibodies and premature atherosclerosis play roles in these processes, but they have not been delineated definitely. Intracranial and intraspinal haemorrhages are much less frequent than ischaemia and are presumably in part due directly to SLE. Vasculitis may cause ischaemia or haemorrhage in the CNS and is involved occasionally, as shown by imaging and histological findings. White matter damage is heterogeneous and ill-understood. It includes white matter degeneration and myelin vacuolation of the spinal cord, and reversible leucoencephalopathy due to oedema. Antibody-induced neuronal dysfunction in the CNS is a realistic hypothesis and may involve anti-ribosomal P antibodies and several other antibodies. Deficiency of psychological reactions forms a separate and entirely different category of mechanisms. Causes have been identified or possible causes have been suggested for most of the CNS syndromes of SLE, thus offering rationales for different forms of prevention and therapy.


Simulated Keratometry Versus Total Corneal Power by Ray Tracing: A Comparison in Prediction Accuracy of Intraocular Lens Power.
To investigate the relationship between corneal powers measured by simulated keratometry (Sim-K) and ray tracing and understand whether their difference influences intraocular lens (IOL) power calculation. In a first sample of healthy eyes, corneal curvature was measured using a rotating Scheimpflug camera (Sirius, CSO). Sim-K was obtained from anterior corneal curvature using the 1.3375 keratometric index. Ray tracing was performed through both corneal surfaces to calculate the total corneal power (TCP). The difference between Sim-K and TCP was correlated with various parameters. In a second sample of patients undergoing cataract surgery, IOL power was subsequently calculated using both Sim-K and TCP. In the first sample (114 eyes), Sim-K (43.64 ± 1.44 D) was higher than TCP (43.07 ± 1.41 D, P < 0.0001); the difference ranged between 0.07 and 1.95 D and correlated with the anterior/posterior (A/P) ratio (r = 0.7292, P < 0.0001), which ranged between 1.10 and 1.30. In the second sample (107 eyes), the A/P ratio influenced the outcomes of the Holladay 1 and SRK/T, but not Haigis and Hoffer Q formulas. However, using TCP, which takes the A/P ratio into account, did not improve the prediction error of any formula. Sim-K provides a higher corneal power compared with TCP. This difference is not constant but depends on the A/P ratio and can influence the refractive outcome of IOL power calculation by theoretical formulas. However, TCP values do not improve the results of these formulas, as they were developed for keratometric values such as Sim-K.


Single-molecule analysis of fluorescent carbon dots towards localization-based super-resolution microscopy.
The advancement of high-resolution bioimaging has always been dependent on the discovery of bright and easily available fluorescent probes. Fluorescent carbon nanodots, an interesting class of relatively new nanomaterials, have emerged as a versatile alternative due to their superior optical properties, non-toxicity, cell penetrability and easy routes to synthesis. Although a plethora of reports is available on bioimaging using carbon dots, single-molecule-based super-resolution imaging is rare in the literature. In this study, we have systematically characterized the single-molecule fluorescence of three carbon dots and compared them with a standard fluorescent probe. Each of these carbon dots showed a long-lived dark state in the presence of an electron acceptor. The electron transfer mechanism was investigated in single-molecule as well as in ensemble experiments. The average on-off rate between the fluorescent bright and dark states, which is one of the important parameters for single-molecule localization-based super-resolution microscopy, was measured by changing the laser power. We report that the photon budget and on-off rate of these carbon dots were good enough to achieve single-molecule localization with a precision of ~35 nm.


[Pathomorphology of membranes appearing in proliferative vitreoretinopathies].
Immunohistochemical techniques were used to investigate the cell content of epiretinal membranes occuring in proliferative vitreoretinopathy (PVR) and proliferative diabetic retinopathy (PDR). Ten epiretinal membranes were obtained during surgery from eyes with PVR and five from eyes with PDR. This material was studied histopathologically and immunohistochemically. Retinal pigment epithelial cells, glial cells, macrophages, T lymphocytes and type IV collagen were identified in these membranes. The findings indicate that the cells mentioned possess a potential role in creating vitreoretinal membranes in PVR and PDR.


Cardiogenic shock: a success story.
When the body is unable to deliver oxygen and perfuse tissues adequately, shock occurs. Cardiogenic shock is the shock syndrome that develops when the heart has lost approximately 40% of its pumping ability. This article reviews the pathophysiology and management of cardiogenic shock.


The Effects of Octanoate and Oleate on Isocitrate Lyase Activity during the Germination of Pinus pinea Seeds.
The changes of isocitrate lyase levels with respect to the catabolism of triglycerides have been studied during the germination of Pinus pinea seeds. We studied the effects of octanoate, oleate, and inhibitors of protein synthesis on isocitrate lyase during germination. Pyruvate kinase, glucose-6-P-dehydrogenase, malate dehydrogenase, and isocitrate dehydrogenase were also assayed. Octanoate and oleate inhibited the isocitrate lyase activity, similarly to cycloheximide, chloramphenicol, and actinomycin, inhibitors of protein biosynthesis. This inhibitory effect is not specific but is strikingly evident with isocitrate lyase. This inhibition was not proportional to the concentration but was proportional to the chain length of oleate and octanoate.


Pylorus resection versus pylorus preservation in pancreatoduodenectomy (PyloResPres): study protocol and statistical analysis plan for a German multicentre, single-blind, surgical, registry-based randomised controlled trial.
Partial pancreatoduodenectomy (PD) is the treatment of choice for various benign and malignant tumours of the pancreatic head or the periampullary region. For reconstruction of the gastrointestinal passage, two stomach-preserving PD variants exist: pylorus preservation PD (ppPD) or pylorus resection PD (prPD) with preservation of the stomach. In pancreatic surgery, delayed gastric emptying (DGE) remains a serious complication after PD with an incidence varying between 4.5% and 45%, potentially delaying hospital discharge or further treatment, for example, adjuvant chemotherapy. Evidence is lacking to assess, which variant of PD entails fewer postoperative DGE. The protocol of a large-scale, multicentre, pragmatic, two-arm parallel-group, registry-based randomised controlled trial (rRCT) using a two-stage group-sequential design is presented. This patient-blind rRCT aims to demonstrate the superiority of prPD over ppPD with respect to the overall incidence of DGE within 30 days after index surgery in a German real-world setting. A total of 984 adults undergoing elective PD for any indication will be randomised in a 1:1 ratio. Patients will be recruited at about 30 hospitals being members of the StuDoQ|Pancreas registry established by the German Society of General and Visceral Surgery. The postoperative follow-up for each patient will be 30 days. The primary analysis will follow an intention-to-treat approach and applies a binary logistic random intercepts model. Secondary perioperative outcomes include overall severe morbidity (Clavien-Dindo classification), blood loss, 30-day all-cause mortality, postoperative hospital stay and operation time. Complication rates and adverse events will be closely monitored. This protocol was approved by the leading ethics committee of the Medical Faculty of the Ludwig-Maximilians-Universität, Munich (reference number 19-221). The results will be published in a peer-reviewed journal and presented at international conferences. Study findings will also be disseminated via the website (http://www.dgav.de/studoq/pylorespres/). DRKS-ID: DRKS00018842.


Comparison of the disposition behavior of organic anions in an animal model for Wilson's disease (Long-Evans Cinnamon rats) with that in normal Long-Evans Agouti rats.
Long-Evans Cinnamon (LEC) rats have an abnormality similar to that observed in Wilson's disease in humans and are therefore a good animal model for the study of Wilson's disease. LEC rats develop hereditary hepatitis and severe jaundice. Mutant animals with hyperbilirubinemia have been widely used as animal models for human diseases. Among these mutant animals, Eisai hyperbilirubinemic rats (EHBR) have defective biliary excretion of organic anions. Thus, biliary excretion of sulfobromophthalein (BSP) and urinary excretion of phenolsulfonphthalein (PSP) in LEC rats were compared with those in Long-Evans Agouti (LEA) rats. In LEC rats, the excretion of BSP, a multidrug resistance-associated protein 2 (Mrp2/Abcc2) substrate, was significantly decreased compared to that in LEA rats. It has been reported that the transport function for organic anions on the kidney is maintained in EHBR. However, the urinary excretion of PSP is impaired in LEC rats. It is possible that organic anion transporters responsible for the urinary excretion of PSP in LEA rats and EHBR are impaired in LEC rats. It is important to elucidate the relationship between organic anion secretion and Wilson's disease.


Poly(L-lactide)-vitamin E TPGS nanoparticles enhanced the cytotoxicity of doxorubicin in drug-resistant MCF-7 breast cancer cells.
Multiple drug resistance (MDR) seriously reduces the efficacy of many chemotherapeutic agents for cancer. P-Glycoprotein, an efflux pump overexpressed on the cell surface, plays an important role in drug resistance, but several surfactants, such as vitamin E TPGS, can inhibit P-glycoprotein. In this study, a polylactide-surfactant block copolymer poly(l-lactide)-vitamin E TPGS (PLA-TPGS) was synthesized using bidentate sulfonamide zinc ethyl complex as an efficient catalyst, and its self-assembled nanoparticles were used as carriers of doxorubicin. We first found that the activity of P-glycoprotein in drug-resistant breast cancer MCF-7/ADR cells was decreased after incubation with PLA-TPGS nanoparticles. In addition, the nuclear accumulation and cytotoxicity of doxorubicin were significantly increased by encapsulation into the nanoparticles. The enhanced efficacy of the doxorubicin-loaded PLA-TPGS nanoparticles may result from the combination of inhibition of efflux and increased entry of doxorubicin into the nucleus in drug-resistant MCF-7/ADR cells. Therefore, this innovative delivery system has potential to act as a nanomedicine for therapy of both drug-sensitive and drug-resistant cancer.


Lyngbyatoxin biosynthesis: sequence of biosynthetic gene cluster and identification of a novel aromatic prenyltransferase.
The lyngbyatoxins are potent skin irritants produced by Lyngbya majuscula and cause a condition known as "Swimmer's Itch" off Honolulu, HI. Reported is the molecular cloning of the lyngbyatoxin (ltx) biosynthetic gene cluster from L. majuscula using a strategy based on its predicted nonribosomal peptide synthetase (NRPS) assembly. The biosynthetic gene cluster spans 11.3 kilobase pairs and encodes for a two-module NRPS (LtxA), a P450 monooxygenase (LtxB), an aromatic prenyltransferase (LtxC), and an oxidase/reductase protein (LtxD). LtxC was heterologously produced and purified from E. coli and shown to catalyze the transfer of a geranyl group to (-)-indolactam V as the final step in the biosynthesis of lyngbyatoxin A.


Additive effects of growth hormone and testosterone on lipolysis in adipocytes of hypophysectomized rats.
The effects of growth hormone (GH) and testosterone, alone or in combination, on the regulation of lipolysis in isolated adipocytes from hypophysectomized rats were investigated. Male Sprague-Dawley rats were hypophysectomized at 50 days of age. One week after operation, hormonal replacement therapy with L-thyroxine and hydrocortisone acetate was given to hypophysectomized rats. Groups of rats were treated with GH (1.33 mg/kg, daily), testosterone (10 mg/kg, once) alone or in combination. After one week of hormonal treatment, adipocytes were isolated from the pooled epididymal and perirenal fat pads and glycerol release after isoproterenol stimulation and 125I-cyanopindolol binding was measured. Hypophysectomy caused a marked decrease in basal and isoproterenol-stimulated lipolysis. There was no effect of testosterone treatment alone on lipolysis, but GH treatment resulted in an increase in isoproterenol-induced lipolysis but not to the levels observed in cells from control rats. Testosterone and GH in combination restored the lipolytic response to isoproterenol. Also 125I-cyanopindolol binding was decreased after hypophysectomy. Testosterone treatment alone and GH treatment alone increased the binding, while in combination the treatment had an additive effect. Affinity was not changed, but the effects seemed to be on receptor number, as determined by Scatchard analysis. Forskolin-stimulated cAMP accumulation in adipocytes was markedly reduced after hypophysectomy. Testosterone treatment alone had no effect. GH treatment alone increased forskolin-stimulated cAMP accumulation, although the level was lower than that found in control rats. The combined treatment resulted in a further increase to levels observed in adipocytes from control rats.(ABSTRACT TRUNCATED AT 250 WORDS)


Melatonin receptor 1b polymorphisms in women with Systemic Lupus Erythematosus.
The pineal hormone melatonin could exert an important influence on the immune system and autoimmunity. Its effect on the immunocompetent cells might be mediated at least partially through specific melatonin receptors. However, the role of melatonin - melatonin receptor 1B (MTNR1B) interrelations in human autoimmune diseases is still unknown. Therefore, the present study aimed to investigate the possible influence of the MTNR1B gene polymorphisms for the development and clinical expression of systemic lupus erythematosus (SLE). 109 female SLE patients and 101 healthy women were genotyped for the MTNR1B rs1562444, rs10830962 and rs10830963 polymorphisms. No genotype distribution differences were found between patients and controls. The presence of MTNR1B rs10830963 C/C genotype was related to increased prevalence of leucopenia compared to genotypes C/G and G/G after Bonferroni correction for multiple comparisons [36.5% vs. 14.5%, p=0.014]. Moreover, the rs10830963 G/G carriers had lower number of lupus criteria in comparison to patients with C/C genotype. The present data suggested that MTNR1B polymorphisms could influence the clinical features in lupus patients, and especially the susceptibility to leucopenia.


Clinical outcomes of negative small-bowel capsule endoscopy for small-bowel bleeding: a systematic review and meta-analysis.
Small-bowel bleeding is the primary indication for capsule endoscopy (CE). Many experts advocate a "watch-and-wait" policy in negative CE. This meta-analysis examines the odds of rebleeding after negative index CE and the impact on long-term follow-up. A comprehensive literature search identified articles examining the rebleeding rate after negative CE. Demographic and clinical information with emphasis on outcomes was retrieved, pooled, and analyzed. Heterogeneity among studies was assessed using the I2 statistic. A random effects model was used as the pooling method because of high heterogeneity. Risk of bias was assessed using the quality assessment of diagnostic accuracy studies (QUADAS-2) tool. The primary outcome evaluated was the pooled odds ratios (ORs) for rebleeding after a negative CE for obscure GI bleeding (OGIB). Twenty-six studies with 3657 patients were included. The pooled rate of rebleeding after negative CE was .19 (95% CI, .14-.25; P < .0001). The pooled OR of rebleeding was .59 (95% CI, .37-.95; P < .001). The effect was more pronounced in studies with a short follow-up (OR, .47; 95% CI, .24-.94; P < .001). There was no statistically significant difference in rebleeding after CE for occult and overt OGIB. Prospective studies showed a lower OR of rebleeding of .24 (95% CI, .08-.73; P = .01). Most studies were high quality. Our analysis shows that negative CE provides adequate evidence of a subsequently low risk of rebleeding. Such patients can therefore be safely managed with watchful waiting. However, patients who rebleed after 2 years may need to be investigated for a new source of blood loss.


Steroid hormones and the pathogenesis of benign prostatic hyperplasia.
The pathogenesis of benign prostatic hyperplasia (BPH) is still poorly understood: there is, however, general acceptance that the condition is not premalignant and that it has an etiology distinct from that of cancer. Interest now focuses on the biochemistry of the target prostate cells and the propensity of the gland for uncontrolled growth. Dihydrotestosterone (DHT) is the active intracellular androgen formed from testosterone by 5 alpha-reductase. DHT concentrations appear a little higher in BPH tissue than in normal tissue, and there is no doubt that DHT-receptor complex modulates gene expression. Current studies suggest that DHT is essential but not sufficient for proliferation, and that other regulatory factors, including peptide growth factors, are prerequisite. The growth responsiveness of prostate tissue to androgens may be dependent on the balance between epithelial and stromal tissues, with biologic processes in the epithelium indirectly controlled by androgen-dependent mediators of stromal origin.


Elevated midtrimester serum methylmalonic acid levels as a risk factor for neural tube defects.
The role of folic acid in the primary prevention of neural tube defects (NTDs) is well established. However, questions related to the protective mechanism remain unanswered. To help answer these questions, we designed a case-control study to assess the role of folate- and cobalamin-related metabolites in the pathogenesis of NTDs. Concentrations of folate, cobalamin, and 14 other related metabolites were measured by gas chromatography/mass spectrometry in midtrimester serum specimens from 32 women with an NTD-affected pregnancy and from 132 control women, and in serum specimens from 46 nonpregnant women who had a history of NTD-affected pregnancy and from 43 nonpregnant control women. Log-transformed means of metabolites were compared between case and control women for both the midtrimester and nonpregnant groups. In the pregnant group, serum methylmalonic acid (MMA) concentrations were higher among case women than among control women (130 vs 105 nM). There was a strong dose-response relationship between midtrimester serum MMA level and the risk for an NTD-affected pregnancy, with the relative risk increasing 13-fold for women with MMA levels > 90th percentile. In the nonpregnant group, there was no difference in serum MMA levels between case and control women (140 vs 140 nM). Thus, the serum MMA levels of women in the midtrimester of pregnancies unaffected by NTDs were significantly lower than the levels of nonpregnant women, whereas the levels of women whose pregnancies were affected by NTDs were similar to those of nonpregnant women. The finding of elevated MMA serum concentrations among women in the midtrimester of NTD-affected pregnancies suggests that cobalamin may be involved in the etiology of NTDs. The possible role of cobalamin in relation to the protective effect of folic acid needs further evaluation.


Easily Created Prediction Model Using Automated Artificial Intelligence Framework (Prediction One, Sony Network Communications Inc., Tokyo, Japan) for Subarachnoid Hemorrhage Outcomes Treated by Coiling and Delayed Cerebral Ischemia.
Introduction Reliable prediction models of subarachnoid hemorrhage (SAH) outcomes and delayed cerebral ischemia (DCI) are needed to decide the treatment strategy. Automated artificial intelligence (AutoAI) is attractive, but there are few reports on AutoAI-based models for SAH functional outcomes and DCI. We herein made models using an AutoAI framework, Prediction One (Sony Network Communications Inc., Tokyo, Japan), and compared it to other previous statistical prediction scores. Methods We used an open dataset of 298 SAH patients, who were with non-severe neurological grade and treated by coiling. Modified Rankin Scale 0-3 at six months was defined as a favorable functional outcome and DCI occurrence as another outcome. We randomly divided them into a 248-patient training dataset and a 50-patient test dataset. Prediction One made the model using training dataset with 5-fold cross-validation. We evaluated the model using the test dataset and compared the area under the curves (AUCs) of the created models. Those of the modified SAFIRE score and the Fisher computed tomography (CT) scale to predict the outcomes. Results The AUCs of the AutoAI-based models for functional outcome in the training and test dataset were 0.994 and 0.801, and those for the DCI occurrence were 0.969 and 0.650. AUCs for functional outcome calculated using modified SAFIRE score were 0.844 and 0.892. Those for the DCI occurrence calculated using the Fisher CT scale were 0.577 and 0.544. Conclusions We easily and quickly made AutoAI-based prediction models. The models' AUCs were not inferior to the previous prediction models despite the easiness.


The DnaC helicase loader is a dual ATP/ADP switch protein.
Helicases are transferred to replication origins by helicase loading factors. The Escherichia coli DnaC and eukaryotic Cdc6/18 helicase loaders contain ATP sites and are both members of the AAA+ family. One might expect that ATP is required for helicase loading; however, this study on DnaC illustrates that ATP is not actually needed for DnaC to load helicase onto single-strand DNA (ssDNA). In fact, it seems to be a paradox that after transfer of helicase to DNA, DnaC-ATP inhibits helicase action. In addition, ATP is required for DnaC function at an early step in oriC replication in which ATP stimulates ssDNA binding by DnaC, leading to expansion of the ssDNA bubble at the origin. Two cofactors, ssDNA and DnaB, trigger hydrolysis of ATP, converting DnaC to the ADP form that no longer inhibits DnaB. These observations have led to the idea that DnaC is a 'dual' switch protein, where both the ATP and the ADP forms are sequentially required for replication. This dual switching process may underlie the sensitivity of DnaB to even small fluctuations in DnaC levels.


Block of calcium-activated potassium channels in mammalian arterial myocytes by tetraethylammonium ions.
The effects of tetraethylammonium ions (TEA+) and tetrapentylammonium ions (TPeA+) on Ca2(+) -activated K+ (KCa) channels were studied in membrane patches from mesenteric arterial myocytes. External TEA+ produced a flickery block. The concentration dependence for reduction in mean unitary current was consistent with 1:1 binding, with dissociation constants (Kd) in rat and rabbit of 196 and 159 microM at 0 mV, and the block was weakly voltage dependent. Rate constants for blocking and unblocking were 380 mM-1.ms-1 and 73 ms-1, respectively. In patches containing several channels TEA+ reduced average current to the same extent as mean unitary current, implying that TEA+ block is independent of the channel state. Block was unaffected by raising external K+ to 120 mM. External TPeA+ blocked with slower kinetics and lower affinity than TEA+ (Kd, 1.49 mM). The sulfonylurea glibenclamide (10-100 microM), the hyperpolarizing vasodilator cromakalim (5 microM), and internal ATP (1 mM) were without effect on channel activity. We conclude that TEA+ is a relatively effective blocker of single KCa channels of arterial smooth muscle and should block macroscopic currents equally well, whereas external TPeA+ is about eight times less effective.


Structural and photoluminescence studies of a europium(III) tetrakis(beta-diketonate) complex with tetrabutylammonium, imidazolium, pyridinium and silica-supported imidazolium counterions.
Tetrakis(naphthoyltrifluoroacetonato)lanthanate(III) complexes (Ln = Eu, Gd) containing the cations tetrabutylammonium, [NBu(4)](+); 1-butyl-3-methylimidazolium, [C(4)mim](+); and 1-butyl-3-methylpyridinium, [C(4)mpyr](+), have been prepared and structurally characterized by single-crystal X-ray diffraction. The {EuO(8)} coordination sphere in [NBu(4)][Eu(NTA)(4)] is best described as a distorted dodecahedron, where the metal ion is located at the 4-fold inversion axis with only one crystallographically independent NTA residue. In [C(4)mim][Eu(NTA)(4)] and [C(4)mpyr][Gd(NTA)(4)], the central Ln(3+) ions are coordinated by eight oxygen atoms from four distinct beta-diketonate ligands, in an overall distorted square-antiprismatic geometry. Besides electrostatic interactions, the crystal packing in all three structures is stabilized by offset pi-pi interactions involving the naphthyl rings of neighboring complexes (and, for [C(4)mim][Eu(NTA)(4)] and [C(4)mpyr][Gd(NTA)(4)], neighboring naphthyl/imidazolium and naphthyl/pyridinium rings) and C-H...pi contacts. The photoluminescence properties of the three Eu(III) complexes were studied at room temperature and -259 degrees C by measuring emission and excitation spectra, (5)D(0) emission decay curves, and absolute emission quantum yields. Under ligand excitation (lambda(ex) = 290-395 nm), the quantum yields (room temperature) were in the range 0.72-0.77 for the 1-butyl-3-methylimidazolium salt. An immobilized analogue of this complex was prepared by supporting [Eu(NTA)(4)](-) on an ordered mesoporous silica derivatized with 1-propyl-3-methylimidazolium groups. The disappearance of the intra-4f(6) lines in the excitation spectrum of the supported material indicated an increase in the ligand's sensitization process of the Eu(3+) ions, relative to direct intra-4f(6) excitation. The emission quantum yield measured for the supported material (0.32-0.40, for excitations between 265 and 360 nm) is the highest so far reported for lanthanide-containing ordered mesoporous silicas.


Prenatal exposure to acid-suppressive drugs and the risk of allergic diseases in the offspring: a cohort study.
Recent studies reported increased risks for the development of asthma in children after prenatal exposure to acid-suppressive drugs. As a result of common pathogenesis, associations could also be present for other allergic diseases. Using the prescription database IADB.nl, we conducted a cohort study amongst 33 536 children in the Netherlands, with a maximum follow-up of 8 years. Maternal exposure was defined as ≥1 dispensed prescription for proton pump inhibitors (PPIs) and/or Histamine 2-antagonists (H2As) during pregnancy. Children were considered to have a drug-treated allergic disease if they received either ≥2 prescriptions for dermal (atopic dermatitis), inhaled (asthma) or nasal (allergic rhinitis) steroids within a 12-month period. Clustered Cox proportional hazard regression was used to estimate crude and adjusted hazard ratios (aHR) with 95% confidence intervals (95% CI). The aHR for the development of any allergic disease was 1.37 (95% CI: 1.14-1.66) for children exposed to PPIs or H2As. Prenatal exposure to PPIs and/or H2As was associated with atopic dermatitis, asthma and allergic rhinitis with aHRs of 1.32 (95% CI 1.06-1.64), 1.57 (95% CI 1.20-2.05) and 2.40 (95% CI 1.42-4.04), respectively. The aHR for the development of two or more (aHR 2.13 95% CI: 1.43-3.19) and three allergic diseases (aHR 5.18 95% CI: 2.16-12.42) were even more elevated after prenatal exposure to PPIs or H2As. Prenatal exposure to PPIs and H2As appeared associated with an increased risk for the development of atopic dermatitis, asthma and allergic rhinitis in the offspring, especially with the development of multiple allergic diseases. Because our study has limitations inherent to observational studies, prospective studies are now warranted to confirm our findings.


Surgical Treatment Versus Conservative Management for Acute Achilles Tendon Rupture: A Systematic Review and Meta-Analysis of Randomized Controlled Trials.
Acute Achilles tendon ruptures can be treated with surgical and nonsurgical treatment. However, the optimal intervention for acute Achilles tendon rupture remains controversial. The aim of the present study was to compare the clinical outcomes of surgical treatment versus conservative management for acute Achilles tendon rupture. Eight randomized controlled studies involving 762 patients were included in the meta-analysis. In general, re-rupture occurred in 14 of 381 surgically treated patients (3.7%) and 37 of 377 nonsurgically treated patients (9.8%). Pooled results showed that the total re-rupture rate was significantly lower in surgical group than that in the nonsurgical group (risk ratio 0.38, 95% confidence interval 0.21 to 0.68; p = .001). No significant differences were found between the 2 treatment groups in the incidence of deep venous thrombosis, the number who returned to sport, ankle range of motion (dorsiflexion, plantarflexion), Achilles tendon total rupture score, or physical activity scale. Surgical treatment can effectively reduce the re-rupture rate and might be a better choice for the treatment of acute Achilles tendon rupture. Multicenter, double-blind randomized controlled trials with stratification and long-term follow-up are needed to obtain a higher level of evidence and to guide clinical practice, especially in the comparison and selection of different treatments.


Octant vectorcardiography and its data basis.
The present communication is a continuation of the previous report published in this journal [Physiol. bohemoslov., 29: 481, 1980]. It presents further information on the practical use and evaluation of the octant vectorcardiography method. The description starts from the elementary table set up according to the Frank lead system. All cases may be classified alpha-numerically into the data bases by means of a newly introduced concept - the octant. The data include 250 cases after myocardial infarction, supplemented by further information on the integration of peaks. In this way, even widely different cases can be compared in general. In advanced cases of coronary artery disease, an octant labelled in our terminology as the fifth octant, has been defined. The method brings new information about cases after myocardial infarction and it helps to determine the progress of the disease.


Patterns of neuropsychological performance in Alzheimer's disease patients with and without extrapyramidal signs.
We investigated the relationship between extrapyramidal signs (EPS) and the cognitive function in 90 patients meeting NINCDS-ADRDA criteria for probable Alzheimer's disease (AD) by dividing them into those showing no evidence of EPS and those showing at least one EPS on the Unified Parkinson's Disease Rating Scale. Cognitive function was assessed by neuropsychological tests of verbal memory, orientation, abstract reasoning, language, and construction. Patients with EPS were significantly more impaired than patients without EPS on tests of short-term learning and memory, orientation, naming, verbal fluency, and construction, but not on tests of long-term memory, abstract reasoning, or verbal comprehension. These results could not be explained by any differences in age, education, or disease duration between the groups. Since this pattern of neuropsychological impairment resembles that of patients with Parkinson's disease and other EPS syndromes, we hypothesize that EPS are associated with neuropathologic and neuropsychological changes that are superimposed over the classic features of AD.


Effect of Enzyme Modified Soymilk on Rennet Induced Gelation of Skim Milk.
In this study, soymilk was hydrolyzed to different degrees with flavourzyme, and then soymilk and enzyme modified soymilk at various levels were added to skim milk respectively, to generate a mixed gel using rennet. Rheological properties, scanning electron microscopy imaging, and physical and chemical indexes were examined to reveal the effect of enzyme modified soymilk on rennet induced gelation of skim milk. Results showed that soymilk inhibited the aggregation of skim milk, led to a decrease in storage modulus (G'), significantly increased moisture content and curd yield, and the resulting network was coarse. Enzyme modified soymilk with a molecular weight below 20 kDa led to a more uniform curd distribution, which counteracted the reduction of G' and allowed for the formation of a stronger gel. Both the moisture content and the curd yield increased with the addition of soymilk and enzyme modified soymilk, and overall the effect of adding a high degree of hydrolysis of enzyme modified soymilk was superior. Compared to untreated soymilk, the addition of a certain amount of enzyme modified soymilk resulted in a new protein structure, which would improve the texture of blend cheese.