Binding of αvβ1 and αvβ6 integrins to tenascin-C induces epithelial-mesenchymal transition-like change of breast cancer cells.
Tenascin-C (TNC), a large hexameric extracellular glycoprotein, is a pleiotropic molecule with multiple domains binding to a variety of receptors mediating a wide range of cellular functions. We earlier reported that TNC induces epithelial-mesenchymal transition (EMT)-like change in breast cancer cells. In the present study, we clarified TNC receptor involvement in this process. Among integrins previously reported as TNC receptors, substantial expression of αv, α2, β1 and β6 subunits was detected by quantitative PCR and immunoblotting in MCF-7 cells. Integrin β6 mRNA was remarkably upregulated by transforming growth factor (TGF)-β1 treatment, and protein expression was prominently increased by additional exposure to TNC. Immunofluorescent labeling demonstrated integrin αvβ6 accumulation in focal adhesions after TNC treatment, especially in combination with TGF-β1. The α2 and β1 subunits were mainly localized at cell-cell contacts, αv being found near cell cluster surfaces. Immunoprecipitation showed increase in αvβ1 heterodimers, but not α2β1, after TNC treatment. Activated β1 subunits detected by an antibody against the Ca(2+)-dependent epitope colocalized with αv in focal adhesion complexes, associated with FAK phosphorylation at tyrosine 925. Neutralizing antibodies against αv and β1 blocked EMT-like change caused by TNC alone. In addition, anti-αv and combined treatment with anti-β1 and anti-αvβ6 inhibited TGF-β1/TNC-induced EMT, whereas either of these alone did not. Integrin subunits αv, β1 and β6, but not α2, bound to TNC immobilized on agarose beads in a divalent cation-dependent manner. Treatments with neutralizing antibodies against β1 and αvβ6 reduced αv subunit bound to the beads. Immunohistochemistry of these receptors in human breast cancer tissues demonstrated frequent expression of β6 subunits in cancer cells forming scattered nests localized in TNC-rich stroma. These findings provide direct evidence that binding of αvβ6 and αvβ1 integrins to TNC as their essential ligand induces EMT-like change in breast cancer cells.


["Hepar"--the clinical computer system and its diagnostic and didactic use].
The possibilities of application of the "Hepar" computer system in medical didactics are discussed. The usefulness of hepatological diagnosis assistance with information++ systems is described and the multistage schema of diagnostic management is presented. In the conclusions the advantages of this system in medicine teaching is stressed and its usefulness in clinical practice is outlined.


Association between prehospital shock index and mortality among patients with COVID-19 disease.
There exists a need for prognostic tools for the early identification of COVID-19 patients requiring intensive care unit (ICU) admission and mortality. Here we investigated the association between a clinical (initial prehospital shock index (SI)) and biological (initial prehospital lactatemia) tool and the ICU admission and 30-day mortality among COVID-19 patients cared for in the prehospital setting. We retrospectively analysed COVID-19 patients initially cared for by a Paris Fire Brigade advanced (ALS) or basic life support (BLS) team in the prehospital setting between 2020, March 08th and 2020, May 30th. We assessed the association between prehospital SI and prehospital lactatemia and ICU admission and mortality using logistic regression model analysis after propensity score matching with Inverse Probability Treatment Weighting (IPTW) method. Covariates included in the IPTW propensity analysis were: age, sex, body mass index (BMI), initial respiratory rate (iRR), initial pulse oximetry without (SpO2i) and with oxygen supplementation (SpO2i.O2), initial Glasgow coma scale (GCSi) value, initial prehospital SI and initial prehospital lactatemia. We analysed 410 consecutive COVID-19 patients [254 males (62%); mean age, 64 ± 18 years]. Fifty-seven patients (14%) deceased on the scene, of whom 41 (72%) were male and were significantly older (71 ± 12 years vs. 64 ± 19 years; P 〈10-3). Fifty-three patients (15%) were admitted in ICU and 39 patients (11%) were deceased on day-30. The mean prehospital SI value was 1.5 ± 0.4 and the mean prehospital lactatemia was 2.0 ± 1.7 mmol.l-1. Multivariate logistic regression analysis on matched population after IPTW propensity analysis reported a significant association between ICU admission and age (adjusted Odd-Ratio (aOR), 0.90; 95% confidence interval (95%CI): 0.93-0.98;p = 10-3), SpO2i.O2 (aOR, 1.10; 95%CI: 1.02-1.20;p = 0.002) and BMI (aOR, 1.09; 95% CI: 1.03-1.16;p = 0.02). 30-day mortality was significantly associated with SpO2i.O2 (aOR, 0.92; 95% CI: 0.87-0.98;p = 0.01 P < 10-3) and GCSi (aOR, 0.90; 95% CI: 0.82-0.99;p = 0.04). Neither prehospital SI nor prehospital lactatemia were associated with ICU admission and 30-day mortality. Neither prehospital initial SI nor lactatemia were associated with ICU admission and 30-day mortality among COVID-19 patients initially cared for by a Paris Fire Brigade BLS or ALS team. Further prospective studies are needed to confirm these preliminary results.


Treatment with oestrogen-receptor agonists or oxytocin in conjunction with exercise protects against myocardial infarction in ovariectomized rats.
What is the central question of this study? Could the activation of oxytocin or oestrogen receptors be protective against myocardial injury after ovariectomy? If so, would exercising have an additional ameliorating effect? What is the main finding and its importance? The results revealed that when accompanied by exercise, both oestrogen receptor agonists and oxytocin improved cardiac dysfunction, inhibited the generation of pro-inflammatory cytokines and reduced myocardial injury in ovariectomized female rats, suggesting a new approach for protecting postmenopausal women against ischaemia-induced myocardial injury. To investigate the putative protective effects of oxytocin or oestrogen receptor agonists against myocardial injury of ovariectomized sedentary or exercised rats, female Sprague-Dawley rats assigned to sham-operated control and ovariectomized (OVX) groups were kept sedentary or undertook swimming exercise for 4 weeks and were treated with saline, an oestrogen receptor (ER) β (DPN) or ERα agonist (PPT) or oxytocin. Ovariectomy increased weight gain and anxiety in sedentary rats, whereas exercise prevented weight gain. When accompanied by exercise, both ER agonists and oxytocin inhibited weight gain and anxiety; oxytocin, in the absence or presence of exercise, increased the left ventricular diastolic dimensions and ejection fraction, whereas ER agonists also increased left ventricular diameter when given to exercised rats. Upon the induction of myocardial ischaemia-reperfusion in the OVX rats, plasma creatine kinase-(muscle-brain) was depressed by PPT and oxytocin, whereas DPN, PPT and OT reduced plasminogen activator inhibitor-1 concentrations. The increased tumour necrosis factor-α concentration in OVX rats was also suppressed by exercise or DPN, PPT or oxytocin treatments, whereas the interleukin-6 concentration was diminished by all the treatments when given in conjunction with exercise. Disorganization of cardiac muscle fibres was reduced in all exercised rats. Oestrogen receptor agonists, as well as oxytocin, in conjunction with exercise may be effective new therapeutics to protect against myocardial ischaemia in postmenopausal women.


Interaction of iodinated human [D-Ala2]beta-endorphin with opitate receptors.
The interaction of beta-endorphin with opiate receptors was studied by using the radioiodinated, metabolically stable D-Ala2 derivative of human beta-endorphin. This analog binds specifically to rat brain membrane preparations with an apparent Kd of about 2.5 x 10-9 M. The ability of various enkephalin analogs, as well as opiate agonists and antagonists, to inhibit the binding of beta-endorphin clearly demonstrates that this peptide can bind to opiate receptors. However, the effects of various cations on the binding of 125I-[D-Ala2]beta-endorphin are markedly different from those found for enkephalin binding. Sodium ion at physiological concentrations decreases substantially the binding of enkephalins but only slightly decreases endorphin binding, whereas manganese enhances enkephalin binding but has no effect on endorphin binding. Moreover, potassium (100 mM) decreases the binding of beta-endorphin but does not affect enkephalin binding. These results suggest that beta-endorphin and enkephalin bind differently to the same receptor or bind to different receptors with overlapping specificity.


A Hyaluronic Acid-Based Formulation with Simultaneous Local Drug Delivery and Antioxidant Ability for Active Viscosupplementation.
Hyaluronic acid (HA) and its derivatives are widely used for intra-articular injection to augment compromised viscoelastic properties of damaged synovial fluid. Combining HA-based devices with anti-inflammatory drugs or bioactive principles in order to provide an additional benefit to the viscosupplementation is emerging as a new promising approach to improve the clinical outcome. Here, we aim to design a novel active viscosupplementation agent that can load and release hydrophobic drugs and at the same time possessing antioxidant properties. Optimized ternary systems named HCV based on HA, (2-hydroxypropyl)-β-cyclodextrin (CD), and vitamin E (VE), without being engaged in formal chemical bonding with each other, showed the best viscoelastic and lubrication properties along with antioxidant capabilities, able to solubilize and release DF. The physical-chemical characterization suggested that the HCV system displayed rheological synergism and higher thermal stability because of the presence of VE and its antioxidant activity, and the loading of hydrophobic drugs was improved by the presence of CD and VE. Cell morphology and viability tests on L929 cells exhibited high biocompatibility of the HCV system with higher level expression of anti-inflammatory interleukin-10.


Evaluation of immune function in mice exposed to Ordram.
The potential effects that the thiocarbamate herbicide Ordram has on the immune system of mice was evaluated following 12 days of acute dosing by oral gavage. Dosages of Ordram ranging from 20 to 320 mg/kg/day had no consistent significant effects on a variety of immune parameters investigated. The immune parameters measured were the following: body and lymphoid organ weights; splenic natural killer (NK) cell activity; lymphoproliferative responses to B and T lymphocyte mitogens and allogeneic spleen cells in a one-way mixed lymphocyte reaction; and delayed-type hypersensitivity and antibody responses to sheep red blood cells (SRBC). The effects that the immunosuppressant cyclophosphamide has on these immune parameters was also examined. The results indicate that Ordram does not appear to affect key parameters of the immune system of mice under the conditions of exposure employed.


Granular cell ameloblastoma: enigmatic cells in an unusual neoplasm.
An unusual granular variant of ameloblastoma presenting as a mandibular mass in a 43-year-old woman is described. These visually striking tumors display unusual and inconsistent immunohistochemical staining patterns although differential diagnosis from other granular cell lesions of the head and neck is usually not problematic.


A transcannula method for subcortical HRP gel implants: inferior olive afferents in the rat.
A technique for using horseradish peroxidase in a polyacrylamide gel form for subcortical implants is described. In order to prevent uptake along the injection tract, the solid HRP gel pellet is delivered through a pre-implanted cannula to the target site. The appreciable advantages of using the HRP-transcannula technique are discussed in conjunction with a description of the afferents to the inferior olivary complex in the rat. The principal sources of afferents were the ipsilateral prerubral parafascicular nucleus, contralateral lateral cerebellar nucleus and dorsal column nuclei.


Development and applications of solid-phase extraction and liquid chromatography-mass spectrometry methods for quantification of microcystins in urine, plasma, and serum.
The protocols for solid-phase extraction (SPE) of six microcystins (MCs; MC-LR, MC-RR, MC-LA, MC-LF, MC-LW, and MC-YR) from mouse urine, mouse plasma, and human serum are reported. The quantification of those MCs in biofluids was achieved using HPLC-orbitrap-MS in selected-ion monitoring (SIM) mode, and MCs in urine samples were also quantified by ultra-HPLC-triple quadrupole-tandem mass spectrometry (UHPLC-QqQ-MS/MS) in multiple reaction monitoring (MRM) mode. Under optimal conditions, the extraction recoveries of MCs from samples spiked at two different concentrations (1 μg/L and 10 μg/L) ranged from 90.4% to 104.3% with relative standard deviations (RSDs) ≤ 4.7% for mouse urine, 90.4-106.9% with RSDs ≤ 6.3% for mouse plasma, and 90.0-104.8% with RSDs ≤ 5.0% for human serum. Matrix-matched internal standard calibration curves were linear with R2 ≥ 0.9950 for MC-LR, MC-RR and MC-YR, and R2 ≥ 0.9883 for MC-LA, MC-LF, and MC-LW. The limits of quantification (LOQs) in spiked urine samples were ∼0.13 μg/L for MC-LR, MC-RR, and MC-YR, and ∼0.50 μg/L for MC-LA, MC-LF, and MC-LW, while the LOQs in spiked plasma and serum were ∼0.25 μg/L for MC-LR, MC-RR, and MC-YR, and ∼1.00 μg/L for MC-LA, MC-LF, and MC-LW. The developed methods were applied in a proof-of-concept study to quantify urinary and blood concentrations of MC-LR after oral administration to mice. The urine of mice administered 50 μg of MC-LR per kg bodyweight contained on average 1.30 μg/L of MC-LR (n = 8), while mice administered 100 μg of MC-LR per kg bodyweight had average MC-LR concentration of 2.82 μg/L (n = 8). MC-LR was also quantified in the plasma of the same mice. The results showed that increased MC-LR dosage led to larger urinary and plasma MC-LR concentrations and the developed methods were effective for the quantification of MCs in mouse biofluids.


Antibiotic supplement in feed can perturb the intestinal microbial composition and function in Pacific white shrimp.
The intestinal microbiota plays crucial roles in host health. The Pacific white shrimp is one of the most profitable aquaculture species in the world. Antibiotic supplement in feed is an optional practice to treat shrimp bacterial diseases. However, little is known about antibiotic effects on intestinal microbiota in pacific white shrimp. Here, shrimps were given feed supplemented with ciprofloxacin (Cip) (40 and 80 mg kg-1) and sulfonamide (Sul) (200 and 400 mg kg-1) to investigate the microbial community by targeting the V4 region of 16S rRNA genes. Within 4 days after feeding with normal feed and with antibiotics, antibiotic concentrations of Cip and Sul groups in the intestine dropped sharply. Significantly, increased abundance of antibiotic resistance genes (ARGs) of ciprofloxacin (qnrB, qnrD, and qnrS) and sulfonamide (sul1, sul2, and sul3) was observed in Cip and Sul groups (P < 0.05). A total of 3191 operational taxonomic units (OTUs) were obtained and 41 phyla were identified from 63 samples in shrimp intestine. The numbers of OTUs and Shannon index decreased rapidly at day 1 (the first day after feeding with antibiotics) and increased at day 3 (the third day after feeding with antibiotics). The relative abundance of dominant phyla and genera in Cip and Sul groups were significantly different from that in the control group (Ctrl). Furthermore, functional potentials that were related to amino acid metabolism, carbohydrate metabolism, and cellular processes and signaling varied significantly in Cip and Sul groups. These results point to an antibiotic-induced shift in shrimp intestinal microbiota, which highlights the importance of considering the microbiota in shrimp health management.


S100A4 gene silencing in oxygen-induced ischemic retinopathy inhibits retinal neovascularization via down-regulation of CREB expression.
To investigate the possible role of S100A4 in retinal neovascularization (RNV), as well as the underlying mechanism, in a mouse model of oxygen-induced retinopathy (OIR). Retinas used in the experiments were obtained from a mouse model of OIR with and without treatment by intravitreal injection of adnoviral-S100A4-RNAi or adenoviral-green fluorescent protein at postnatal day 12 (P12). At P17, the efficacy of adenoviral gene transfer was assessed with fluorescence microscopy. RNV was evaluated by whole-mount immunofluorescent staining of the mouse retina with Griffonia simplicifolia isolectin B4 conjugated to Alexa Fluor 594 and quantified by counting the number of pre-retinal neovascular cells. Protein and mRNA expression levels of S100A4, cyclic AMP (cAMP) response element-binding protein (CREB), B-cell lymphoma 2 (Bcl-2), and caspase-3 were determined using Western blot analysis and real-time PCR. Retinal S100A4 levels were positively correlated with the progression of RNV. In the intravitreal Ad-S100A4-RNAi transfer mice, both S100A4 protein and mRNA expression levels in the retina were significantly decreased at P17. Ad-S100A4-RNAi transfer was clearly demonstrated by the green fluorescent protein (GFP) in many layers of the retina, including the ganglion cell layer (GCL), inner plexiform layer (IPL), inner nuclear layer (INL), outer plexiform layer (OPL), and outer nuclear layer (ONL), 3 days after intravitreal injection. Whole-mount immunofluorescent staining of the retina and quantification of the pre-retinal neovascular cells demonstrated that RNV was significantly ameliorated. At the same time, the expression of CREB and Bcl-2 were significantly decreased at the transcriptional and translational levels in the OIR-S100A4 group, whereas caspase-3 expression was increased. Our results indicated that RNV was ameliorated by Ad-S100A4-RNAi transfer in a mouse model of OIR through mediation of the anti-apoptotic effect of Bcl-2 by reducing the expression of CREB, and that S100A4 may be a novel therapeutic target for ocular neovascularization diseases.


Is Sadness Only One Emotion? Psychological and Physiological Responses to Sadness Induced by Two Different Situations: "Loss of Someone" and "Failure to Achieve a Goal".
We investigated whether sadness elicited by two different situations-loss of someone (loss) and failure to achieve a goal (failure)-had different physiological responses. Seventy-four participants were randomly assigned to one of three conditions (loss, failure, and neutral). Physiological responses were recorded during an imagery task that was designed to evoke sadness. The results of characteristics in the subjective ratings indicated that loss-evoked sadness was only associated with expressive words relating to tears. For the results of physiological measures, skin conductance levels (SCLs) increased during the imagery task across all conditions and differed between conditions during the post-task. For the loss condition, restoration to baseline level took longer, while in the failure the SCL decreased sharply back to baseline. Furthermore, tear ratings correlated with blood pressures in the loss condition, while sadness intensity correlated with blood pressures in the failure condition. These results suggest that sadness includes at least two subtypes that produce different responses in subjective ratings and physiological measures.


Alteration in methylation pattern of GATA-4 promoter region in vitamin A-deficient offspring's heart.
Epigenetics might explain correlations between lifestyle and risk of disease. Maternal diet has been shown to dynamically alter epigenetic regulation, including affecting DNA methylation status. This study was designed to test the hypothesis that GATA-4 gene methylation would lead to congenital heart defects in vitamin A-deficient offspring. Ten weaning female rats (VAN group) were fed with a diet which contents 4 IU vitamin A/g diet, while 20 rats (VAD group) were maintained on a diet without vitamin A. After 10 weeks of feeding, all the female rats were mated with normal male rats. The VAN group and a portion of VAD group rats were still given the same diet as before mating, while the rest of the rats from the VAD group (VADS group) were transferred to a diet with enough added vitamin A (10 IU/g diet) for the pregnancy cycle. The embryo hearts were dissected out at embryonic day 13.5 (E13.5) for observation of cardiac development, GATA-4 gene methylation status and the expression of DNA methyltransferases (DNMTs). Embryos from vitamin A-deficient group exhibited a high incidence of cardiac defects. High methylation was present in the CpG loci of GATA-4 gene with a low expression of GATA-4 mRNA from vitamin A-deficient group embryos. Moreover, up-regulation of DNMT1 and down-regulation of DNMT3a and DNMT3b expression were found in this group embryo. These findings show that aberrant methylation is one of key mechanisms to heart defects in vitamin A-deficient offspring. DNMTs play a critical role in this process.


Targeting oxidative stress in surgery: effects of ageing and therapy.
In the current era cardiac surgeons are being called upon to operate upon older, sicker patients. The effect is to augment oxidative stress and increase the rate of post-operative complications and ultimately mortality. We have developed antioxidant-based pre-treatment regimes initially based on coenzyme Q(10). A randomised trial of coenzyme Q(10) in elective cardiac surgery patients demonstrated augmented plasma and cardiac mitochondrial membrane coenzyme Q(10) content, improved mitochondrial respiration and increased myocardial tolerance of oxidative stress. The addition of omega-3 polyunsaturated fatty acids, alpha-lipoic acid, selenium and magnesium orotate in a second clinical trial, improved post-operative recovery with demonstrable reductions in myocardial damage, rate of atrial fibrillation and length of hospital stay. Finally we performed a pilot study of this combined metabolic therapy regimen to which we added preoperative physical exercise and mental stress reduction with indications of further improvements in post-operative recovery. We conclude that simultaneously targeting a number of key deficiencies with a metabolic formulation prior to surgery results in peri- and post-operative clinical and economic benefits.


Spectra of central nervous system melioidosis.
Burkholderia pseudomallei infection of the central nervous system (CNS) is rare with less than 50 cases reported over the last 30 years. The retrospective melioidosis study at University Malaya Medical Centre has documented three cases of CNS melioidosis out of more than 160 cases of melioidosis since 1978. There were two patients with brain abscess and one with spinal epidural abscess. The predisposing factors were: one patient was an aboriginal farmer and the other two were diabetic. Their age ranged from 17 to 45 years. Prominent neurological features were limb weakness, cranial nerve palsy (6th and 7th) and visual disturbance. CT brain scan and MRI spine showed abscess formation, subdural collection, and spinal epidural collection, osteomyelitis of vertebra and occipital bone and also sagital sinus thrombosis. All these patients underwent surgical drainage leading to bacteriological diagnosis as well as appropriate long-term antibiotic therapy. All had good recovery at 6 months after completion of treatment.


Contribution of iron and protein contents from rat brain subcellular fractions to MR phase imaging.
Investigation of magnetic susceptibility and chemical exchange as sources of MRI phase contrast between gray and white matter resulting from protein and iron content from subcellular fractions. This study analyzes the iron and macromolecule content of different subcellular fractions from rat brain and their relation to the water-resonance frequency by NMR spectroscopy. Additionally, the contributions of susceptibility and exchange were determined with different NMR reference substances. Only weak correlations between iron (r = 0.4318, P = 0.76) or protein content (r = 0.4704, P = 0.70) and frequency shift were observed. After membrane depletion, the correlation for iron increased to r = -0.9006 (P = 0.0009), whereas the shift relative to protein content increased much less (r = -0.4982, P = 0.64). Exchange-driven frequency shifts were 1.283 ppb/(mg/ml) for myelin and 0.775 ppb/(mg/ml) for synaptosomes; susceptibility-driven shifts were -1.209 ppb/(mg/ml) for myelin and -0.368 ppb/(mg/ml) for synaptosomes. The ratios between susceptibility and exchange differ significantly from simple protein solutions. As a result of counteracting susceptibility and exchange and increased relative shifts in membrane-depleted fractions, we conclude that tissue microstructure accounts more for the in vivo phase contrast than in the situation of homogenized tissue. Thus, membranes may generate much of the in vivo MR phase contrast resulting from anisotropy. Magn Reson Med 77:2028-2039, 2017. © 2016 International Society for Magnetic Resonance in Medicine.


Heat and moisture exchangers and heated humidifiers in acute lung injury/acute respiratory distress syndrome patients. Effects on respiratory mechanics and gas exchange.
To compare, in acute lung injury/acute respiratory distress syndrome (ALI/ARDS) patients, the short-term effects of heat and moisture exchangers (HME) and heated humidifiers (HH) on gas exchange, and also on respiratory system mechanics when isocapnic conditions are met. Prospective open clinical study. Intensive Care Service. Seventeen invasively ventilated ALI/ARDS patients. The study was performed in three phases: (1) determinations were made during basal ventilatory settings with HME; (2) basal ventilatory settings were maintained and HME was replaced by an HH; (3) using the same HH, tidal volume (Vt) was decreased until basal PaCO2 levels were reached. FiO2, respiratory rate and PEEP were kept unchanged. Respiratory mechanics, Vdphys, gas exchange and hemodynamic parameters were obtained at each phase. By using HH instead of HME and without changing Vt, PaCO2 decreased from 46+/-9 to 40+/-8 mmHg (p<0.001) and Vdphys decreased from 352+/-63 to 310+/-74 ml (p<0.001). Comparing the first phase with the third, Vt decreased from 521+/-106 to 440+/-118 ml (p<0.001) without significant changes in PaCO2, Vd/Vt decreased from 0.69+/-0.11 to 0.62+/-0.12 (p<0.001), plateau airway pressure decreased from 25+/-6 to 21+/-6 cmH2O (p<0.001) and respiratory system compliance improved from 35+/-12 to 42+/-15 ml/cmH2O (p<0.001). PaO2 remained unchanged in the three phases. Reducing dead space with the use of HH decreases PaCO2 and more importantly, if isocapnic conditions are maintained by reducing Vt, this strategy improves respiratory system compliance and reduces plateau airway pressure.


Comparing the Health Workforce Provider Mix and the Distance Travelled for Mental Health Services by Rural and Urban Medicare Beneficiaries.
To describe the mix of health professionals who care for rural and urban seniors suffering from mood and/or anxiety disorders, the quantity of services they receive, and to understand where beneficiaries receive care for mood and/or anxiety disorders and the distance and time they travel for care. We used 2014 Medicare administrative claims data to examine access to health care for fee-for-service Medicare beneficiaries aged ≥ 65 years who received outpatient services for mood and anxiety disorders. We classified providers into 9 categories: (1) family physicians/general practice, (2) internists, (3) nurse practitioners (NPs) and physician assistants (PAs), (4) psychiatrists, (5) psychologists, (6) clinical social workers, (7) emergency medicine physicians, (8) other physicians, and (9) other providers. We calculated the 1-way driving distance and travel time between the beneficiary residence and provider location. We classified beneficiaries into 1 of 4 geographic categories based on their residence ZIP Code. Urban beneficiaries had an average of 2.7 visits for mood and anxiety disorders, while rural beneficiaries had 2.4. Generalist physicians and NPs/PAs provided 50.8% of all visits. Urban beneficiaries saw more behavioral health specialists (34.3%) than rural beneficiaries (16.1%). NPs and PAs provided more than twice as much of the care for rural beneficiaries (14.8%) as for urban beneficiaries (6.4%). Rural beneficiaries travelled about twice as far as urban beneficiaries. Rural and urban Medicare beneficiaries received care for mood/anxiety disorders from different mixes of health care providers, and ensuring access for rural populations will require innovative solutions.


Repeatability of Junctional Zone Measurements Using 3-Dimensional Transvaginal Sonography in Healthy Fertile Women.
To describe the junctional zone and determine the intraobserver and interobserver repeatability of junctional zone measurements using 3-dimensional (3D) transvaginal sonography in healthy fertile women. We examined 82 consecutive women with 3D transvaginal sonography. The maximum and minimum junctional zone thickness was measured in all uterine walls. The difference between maximum and minimum thickness and average measurements (maximum thickness + minimum thickness/2) of the anterior, posterior, fundal, and lateral walls were calculated. Among the first 40 consecutive women, intraobserver and interobserver repeatability was evaluated according to the Bland-Altman method and expressed as a coefficient of repeatability. Using 3D transvaginal sonography, we visualized a thin and regular junctional zone in most women. The posterior uterine wall had the largest median maximum junctional zone thickness value of 5.2 (interquartile range, 3.8-6.5) mm. Ten women (12%) had maximum thickness values of 8.0 to 12.0 mm. The maximum thickness in each uterine wall had intraobserver and interobserver coefficients of repeatability of ±2.1 to ±3.4 and ±2.6 to ±3.9 mm, respectively, which were reduced by average measurements: ±1.9 and ±2.0 mm (anterior and posterior walls) and ±1.5 mm (fundal and lateral walls) for intraobserver and interobserver values. Correlations between measurements were poor in the narrow range of junctional zone thickness. The junctional zone has an indistinct outline on 3D transvaginal sonography, resulting in measurement errors within a broad range of ±2 to ±4 mm, which were only reduced to some extent by average measurements. The thickness of the junctional zone varied within a narrow range in this healthy fertile population, and reliability measurements of junctional zone thickness have to be evaluated in women with a wider range of thickness. The observer repeatability and reliability of junctional zone measurements need to be further evaluated and refined before applying this method in clinical practice.


Inverted papillomas of nasal cavity and paranasal sinuses. Ultrastructural investigations on epithelial-stromal interface.
10 cases of inverted papillomas of nasal cavity and paranasal sinuses were examined electron microscopically with particular regard to the epithelial-stromal interface. The papilloma cells were clearly demarcated from the stromal tissue by a basement membrane-like material. However, this structure mainly consisted of two or more layers, occasional breaks in the basement membrane were visible. Within the basal epithelial cells resting on the basement membrane accumulations of actin-like microfilaments could sometimes be observed. Furthermore, in the stromal tissue some myofibroblasts were present in the vicinity of the papillomatous cell complexes and capillary vessels also demonstrated several layers of basement membrane. All the changes presented here are considered to be the result of a permanent interaction between aggressive forces of papilloma cells and the defence mechanism of stromal tissue. The results allow the conclusion that inverted papillomas of nasal cavity and paranasal sinuses are true neoplasms with very low malignancy.


Role of RPTPβ/ζ in neuroinflammation and microglia-neuron communication.
Pleiotrophin (PTN) is a cytokine that is upregulated in different neuroinflammatory disorders. Using mice with transgenic PTN overexpression in the brain (Ptn-Tg), we have found a positive correlation between iNos and Tnfα mRNA and Ptn mRNA levels in the prefrontal cortex (PFC) of LPS-treated mice. PTN is an inhibitor of Receptor Protein Tyrosine Phosphatase (RPTP) β/ζ, which is mainly expressed in the central nervous system. We aimed to test if RPTPβ/ζ is involved in the modulation of neuroinflammatory responses using specific inhibitors of RPTPβ/ζ (MY10 and MY33-3). Treatment with MY10 potentiated LPS-induced microglial responses in the mouse PFC. Surprisingly, MY10 caused a decrease in LPS-induced NF-κB p65 expression, suggesting that RPTPβ/ζ may be involved in a novel mechanism of potentiation of microglial activation independent of the NF-κB p65 pathway. MY33-3 and MY10 limited LPS-induced nitrites production and iNos increases in BV2 microglial cells. SH-SY5Y neuronal cells were treated with the conditioned media from MY10/LPS-treated BV2 cells. Conditioned media from non-stimulated and from LPS-stimulated BV2 cells increased the viability of SH-SY5Y cultures. RPTPβ/ζ inhibition in microglial cells disrupted this neurotrophic effect of microglia, suggesting that RPTPβ/ζ plays a role in the neurotrophic phenotype of microglia and in microglia-neuron communication.


Ascaris lumbricoides suum: morphological characterization of apparent cuticular pores by ionic permeability and electron microscopy.
The cuticle of the parasitic roundworm Ascaris lumbricoides suum has been found to contain apparent pores, using scanning and transmission electron microscopy. However, X-ray spectral analysis and dot mapping analysis of diffusing divalent cations in the cuticle found these ions to be randomly distributed on the pseudocoelomic surface of the cuticle. This indicates that the pores seen with the electron microscope were not true structural pores and that the cuticle is homogeneously permeable to ions.


Surgical Techniques for the Correction of Postrhinoplasty Depressed Scars on the Nasal Tip.
Augmentation rhinoplasty is common in the Asian population and the lack of suitable autologous material for augmentation has led to the use of alloplastic materials. Many of these patients develop complications, including a depressed dimple-like scar of the nasal tip. Causes of such dimpling include the use of large implants, infection, extrusion, and surgeon incompetence. To describe the various techniques that can be used to correct dimpling of the nasal tip. Data were retrospectively reviewed from 28 patients who had undergone surgical procedures from January 1, 2013, through July 31, 2014, in a rhinoplasty clinic in Seoul, Korea, for the correction of a contracted nose with nasal scars secondary to previous rhinoplasties. Data analysis was conducted from August 1, 2014, through February 16, 2015. Before surgery, a complete rhinological examination was conducted, digital photographs were taken, and an assessment was made regarding the severity of the dimpling, the condition of the nasal skin, and the underlying supporting structures. Based on these factors, the appropriate type of procedure was planned. Patient satisfaction with change after surgery was assessed using a 3-point Likert scale (3 indicates satisfied; 2, fairly satisfied; and 1, dissatisfied). Outcomes were also reviewed by 2 surgeon-peers who gave an outcome score ranging from 1 to 10 (1 indicates a poor cosmetic outcome; 10, the best possible outcome). The nasal contour and tip symmetry were restored to aesthetic standards with these relatively simple techniques. Eleven patients (39.2%) were treated with unilobed flap, 4 (14.2%) with a bilobed flap, 3 (10.7%) with Z-plasty, 9 (32.1%) with soft-tissue interposition, and 1 (3.5%) with a transposition flap. Twenty-four of the 28 patients (85.7%) were satisfied with their outcomes and 4 (14.3%) patients were dissatisfied and were given a revision procedure, following which they were satisfied with their outcomes. The follow-up period ranged between 6 to 32 months (mean, 12.3 months). The mean surgeon-reviewed outcome score for soft-tissue interposition procedure was 8.0 of 10; for the unilobed flap, 8.0 of 10; for the bilobed flap, 7.5 of 10; for the transposition flap, 8.0 of 10; and for Z-plasty, 7.8 of 10. These techniques are reproducible and the choice of the technique depends on the shape, size, and location of the scar; skin condition; patient expectations; and the surgeon's experience and comfort level with the procedure. 4.


Morphofunctional Changes in the Spinal Cord of Rats after Contusion Injury with Local Delivery of Methylprednisolone in Combination with a Copolymer.
We studied the neuroprotective effect of local application of methylprednisolone in combination with a block copolymer after contusion spinal cord injury in rats. Histological analysis of the spinal cord showed that delivery of a complex of methylprednisolone with a block copolymer reduced the volume of white and gray matter lesions. An increase in the amplitude of the evoked response of the gastrocnemius muscle was observed during epidural stimulation of the spinal cord 6 h after the injury. The maximum amplitude of the muscle response was greater in the group with local delivery of the methylprednisolone complex with the polymer 72 h after the injury. The obtained results demonstrate the neuroprotective effect of the local administration of the complex and allow to make positive prognosis for the recovery of the sensorimotor functions in rats.


Porous titanium oxynitride sheets as electrochemical electrodes for energy storage.
A flexible, porous TiOxNy sheet consisting of numerous conductive fibers was synthesized by nitridation of titanate and further used as an electrochemical electrode. The high surface area and mixed-oxidation state of titanium make TiOxNy sheets to be promising candidates for a good supercapacitor.


Genetic diversity in natural populations: a fundamental component of plant-microbe interactions.
Genetic diversity for plant defense against microbial pathogens has been studied either by analyzing sequences of defense genes or by testing phenotypic responses to pathogens under experimental conditions. These two approaches give different but complementary information but, till date, only rare attempts at their integration have been made. Here we discuss the advances made, because of the two approaches, in understanding plant-pathogen coevolution and propose ways of integrating the two.


The development of plate osteosynthesis for the treatment of fractures of the mandibular body - a literature review.
Today plate and screw osteosynthesis of mandibular fractures is a standard procedure in routine clinical practice. In this review, the breakthroughs and drawbacks of the development of this important aspect of maxillofacial surgery are followed-up. Medline search of relevant English and German literature. In 1886, Carl Hansmann was the first who applied steel screws and plates. Until today the material, the types of plates and applications have been continually improved. Over the last two decades miniplate osteosynthesis has induced a revolution in mandibular fracture treatment. The modern systems provide better handling, higher stability and less pressure on the bone. Modern miniplates have great advantages, like the intra-oral approach and the easy adaptability. In addition, it is no longer necessary to expose bone as extensively.


Relapsed Acute Lymphoblastic Leukemia.
Outcomes for children with acute lymphoblastic leukemia (ALL) have improved worldwide to >85%. For those who relapse, outcomes have remained static at ~50% making relapsed acute lymphoblastic leukemia one of the leading causes of death in childhood cancers. Those relapsing within 18 mo in the bone marrow have a particularly dismal outcome. The mainstay of treatment is chemotherapy, local radiotherapy with or without hematopoietic stem cell transplantation (HSCT). Improved biological understanding of mechanisms of relapse and drug resistance, use of innovative strategies to identify the most effective and least toxic treatment regimens and global partnerships are needed to improve outcomes in these patients. Over the last decade, new therapeutic options and strategies have been developed for relapsed ALL including immunotherapies and cellular therapies. It is imperative to understand how and when to use these newer approaches in relapsed ALL. Increasingly, integrated precision oncology strategies are being used to individualize treatment of patients with relapsed ALL, especially in patients with poor response disease.


Systemic administration of insulin-like growth factor decreases motor neuron cell death and promotes muscle reinnervation.
Neonatal sciatic nerve axotomy causes motoneuron death and muscle denervation atrophy. The aim of the present study was to determine whether insulin-like growth factor-I (IGF-I) administration promotes muscle reinnervation and counteracts motor neuron loss after such an injury. Six weeks after sciatic nerve axotomy performed in 2-day-old pups, the number of motor neurons, as assessed by retrograde transport of horseradish peroxidase injected into the extensor digitorum longus (EDL) muscle, was reduced from 52 +/- 3 to 26 +/- 3. Subsequent administration of IGF-I at the doses of 0.02 mg/kg or 1 mg/kg increased the number of motor neurons to 35 +/- 2 and 37 +/- 5, respectively. The effect on motoneuron survival was accompanied by improved muscle fibre morphometry and restoration of indirect EDL muscle isometric twitch tension, which was about 80 % of control values for both doses of IGF-I compared with 60% observed with saline treatment. Reinnervated EDL muscle from saline-treated rats cannot hold tetanic tension, which is, however, achieved after IGF-I treatment at either dose. Thus, both high and low doses of IGF-I counteracted motoneuron death and improved muscle reinnervation following neonatal sciatic nerve axotomy. IGF-I at 5 microg/kg failed to increase muscle reinnervation.


PEGylated polylysine dendrimers increase lymphatic exposure to doxorubicin when compared to PEGylated liposomal and solution formulations of doxorubicin.
Improved delivery of chemotherapeutic drugs to the lymphatic system has the potential to augment outcomes for cancer therapy by enhancing activity against lymph node metastases. Uptake of small molecule chemotherapeutics into the lymphatic system, however, is limited. Nano-sized drug carriers have the potential to promote access to the lymphatics, but to this point, this has not been examined in detail. The current study therefore evaluated the lymphatic exposure of doxorubicin after subcutaneous and intravenous administration as a simple solution formulation or when formulated as a doxorubicin loaded PEGylated poly-lysine dendrimer (hydrodynamic diameter 12 nm), a PEGylated liposome (100 nm) and various pluronic micellar formulations (~5 nm) to thoracic lymph duct cannulated rats. Plasma and lymph pharmacokinetics were analysed by compartmental pharmacokinetic modelling in S-ADAPT, and Berkeley Madonna software was used to predict the lymphatic exposure of doxorubicin over an extended period of time. The micelle formulations displayed poor in vivo stability, resulting in doxorubicin profiles that were similar to that observed after administration of the doxorubicin solution formulation. In contrast, the dendrimer formulation significantly increased the recovery of doxorubicin in the thoracic lymph after both intravenous and subcutaneous dosing when compared to the solution or micellar formulation. Dendrimer-doxorubicin also resulted in increases in lymphatic doxorubicin concentrations when compared to the liposome formulation, although liposomal doxorubicin did increase lymphatic transport when compared to the solution formulation. Specifically, the dendrimer formulation increased the recovery of doxorubicin in the lymph up to 30 h post dose by up to 685 fold and 3.7 fold when compared to the solution and liposomal formulations respectively. Using the compartmental model to predict lymphatic exposure to longer time periods suggested that doxorubicin exposure to the lymphatic system would ultimately be 9796 times and 6.1 times greater after administration of dendrimer doxorubicin when compared to the solution and liposome formulations respectively. The recovery of doxorubicin in the sentinel lymph nodes draining the subcutaneous injection site was also quantified directly, and consistent with the lymph pharmacokinetic data, lymph node recovery was greatest for the dendrimer formulation (12% of dosed doxorubicin/g node) when compared to the liposome (1.4%/g node) and solution (<1%/g node) formulations. The data suggest that dendrimer-based drug delivery systems have the potential to enhance drug exposure to lymph-based drug targets such as lymphatic metastases.


Systematic reviews of complex interventions: framing the review question.
The first and most important decision in preparing any systematic review is to clearly frame the question the review team seeks to answer. However, this is not always straightforward, particularly if synthesis teams are interested in the effects of complex interventions. In this article, we discuss how to formulate good systematic review questions of complex interventions. We describe the rationale for developing well-formulated review questions and review the existing guidance on formulating review questions. We discuss that complex interventions can contain a mix of effective and ineffective (or even harmful) actions, which may interact synergistically or dysynergistically or be interdependent, and how these interactions and interdependencies need to be considered when formulating systematic review questions. We discuss complexity specifically in terms of how it relates to the type of question, the scope of the review (i.e., lumping vs. splitting debate), and specification of the intervention. We offer several recommendations to assist review authors in developing a definition for their complex intervention of interest, which is an essential first step in formulating the review question. We end by identifying areas in which future methodological research aimed at improving question formulation, especially as it relates to complex interventions, is needed.


Single-stranded DNA scanning and deamination by APOBEC3G cytidine deaminase at single molecule resolution.
APOBEC3G (Apo3G) is a single-stranded (ss)DNA cytosine deaminase that eliminates HIV-1 infectivity by converting C → U in numerous small target motifs on the minus viral cDNA. Apo3G deaminates linear ssDNA in vitro with pronounced spatial asymmetry favoring the 3' → 5' direction. A similar polarity observed in vivo is believed responsible for initiating localized C → T mutational gradients that inactivate the virus. When compared with double-stranded (ds)DNA scanning enzymes, e.g. DNA glycosylases that excise rare aberrant bases, there is a paucity of mechanistic studies on ssDNA scanning enzymes. Here, we investigate ssDNA scanning and motif-targeting mechanisms for Apo3G using single molecule Förster resonance energy transfer. We address the specific issue of deamination asymmetry within the general context of ssDNA scanning mechanisms and show that Apo3G scanning trajectories, ssDNA contraction, and deamination efficiencies depend on motif sequence, location, and ionic strength. Notably, we observe the presence of bidirectional quasi-localized scanning of Apo3G occurring proximal to a 5' hot motif, a motif-dependent DNA contraction greatest for 5' hot > 3' hot > 5' cold motifs, and diminished mobility at low salt. We discuss the single molecule Förster resonance energy transfer data in terms of a model in which deamination polarity occurs as a consequence of Apo3G binding to ssDNA in two orientations, one that is catalytically favorable, with the other disfavorable.


Correlation between arginyl residue modification and benzodiazepine binding to human serum albumin.
Human serum albumin (HSA) has been chemically modified with 1,2-cyclohexanedione and N-acetylimidazole under nondenaturing conditions. Derivatives, in which 10 arginine residues (1,2-dihydroxycyclohex-1,2-ylene (DHCH)-HSA), 56 to 57 lysine and 5 tyrosine residues (acetyl-HSA), or 56 to 57 lysine residues alone (O-deacetyl-HSA) are modified, have been isolated. Their conformation has been tested by circular dichroism measurement, gel filtration on Sephadex G-150, and ultracentrifugation. From these analyses and binding studies it is concluded that only insignificant changes of conformation have occurred. The binding properties of the HSA derivatives have been tested with bilirubin, diazepam (a benzodiazepine drug), phenylbutazone, and indomethacin by circular dichroism. The binding of diazepam to DHCH-HSA is almost completely inhibited and that of phenylbutazone and indomethacin decreased, while the binding of bilirubin is essentially unaffected. In acetyl-HSA and O-deacetyl-HSA, the bilirubin binding is significantly decreased, while the binding of the drugs mentioned is less affected. It is concluded that bilirubin and the drugs bind to two separate sites which contain positive charges essential for the binding properties. The charge(s) in the benzodiazepine site from arginine.


Neoadjuvant chemoradiotherapy of rectal carcinoma : Baseline hematologic parameters influencing outcomes.
The link between the blood count and a systemic inflammatory response (SIR) is indisputable and well described. Pretreatment hematological parameters may predict the overall clinical outcomes in many types of cancer. Thus, this study aims to systematically evaluate the relationship between baseline blood count levels and treatment response in rectal cancer patients treated with neoadjuvant chemoradiotherapy. From 2009-2015, 173 patients with locally advanced rectal cancer were retrospectively enrolled in the study and analyzed. The baseline blood count was recorded in all patients 1 week before chemoradiation. Tumor response was evaluated through pathologic findings. Blood count levels which included RBC (red blood cells), Hb (hemoglobin), PLT (platelet count), neutrophil count, WBC (white blood cells), NLR (neutrophil-to-lymphocyte ratio), and PLR (platelet-to-lymphocyte ratio) were analyzed in relation to tumor downstaging, pCR (pathologic complete response), OS (overall survival), and DFS (disease-free survival). Hb levels were associated with a response in logistic regression analysis: pCR (p = 0.05; OR 1.04, 95 % CI 1.00-1.07); T downstaging (p = 0.006; OR 1.03, 95 % CI 1.01-1.05); N downstaging (p = 0.09; OR 1.02, 95 % CI 1.00-1.04); T or N downstaging (p = 0.007; OR 1.04, 95 % CI 1.01-1.07); T and N downstaging (p = 0.02; OR 1.02, 95 % CI 1.00-1.04); Hb and RBC were the most significant parameters influencing OS; PLT was a negative prognostic factor for OS and DFS (p = 0.008 for OS); an NLR value of 2.8 was associated with the greatest significance for OS (p = 0.03) and primary tumor downstaging (p = 0.02). Knowledge of pretreatment hematological parameters appears to be an important prognostic factor in patients with rectal carcinoma.


Effects of adaptive servo-ventilation therapy on cardiac function and remodeling in patients with chronic heart failure (SAVIOR-C): study protocol for a randomized controlled trial.
Adaptive servo-ventilation (ASV) therapy, which is a form of noninvasive positive pressure ventilation therapy and uses an innovative ventilator that has simple operability and provides good patient adherence, potentially has therapeutic benefits-suppression of the deterioration and progression of chronic heart failure (CHF) and a reduction in the number of repeated hospitalizations. Therefore, ASV therapy draws attention as a novel, noninvasive nonpharmacotherapy for patients with CHF owing to its hemodynamics-improving effect, and it is currently being accepted in real-world clinical settings in Japan. However, clinical evidence sufficient for treatment recommendation is lacking because a multicenter, randomized, controlled study of ASV therapy has never been conducted. The present study is a confirmatory, prospective, multicenter, collaborative, open-label, blinded-endpoint, parallel-group, randomized, controlled study. At 40 medical institutions in Japan, 200 Japanese outpatients with mild to severe CHF (age: ≥ 20 years; New York Heart Association classification: greater than or equal to class II) will be randomly assigned to either of the following two study groups: the ASV group, in which 100 outpatients undergo guideline-directed medical therapy and ASV therapy for 24 weeks; and the control group, in which 100 outpatients undergo only guideline-directed medical therapy for 24 weeks. The objective of the present study is to confirm whether the ASV group is superior to the control group concerning the improvement of left ventricular contractility and remodeling, both assessed by two-dimensional echocardiography. Furthermore, the present study will also secondarily examine the effects of ASV therapy on the prognosis and quality of life of patients with CHF. ASV therapy using the device has the potential to provide therapeutic benefits based on its simple operability and good patient adherence and possesses the potential to improve left ventricular contractility and remodeling. Therefore, the present study is expected to afford more solid scientific evidence regarding ASV therapy as a novel, noninvasive, nonpharmacological, in-home, long-term ventilation therapy for patients with mild to severe CHF. UMIN identifier: UMIN000006549 , registered on 17 October, 2011.


Children's Bonding with Parents and Grandparents and Its Associated Factors.
Previous literature has focused on the importance of both parental and grandparental bonding. However, few studies have been conducted to measure children's bonding with parents and grandparents simultaneously, especially tested by the same instrument that offers more comparable results. Therefore, we studied the relationships between parental and grandparental bonding using the Parental Bonding Instrument (PBI), and possible associations between these bonds and sociodemographic variables in 905 Chinese children aged 10-14 years. Children's bonding with mother, father, and grandparents were positively correlated, and the final mixed-effect model showed that several sociodemographic factors (e.g., gender, only children, parents' marital status, and mother's occupation) were associated with parental and grandparental bonding.


Ranging patterns of critically endangered colobine, Presbytis chrysomelas chrysomelas.
Presbytis chrysomelas chrysomelas endemic only in Sarawak and Kalimantan was categorized by IUCN as a critically endangered primate that require special attention from research and conservation perspectives. A qualitative study on ranging patterns of P. c. chrysomelas was conducted in the Samunsam Wildlife Sanctuary, Sarawak. The study was conducted over a period of 13 months from December 2004 to December 2005 with 213 days of observation. Behavioural observation covered 17 groups with special emphasis on two main groups and 1 subadult group. Scanning and focal sampling were employed as the observation methods. Results indicated that P. c. chrysomelas had vertical, straight horizontal, and cross-horizontal types of movement patterns. P. c. chrysomelas was recorded to have a short movement distance (31.8-54.3 m). Distribution, abundance types, and food resources might be the factors that shaped the patterns of movement and distance in P. c. chrysomelas.


A new stereotactic X-ray knife.
For many years, the irradiation of small volumes of tissue in the brain to necrotizing doses has been investigated as a "non-invasive" alternative to neurosurgery. We propose a new system in which a precisely machined helmet serves as a multi-port focussed X-ray collimator when it is itself irradiated by a conventional medical linear accelerator run in the electron mode. When the collimator is attached to a stereotactic frame, the geometric accuracy of delivering small radiation fields to the brain is limited primarily by the accuracy of the stereotactic localization, and is relatively independent of the positional stability of the accelerator. Field sizes as small as two millimeters are readily achievable. The problem of low dose rate associated with these small fields is overcome by the use of high electron beam currents.


Angulation Toward Coronal Convexity Measure and Catheter Length Indication Improves the Quality of Ventricular Catheter Placement-A Smartphone-Assisted Guidance Technique.
Accurate placement of a ventricular catheter (VC) is crucial to reduce the risk of shunt failure. In the randomized, prospective, multicenter GAVCA (guided application of ventricular catheters) trial, which evaluated the quality of VC placement, the subgroup of patients with detailed length-marked VCs (dVCs) reflected a difference in the primary endpoint of optimal VC placement compared with the subgroup of patients with simplified length-marked VCs (sVCs). The objective of the present analysis was to compare the dVC and sVC groups and the smartphone-assisted guidance technique (GA) with the standard freehand technique (FH) for VC placement. We performed a further analysis of the GAVCA trial in 2 steps. First, we compared the dVCs, which provided a detailed distance from the tip to the base (3-13 cm) in 0.5-cm intervals, with the sVCs with a length indication at 5 and 10 cm from the tip to base. Second, we compared the GA technique with the FH in the dVC group. The data from 137 patients (104 dVC patients vs. 33 sVC patients) were eligible for the present analysis. Optimal VC placement was achieved in 72.1% of the dVC group and 39.4% of the sVC group (odds ratio, 3.9; 95% confidence interval, 1.7-9.3; P ≤ 0.001). In addition, we performed a subgroup analysis of the 104 dVC patients concerning the accuracy of catheter placement using 2 different techniques (GA, n = 54; and FH, n = 50). Optimal catheter placement was achieved in 81.5% of the GA group and 62.0% of the FH group (odds ratio, 2.7; 95% confidence interval, 1.1-6.8; P = 0.03). VC placement using the GA technique was successful in all patients at the primary puncture. In contrast, for 8.7% of the patients in the FH group, multiple attempts were necessary (P = 0.03). The results from the present analysis suggest that the combination of a GA technique and the use of a dVC will improve the rate of accurate VC placement. Compared with the FH technique, patient safety was increased by the reduction of unsuccessful VC placement attempts using the GA technique and dVCs.


Effects of strain, sex, route of administration and partial hepatectomy on the induction by chemical carcinogens of gamma-glutamyltranspeptidase foci in rat liver.
The incidence of gamma-glutamyltranspeptidase (GGT)-positive foci induced by 0.3 mmol/kg diethylnitrosamine (DENA) followed by promotion with 500 ppm sodium phenobarbital in drinking water and was the same in Fischer 344, Sprague-Dawley and Wistar-Lewis rats. There was no difference in the level of GGT-foci initiated by DENA, 7,12-dimethylbenz[a]-anthracene (DMBA), or 1,2-dimethylhydrazine (DMH) followed by promotion with phenobarbital with respect to sex or route of administration including gavage and intraperitoneal injection. Maximal stimulation by partial hepatectomy of DENA initiation of GGT-foci occurred when the DENA was administered 18 h after the operation. Our results indicate that the optimal protocol for the rat liver foci assay consists of using partial hepatectomized rats of 1 of the 3 strains and of either sex. The test substance should be administered by either gavage or intraperitoneal injection so that maximal DNA binding coincides with the maximal rate of DNA replication resulting from partial hepatectomy.


Treatment and survival of lymphoid malignancy in the north-west of England: a population-based study.
Classification of lymphoid malignancy has changed markedly in recent years and advances have been made in therapy. This study investigated the variations in treatment and survival of 1622 patients in a population-based registry. A total of 1009 cases of malignant lymphoma (ML) were classified according to the Kiel classification. Pathology review resulted in major diagnostic changes for 24% of cases. Of the ML cases, 39% had not had full staging procedures. Younger patients were more likely to have been treated with multiagent chemotherapy regimens, as were patients with B symptoms. Median survival for ML patients was 12 months for high-grade patients and more than 60 months for low-grade patients. Significant factors affecting the survival of ML patients were performance status, whether treatment had followed a recognised protocol, whether treatment had been carried out at a specialist oncology centre (SOC), grade of disease, stage, gender and age. The same factors had a significant effect on survival of the leukaemia patients, except for treatment at an SOC, which had a significant favourable effect on survival of acute lymphoblastic leukaemia (ALL) patients only. Median survival for patients with chronic lymphocytic leukaemia was 43 months and 7 months for ALL patients.


Quantitative determination of the lateral diffusion coefficients of the hormone-receptor complexes of insulin and epidermal growth factor on the plasma membrane of cultured fibroblasts.
Fluorescent derivatives of insulin and epidermal growth factor bound to 3T3 mouse fibroblasts are mobile on the cell surface, with similar diffusion coefficients, D approximately (3--5) x 10(-10) cm2/sec at 23 degrees C. Increasing the temperature to 37 degrees C results in rapid receptor immobilization. The immobilization is attributed to aggregation of hormone-receptor complexes, their internalization, or a combination of both processes.


STR data for 15 autosomal STR markers from Paraná (Southern Brazil).
Allelic frequencies for 15 STR autosomal loci, using AmpFℓSTR® Identifiler™, forensic, and statistical parameters were calculated. All loci reached the Hardy-Weinberg equilibrium. The combined power of discrimination and mean power of exclusion were 0.999999999999999999 and 0.9999993, respectively. The MDS plot and NJ tree analysis, generated by FST matrix, corroborated the notion of the origins of the Paraná population as mainly European-derived. The combination of these 15 STR loci represents a powerful strategy for individual identification and parentage analyses for the Paraná population.


Length impairments of the axon initial segment in rodent models of attention-deficit hyperactivity disorder and autism spectrum disorder.
The axon initial segment (AIS) is a structural neuronal compartment of the proximal axon that plays key roles in sodium channel clustering, action potential initiation, and signal propagation of neuronal outputs. Mutations in constitutive genes of the AIS, such as ANK3, have been identified in patients with neurodevelopmental disorders. Nevertheless, morphological changes in the AIS in neurodevelopmental disorders have not been characterized. In this study, we investigated the length of the AIS in rodent models of attention-deficit hyperactivity disorder (ADHD) and autism spectrum disorder (ASD). We observed abnormalities in AIS length in both animal models. In ADHD model rodents, we observed shorter AIS length in layer 2/3 (L2/3) neurons of the medial prefrontal cortex (mPFC) and primary somatosensory barrel field (S1BF). Further, we observed shorter AIS length in S1BF L5 neurons. In ASD model mice, we observed shorter AIS length in L2/3 and L5 neurons of the S1BF. These results suggest that impairments in AIS length are common phenomena in neurodevelopmental disorders such as ADHD and ASD and may be conserved across species. Our findings provide novel insight into the potential contribution of the AIS to the pathophysiology and pathogenesis of neurodevelopmental disorders.


The validity of the severity-psychosis hypothesis in depression.
Psychotic depression (PD) is classified as a subtype of severe depression in the current diagnostic manuals. Accordingly, it is a common conception among psychiatrists that psychotic features in depression arise as a consequence of depressive severity. The aim of this study was to determine whether the severity of depressive and psychotic symptoms correlate in accordance with this "severity-psychosis" hypothesis and to detect potential differences in the clinical features of PD and non-psychotic depression (non-PD). Quantitative analysis of Health of the Nation Outcome Scales (HoNOS) scores from all patients admitted to a Danish general psychiatric hospital due to a severe depressive episode in the period between 2000 and 2010 was performed. A total of 357 patients with severe depression, of which 125 (35%) were of the psychotic subtype, formed the study sample. Mean HoNOS scores at admission differed significantly between patients with non-PD and PD on the items hallucinations and delusions (non-PD=0.33 vs. PD=1.37, p<0.001), aggression (non-PD=0.20 vs. PD=0.36, p=0.044) and on the total score (non-PD=10.55 vs. PD=11.87, p=0.024). The HoNOS scores on the two items "depression" and "hallucinations and delusions" were very weakly correlated. Diagnoses were based on normal clinical practice and not formalized research criteria. The symptomatology of PD and non-PD differs beyond the mere psychosis. Furthermore, severity ratings of depressive and psychotic symptoms are very weakly correlated. These findings offer further support to the hypothesis stating that the psychotic- and non-psychotic subtypes of depression may in fact be distinct clinical syndromes.


Cytoplasmic protein mobility in osmotically stressed Escherichia coli.
Facile diffusion of globular proteins within a cytoplasm that is dense with biopolymers is essential to normal cellular biochemical activity and growth. Remarkably, Escherichia coli grows in minimal medium over a wide range of external osmolalities (0.03 to 1.8 osmol). The mean cytoplasmic biopolymer volume fraction ((phi)) for such adapted cells ranges from 0.16 at 0.10 osmol to 0.36 at 1.45 osmol. For cells grown at 0.28 osmol, a similar phi range is obtained by plasmolysis (sudden osmotic upshift) using NaCl or sucrose as the external osmolyte, after which the only available cellular response is passive loss of cytoplasmic water. Here we measure the effective axial diffusion coefficient of green fluorescent protein (D(GFP)) in the cytoplasm of E. coli cells as a function of (phi) for both plasmolyzed and adapted cells. For plasmolyzed cells, the median D(GFP) (D(GFP)(m)) decreases by a factor of 70 as (phi) increases from 0.16 to 0.33. In sharp contrast, for adapted cells, D(GFP)(m) decreases only by a factor of 2.1 as (phi) increases from 0.16 to 0.36. Clearly, GFP diffusion is not determined by (phi) alone. By comparison with quantitative models, we show that the data cannot be explained by crowding theory. We suggest possible underlying causes of this surprising effect and further experiments that will help choose among competing hypotheses. Recovery of the ability of proteins to diffuse in the cytoplasm after plasmolysis may well be a key determinant of the time scale of the recovery of growth.


Potential for promoting recurrent laryngeal nerve regeneration by remote delivery of viral gene therapy.
The aims of this study were to demonstrate the ability to enhance nerve regeneration by remote delivery of a viral vector to the crushed recurrent laryngeal nerve (RLN), to demonstrate the usefulness of a crushed RLN model to test the efficacy of viral gene therapy, and to discuss future potential applications of this approach. Animal study. Adult Sprague-Dawley rats were assigned to two groups. In the experimental group, an adeno-associated viral (AAV) vector carrying a zinc-finger transcription factor, which stimulates endogenous insulinlike growth factor I production (AAV2-TO-6876vp16), was injected into the crushed RLN. In the control group, an AAV vector carrying the gene for green fluorescent protein was injected into the crushed RLN. Unilateral RLN paralysis was confirmed endoscopically. At 1 week, laryngeal endoscopies were repeated and recorded. Larynges were cryosectioned in 15-μm sections and processed for acetylcholine histochemistry (motor endplates) followed by neurofilament immunoperoxidase (nerve fibers). Percentage nerve-endplate contact (PEC) was determined and compared. Vocal fold motion was evaluated by blinded reviewers using a visual analogue scale (VAS). The difference between PEC on the crushed and uncrushed sides was statistically less in the experimental group (0.54 ± 0.18 vs. 0.30 ± 0.26, P = .0006). The VAS score at 1 week was significantly better in the experimental group (P = .002). AAV2-TO-6876vp16 demonstrated a neurotrophic effect when injected into the crushed RLN. The RLN offers a conduit for viral gene therapy to the brainstem that could be useful for the treatment of RLN injury or bulbar motor neuron disease.


Dialysis cultures with immobilized hybridoma cells for effective production of monoclonal antibodies.
An industrial scale reactor concept for continuous cultivation of immobilized animal cells (e.g. hybridoma cells) in a radial-flow fixed bed is presented, where low molecular weight metabolites are removed via dialysis membrane and high molecular products (e.g. monoclonal antibodies) are enriched. In a new "nutrient-split" feeding strategy concentrated medium is fed directly to the fixed bed unit, whereas a buffer solution is used as dialysis fluid. This feeding strategy was investigated in a laboratory scale reactor with hybridoma cells for production of monoclonal antibodies. A steady state monoclonal antibody concentration of 478 mg l(-1) was reached, appr. 15 times more compared to the concentration reached in chemostat cultures with suspended cells. Glucose and glutamine were used up to 98%. The experiments were described successfully with a kinetic model for immobilized growing cells. Conclusions were drawn for scale-up and design of the large scale system. c(Glc) - glucose concentration, mmol l(-1); c(Gln) - glutamine concentration, mmol l(-1); c(Amm) - ammonia concentration, mmol l(-1); c(Lac) - lactate concentration, mmol l(-1); c(MAb) - MAb concentration, mg l(-1); D - dilution rate, d(-1); D(i) - dilution rate in the inner chamber of the membrane dialysis reactor, d(-1); D(0) - dilution rate in the outer chamber of the membrane dialysis reactor, d(-1); q*(FB,Glc) - volume specific glucose uptake rate related to the fixed bed volume, mmol l(FB) (-1) h(-1); q*(FB,Gln) - volume specific glutamine uptake rate related to the fixed bed volume, mmol l(FB) (-1) h(-1).


Myo-inositol supplementation to prevent gestational diabetes in overweight non-obese women: bioelectrical impedance analysis, metabolic aspects, obstetric and neonatal outcomes - a randomized and open-label, placebo-controlled clinical trial.
This study aims to evaluate the effects of myo-inositol supplementation on gestational diabetes mellitus (GDM) rates and body water distribution in overweight non-obese women. 223 overweight non-obese women pregnant were randomly assigned to the treatment group (2 g of myo-inositol plus 200 µg of folic acid) or to the placebo one (200 µg of folic acid). The treatment lasted until three weeks after delivery. A tetrapolar impedance analyser was used to study body composition. The incidence of GDM was significantly reduced in the myo-inositol group compared with the placebo group. There was a significant increase in TBW, ECW and ICW values in the placebo group compared to the myo-inositol group. We have recorded a significant reduction in the overall incidence of pregnancy-induced hypertension in the myo-inositol group compared with the placebo group. Our results demonstrate the effectiveness of myo-inositol supplementation in preventing GDM in overweight non-obese pregnant women.


Impact of source position on high-dose-rate skin surface applicator dosimetry.
Skin surface dosimetric discrepancies between measured and treatment planning system predicted values were traced to source position sag inside the applicator and to source transit time. We quantified their dosimetric impact and propose corrections for clinical use. We measured the dose profiles from the Varian Leipzig-style high-dose-rate (HDR) skin applicator, using EBT3 film, photon diode, and optically stimulated luminescence dosimeter for three different GammaMedplus HDR afterloaders. The measured dose profiles at several depths were compared with BrachyVision Acuros calculated profiles. To assess the impact of the source sag, two different applicator orientations were considered. The dose contribution during source transit was assessed by comparing diode measurements using an HDR timer and an electrometer timer. Depth doses measured using the three dosimeters were in good agreement, but were consistently higher than the Acuros dose calculations. Measurements with the applicator face up were significantly (exceeding 10%) lower than those in the face down position, due to source sag inside the applicator. Based on the inverse square law, the effective source sag was evaluated to be about 0.5 mm from the planned position. The additional dose during source transit was evaluated to be about 2.8% for 30 seconds of treatment with a 40700 U (10 Ci) source. With a very short source-to-surface distance, the small source sag inside the applicator has a significant dosimetric impact. This effect is unaccounted for in the vendor's treatment planning template and should be considered before the clinical use of the applicator. Further investigation of other applicators with large source lumen diameter may be warranted.


The radiographic evaluation of shoulder and elbow pain in the athlete.
This article presents the important anatomic features of the shoulder joint and the abnormalities to which the athlete is most susceptible leading to pain and disability. It also reviews the pertinent elbow anatomy and discusses the radiographic techniques currently employed in diagnosing elbow problems in the athlete, including tomography, arthrography, and computed tomography.


The five-item modified frailty index predicts long-term outcomes in elderly patients undergoing colorectal cancer surgery.
Frailty has been globally recognized as a predictor of adverse postoperative outcomes. Frailty assessment using the five-factor modified frailty index (5-mFI) has recently gained traction; however, long-term outcomes are unknown in colorectal cancer (CRC) surgery. This study aimed to investigate whether the 5-mFI predicted long-term survival and cause of death on the basis of frailty severity in elderly patients who underwent CRC surgery and to determine the risk factors for mortality. A total of 299 patients underwent CRC surgery with curative intent between January 2013 and December 2017. Patients were divided into three groups by the 5-mFI score: group 1 (5-mFI: 0 or 1; n = 164): no frailty; group 2 (5-mFI: 2; n = 91): moderate frailty; and group 3 (5-mFI: ≥ 3; n = 44): severe frailty. Clinicopathological variables, namely comorbidities, 5-mFI, prognostic nutrition index, operative/postoperative data, and outcome, including cause of death, were compared between the three groups. To identify factors associated with death from CRC- and non-CRC-related causes, univariate and multivariate analyses using a Cox regression model were performed. The immediate postoperative morbidity of patients with Clavien-Dindo grade ≥ III complications (9.1%) in group 3 was not significantly different from that in group 1 (9.1%) or group 2 (14.3%); however, the 30-day mortality rate (4.5%) in group 3 was significantly higher. Long-term disease-free survival was similar between frailty groups, suggesting that CRC surgery provides oncological benefit to patients irrespective of frailty. The 5-year survival rates in groups 1, 2, and 3 were 83.5%, 71.2%, and 47.9%, respectively, showing a significantly lower survival rate as frailty advanced. Sixty percent of the deaths in frail patients were due to respiratory failure and cardiovascular diseases. Multivariate analysis demonstrated that advanced age, higher 5-mFI score, and longer postoperative hospital stay were risk factors for mortality unrelated to CRC. Multivariate analysis also revealed that advanced tumor stage, carcinoembryonic antigen ≥ 5 ng/ml, undifferentiated tumor, and R1 resection were risk factors for CRC-related mortality. The 5-mFI score can predict postoperative short- and long-term outcomes and risk factors for mortality unrelated to CRC. Additionally, long-term survival was negatively associated with the 5-mFI score.


A nuclear factor for IL-6 expression (NF-IL6) is a member of a C/EBP family.
NF-IL6 is a nuclear factor that specifically binds to an IL1-responsive element in the IL-6 gene. In this study the gene encoding NF-IL6 has been cloned by direct screening of a lambda gt11 library using NF-IL6 binding sequence as a ligand. The full-length cDNA encoded a 345 amino acid protein with a potential leucine zipper structure and revealed a high degree of homology to a liver-specific transcriptional factor, C/EBP, at the C-terminal portion. The bacterial fusion protein bound to the CCAAT homology as well as the viral enhancer core sequences as in the case of C/EBP. Recombinant NF-IL6 activated the human IL-6 promoter in a sequence-specific manner. Southern blot analysis demonstrated the high-degree conservation of the NF-IL6 gene through evolution and the existence of several other related genes sharing the DNA-binding domain. NF-IL6 mRNA was normally not expressed, but induced by the stimulation with either LPS, IL-1 or IL-6. Interestingly, NF-IL6 was shown to bind to the regulatory regions for various acute-phase protein genes and several other cytokine genes such as TNF, IL-8 and G-CSF, implying that NF-IL6 has a role in regulation not only for the IL-6 gene but also for several other genes involved in acute-phase reaction, inflammation and hemopoiesis.


Hand disability and related variables in patients with rheumatoid arthritis.
To carry out a cross-sectional study of patients with rheumatoid arthritis (RA) for hand disability, articular damage and to define their relation with demographic, laboratory and clinical parameters. The study included 105 RA patients with a mean age of 49.4 years. Demographic parameters of the patients were recorded. Clinical parameters including disease duration, duration of morning stiffness, pain assessed by visual analog scale, Ritchie Articular Index, grip strength, lateral, tip and three-fingered pinch, and laboratory parameters comprising C-reactive protein, erythrocyte sedimentation rate and rheumatoid factor were evaluated in all patients. The Rheumatoid Arthritis Articular Damage (RAAD) score was used to assess the irreversible articular damage and deformities of the hand. Hand disability was assessed by the special hand disability index of Standford Health Assessment Questionnaire (HAQ). Hand disabilities of various levels were detected in 81% of the patients. Disease duration, grip strength, pinch measurements, clinical and laboratory activity parameters were strongly correlated with hand disability (p<0.01). Hand disability was more related to disease activity parameters than articular damage (p<0.01 and p<0.05, respectively). Grip strength and pinch measurements were the most related parameters with hand disability. The disability scores were significantly higher in female patients (p<0.01). The RAAD score was correlated with disease duration and grip strength (p<0.01). The clinical and laboratory parameters and seropositivity were not correlated with articular damage assessed by RAAD score (p>0.05). Our data suggest that grip strength and pinch measurements seem to be the most related variables with hand disability and articular damage. Therefore, grip strength and pinch measurement should be included in the evaluation and follow-up of the patients with RA in hand rehabilitation units.


Lack of effect of combining different stressors on innate immune responses of seabream (Sparus aurata L.).
A complex stressful event, which commonly occurs in modern aquacultural practices, was broken down into factors that were analysed both individually and jointly to assess their effect on two stress indicators (blood glucose and serum cortisol levels) and two activities of the innate immune response (serum complement and head-kidney leukocyte respiratory burst). For this, gilthead seabream (Sparus aurata L.) specimens were exposed to the following stressors: physical disturbance, crowding, anaesthesia with 2-phenoxyethanol and air exposure. At 0, 1, 2, 3 and 4 days post-stress, blood and serum samples were collected to measure glucose concentration and cortisol and complement levels, respectively. Head-kidney leukocytes were isolated and assayed to evaluate respiratory burst activity. The results show that physical disturbance, crowding and anaesthesia produced an occasional increase in glucose and cortisol concentrations. Crowding and anaesthesia induced a depression in complement activity, while hypoxia by air exposure caused a reduction in the respiratory burst. When all factors were jointly applied both humoral and cellular defences were compromised and cortisol values remained high throughout the experimental period. Any long-term effects of this abnormal serum cortisol levels on the immune system remain unknown.


Modeling gamma knife radiosurgical toxicity for multiple brain metastases.
Radiation oncology protocols for single fraction radiosurgery recommend setting dosing criteria based on assumed risk of radionecrosis, which can be predicted by the 12 Gy normal brain volume (V12). In this study, we show that tumor surface area (SA) and a simple power-law model using only preplan variables can estimate and minimize radiosurgical toxicity. A 245-patient cohort with 1217 brain metastases treated with single or distributed Gamma Knife sessions was reviewed retrospectively. Univariate and multivariable linear regression models and power-law models determined which modeling parameters best predicted V12. The V12 power-law model, represented by a product of normalized Rx dose Rxn, and tumor longest axial dimension LAD (V12 ∼ Rxn1.5*LAD2), was independently validated using a secondary 63-patient cohort with 302 brain metastases. Surface area was the best univariate linear predictor of V12 (adjR2 = 0.770), followed by longest axial dimension (adjR2 = 0.755) and volume (adjR2 = 0.745). The power-law model accounted for 90% variance in V12 for 1217 metastatic lesions (adjR2 = 0.906) and 245 patients (adjR2 = 0.896). The average difference ΔV12 between predicted and measured V12s was (0.28 ± 0.55) cm3 per lesion and (1.0 ± 1.2) cm3 per patient. The power-law predictive capability was validated using a secondary 63-patient dataset (adjR2 = 0.867) with 302 brain metastases (adjR2 = 0.825). Surface area was the most accurate univariate predictor of V12 for metastatic lesions. We developed a preplan model for brain metastases that can help better estimate radionecrosis risk, determine prescription doses given a target V12, and provide safe dose escalation strategies without the use of any planning software.


High abundance of viruses found in aquatic environments.
The concentration of bacteriophages in natural unpolluted waters is in general believed to be low, and they have therefore been considered ecologically unimportant. Using a new method for quantitative enumeration, we have found up to 2.5 x 10(8) virus particles per millilitre in natural waters. These concentrations indicate that virus infection may be an important factor in the ecological control of planktonic micro-organisms, and that viruses might mediate genetic exchange among bacteria in natural aquatic environments.


High fat diet induces hypermethylation of the hypothalamic Pomc promoter and obesity in post-weaning rats.
Impaired response of the brain to the leptin signal leads to a persisting dysregulation of food intake and energy balance. High plasma leptin or insulin should activate proopiomelanocortin (POMC), the precursor of the anorexigenic neuropeptide α-melanocyte-stimulating hormone (α-MSH) in the hypothalamic arcuate nucleus (ARC). Nevertheless, in obesity, this signal transduction pathway might be impaired. In this study we investigated whether chronic high fat (HF) diet consumption from post-weaning to adulthood increases CpG methylation of the Pomc promoter. The hypothesis that this would disrupt the essential binding of the transcription factor Sp1 to the Pomc promoter was tested. Male rats were raised from postnatal day 21 till 90 on either HF or standard diet. As a result HF fed rats were significantly heavier, with high leptin and insulin levels in their plasma but almost no changes in ARC mRNA expression levels of Pomc. The Pomc promoter area in the HF-treated rats was found to be hypermethylated. Furthermore, there was a direct correlation in individual rats between CpG methylation at specific sites that affect Sp1 binding and plasma leptin levels and/or body weight. Although, as expected the HF diet resulted in up-regulation of Sp1, the binding of Sp1 to the hypermethylated Pomc promoter was significantly reduced. Therefore, we suggest that hypermethylation on the promoter region of the Pomc gene can emerge at post-lactation periods and interfere with transcription factor binding, thus blocking the effects of high leptin levels, leading to obesity.


Zeolite X from coal fly ash inhibits proliferation of human breast cancer cell lines (MCF-7) via induction of S phase arrest and apoptosis.
In this study, zeolite X synthesised from coal fly ash (CFA) was examined for its anti-proliferative effects on human breast cancer MCF-7 cells by in vitro studies and allied probable molecular mechanism were also assessed. MTT assay exposed that zeolite X showed a concentration-dependent inhibitory result on the proliferation of MCF-7 cells and caused early apoptotic death and does not induce cytotoxicity in non-cancerous cells (MCF-12A). Flow cytometric study specified the accrual of cells at S phase suggest induction of apoptosis which was avowed through fluorescence and confocal microscopy following annexin V-FITC/propidium iodide (PI). MCF-7 cells treated by 10, 15 and 20 µg/ml concentrations of zeolite X showed internucleosomal DNA fragments in ladder form, thereby indicates cell death is associated with apoptosis. Mechanistically, our data support the induction of apoptosis through the activation of mitochondrial dependent pathway as indicated by an up-regulation of Bax and downregulation of Bcl-2 ratio, the expulsion of cytochrome c from the mitochondria to the cytosol and cleavage of pro-caspase-3 into activation of caspase-3 in MCF-7 cells. Based on these outcomes zeolite X induced early apoptosis and strongly provided experimental evidence for the usage of zeolite X as an essential therapeutic agent in the prevention and therapy of human breast cancer.


The effect of glucagon on intestinal lymph flow in rats.
Intravenous injection of a bolus of 5 microgram of glucagon to rats with an intestinal lymph fistula produces a marked, transient rise in lymph flow and protein output from the fistula. The effect is presumably the result of brief enhancement of splanchnic blood flow.


DNA-support coupling for transcription factor purification. Comparison of aldehyde, cyanogen bromide and N-hydroxysuccinimide chemistries.
Purification of transcription factor IIIA on internal control region DNA coupled to aldehyde-silica is described and compared with purification on cyanogen bromide-activated Sepharose and Bio-Rad Affi-Gel-10. The Affi-Gel support results in mixed-mode chromatography; both ion-exchange and affinity modes contribute. Coupling DNA to aldehyde-silica is advantageous in that it has no ion-exchange properties and performs as well as DNA coupled to CNBr-activated Sepharose. Purification of lac repressor on aldehyde-silica, and CAAT enhancer binding protein on Affi-Gel also shows the advantages of a neutral support and the disadvantages of mixed-mode chromatography for transcription factor purification. Aldehyde-silica couples to alkylamines and to the amines of adenine, guanine, and cytosine nucleoside bases. Reaction occurs with either single- or double-stranded DNA, although it is less efficient with the latter. Overall, the results demonstrate that predominantly neutral coupling chemistries, such as aldehyde or CNBr-mediated coupling, have distinct advantages for transcription factor purification. Since the CNBr chemistry has not yet been applied to silica supports, aldehyde-silica coupling is currently the most attractive method for DNA affinity HPLC.


Adaptation and validation of the Physical Activity Questionnaire for Adolescents (PAQ-A) among Polish adolescents: cross-sectional study.
The lack of a widely accepted questionnaire used to assess the physical activity (PA) of adolescents in the Polish language creates a need to introduce such a valid, reliable, inexpensive and quick tool for assessment. This study was designed to culturally adapt and validate the Physical Activity Questionnaires for Adolescents (PAQ-A) in the Polish language. Cross-sectional study. Cultural adaptation of the Polish version of the PAQ-A was performed following the standardised questionnaires cultural adaptation process. In a sample of 78 adolescents aged 14-19 years, the PAQ-A test-retest was administered within a 1-week interval. Reliability was analysed by the intraclass correlation coefficient (ICC) and internal consistency with Cronbach's α. Participants completed the PAQ-A, and wore an accelerometer for 7 consecutive days. The PAQ-A was compared with PA parameters obtained using the accelerometer. Test-retest reliability showed ICC=0.97 for the total score of PAQ-A. Internal consistency was excellent (α=0.93). The PAQ-A was very strongly correlated with steps per day (r=0.94) and with moderate-to-vigorous PA (r=0.81) assessed by the accelerometer. The Polish version of the PAQ-A is a valuable tool to estimate general levels of PA among adolescents from 14 to 19 years old.


The prebiotics (Fructo-oligosaccharides and Xylo-oligosaccharides) modulate the probiotic properties of Lactiplantibacillus and Levilactobacillus strains isolated from traditional fermented olive.
This study aimed to examine the influence of two prebiotics, fructo-oligosaccharides (FOS) and xylo-oligosaccharides (XOS), on probiotic properties (resistance to low pH and bile salt, hydrophobicity and auto-aggregation), metabolites production, and antimicrobial activity of probiotic Lactiplantibacillus (L. pentosus S42 and L. plantarum S61) and Levilactobacillus (L. brevis S27) strains isolated from fermented olive. The results demonstrated the ability of strains to ferment XOS more than FOS as a sole carbon source, resulting in pH reduction. The prebiotics (FOS and XOS) significantly increased (p < 0.05) their survival in gastro-intestinal conditions (low pH and 0.3% of bile salts), as well as their hydrophobicity, auto-aggregation and production of proteins, compared to glucose (control). The major organic acids produced by Lactiplantibacillus and Levilactobacillus strains, were oxalic, malic and lactic acids from FOS, XOS and glucose, respectively. No antimicrobial activity was observed from cell-free supernatant (CFS) of Lactiplantibacillus and Levilactobacillus strains obtained from FOS. In the presence of XOS the organic acids, produced by Lactiplantibacillus and Levilactobacillus strains, were more diverse with high contents, and exhibited higher antifungal and antibacterial activities, more than that of FOS and glucose. The combination of L. plantarum S61 and XOS demonstrated the highest inhibition zones ranges of 20.7-22.2 mm against pathogenic bacteria and 29.2-30 mm against yeasts. This combination can be used in production of antifungal preservatives and pharmaceuticals, against pathogenic and spoilage yeasts.


Compensatory lung growth: protein, DNA and RNA lung contents in undernourished trilobectomized rats.
To demonstrate compensatory lung growth (CLG) by lung contents of proteins, DNA, and RNA in undernourished young adult rats, submitted to pulmonary trilobectomy. We used 137 male Wistar rats, randomly distributed into 9 groups; they were submitted to three treatments (control, thoracotomy, and trilobectomy), and sacrificed at three different times (7, 30, and 90 days). In trilobectomy we removed the right median, accessory, and caudal lobes. We studied lung proteins, DNA, and RNA contents. In the cranial lobe and left lung, protein content was higher in trilobectomized rats however there was insufficient CLG to make up for the loss. The increase of DNA in the cranial lobe and left lung of trilobectomized rats was sufficient to compensate for this loss, resulting in a similar content to controls. RNA content in trilobectomized rats, was higher in the cranial lobe and left lung, more efficient in the cranial lobe, but less than in the other groups. CLG occurred in trilobectomized rats, probably with cell hyperplasia and little hypertrophy, due to the large DNA compensation and small RNA compensation. This was markedly different to well-nourished animals, who had pronounced hypertrophy.


Crop effect and mechanism of amino acid-modified biomass ash in remediation of cadmium-contaminated soil.
Lignocellulosic biomass ash (BA) has certain adsorption and passivation effects on heavy metals, but its function is generally weak. Amino acid salt can facilitate the leaching of heavy metals in soil. Therefore, modification of BA with amino acid salt may realize a higher leaching rate and better passivation of heavy metals in soil. In this study, BA was modified by amino acid hydrolysate obtained from the hydrolysis of chicken feathers by sulfuric acid. The physicochemical properties of BA and modified BA (MBA), their effects on Chinese cabbage (CC) yield and nutritional quality, and passivation effects on soil cadmium (Cd) were compared, and the related mechanisms were investigated. SEM-EDS, XRD, and FTIR demonstrated that BA was a CaCO3-type soil amendment, while MBA was a CaSO4-type soil amendment with the loading of amino acid. Compared with BA, MBA significantly increased the fresh weight, soluble sugar, vitamin C (Vc), and protein contents of CC in both non-Cd contaminated soil and Cd contaminated soil, and obviously decreased the nitrate content and Cd uptake of CC in Cd-contaminated soil. After the application of MBA, cadmium species in potted soil were transformed from higher plant availability, representing by exchangeable and carbonate-bound, into lower plant availability, representing by iron-manganese oxide bound, which was identified as the key reason for the significant reduction of Cd content in CC under MBA application.


10-year single center experience of catheter ablation of focal atrial tachycardia.
Focal atrial tachycardias (ATs) represent 5-15% of sustained supraventricular tachycardias (SVTs). Characteristic distribution of sites of origin and detailed electrophysiologic characterization of AT from specific sites of origin (SOO) have been described. Acute success and recurrence are less favorable than for other SVTs. In this series, we present our experience of focal AT ablation over a 10-year period. We undertook a retrospective review of an electronically maintained database of all patients undergoing AT ablation at our institution between January 2011 and December 2020. Demographic, procedural, and outcomes data were reviewed. A total of 293 distinct atrial tachycardias were treated during 279 procedures in 256 patients, including 207 first AT ablations. Acutely successful AT suppression was achieved in 91% of first-time ablations. Acute success was dependent on SOO of AT with lowest rates of acute suppression in the para-Hisian region and the crista terminalis (CT). The most common reason for failure to acutely suppress the AT was proximity to a critical structure (phrenic nerve, sinus node, and AV node). 8.9% of patients in this series presented with a tachycardia-mediated cardiomyopathy (TCM). 48% of TCM patients underwent an ablation attempt during an acute medical admission. Among the TCM group, median LV ejection fraction increased from 25% (range 10-50%) to 55% (range 35-65%) with successful treatment of AT. Five patients undergoing a repeat procedure had planned pericardial access for displacement of the phrenic nerve to permit ablation of the AT, which was successful in all cases. Among patients without a pre-existing diagnosis of AF, peri-procedural AF was not associated with a higher incidence of a subsequent diagnosis of AF (odds ratio 1.169, 95% CI 0.4058-3.475, p = 0.7628). Median duration of follow-up was 832 days. By Kaplan-Meier estimate, recurrence-free survival was 78% (95% CI 67-88%). In this series, focal AT ablation is associated with good acute results and a low rate of complications, but outcomes remain less favorable than previously reported for other forms of SVT.


Evaluation of the nephron-sparing surgery formula in Wilms tumors.
Definitions of nephron-sparing surgery (NSS) procedures in Wilms tumor (WT) are not clear. The new UMBRELLA protocol offers a formula (NSS(X)-SRM(n)-PRM(n)-RRP(n%)) to better define the different NSS parameters. We aimed to assess the advantages and limits of this new formula. This retrospective monocentric study included patients operated by NSS for WT from 1975 to 2018. We reviewed the medical records and applied the NSS formula to all included patients. Eighty kidneys were operated on 56 patients at a mean age of 19.2 months (4 days-7.5 years), with 49 partial nephrectomies and 31 tumorectomies. The assessment of the surgical resection margins (SRM) showed a doubt in six cases and one tumor breach. An intact pseudocapsule along the resection margin with no renal parenchyma was found in four cases at pathological resection margins (PRM) assessment, whereas a tumor breach was described in seven cases. Among the six patients with a surgical doubt, only one had a pathological stage III. There were no surgical doubts in the seven patients with tumor breach at pathology. At a mean follow-up of eight years (15 days-28.6 years), eight patients had elevated blood pressure levels. Ten had proteinuria. These two parameters were significantly increased in patients with a remaining renal parenchyma (RRP) of less than half of the initial total renal parenchyma. The serum creatinine level was normal for all except two patients. The new NSS formula described all the crucial elements of NSS. RRP seemed essential for the evaluation of long-term renal function.