A comparison between short-interval and regular-interval follow-up for BI-RADS category 3 lesions.
The aim of this study was to compare the short-interval and regular-interval follow-up in women with Breast Imaging Reporting and Data System (BI-RADS) category 3 screen imaging studies. The image stability, rate of image-detected breast cancer and stage of cancer are studied. Women who had BI-RADS 3 screen imaging studies (mammography and ultrasonography) conducted between the period January 2003 and December 2005 were retrospectively identified using the computerised database at the Department of Radiology, Ramathibodi Hospital, Thailand. Women who had known breast cancer status at two years after screening were included in the study and divided into two groups: short-interval (six months after screening) or regular-interval (one year after screening) follow-up. The two groups were compared in terms of the baseline clinicoradiologic characteristics and outcomes, including the image stability, image-detected breast cancer and the cancer stage at detection. A total of 10,086 women underwent screen imaging studies within the study period. Of these, 1,541 (15 percent) were categorised as BI-RADS 3. Only 1,036 women (67 percent) had follow-up images done six to 12 months after screening, and 846 (82 percent of 1,036 women) also had known cancer status two years after the screening. Breast cancer was noted in seven women (a positive predictive value of 0.7 percent). There were no significant differences between the two groups of women in terms of their baseline characteristics, image stability at the initial follow-up, the rate of image-detected breast cancer and the stage of cancer at detection. There were no significant differences in the effectiveness of short-interval versus regular-interval follow-up in women with BI-RADS 3 screen imaging studies.


Occurrence of ungulic acid in some epidermal tissues.
The lipids isolated from different animal tissues have been studied qualitatively, by TLC, for the occurrence of the ungulic acid fraction. This fraction was found in considerable amounts only in epidermal tissues and its keratinized derivatives. In the present study it was isolated from human keratinous epidermis, hair, and nails, pig bristles, wool, and feathers. The analytical results indicated that a lipid fraction from all of these sources contained ceramide, galactose, galactosamine, sulfate, and sialic acid in equimolar amounts, and that the fractions were similar to the ungulic acid isolated earlier from a horse's hoof.


[Hydrocepahlia and subarachnoid cyst due to neurocysticercosis. A new case from rural Extremadura].
Neurocystercicosis (NCC) is a disorder caused by the Taenia solium larva. It is the commonest parasitosis of the CNS. In Spain most patients are from countries where the condition is endemic. However, sporadic cases still occur amongst the non immigrant population of rural regions. We report a new case of NCC from the Valle del Jerte in rural Extremadura. A 51 year old man presented with intellectual impairment, motor aphasia, apraxia, right hemiparesia and sphincter incontinence for the past six months. On cranial CAT and MR hydorcephalia was seen together with a subarachnoid cyst in the left Sylvan fissure. Serology was positive for cysticercosis in both plasma and CSF. The patient was treated with albendazol after insertion of a ventriculo peritoneal shunt. Three months later there was both clinical and radiological improvement. NCC is still a condition which must be included in the differential diagnosis of patients in Spain who present with CNS involvement and cystic lesions on neuro imaging investigations, even when they are not from countries where the disease is endemic. Treatment with albendazol and steroids given after insertion of a ventriculo peritoneal shunt was effective in our patient.


Effects of preeclampsia-like symptoms at early gestational stage on feto-placental outcomes in a mouse model.
Early and late-onset preeclampsia is thought to be different disease entities. This study aimed to determine the effects of early-onset preeclampsia-like symptoms on feto-placental outcomes and the adverse impacts of various factors on placental and fetal growth and development at different gestational stages in a mouse model. Pregnant C57BL/6J mice were divided into control and preeclampsia (PE) groups, and injected subcutaneously with the nitric oxide synthase inhibitor L-arginine methyl ester (L-NAME) 50 mgxkg(-1)d(-1). The PE group was divided into early-, mid- and late-PE groups with L-NAME injections starting on days 7, 11 and 16 of pregnancy, respectively. Corresponding control groups were injected with saline at the same time points. Blood pressure was measured until days 14 and 18, when the fetuses and placentas were removed under anesthesia. Blood pressure, urinary protein, and fetal and placental conditions were analyzed. Blood pressure and urinary protein increased following L-NAME injection. The fetal survival rate and fetal weight were reduced and the fetal absorption rate was increased in the early-PE group on days 14 and 18 of pregnancy, compared with the control group. There were no significant differences in these parameters between the late-PE group and the respective control group. Placental weights in the early- and mid-PE groups were significantly reduced at days 14 and 18 of pregnancy compared with the control groups, but there was no significant difference in placental weight between the late-PE group and the respective control group. Morphologic examination of placentas from the early- and mid-PE groups showed varying degrees of fibrinoid necrosis and villous interstitial edema, but no significant pathologic changes were found in the placentas from the late-PE or control groups. Preeclampsia-like symptoms occurring during the early stage of pregnancy are more likely to affect placental and fetal development, whereas late onset preeclampsia-like symptoms have a direct impact on the mothers.


Adaptive Peptide Molecule as the Promising Highly-Efficient Gas-Sensor Material: In Silico Study.
Gas sensors are currently employed in various applications in fields such as medicine, ecology, and food processing, and serve as monitoring tools for the protection of human health, safety, and quality of life. Herein, we discuss a promising direction in the research and development of gas sensors based on peptides-biomolecules with high selectivity and sensitivity to various gases. Thanks to the technique developed in this work, which uses a framework based on the density-functional tight-binding theory (DFTB), the most probable adsorption centers were identified and used to describe the interaction of some analyte molecules with peptides. The DFTB method revealed that the physical adsorption of acetone, ammonium, benzene, ethanol, hexane, methanol, toluene, and trinitrotoluene had a binding energy in the range from -0.28 eV to -1.46 eV. It was found that peptides may adapt to the approaching analyte by changing their volume up to a maximum value of approx. 13%, in order to confine electron clouds around the adsorbed molecule. Based on the results obtained, the prospects for using the proposed peptide configurations in gas sensor devices are good.


Application of a new robust ECG T-wave delineation algorithm for the evaluation of the autonomic innervation of the myocardium.
T-wave amplitude (TWA) is a well know index of the autonomic innervation of the myocardium. However, until now it has been evaluated only manually or with simple and inefficient algorithms. In this paper, we developed a new robust single-lead electrocardiogram (ECG) T-wave delineation algorithm that is able to detect the T-wave with a wavelet based method and automatically calculate the TWA. We evaluated the algorithm on the QT database, achieving a sensitivity of 99.92% for the T wave peak and 99.38% for the T wave end. In addition, the percentage of records automatically delineated with high precision was higher than previous published works. Finally, the algorithm was applied to study the influence of anticholinergic and antiadrenergic drugs (i.e. atropine and metoprolol) on the TWA. It was observed that atropine significantly decreased the TWA when compared to baseline level, that head-up tilt caused a decrease of TWA and that metoprolol blunted this decrease. Through the development of a robust algorithm, this study opens the way for further research on the T-wave analysis for the assessment of the autonomic innervation of the ventricular myocardium.


Stability of the contractile assembly and Ca2+-activated tension in adenovirus infected adult cardiac myocytes.
Adenovirus-mediated gene transfer into adult cardiac myocytes in primary culture is a potentially useful method to study the structure and function of the contractile apparatus. However, the consequences of adenovirus infection on the highly differentiated state of the cultured myocyte have not been determined. We report here a detailed analysis of myofilament structure and function over time in primary culture and after adenovirus infection. Adult rat ventricular myocytes in primary culture were infected with a recombinant adenovirus vector expressing either the LacZ or alkaline phosphatase reporter gene. Control and infected myocytes were collected at days 0-7 post-isolation/infection, and myofilament isoform expression was determined by SDS-PAGE and Western blot. Laser scanning densitometry showed that the alpha- to beta-myosin heavy chain ratio, the stoichiometry of the myosin light chains and the expression of the adult troponin T isoform did not change over time in culture or with adenovirus treatment. Importantly, examination of Ca2+-activated tension in single myocytes showed no change in the shape or position of the tension-pCa relationship in the control and adenovirus infected myocytes during primary culture. These results indicate that the structure and function of adult cardiac myocytes are stable in short term primary culture and are not affected by adenovirus infection per se, and therefore provide the foundation for the use of adenovirus-mediated myofilament gene transfer to study contractile apparatus structure and function in adult cardiac myocytes.


HMG-CoA reductase inhibitors in kidney transplant recipients receiving tacrolimus: statins not associated with improved patient or graft survival.
The beneficial effects of early statin use in kidney transplant recipients, especially those on tacrolimus-based immunosuppression, are not well established. We evaluated the predictors of statin use following kidney transplantation and examined its association with patient and allograft survival. We examined 615 consecutive patients who underwent kidney transplant at our institution between January 1998 and January 2002. Statin use was assessed at baseline and 3, 6, 9, and 12 months following kidney transplant. Patients were followed for allograft and patient survival. 36% of the 615 kidney transplant recipients were treated with statin treatment. Statin use increased over the course of the study period. Older age, elevated body mass index, higher triglyceride levels, hypercholesterolemia, diabetes, history of myocardial infarction were associated with higher rates of statin use; elevated alkaline phosphatase levels and CMV IgG seropositivity were associated with less statin use. Older age, elevated BMI and hypercholesterolemia remained significant predictors of increased statin use after accounting for covariates using multiple regression. The early use of statins was not associated with improvements in unadjusted patient survival [HR 0.99; 95%CI 0.72-1.37] or graft survival [HR 0.97; 95% CI 0.76-1.24]. The risks of death and graft survival were not consistently reduced with exposure to statin using either adjusted models or propensity scores in Cox Proportional Hazards models. In a kidney transplant population primarily receiving tacrolimus-based immunosuppression, early statin use was not associated with significantly improved graft or patient survival.


Exploring Self-Healing and Switchable Adhesives based on Multi-Level Dynamic Stable Structure.
It is a challenge to develop adhesives simultaneously capable of strong adhesion and efficient switchable ability. Herein, the authors report multifunctional switchable adhesives named Cu2+ -curcumin-imidazole-polyurethane (CIPUs:Cu2+ ) by introducing 1-(3-aminopropyl) imidazole and curcumin into polyurethane system crossed by Cu2+ forming dynamic metal-ligand bonds. This CIPUs:Cu2+ has strong adhesion (up to 2.46 MPa) on various material surfaces due to their specially designed functional groups alike the secretions from mussels. It can achieve fast switching speed (30 s) and high switch efficiency through multiple contactless remote stimulations. Importantly, density functional theory (DFT) calculation reveals that such metal-ligand bonds consisting of two components: stronger Cu2+ -curcumin complexes and weaker Cu2+ -imidazole complexes can aggregate to form multi-level dynamic stable structure . The special structure can not only be acted as sacrificial sites for easily broken and reformed, allowing efficient switchable adhesion and enormous energy dissipation but also acted as firm sites to maintain the cohesion of the adhesive and the reversible reconstruction network. Intriguingly, the CIPUs:Cu2+ can achieve self-healing at room temperature without needing external stimuli. Overall, this strategy can further broaden the design of switchable adhesives in the fields of intelligent gadgets, wearable bio-monitoring devices, etc.


[Safety assessment of nanomaterials using toxicokinetics and toxicoproteome analysis].
With recent development of the nanotechnology, nanomaterials have been successfully employed in various industrial applications such as medicine and cosmetics. Nanomaterials show the useful properties such as electronic reactivity and the tissue permeability that were not provided by micromaterials. Thus, nanomaterials are expected as innovative materials for the development of medicine and cosmetics. However, these innovative properties may show unknown biological responses that could not been detected by the conventional toxicity assay. For industrial development and affluent society establishment that enjoyed only a benefit of nanomaterials, it is urgent to gather information of the properties and the biological effects, and to establish the standard safety evaluation method of nanomaterials. So, we are analyzing association among property, biodistribution and biological effects of nanomaterials to search for the safety biomarker (functional-, molecular- and biochemical-biomarker) using nanosilicas (nSP) as a standard nanomaterial. Because nSP shows high uniform dispersibility and is already used in medicine, cosmetics and food additive, the results of this study are useful to extrapolate it to other nanomaterials and to make practicable as safety biomarker. In this report, we show the latest knowledge about the linkage information among property, biodistribution and biological effects of nSP by toxicokinetics and toxicoproteomics, and the search study of safety biomarker based on these basic information.


Ulcerative Colitis Concomitant with Cytomegalovirus Infection, Bullous Sweet's Syndrome, and Acute Myeloid Leukemia: A Case Report and Literature Review.
Ulcerative colitis (UC) is a chronic, relapsing progressive inflammatory immune disease. There is still no cure for it. Even worse, UC may predispose patients to opportunistic infections, and several extra-intestinal manifestations (EIMs) and comorbidities may antedate, occur with, or postdate the onset of UC, which may increase the mortality risk. But case reports of UC patients simultaneously concomitant with opportunistic infection, EIM, and comorbidity are extremely rare. We report a case of 51-year-old male patient with incipient UC accompanied by cytomegalovirus (CMV) infection and bullous Sweet's syndrome (bSS, a cutaneous EIM of UC) after treatment with oral mesalazine and prednisolone for 3 weeks. After clearance of the CMV infection by using ganciclovir, the patient was administered two cycles of infliximab to cure UC and bSS; however, he developed acute myeloid leukemia (AML) a month later and died after two cycles of chemotherapy. Based on this rare case of UC concomitant with CMV infection, bSS and AML, we recommend that it is important to distinguish between an acute UC flare and opportunistic infections, especially in patients receiving immunosuppressive therapy, and monitor EIMs and comorbidities timely. Particular attention should be paid to cancer surveillance. Clinicians should be mindful of these facts to adopt optimal therapeutic options to address all aspects of UC. Early initiation of biological therapy may be of benefit to patients with newly diagnosed severe UC.


Unveiling Extreme Anisotropy in Elastic Structured Media.
Periodic structures can be engineered to exhibit unique properties observed at symmetry points, such as zero group velocity, Dirac cones, and saddle points; identifying these and the nature of the associated modes from a direct reading of the dispersion surfaces is not straightforward, especially in three dimensions or at high frequencies when several dispersion surfaces fold back in the Brillouin zone. A recently proposed asymptotic high-frequency homogenization theory is applied to a challenging time-domain experiment with elastic waves in a pinned metallic plate. The prediction of a narrow high-frequency spectral region where the effective medium tensor dramatically switches from positive definite to indefinite is confirmed experimentally; a small frequency shift of the pulse carrier results in two distinct types of highly anisotropic modes. The underlying effective equation mirrors this behavior with a change in form from elliptic to hyperbolic exemplifying the high degree of wave control available and the importance of a simple and effective predictive model.


Ascorbic acid, catalase and chlorpromazine reduce cryopreservation-induced damages to crossbred bull spermatozoa.
The present study evaluated the effectiveness of ascorbic acid, catalase, chlorpromazine and their combinations in reducing the cryodamages to crossbred bull (Bos taurus x Bos indicus) spermatozoa. A total of 32 ejaculates (eight each from four bulls) were diluted in Tris-citric acid-fructose-egg yolk-glycerol extender. Each ejaculate was split into six parts (five treatment and one control). Treatment groups included 10 mm ascorbic acid, 0.1 mm chlorpromazine, 200 IU/ml catalase, 10 mm ascorbic acid + 0.1 mm chlorpromazine or 200 IU/ml catalase + 0.1 mm chlorpromazine in the extender. Fluorescent probes (Fluorescein isothiocyanate--Pisum sativum agglutinin + Propidium iodide) were used for the assessment of spermatozoa viability and acrosomal status. The proportion of acrosome intact live (AIL), acrosome intact dead, acrosome reacted live and acrosome reacted dead sperm was assessed in fresh, equilibrated and frozen-thawed semen. The functional status of the sperm was assessed using hypo-osmotic sperm swelling test (HOSST). Activities of acrosin and hyaluronidase enzyme were also determined. Lipid peroxidation level was assayed based on the melonaldehyde (MDA) production. In cryopreserved semen, the values of AIL spermatozoa, HOSST response, hyaluronidase and acrosin activity were reduced by 53%, 47%, 34% and 54%, respectively from their initial values in fresh semen. However, MDA level was threefold higher in the frozen-thawed sperm compared with fresh sperm. Significant (p < 0.05) improvement in motility, viability, HOSST response, retention of hyaluonidase and acrosin and reduction in MDA was recorded in ascorbic acid, catalase, ascorbic acid + chlorpromazine and catalase + chlorpromazine incorporated groups. The percentage of AIL sperm was significantly (p < 0.05) higher in ascorbic acid, catalase and ascorbic acid + chlorpromazine incorporated groups compared with the control. Chlorpromazine alone did not improve the post-thaw semen quality but when combined with either ascorbic acid or catalase, improvement in semen quality was noticed. It was inferred that incorporation of ascorbic acid, catalase and ascorbic acid + chlorpromazine in semen extender improved the post-thaw semen quality in crossbred bulls.


Effect of annealing on morphology and photoluminescence of beta-Ga2O3 nanostructures.
A novel method was applied to prepare one-dimensional beta-Ga2O3 nanostructure films. In this method, beta-Ga2O3 nanostructures have been successfully synthesized on Si(111) substrates through annealing sputtered Ga22O3/Mo films for differernt time under flowing ammonia. The as-synthesized beta-Ga2O3 nanostructures were characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM) and photoluminescence (PL) spectrum. The results show that the formed nanostructures are single-crystalline Ga2O3 with monoclinic structure. The annealing time of the samples has an evident influence on the morphology and optical property of the nanostructured beta-Ga2O3 synthesized. The representative photoluminescence spectrum at room temperature exhibits a strong and broad emission band centered at 411.5 nm and a relatively weak emission peak located at 437.6 nm. The growth mechanism of the beta-Ga2O3 nanostructured materials is also discussed briefly.


Editor's Highlight: PlacentalDisposition and Effects of Crizotinib: An Ex Vivo Study in the Isolated Dual-Side Perfused Human Cotyledon.
Tyrosine kinase inhibitors (TKIs) play an important role in cancer pharmacotherapy, yet there is limited data on their use during pregnancy. We studied placental disposition and placental toxicity of crizotinib, a TKI used to treat nonsmall cell lung cancer. Term placentas were perfused for 3 h with crizotinib (1 µM) using the ex vivo dual-side cotyledon perfusion technique. Interference of TKIs with trophoblast viability was studied using BeWo cells. Expression of P-glycoprotein (P-gp) and Breast Cancer Resistance Protein (BCRP) in placental tissue was assessed by immunohistochemistry and inhibition of these transporters was determined in vitro by transport studies with membrane vesicles overexpressing human P-gp or BCRP. We found that crizotinib rapidly and strongly accumulates in cotyledon perfusion experiments, reaching a concentration of 3.1 ± 0.4 µM in placental tissue. Final drug concentrations in the maternal and foetal reservoirs were 0.2 ± 0.05 and 0.08 ± 0.01 µM, respectively. Furthermore, crizotinib inhibited BeWo cell viability (IC50: 234 nM, 95% CI: 167-328 nM) 10 times more potently than other TKIs tested. In vitro transport studies revealed that crizotinib is a potent inhibitor of the transport activities of BCRP (IC50: 5.7 µM, 95% CI: 2.7-11.8 µM) and P-gp (IC50: 7.8 µM, 95% CI: 3.4-18.0 µM). In conclusion, crizotinib strongly accumulated in placental tissue at clinically relevant concentrations. IC50 values for transporter inhibition and trophoblast cell viability were similar to the tissue concentrations reached, suggesting that crizotinib can inhibit placental BCRP and P-gp function and possibly affect trophoblast viability.


Developmentally regulated telomerase activity is correlated with chromosomal healing during chromatin diminution in Ascaris suum.
Telomerase is the ribonucleoprotein complex responsible for the maintenance of the physical ends, or telomeres, of most eukaryotic chromosomes. In this study, telomerase activity has been identified in cell extracts from the nematode Ascaris suum. This parasitic nematode is particularly suited as a model system for the study of telomerase, because it shows the phenomenon of chromatin diminution, consisting of developmentally programmed chromosomal breakage, DNA elimination, and new telomere formation. In vitro, the A. suum telomerase is capable of efficiently recognizing and elongating nontelomeric primers with nematode-specific telomere repeats by using limited homology at the 3' end of the DNA to anneal with the putative telomerase RNA template. The activity of this enzyme is developmentally regulated, and it correlates temporally with the phenomenon of chromatin diminution. It is up-regulated during the first two rounds of embryonic cell divisions, to reach a peak in 4-cell-stage embryos, when three presomatic blastomeres prepare for chromatin diminution. The activity remains high until the beginning of gastrulation, when the last of the presomatic cells undergoes chromatin diminution, and then constantly decreases during further development. In summary, our data strongly argue for a role of this enzyme in chromosome healing during the process of chromatin diminution.


Virgil Sydenstricker: special reference to niacin deficiency encephalopathy.
Virgil Sydenstricker was a member of a notable American family which included authoress Pearl S. Buck and the eminent epidemiologist Edgar Sydenstricker. Dr. Sydenstricker's contributions in the fields of hematology and nutritional disease are legion. His landmark work in sickle cell anemia characterized a definite symptom complex with specific hematologic findings and inheritance pattern. He wrote on the complications of malnutrition and attempted to delineate the specific effects of individual nutritional factors. Dr. Sydenstricker and his associate H. M. Cleckley first described the syndrome of niacin deficiency encephalopathy. Today, the syndrome is still occasionally reported. Niacin deficiency should be considered when unexplained acute confusional states or neurologic deficits occur in the setting of malnutrition, antituberculous drug use, or chronic partial nutritional deficiency with acute increase in metabolic demand.


Effect of duplicate isolates of methicillin-susceptible and methicillin-resistant Staphylococcus aureus on antibiogram data.
Duplicate Staphylococcus aureus isolates were analyzed to determine the impact of multiple isolates from the same patient on annual antibiogram data. During a 6-year period (1996 to 2001), 3,227 patients with 4,844 S. aureus isolates were evaluated. A total of 39% of patients with methicillin-resistant S. aureus (MRSA) (n = 860) and 23% of patients with methicillin-susceptible S. aureus (MSSA) (n = 2,367) infections had duplicate isolates. Cumulative data show that 91% of the patients during this 6-year period with duplicate isolates (2 to 13 duplicates/year) did not switch between MSSA and MRSA but retained the original S. aureus strain whether it was MSSA or MRSA. Rates of MRSA were calculated for each year by using all isolates and then eliminating duplicates. The impact of duplicate MRSA and MSSA isolates was evaluated by using the ratio of isolates per patient such that ratios of >1.0 indicate >1 isolate per patient. The 6-year ratio for MRSA was 1.90 isolates/patient, and the ratio for MSSA was 1.35. A significant difference (P < 0.05) was noted in the MRSA rates in 4 of 6 years when duplicate isolates were removed. Common phenotypic antibiogram patterns were compared for all MRSA isolates during the 6-year period, and 64% were of a single antibiogram phenotype. Eighty-eight percent of patients with duplicate MRSA isolates had phenotypically identical multiple isolates. The rate of MRSA differs when duplicate isolates are removed from the antibiogram data.


Aseptic meningitis in systemic lupus erythematosus. Report of three cases.
Three patients with aseptic meningitis were subsequently diagnosed as having lupus erythematosus. One patient had a single meningitic episode, another had chronic meningitis, and the third two acute episodes 5 years apart. All 3 patients developed further neurophychiatric manifestations of SLE, leading to death in 1. Aseptic meningitis appears to be an early manifestation of SLE and may herald more serious brain damage. No new cases of aseptic meningitis occurred in this series after initiation of therapy for SLE. In contrast, bacterial meningitis did occur as a late complication of the disease.


Inherited interstitial nephritis in kdkd mice.
kdkd mice, a mutant subline of CBA/Ca mice, develop a progressive, T cell-mediated, autoimmune interstitial nephritis which leads to renal failure and death of all mice at 20-28 weeks of age. This disease is inherited in an autosomal recessive manner, with complete penetrance, and has been linked to grizzled and waltzer on mouse chromosome 10. Immunologic evaluation of this lesion has demonstrated that histologic disease is initiated by a population of CD8+, H-2Kk-restricted T cells, which recognize an antigen in collagenase-solubilized syngeneic renal tubules. These nephritogenic effector cells can also be demonstrated in non-disease prone CBA/Ca mice. Susceptibility to autoimmune nephritis correlates with distinct expression of regulatory, rather than effector, T cells. Interstitial nephritis in kdkd mice can be inhibited by protein-calorie restriction, infusions of CBA/Ca CD8+ T cells, or monoclonal antibodies of ICAM-1. This murine model most closely resembles medullary cystic disease in humans, which has not historically been considered an autoimmune disease. Mapping of the genes for both medullary cystic disease and the defect in kdkd mice should augment our understanding of mechanisms of organ-specific autoimmunity.


Comparison and validation of ELISA assays for plasma insulin-like growth factor-1 in the horse.
Insulin-like growth factor-1 (IGF-1) plays several important physiological roles, and IGF-related pathways have been implicated in developmental osteochondral disease and endocrinopathic laminitis. This factor is also a downstream marker of growth hormone activity and its peptide mimetics. Unfortunately, previously used assays for measuring equine IGF-1 (radioimmunoassays and ELISAs) are no longer commercially available, and many of the kits on the market give poor results when used on horse samples. The aim of the present study was to compare three different ELISA assays (two human and one horse-specific). Plasma samples from six Standardbreds, six ponies and six Andalusians were used. The human IGF-1 ELISA kit from Immunodiagnostic Systems (IDS) proved to be the most accurate and precise of the three kits; the other two assays gave apparently much lower concentrations, with poor recovery of spiked recombinant human IGF-1 and unacceptably poor intra-assay coefficients of variation (CV). The IDS assay gave an intra-assay CV of 3.59 % and inter-assay CV of 7.31%. Mean percentage recovery of spiked IGF-1 was 88.82%, and linearity and dilutional parallelism were satisfied. The IGF-1 plasma concentrations were 123.21 ±8.24 ng/mL for Standardbreds, 124.95 ±3.69 ng/mL for Andalusians and 174.26 ±1.94 ng/mL for ponies. Therefore of the three assays assessed, the IGF-1 ELISA manufactured by IDS was the most suitable for use with equine plasma samples and may have many useful applications in several different research areas. However, caution should be used when comparing equine studies where different analytical techniques and assays may have been used to measure this growth factor.


Characteristics of uptake and cytotoxicity of a low-density lipoprotein-daunomycin complex in P388 leukemic cells.
We have investigated the in vitro drug-cell interaction and therapeutic effects of a low-density lipoprotein (LDL)-daunomycin complex as compared to free daunomycin. Uptake and retention of daunomycin were significantly enhanced in sensitive and resistant P388 leukemia cells when they were perfused with LDL-daunomycin complex relative to free drug. The interaction of the LDL-daunomycin complex with P388 cells appeared to be through a LDL-specific pathway since competition with free LDL but not high-density lipoprotein significantly reduced daunomycin uptake. The LDL-daunomycin complex was cytotoxic to both daunomycin-sensitive and daunomycin-resistant P388 sublines. Colony growth studies showed the LDL-daunomycin complex to be more cytotoxic at shorter exposure times relative to free drug. Resistant cells showed 55 and 12% colony growth with 10-min and 2-hr exposures, respectively, to 0.4 microgram daunomycin/ml of the LDL-daunomycin complex. Free drug at 0.4 microgram/ml drug concentration and similar exposure times resulted in no loss in colony growth. The resistant cells were subjected to cell cycle analysis based on DNA content and showed an accumulation of cells at the G2-M phase of the cell cycle when treated with the LDL-daunomycin complex, with no effects being observed with free daunomycin.


[Databases in neurological diseases: overview of international examples in multiple sclerosis].
Vast amounts of data are created during routine patient care which are stored in unstructured digital and hardcopy formats in healthcare institutions. Analysis of large databases help to define the healthcare needs of the population and to organize healthcare services for specific diseases. As a model, we selected multiple sclerosis (MS), a disease with well-defined diagnostic criteria, a usually inpatient initial diagnosis, and a need for regular outpatient check-up. Using multiple sclerosis as an example, we set forth to screen and analyze international and Hungarian databases. In the framework of the initiation of the data lake system of Semmelweis University, we aim to define features of the data system needed for disease-specific databases for future applications. To determine essential data-entry criteria for such a database, we review the most important multiple sclerosis registries. We evaluate the type of registered data, structure of database, privacy issues, the availability and ways of application of the databases. Initially, the MS databases were created locally, aiming for better care of patients. As a further step, data were collected for scientific research by national and international co-operations. Disease-specific databases have become of high priority for national healthcare providers, and long-term information on a population ("real-world" data) is extremely important to assess the effectivity and safety of a treatment at the population level. Our analysis contributes to a project which focuses on the aspects of developing a data lake at a service provider level including clinical, diagnostic and digital healthcare departments of Semmelweis University, Budapest, Hungary. Orv Hetil. 2019; 160(4): 123-130.


Estimates of lateral and longitudinal bond energies within the microtubule lattice.
We developed a stochastic model of microtubule (MT) assembly dynamics that estimates tubulin-tubulin bond energies, mechanical energy stored in the lattice dimers, and the size of the tubulin-GTP cap at MT tips. First, a simple assembly/disassembly state model was used to screen possible combinations of lateral bond energy (DeltaG(Lat)) and longitudinal bond energy (DeltaG(Long)) plus the free energy of immobilizing a dimer in the MT lattice (DeltaG(S)) for rates of MT growth and shortening measured experimentally. This analysis predicts DeltaG(Lat) in the range of -3.2 to -5.7 k(B)T and DeltaG(Long) plus DeltaG(S) in the range of -6.8 to -9.4 k(B)T. Based on these estimates, the energy of conformational stress for a single tubulin-GDP dimer in the lattice is 2.1-2.5 k(B)T. Second, we studied how tubulin-GTP cap size fluctuates with different hydrolysis rules and show that a mechanism of directly coupling subunit addition to hydrolysis fails to support MT growth, whereas a finite hydrolysis rate allows growth. By adding rules to mimic the mechanical constraints present at the MT tip, the model generates tubulin-GTP caps similar in size to experimental estimates. Finally, by combining assembly/disassembly and cap dynamics, we generate MT dynamic instability with rates and transition frequencies similar to those measured experimentally. Our model serves as a platform to examine GTP-cap dynamics and allows predictions of how MT-associated proteins and other effectors alter the energetics of MT assembly.


Roles of motor and cortical activity in sleep rebound in rat.
Sleep pressure that builds up gradually during the extended wakefulness results in sleep rebound. Several lines of evidence, however, suggest that wake per se may not be sufficient to drive sleep rebound and that rapid eye movement (REM) and non-rapid eye movement (NREM) sleep rebound may be differentially regulated. In this study, we investigated the relative contribution of brain versus physical activities in REM and NREM sleep rebound by four sets of experiments. First, we forced locomotion in rats in a rotating wheel for 4 hr and examined subsequent sleep rebound. Second, we exposed the rats lacking homeostatic sleep response after prolonged quiet wakefulness and arousal brain activity induced by chemoactivation of parabrachial nucleus to the same rotating wheel paradigm and tested if physical activity could rescue the sleep homeostasis. Third, we varied motor activity levels while concurrently inhibiting the cortical activity by administering ketamine or xylazine (motor inhibitor), or ketamine + xylazine mixture and investigated if motor activity in the absence of activated cortex can cause NREM sleep rebound. Fourth and finally, we manipulated cortical activity by administering ketamine (that induced active wakefulness and waking brain) alone or in combination with atropine (that selectively inhibits the cortex) and studied if cortical inhibition irrespective of motor activity levels can block REM sleep rebound. Our results demonstrate that motor activity but not cortical activity determines NREM sleep rebound whereas cortical activity but not motor activity determines REM sleep rebound.


Characteristics of Transmissible CTX-M- and CMY-Type β-Lactamase-Producing Escherichia coli Isolates Collected from Pig and Chicken Farms in South Korea.
The rapid dissemination of extended-spectrum β-lactamase (ESBL)-producing Escherichia coli has significantly contributed to public health hazard globally. A total of 281 E. coli strains recovered from pigs and chickens between 2009 and 2015 in South Korea were analyzed for ESBL production. ESBL phenotypes were recognized in 14 E. coli isolates; ten and three ESBLproducing isolates carried only blaCTX-M and blaCMY genes, respectively, and one isolate harbored both genes. The predominant CTX-M and CMY types were CTX-M-15 (n = 8) and CMY-2 (n = 3). We also detected ESBL-producing isolates harboring blaCTX-M-65, blaCTX-M-14, blaCMY-6, blaDHA-1, and blaTEM-1 genes. All ESBL-producing isolates showed resistance to the extent of the fourth generation cephalosporins, along with multidrug resistance. CTX-M-15- producing isolates showed higher MIC values than CTX-M-14- and CTX-M-65-producing isolates. The blaCTX-M and blaCMY genes have the potential to be transferable. The spreading of blaCMY and blaCTX-M genes was arbitrated mainly v ia F rep a nd I ncI1 plasmids. Our i solates showed clonal diversity in PFGE analysis. This is the first report of E. coli isolates carrying blaCMY-6 in chicken from South Korea. The emergence of CMY-6 ESBLs in a population of poultry suggests that extensive screening with long-term surveillance is necessary to prevent the dissemination of ESBL from chicken to human.


Trichotillomania and non-epileptic seizures as sleep-related dissociative phenomena.
The occurrence of non-epileptic seizures (NES) and trichotillomania during sleep is rare. We describe the case of an adult woman with a personal history of childhood maltreatment and psychiatric morbidity (major depression, trichotillomania, and conversion disorder), who was referred to the sleep unit because of nocturnal hair-pulling and psychomotor agitation during sleep. An all-night PSG recording with audiovisual monitoring documented seven episodes of trichotillomania and one NES, all of which arose from unequivocal wakefulness. Improvement of nocturnal behaviors was observed after long-term psychotherapy. This case illustrates that nocturnal trichotillomania and NES may be symptoms of a sleep-related dissociative disorder.


Modeling of multivariate longitudinal phenotypes in family genetic studies with Bayesian multiplicity adjustment.
Genetic studies often collect data on multiple traits. Most genetic association analyses, however, consider traits separately and ignore potential correlation among traits, partially because of difficulties in statistical modeling of multivariate outcomes. When multiple traits are measured in a pedigree longitudinally, additional challenges arise because in addition to correlation between traits, a trait is often correlated with its own measures over time and with measurements of other family members. We developed a Bayesian model for analysis of bivariate quantitative traits measured longitudinally in family genetic studies. For a given trait, family-specific and subject-specific random effects account for correlation among family members and repeated measures, respectively. Correlation between traits is introduced by incorporating multivariate random effects and allowing time-specific trait residuals to correlate as in seemingly unrelated regressions. The proposed model can examine multiple single-nucleotide variations simultaneously, as well as incorporate familyspecific, subject-specific, or time-varying covariates. Bayesian multiplicity technique is used to effectively control false positives. Genetic Analysis Workshop 18 simulated data illustrate the proposed approach's applicability in modeling longitudinal multivariate outcomes in family genetic association studies.


Effects of riboflavin and fatty acid methyl esters on cholesterol oxidation during illumination.
The effect of riboflavin or fatty acid methyl esters on cholesterol photooxidation was studied. Samples containing cholesterol, either alone or in combination with riboflavin or fatty acid methyl esters, were illuminated at 25 degrees C in an incubator for 28 days. The various cholesterol oxidation products (COPs) and cholesterol were analyzed by gas chromatography-mass spectrometry (GC-MS), and riboflavin was determined by HPLC. Results showed that the presence of riboflavin or fatty acid methyl esters facilitated production of COPs and degradation of cholesterol, and the degradation fits a first-order model. The COPs formed during light storage included 7 alpha-OH, 7 beta-OH, 7-keto, 3,5-cholestadien-7-one, 5,6alpha-EP, and 5,6beta-EP. The addition of riboflavin caused formation of 3,5-cholestadien-7-one through dehydration of 7-keto, whereas in the presence of docosahexaenoic acid methyl ester, the formation of 5,6alpha-EP or 5,6beta-EP was favored. Riboflavin was more effective for generation of COPs than fatty acid methyl esters.


Unequal distribution of penicillin-binding proteins among inner membrane vesicles of Escherichia coli.
Escherichia coli penicillin-binding proteins (PBPs) were associated only with inner membrane vesicles when separated on 30 to 65% or 19 to 49% (wt/wt) sucrose gradients. Fractionation of vesicles through the low-density gradient revealed at least two classes of PBP-inner membrane associations. The first class consisted of PBPs 1 through 4, and the second class consisted of PBPs 5 through 8. These classes were distinguished by the density of vesicles with which they were associated; class 1 PBPs migrated with vesicles of higher density than did class 2 PBPs. Such combinations suggest that PBPs are nonrandomly distributed within the inner membrane, implying potential functional relationships among the PBPs themselves and with particular membrane domains. In addition, in cell lysates and in vesicle fractions, a 60,000-dalton aztreonam-insensitive PBP or protein fragment was observed which could potentially be confused with PBP3.


Malignant lymphoma of the posterior mediastinum with transverse myelopathy.
A case of malignant lymphoma in the thoracic paravertebral region with a severe spinal cord compression syndrome is presented. The patient was a 59-year-old female who had had right hypochondralgia as initial symptoms for more than one year. Chest radiograph and computed tomographic (CT) scanning on admission to another hospital had demonstrated a mass lesion in the thoracic paravertebral area. Due to the subsequent occurrence of progressive cord compression syndrome, she was transferred to our institution for the purpose of surgical treatment. Myelography demonstrated an obstruction of the spinal canal at the level of the 8th thoracic vertebra. CT-myelography showed a paravertebral tumour which extended through the intervertebral foramen into the epidural space and also spread to the posterior thoracic wall in succession. Tumour extirpation and decompressive laminectomy by single stage operation were performed. The pathological diagnosis was extranodal malignant lymphoma, diffuse, medium-sized cell type, with immunohistological demonstration of B-cell markers. Postoperatively, she was given a combination chemotherapy with VEPA (vincristine, cyclophosphamide, prednisone and doxorubisin), and the functional disorder has improved to some extent. To our knowledge, this is the first case of soft tissue malignant lymphoma in the posterior mediastinum.


The social and environmental dimensions of nutrition science.
To emphasise the importance of defining a new nutrition science and food policy that includes social and environmental dimensions. Nutrition science and food policy is put in the context of sustainable development. Examples are presented to show that a number of factors including exploitation of resources, disrespect for land and food insecurity contribute to the decline of a culture. The fate of cultures that lack implemented sustainable development strategies is discussed. Pressure from low-income and economically challenged countries combined with the efforts of not-for-profit private institutions is proposed. The goal is to produce and provide science-based evidence and guidelines to be used as a tool to encourage institutions and organisations to redefine their policies to deal effectively with global issues.


The detection of higher-order acoustic transitions is reflected in the N1 ERP.
The auditory system features various types of dedicated change detectors enabling the rapid parsing of auditory stimulation into distinct events. The activity of such detectors is reflected by the N1 ERP. Interestingly, certain acoustic transitions show an asymmetric N1 elicitation pattern: whereas first-order transitions (e.g., a change from a segment of constant frequency to a frequency glide [c-to-g change]) elicit N1, higher-order transitions (e.g., glide-to-constant [g-to-c] changes) do not. Consensus attributes this asymmetry to the absence of any available sensory mechanism that is able to rapidly detect higher-order changes. In contrast, our study provides compelling evidence for such a mechanism. We collected electrophysiological and behavioral data in a transient-detection paradigm. In each condition, a random (50%-50%) sequence of two types of tones occurred, which did or did not contain a transition (e.g., c-to-g and constant stimuli or g-to-c and glide tones). Additionally, the rate of pitch change of the glide varied (i.e., 10 vs. 40 semitones per second) in order to increase the number of responding neural assemblies. The rate manipulation modulated transient ERPs and behavioral detection performance for g-to-c transitions much stronger than for c-to-g transitions. The topographic and tomographic analyses suggest that the N1 response to c-to-g and also to g-to-c transitions emerged from the superior temporal gyrus. This strongly supports a sensory mechanism that allows the fast detection of higher-order changes.


An artificial beta-sheet that dimerizes through parallel beta-sheet interactions.
This Article introduces a simple chemical model of a beta-sheet (artificial beta-sheet) that dimerizes by parallel beta-sheet formation in chloroform solution. The artificial beta-sheet consists of two N-terminally linked peptide strands that are linked with succinic or fumaric acid and blocked along one edge with a hydrogen-bonding template composed of 5-aminoanisic acid hydrazide. The template is connected to one of the peptide strands by a turn unit composed of (S)-2-aminoadipic acid (Aaa). 1H NMR spectroscopic studies show that these artificial beta-sheets fold in CDCl3 solution to form well-defined beta-sheet structures that dimerize through parallel beta-sheet interactions. Most notably, all of these compounds show a rich network of NOEs associated with folding and dimerization. The compounds also exhibit chemical shifts and coupling constants consistent with the formation of folded dimeric beta-sheet structures. The aminoadipic acid unit shows patterns of NOEs and coupling constants consistent with a well-defined turn conformation. The present system represents a significant step toward modeling the type of parallel beta-sheet interactions that occur in protein aggregation.


Targeting Quorum Sensing: High-Throughput Screening to Identify Novel LsrK Inhibitors.
Since quorum sensing (QS) is linked to the establishment of bacterial infection, its inactivation represents one of the newest strategies to fight bacterial pathogens. LsrK is a kinase playing a key role in the processing of autoinducer-2 (AI-2), a quorum-sensing mediator in gut enteric bacteria. Inhibition of LsrK might thus impair the quorum-sensing cascade and consequently reduce bacterial pathogenicity. Aiming for the development of a target-based assay for the discovery of LsrK inhibitors, we evaluated different assay set-ups based on ATP detection and optimized an automation-compatible method for the high-throughput screening of chemical libraries. The assay was then used to perform the screening of a 2000-compound library, which provided 12 active compounds with an IC50 ≤ 10 µM confirming the effectiveness and sensitivity of our assay. Follow-up studies on the positive hits led to the identification of two compounds, harpagoside and rosolic acid, active in a cell-based AI-2 QS interference assay, which are at the moment the most promising candidates for the development of a new class of antivirulence agents based on LsrK inhibition.


[Quality of life after total gastrectomy depending on the method of alimentary tract reconstruction].
The quality of life was subjected to evaluation in two groups of patients after total gastrectomy depending on the method of alimentary tract reconstruction within at least nine months after the operation. One group included patients in whom the alimentary tract reconstruction was performed without creation of a reservoir according to Roux-Y (n = 10). The second group consisted of patients in whom a reservoir was created from the small intestine (jejunal pouch) according to Hunt-Lawrence-Rodino in Herfarth modification (n = 13). In both groups no statistically significant differences were found with respect to sex, age and time after the operation. The symptoms, body weight gain, amount of consumed food and the level of certain substances in blood were analysed. A higher quality of life was found in patients in whom in place of removed stomach, jejunal pouch was created.


Sliding of proteins non-specifically bound to DNA: Brownian dynamics studies with coarse-grained protein and DNA models.
DNA binding proteins efficiently search for their cognitive sites on long genomic DNA by combining 3D diffusion and 1D diffusion (sliding) along the DNA. Recent experimental results and theoretical analyses revealed that the proteins show a rotation-coupled sliding along DNA helical pitch. Here, we performed Brownian dynamics simulations using newly developed coarse-grained protein and DNA models for evaluating how hydrodynamic interactions between the protein and DNA molecules, binding affinity of the protein to DNA, and DNA fluctuations affect the one dimensional diffusion of the protein on the DNA. Our results indicate that intermolecular hydrodynamic interactions reduce 1D diffusivity by 30%. On the other hand, structural fluctuations of DNA give rise to steric collisions between the CG-proteins and DNA, resulting in faster 1D sliding of the protein. Proteins with low binding affinities consistent with experimental estimates of non-specific DNA binding show hopping along the CG-DNA. This hopping significantly increases sliding speed. These simulation studies provide additional insights into the mechanism of how DNA binding proteins find their target sites on the genome.


[Dynamics of some serum enzymes in the postnatal development of calves].
The activity of the following enzymes in clinically normal newborn calves was investigated: glutamate-oxalacetate transaminase (GOT), glutamate-pyruvate transaminase (GPT), alkaline phosphatase (APh), creatine phosphokinase (CPhK), lactate dehydrogenase (LDH), leucine-aminopeptidase (LAP), aldolase (A), and cholinesterase (ChE). The studies were carried out at the first hour prior to offering colostrum as well as at the 6th, 12th, 24th hr and on the 2nd, 3rd, 4th, 5th, 7th, 10th, 15th, and 20th day following it first intake. Regularly rising values of the enzyme activity up to the 24th hour were observed with APh, GOT, GPT, CPhK, and LAP. The aldolase enzyme (after colostrum had been given for the first time) in all animals showed a statistically significant drop of activity at the 6th hour. The activity of LDH displayed a consistently rising trend up to the end of the experimental period. The cholinesterase activity showed high values immediately following birth, reaching those found in the dams by the end of the observation period.


Actin branches out to link pathogen perception and host gene regulation.
Cellular functions of actin, and associated actin binding proteins (ABPs), have been well characterized with respect to their dynamic cytosolic role as components of the complex cytoskeletal network. In this regard, the collective research in this field has vastly expanded our knowledge of the role of actin to more recently identify a key role within the nucleus as an integral part gene organization and expression. Herein, we describe the requirement of the ABP actin depolymerizing factor-4 (ADF4) as a regulator of resistance to Pseudomonas syringae DC3000 expressing the effector AvrPphB via ADF4's cytosolic and nuclear functions. In total, our work has identified significant alterations in the expression of the resistance protein RPS5 in an ADF4 phosphorylation dependent manner. In this mini-review, we provide compelling evidence in support of both a nuclear function for ADF4, as well as potential targeting of the actin cytoskeleton by the bacterial effector AvrPphB.


Immunohistochemical localization of rhamnose-binding lectins in the steelhead trout (Oncorhynchus mykiss).
The localization of three -rhamnose-binding lectins named STL1, STL2, and STL3 from eggs of steelhead trout (Oncorhynchus mykiss) was analyzed by indirect immunohistochemical staining with specific antisera against individual lectins. In early previtellogenic oocyte, STL1 was localized not only in the cortical vesicles, but also in the plasma membrane and germinal vesicle. On the other hand, STL2 and STL3 were localized only in the cortical vesicles. In pre-fertilization mature egg, STLs were localized in a thin layer of cortical granules at the cytoplasmic side of the plasma membrane. STLs were accumulated on the surface of cytoplasm and inner membrane 30 min after fertilization. The strong staining with anti-STL1 antiserum was observed in several tissues and cells of the steelhead trout, such as spleen, thrombocytes, and blood leukocytes, but not erythrocytes. STL1 was also identified in exocrine cells, such as goblet cells of intestine and mucous cells of gill. These results indicate that the multiple lectins found in eggs of the steelhead trout play physiological roles not only in eggs, but also in various cells related to the innate immunity.


Gold Nanocluster-Decorated Nanocomposites with Enhanced Emission and Reactive Oxygen Species Generation.
Ligand-protected gold nanoclusters (AuNCs) show promise for high performance in biological applications, such as imaging and therapeutics. The assembly of AuNCs with biological macromolecules represents a simple but effective approach to fine-tuning of material functionalities. Thus, these materials might enable intracellular applications of AuNCs. Herein, we prepared a new AuNC-based nanometric system through a self-assembly approach mediated by hydrophobic and electrostatic effects. We show that hydrophobic and electrostatic effects between fluorescent AuNCs with protamine and hyaluronic acid contribute to the formation of small nanocomposites with acceptable colloidal stability. More importantly, the AuNC-decorated nanocomposites show assembly enhanced emission and singlet oxygen generation. In vitro experiments showed that our nanocomposites labeled specific cells by targeting CD44 and induced cell death by producing singlet oxygen. Hence, our AuNC-decorated nanocomposites show great potential as theranostic fluorescent nanomaterials.


MicroRNA-219-5p Inhibits Morphine-Induced Apoptosis by Targeting Key Cell Cycle Regulator WEE1.
BACKGROUND To identify the effects of microRNA (miR)-219-5p on morphine-induced apoptosis by targeting WEE1. MATERIAL AND METHODS Forty Balb/C mice (Toll-like receptor 9, TLR9 knockout) were randomly allocated to the experimental and control groups (20 in each group). The baseline miR-219-5p expression was detected using quantitative real-time PCR (qRT-PCR). After morphine was injected at 6 h on the 2nd and 6th days, experimental and control groups received miR-219-5p mimics or miRNA-negative control (NC), respectively, compound injection. Tissues and cells were later obtained from subjects in each group separately after mice were killed. TUNEL assay was used to investigate apoptosis in both groups. RAW264.7 cells were treated with miR-219-5p mimics and controls, respectively. After 24 h, 10 μM of morphine was added at 24 h. Cell apoptosis was assessed by flow cytometer. The WEE1 and Phospho-cdc2 (Tyr15) expressions were examined by Western blotting. RESULTS MiR-219-5p expression in the experimental group was significantly lower than that in the control group (P<0.05). Mice injected with miR-219-5p mimic experienced an evident increase in apoptosis rate compared with the control group (P<0.05). The miR-219-5p NC group and the morphine group both presented an elevated apoptosis rate compared with the blank control group (both, P<0.05). The apoptosis rate in the miR-219-5p mimic group was 10.06%, remarkably lower than in the miR-219-5p NC group and blank control group (both P<0.05). WEE1 and Tyr15 protein expressions in the miR-219-5p NC group and morphine group were obviously stronger than those in the blank control group (all P<0.05). In the miR-219-5p mimic group, WEE1 and Tyr15 protein expressions were significantly lower compared with those in the miR-219-5p NC group and morphine group (all P<0.05). CONCLUSIONS Morphine significantly downregulated the expression of miRNA-219-5p, which targets WEE1 to suppress Tyr15 expressions and activate Cdc2, thus inhibiting the morphine-induced macrophage apoptosis.


Mid-term outcomes and hemodynamic performance of transcatheter aortic valve implantation in bicuspid aortic valve stenosis: Insights from the bicuSpid TAvi duraBILITY (STABILITY) registry.
Limited data are available on transcatheter heart valves (THVs) durability in bicuspid aortic valve (BAV) stenosis. To evaluate evaluating 4-year clinical and echocardiographic outcomes of patients with BAV undergoing transcatheter aortic valve implantation (TAVI). The bicuSpid TAvi duraBILITY (STABILITY) registry is an Italian multicentre registry including all consecutive patients with BAV and severe aortic stenosis (AS), treated by means of TAVI between January 2011 and December 2017. Outcomes of interest were all-cause death at 4-year, over time changes in echocardiographic measurements, and THV durability according to the valve aortic research consortium (VARC)-3 update definitions. Study population included 109 patients (50% females; mean age 78 ± 7.5 years) with a mean Society of Thoracic Surgeons Predicted Risk of Mortality score of 5.1 ± 4.3%. Median follow-up (FU) duration was 4.1 years [interquartile range: 2.8-5.1]. The overall cumulative incidence of all-cause death by Kaplan-Meier estimates at 4 years was 32%. Compared to baseline, a significant decrease in transprosthetic mean gradient was obtained after TAVI (54 ± 16 vs. 10 ± 5 mmHg; p < 0.001), whereas a significant increase was observed at 4-year (13 ± 6.4 mmHg, p = 0.03). Cumulative incidence of hemodynamic valve dysfunction (HVD) was 4%. Six patients met HVD criteria: three moderate and three severe HVD. All three cases of severe HVD were clinically relevant (bioprosthetic valve failure [BVF]) with two patients receiving a reintervention (TAVI in TAVI), and one patient experiencing a valve-related death due to endocarditis. The STABILITY registry suggests that in patients with severe AS and BAV undergoing TAVI, postprocedural clinical benefits might last, over time, up to 4-year FU. The low rates of severe HVD and BVF may support the hypothesis of good THV durability also in BAV recipient.


Exploring Spatial Variability in the Relationship between Long Term Limiting Illness and Area Level Deprivation at the City Level Using Geographically Weighted Regression.
Ecological influences on health outcomes are associated with the spatial stratification of health. However, the majority of studies that seek to understand these ecological influences utilise aspatial methods. Geographically weighted regression (GWR) is a spatial statistics tool that expands standard regression by allowing for spatial variance in parameters. This study contributes to the urban health literature, by employing GWR to uncover geographic variation in Limiting Long Term Illness (LLTI) and area level effects at the small area level in a relatively small, urban environment. Using GWR it was found that each of the three contextual covariates, area level deprivation scores, the percentage of the population aged 75 years plus and the percentage of residences of white ethnicity for each LSOA exhibited a non-stationary relationship with LLTI across space. Multicollinearity among the predictor variables was found not to be a problem. Within an international policy context, this research indicates that even at the city level, a "one-size fits all" policy strategy is not the most appropriate approach to address health outcomes. City "wide" health polices need to be spatially adaptive, based on the contextual characteristics of each area.


The myelodysplastic syndromes: biology and implications for management.
Since the initial efforts to characterize the myelodysplastic syndromes in 1976, an extensive body of information has accumulated defining biologic features and the relation to clinical aspects of disease. While the pathogenesis of these disorders remains incompletely understood, laboratory investigations indicate that they are clonal disorders affecting hematopoietic stem cells characterized by a progressive imbalance between self-renewal and differentiation. Despite karyotypic resemblance to acute myeloid leukemia, fundamental biologic differences may underly the disappointing results achieved to date with intensive chemotherapy. The recent availability of recombinant hematopoietic growth factors for use in clinical trials has shown that the maturation defect in many instances can be overcome with administration of lineage-restricted recombinant hematopoietins. Routine use of these promising agents must await results of randomized clinical trials to determine the impact of prolonged treatment on leukemic evolution and disease-related morbidity.


Enhancing radiosensitisation of BRCA2-proficient and BRCA2-deficient cell lines with hyperthermia and PARP1-i.
Poly(ADP-ribose)polymerase1 (PARP1) is an important enzyme in regulating DNA replication. Inhibition of PARP1 can lead to collapsed DNA forks which subsequently causes genomic instability, making DNA more susceptible in developing fatal DNA double strand breaks. PARP1-induced DNA damage is generally repaired by homologous recombination (HR), in which BRCA2 proteins are essential. Therefore, BRCA2-deficient tumour cells are susceptible to treatment with PARP1-inhibitors (PARP1-i). Recently, BRCA2 was shown to be down-regulated by hyperthermia (HT) temporarily, and this consequently inactivated HR for several hours. In this study, we investigated whether HT exclusively interferes with HR by analysing thermal radiosensitisation of BRCA2-proficient and deficient cells. After elucidating the equitoxicity of PARP1-i on BRCA2-proficient and deficient cells, we studied the cell survival, apoptosis, DNA damage (γ-H2AX foci and comet assay) and cell cycle distribution after different treatments. PARP1-i sensitivity strongly depends on the BRCA2 status. BRCA2-proficient and deficient cells are radiosensitised by HT, indicating that HT does not exclusively act by inhibition of HR. In all cell lines, the addition of HT to radiotherapy and PARP1-i resulted in the lowest cell survival, the highest levels of DNA damage and apoptotic levels compared to duo-modality treatments. Concluding, HT not only inhibits HR, but also has the capability of radiosensitising BRCA2-deficient cells. Thus, in case of BRCA2-mutation carriers, combining HT with PARP1-i may boost the treatment efficacy. This combination therapy would be effective for all patients with PARP1-i regardless of their BRCA status.


Deconstructing magnetization noise: Degeneracies, phases, and mobile fractionalized excitations in tetris artificial spin ice.
Direct detection of spontaneous spin fluctuations, or "magnetization noise," is emerging as a powerful means of revealing and studying magnetic excitations in both natural and artificial frustrated magnets. Depending on the lattice and nature of the frustration, these excitations can often be described as fractionalized quasiparticles possessing an effective magnetic charge. Here, by combining ultrasensitive optical detection of thermodynamic magnetization noise with Monte Carlo simulations, we reveal emergent regimes of magnetic excitations in artificial "tetris ice." A marked increase of the intrinsic noise at certain applied magnetic fields heralds the spontaneous proliferation of fractionalized excitations, which can diffuse independently, without cost in energy, along specific quasi-1D spin chains in the tetris ice lattice.


Cumulative meta-analysis and trial sequential analysis of correlation between hOGG1 Ser326Cys polymorphism and the risk of head and neck squamous cell carcinoma.
The human 8-oxoguanine glycosylase 1 (hOGG1) Ser326Cys polymorphism has been involved in the risk of head and neck squamous cell carcinoma (HNSCC), but the results of published studies on this topic still inconsistent. Finally 11 qualified publications with 13 independent case-control studies were yielded. Overall, we observed significant differences in CysCys vs. SerSer [odds ratio (OR) = 1.55, 95% confidence interval (95% CI) = 1.01-2.38] and CysCys vs. SerCys+SerSer (OR = 1.42, 95% CI = 1.005-1.99) genetic models. Sensitivity analyses showed the results were not robust, cumulative meta-analyses and trial sequential analysis indicated the results didn't not need more studies to identification. Subgroup analyses showed there was a significant association in Caucasian, laryngeal squamous cell carcinoma, studies agreement with Hardy-Weinberg equilibrium, and alcohol drinkers subgroups under the corresponding contrasts. In addition, the results of Egger's test were contradictory. All eligible studies were searched from the online databases including PubMed, Web of Science, China Knowledge Resource Integrated Database, and Wanfang databases up to February 10, 2017. After study selection and data extraction, the meta-analysis was performed using STATA 12.0 software and TSA software version 0.9 Beta. Our meta-analysis results indicated that hOGG1 Ser326Cys polymorphism may be associated with increased risk of HNSCC, especially in Caucasians, alcohol drinkers and the patients with laryngeal squamous cell carcinoma.


Investigation and Functional Enrichment Analysis of the Human Host Interaction Network with Common Gram-Negative Respiratory Pathogens Predicts Possible Association with Lung Adenocarcinoma.
Haemophilus influenzae (Hi), Moraxella catarrhalis (MorCa) and Pseudomonas aeruginosa (Psa) are three of the most common gram-negative bacteria responsible for human respiratory diseases. In this study, we aimed to identify, using the functional enrichment analysis (FEA), the human gene interaction network with the aforementioned bacteria in order to elucidate the full spectrum of induced pathogenicity. The Human Pathogen Interaction Database (HPIDB 3.0) was used to identify the human proteins that interact with the three pathogens. FEA was performed via the ToppFun tool of the ToppGene Suite and the GeneCodis database so as to identify enriched gene ontologies (GO) of biological processes (BP), cellular components (CC) and diseases. In total, 11 human proteins were found to interact with the bacterial pathogens. FEA of BP GOs revealed associations with mitochondrial membrane permeability relative to apoptotic pathways. FEA of CC GOs revealed associations with focal adhesion, cell junctions and exosomes. The most significantly enriched annotations in diseases and pathways were lung adenocarcinoma and cell cycle, respectively. Our results suggest that the Hi, MorCa and Psa pathogens could be related to the pathogenesis and/or progression of lung adenocarcinoma via the targeting of the epithelial cellular junctions and the subsequent deregulation of the cell adhesion and apoptotic pathways. These hypotheses should be experimentally validated.


2'-Fluorinated arabinonucleosides of 5-(2-haloalkyl)uracil: synthesis and antiviral activity.
The synthesis of 5-(2-fluoroethyl)-2'-deoxyuridine (FEDU, 4b), its 2'-fluoro analogue 1-(2-deoxy-2-fluoro-beta-D-arabinofuranosyl)-5-(2-fluoroethyl)-1H,3H- pyrimidine-2,4-dione (FEFAU, 4k), and the 2'-fluoro analogue of the potent antiherpes virus compound 5-(2-chloroethyl)-2'-deoxyuridine (CEDU), 5-(2-chloroethyl)-1-(2-deoxy-2-fluoro-beta-D-arabinofuranosyl)-1H,3H-pyr imidine - 2,4-dione (CEFAU, 4i), is described. The antiviral activities of these compounds were determined in cell culture against herpes simplex virus (HSV) types 1 and 2 and varicella zoster virus (VZV). All compounds were shown to possess significant and selective antiviral activity. FEDU proved less potent than CEDU against VZV replication; however, it was more active against HSV-2. CEFAU showed marked activity against HSV-1, HSV-2, and VZV. The compound containing fluorine at both positions, FEFAU, exhibited the strongest antiviral potency against HSV-1, HSV-2, and VZV. It inhibited HSV-1 at a concentration of 0.03-0.2 microgram/mL, HSV-2 at 0.1-0.3 microgram/mL, and VZV at 0.03 microgram/mL. Neither FEDU nor CEFAU or FEFAU exerted a significant inhibitory effect on cell proliferation at a concentration of 100 micrograms/mL. Thus, the cytotoxicity of these compounds is as low as that of CEDU and compares favorably to that of previously described 2'-fluoroarabinosyl nucleoside analogues.


Stanhope Speer, Physician and Alpinist: In 1853, First to Describe Mountain Sickness?
In 1853, Stanhope Templeman Speer published a two-part paper in The Association Medical Journal on Mountain Sickness. Speer was a physician who had worked at the Brompton Hospital for Chest Diseases in London and had been Professor of Medicine in Dublin. He was also an Alpine climber and had made the first ascent of one of the Wetterhorn peaks. His article ran to ten and a half pages in the Journal and to 50 pages in a reprint. It consists of anecdotal accounts of symptoms suffered at altitude from the literature and from his own experiences in the European Alps. He asks three pertinent questions. Is there a condition of mountain sickness? Are these symptoms felt by all persons alike and at the same height? What are the causes, and whence the explanation of such phenomena? In the course of the article, he answers the first two questions but, like us, 162 years later, is unable to answer the third. This article seeks to present Speer's original work and such facts about his life as I have been able to discover.


Resveratrol suppresses TPA-induced matrix metalloproteinase-9 expression through the inhibition of MAPK pathways in oral cancer cells.
Naturally occurring agents, such as resveratrol, have been determined to benefit health. Numerous studies have demonstrated that resveratrol has antioxidative, cardioprotective, and neuroprotective properties. However, the effect of resveratrol exerts on the metastasis of oral cancer cells remains unclear. In this study, we investigated the effect the anti-invasive activity of resveratrol on a human oral cancer cell line (SCC-9) in vitro and the underlying mechanisms. Cell viability was examined by MTT assay, whereas cell motility was measured by migration and wound-healing assays. Zymography, reverse-transcriptase polymerase chain reaction (PCR), and promoter assays confirmed the inhibitory effects of resveratrol on matrix metalloproteinase-9 (MMP-9) expression in oral cancer cells. We established that various concentrations (0-100 μM) of resveratrol inhibited the 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced migration capacities of SCC-9 cells and caused no cytotoxic effects. Zymography and Western blot analyses suggested that resveratrol inhibited TPA-induced MMP-9 gelatinolytic activity and protein expression. In addition, the results indicated that resveratrol inhibited the phosphorylation of c-Jun N-terminal kinase (JNK)1/2 and extracellular-signal-regulated kinase (ERK)1/2 involved in downregulating protein expression and the transcription of MMP-9. In summary, resveratrol inhibited MMP-9 expression and oral cancer cell metastasis by downregulating JNK1/2 and ERK1/2 signals pathways and, thus, exerts beneficial effects in chemoprevention.


Tongue protrusion dystonia: treatment with botulinum toxin.
We report the treatment experience in a series of patients with involuntary tongue protrusion resulting from oromandibular dystonia (OMD) or Meige's syndrome. A retrospective analysis of clinical findings and results of treatment was conducted on patients treated at Vanderbilt University Medical Center between 1989 and 1995. After unsuccessful treatment with conventional oral medications, nine patients having involuntary tongue protrusion resulting from OMD or Meige's syndrome were treated with botulinum toxin type A (BTX-A) injected into the genioglossus muscle at four sites via a submandibular approach. A marked reduction in tongue protrusion was achieved in six patients (67%). Of 35 consecutive injections, 83% were successful at reducing tongue protrusion. Mild dysphagia complicated 14% of the injections. The average dose injected was 34 (+/- 3) units producing a 15 (+/- 2) week average duration of effect. Injection of the genioglossus with BTX-A may prove to be a valid treatment option for involuntary tongue protrusion related to OMD or Meige's syndrome. A double-blind, placebo-controlled trial is needed to better define efficacy and adverse events.


Reversible unilateral nigrostriatal pathway inhibition induced through expression of adenovirus-mediated clostridial light chain gene in the substantia nigra.
Clostridial light chain (LC) inhibits synaptic transmission by digesting a vesicle-docking protein, synaptobrevin, without killing neurons. We here report the feasibility of creating a rat hemiparkinsonism model through LC gene expression in the substantia nigra (SN), inhibiting nigrostriatal transmission. 40 adult Sprague Dawley rats were divided into four groups for SN injections of PBS, 6-hydroxydopamine (6-OHDA), or adenoviral vectors for the expression of LC (AdLC), or GFP (AdGFP). Amphetamine and apomorphine induced rotations were assessed before and after SN injection, revealing significant rotational alterations at 8 or 10 days after injection in both AdLC and 6-OHDA but not PBS and AdGFP groups. Induced rotation recovered by one month in AdLC rats but persisted in 6-OHDA rats. Histological analysis of the SN revealed LC and GFP expression with corresponding synaptobrevin depletion in the LC, but not the GFP groups. Tyrosine hydroxylase (TH) and dopamine transporter (DAT) immunohistochemistry (IHC) showed markedly decreased staining in ipsilateral SN and striatum in 6-OHDA but not AdLC or AdGFP rats. Similarly, compared with contralateral, ipsilateral striatal dopamine level only decreased in 6-OHDA but not AdLC, AdGFP, or PBS treated rats. Thus, LC expression induces nigral synaptobrevin depletion with resulting inhibition of nigrostriatal synaptic transmission. Unlike 6-OHDA, LC expression inhibits synaptic activity without killing neurons. This approach, therefore, represents a potentially reversible means of nigrostriatal pathway inhibition as a model for Parkinson's disease. Such a model might facilitate transient and controlled nigral inhibition for studying striatal recovery, dopaminergic re-innervation, and normalization of striatal receptors following the recovery of nigrostriatal transmission.


Effectiveness of an oral health program for mothers and their infants.
The baby oral health program (bOHP) provides pregnant women and their future babies with oral care. To assess the bOHP effectiveness by comparing caries prevalence in infants enrolled and not enrolled in the oral health program (OHP). Mothers who had been invited to participate in the bOHP from 2006 to 2009 were contacted. Two groups were formed: 87 pairs of mothers and infants who effectively participated in the OHP (G1) and 107 pairs who did not (G2). Mothers and infants were given a dental examination. Socio-economic status (SES) and education level (EL) questionnaires were completed. t-tests and multivariate logistic regression were used in analyzing data. Statistically significant differences were observed in the mean age of mothers (G1 = 33.8 years; G2 = 35.6 years; P = 0.015) and mean decayed, missed and filled surface (DMFS) score (G1 = 24.71; G2 = 32.58; P < 0.001), not in SES (P = 0.758) and EL (P = 0.109). Mean age and mean dmfs scores of G1 and G2 children were 4.2 and 4.4 years (P = 0.068), and 0.25 and 4.12 (P < 0.001), respectively. The odds ratio (OR) for children in G2 to develop dentine lesions, as opposed to those in G1, was 48.56. The bOHP was effective in preventing caries in infants enrolled in the program.


Parathyroid hormone-related protein induced coupled increases in bone formation and resorption markers for 7 years in a patient with malignant islet cell tumors.
Parathyroid hormone-related protein (PTHrP) and PTH share the common PTH/PTHrP receptor. Although an elevated level of circulating PTHrP in patients with malignancies causes hypercalcemia as does PTH, chronic and systemic effects of PTHrP on bone metabolism in humans are not well understood because tumor-burden patients showing hypercalcemia usually have a poor prognosis. We investigated bone and calcium metabolism in a patient with malignant islet cell tumors showing hypercalcemia due to the elevated plasma PTHrP level for 7 years. Hypercalcemia and hypercalciuria continued throughout the clinical course in spite of frequent infusions of bisphosphonates. Bone resorption markers and a bone formation marker were consistently elevated as seen in primary hyperparathyroidism, a disease caused by an autonomous hypersecretion of PTH. Based on biochemical measurements including bone markers and serum 1,25-dihydroxyvitamin D, the clinical features of this case essentially are the same as those of primary hyperparathyroidism except for the elevated level of plasma PTHrP with suppressed intact PTH level. Therefore, it is suggested that chronic and systemic effects of PTHrP on bone as well as calcium metabolism are indistinguishable from those of PTH in human.


Spontaneous healing of iatrogenic direct carotid cavernous fistula.
Direct carotid-cavernous fistula (CCF) by selective navigation using a microcatheter or microwire is a rare complication, and its timing of treatment has not been elucidated. We report two cases of direct CCFs resulting from injury to the cavernous posterior segment of the internal carotid artery during selective navigation. We did not plan to perform emergent endovascular treatment for these direct CCFs because no symptoms related to direct CCFs developed. Follow-up angiography revealed spontaneous healing of both direct CCFs. Close observation rather than emergent treatment may represent another option for direct CCF by selective navigation during the endovascular procedure.


Covalent protein modification with ISG15 via a conserved cysteine in the hinge region.
The ubiquitin-like protein ISG15 (interferon-stimulated gene of 15 kDa) is strongly induced by type I interferons and displays antiviral activity. As other ubiquitin-like proteins (Ubls), ISG15 is post-translationally conjugated to substrate proteins by an isopeptide bond between the C-terminal glycine of ISG15 and the side chains of lysine residues in the substrates (ISGylation). ISG15 consists of two ubiquitin-like domains that are separated by a hinge region. In many orthologs, this region contains a single highly reactive cysteine residue. Several hundred potential substrates for ISGylation have been identified but only a few of them have been rigorously verified. In order to investigate the modification of several ISG15 substrates, we have purified ISG15 conjugates from cell extracts by metal-chelate affinity purification and immunoprecipitations. We found that the levels of proteins modified by human ISG15 can be decreased by the addition of reducing agents. With the help of thiol blocking reagents, a mutational analysis and miRNA mediated knock-down of ISG15 expression, we revealed that this modification occurs in living cells via a disulphide bridge between the substrates and Cys78 in the hinge region of ISG15. While the ISG15 activating enzyme UBE1L is conjugated by ISG15 in the classical way, we show that the ubiquitin conjugating enzyme Ubc13 can either be classically conjugated by ISG15 or can form a disulphide bridge with ISG15 at the active site cysteine 87. The latter modification would interfere with its function as ubiquitin conjugating enzyme. However, we found no evidence for an ISG15 modification of the dynamin-like GTPases MxA and hGBP1. These findings indicate that the analysis of potential substrates for ISG15 conjugation must be performed with great care to distinguish between the two types of modification since many assays such as immunoprecipitation or metal-chelate affinity purification are performed with little or no reducing agent present.


PADGEM-dependent adhesion of platelets to monocytes and neutrophils is mediated by a lineage-specific carbohydrate, LNF III (CD15).
PADGEM (platelet activation-dependent granule-external membrane protein) is a leukocyte receptor of activated platelets that mediates cellular adhesion of platelets to neutrophils and monocytes. To identify the natural ligand on neutrophils and monocytes that interacts with PADGEM, we have evaluated anti-leukocyte antibodies for their ability to block leukocyte-PADGEM binding. Only anti-CD15 antibodies were able to inhibit the binding of neutrophils, monocytes, HL60 cells, and U937 cells to platelets. Anti-CD15 antibodies inhibited the binding of U937 cells to PADGEM-expressing COS cells and to purified PADGEM incorporated into phospholipid vesicles. The CD15 antigen, lacto-N-fucopentaose III (Gal beta 1----4[Fuc alpha 1----3]NAcGlc beta 1----3Gal-beta 1----4Glc), inhibited the interaction of neutrophils or HL60 cells with platelets, whereas lacto-N-fucopentaose I did not; lacto-N-fucopentaose II demonstrated minimal inhibition. Lacto-N-fucopentaose III, and to a lesser extent lacto-N-fucopentaose II, but not lacto-N-fucopentaose I, inhibited the interaction of HL60 cells with COS cells transfected with PADGEM cDNA. CD15, lacto-N-fucopentaose III or Lex, is a component of the PADGEM ligand on neutrophils and monocytes.


Amorphous silica modeled with truncated and screened Coulomb interactions: a molecular dynamics simulation study.
We show that finite-range alternatives to the standard long-range pair potential for silica by van Beest et al. [Phys. Rev. Lett. 64, 1955 (1990)] might be used in molecular dynamics simulations. We study two such models that can be efficiently simulated since no Ewald summation is required. We first consider the Wolf method, where the Coulomb interactions are truncated at a cutoff distance rc such that the requirement of charge neutrality holds. Various static and dynamic quantities are computed and compared to results from simulations using Ewald summations. We find very good agreement for rc approximately 10 A. For lower values of rc, the long-range structure is affected which is accompanied by a slight acceleration of dynamic properties. In a second approach, the Coulomb interaction is replaced by an effective Yukawa interaction with two new parameters determined by a force fitting procedure. The same trend as for the Wolf method is seen. However, slightly larger cutoffs have to be used in order to obtain the same accuracy with respect to static and dynamic quantities as for the Wolf method.


Boron neutron capture therapy in cancer: past, present and future.
Undifferentiated thyroid cancer (UTC) is a very aggressive tumor with no effective treatment, since it lacks iodine uptake and does not respond to radio or chemotherapy. The prognosis of these patients is bad, due to the rapid growth of the tumor and the early development of metastasis. Boron neutron capture therapy (BNCT) is based on the selective uptake of certain boron non-radioactive compounds by a tumor, and the subsequent irradiation of the area with an appropriate neutron beam. 10B is then activated to 11B, which will immediately decay releasing alpha particles and 7Li, of high linear energy transfer (LET) and limited reach. Clinical trials are being performed in patients with glioblastoma multiforme and melanoma. We have explored its possible application to UTC. Our results demonstrated that a cell line of human UTC has a selective uptake of borophenylalanine (BPA) both in vitro and after transplantation to nude mice. Treatment of mice by BNCT led to a complete control of growth and cure of 100% of the animals. Moreover dogs with spontaneous UTC also have a selective uptake of BPA. At the present we are studying the biodistribution of BPA in patients with UTC before its application in humans.


Temple-Baraitser Syndrome and Zimmermann-Laband Syndrome: one clinical entity?
KCNH1 encodes a voltage-gated potassium channel that is predominantly expressed in the central nervous system. Mutations in this gene were recently found to be responsible for Temple-Baraitser Syndrome (TMBTS) and Zimmermann-Laband syndrome (ZLS). Here, we report a new case of TMBTS diagnosed in a Lebanese child. Whole genome sequencing was carried out on DNA samples of the proband and his parents to identify mutations associated with this disease. Sanger sequencing was performed to confirm the presence of detected variants. Whole genome sequencing revealed three missense mutations in TMBTS patient: c.1042G > A in KCNH1, c.2131 T > C in STK36, and c.726C > A in ZNF517. According to all predictors, mutation in KCNH1 is damaging de novo mutation that results in substitution of Glycine by Arginine, i.e., p.(Gly348Arg). This mutation was already reported in a patient with ZLS that could affect the connecting loop between helices S4-S5 of KCNH1 with a gain of function effect. Our findings demonstrate that KCNH1 mutations cause TMBTS and expand the mutational spectrum of KCNH1 in TMBTS. In addition, all cases of TMBTS were reviewed and compared to ZLS. We suggest that the two syndromes are a continuum and that the variability in the phenotypes is the result of the involvement of genetic modifiers.


Circulating Actin-Binding Proteins in Laryngeal Cancer: Its Relationship with Circulating Tumor Cells and Cells of the Immune System.
We previously exposed the role of actin-binding proteins (ABPs) in cancer development and progression. In this paper, we studied the relationship between circulating ABPs and the number of ABP-expressing leukocytes and circulating tumor cells (CTCs) in patients with highly aggressive laryngeal squamous cell carcinoma (LSCC). The levels of cofilin (CFL1), profilin (PFN1), ezrin (EZR), fascin (FSCN1), and adenylate cyclase-associated protein 1 (CAP1) were determined using enzyme immunoassay. The ABP expression by the cellular pools was analyzed by flow cytometry. The highest levels of FSCN1 and EZR were found in the blood serum of LSCC patients. There was a difference in ABP expression between the pools of leukocytes and CTCs. Leukocytes were mainly represented by CAP1+ and FSCN1+ pools, and CTCs contained CAP1+, FSCN1+, and EZR+ cells. The serum FSCN1 level correlated with the number of FSCN1-containing and CFL1-containing leukocytes. Thus, the level of circulating EZR is likely related to its expression in CTCs. The levels of CFL1 and PFN1 are likely to be supported by the expression of these proteins by leukocytes. Both CTCs and leukocytes can be a source of FSCN1 and CAP1 in blood serum. The results suggest that serum proteins can be produced by various cells, thus indicating both cancer development and the response of the immune system to this process.


Reporting of suicide in the Australian media.
The media monitoring project aimed to establish a baseline picture of the extent, nature and quality of reporting of suicide by the Australian media, with a view to informing future strategies intended to optimize reporting of suicide. Newspaper, television and radio items on suicide were retrieved over 12 months. Identifying and descriptive information were extracted for each item. Approximately 10% of items were rated for quality, using a rating scale based on criteria from Achieving the Balance, a kit designed to promote awareness among media professionals of issues relating to suicide. The scale ranged from 0 (poor quality) to 100 (good quality). Reporting of suicide was extensive (with 4813 items retrieved). The nature of reporting was variable. Items tended to be about completed suicide (rather than attempted suicide or suicidal ideation), and most commonly involved content related to an individual's experiences, policy/programme initiatives and/or suicide statistics, although there were differences across media types. Items showed variability across dimensions of quality. The majority of suicide items did not have examples of inappropriate language, were not inappropriately located, did not use the word 'suicide' in the headline, and did not use explicit photographs/diagrams or footage. However, around half of the suicide items provided a detailed discussion of the method of self-harm and portrayed suicide as merely a social phenomenon. Where items concerned the suicide of a celebrity, reference was commonly made to that person's celebrity status. Most items failed to provide information on help services. The median total quality score was 57.1%. The reporting of suicide is extensive across all media types, and varies in nature and quality. In general, good items outnumber poorer items. However, there are still opportunities for improving media reporting of suicide.


The effects of bacterial volatile emissions on plant abiotic stress tolerance.
Plant growth-promoting rhizobacteria (PGPR) are beneficial plant symbionts that have been successfully used in agriculture to increase seedling emergence, plant weight, crop yield, and disease resistance. Some PGPR strains release volatile organic compounds (VOCs) that can directly and/or indirectly mediate increases in plant biomass, disease resistance, and abiotic stress tolerance. This mini-review focuses on the enhancement of plant abiotic stress tolerance by bacterial VOCs. The review considers how PGPR VOCs induce tolerance to salinity and drought stress and also how they improve sulfur and iron nutrition in plants. The potential complexities in evaluating the effects of PGPR VOCs are also discussed.


Medical ethics, bioethics and research ethics education perspectives in South East Europe in graduate medical education.
Ethics has an established place within the medical curriculum. However notable differences exist in the programme characteristics of different schools of medicine. This paper addresses the main differences in the curricula of medical schools in South East Europe regarding education in medical ethics and bioethics, with a special emphasis on research ethics, and proposes a model curriculum which incorporates significant topics in all three fields. Teaching curricula of Medical Schools in Bulgaria, Bosnia and Herzegovina, Croatia, Serbia, Macedonia and Montenegro were acquired and a total of 14 were analyzed. Teaching hours for medical ethics and/or bioethics and year of study in which the course is taught were also analyzed. The average number of teaching hours in medical ethics and bioethics is 27.1 h per year. The highest national average number of teaching hours was in Croatia (47.5 h per year), and the lowest was in Serbia (14.8). In the countries of the European Union the mean number of hours given to ethics teaching throughout the complete curriculum was 44. In South East Europe, the maximum number of teaching hours is 60, while the minimum number is 10 teaching hours. Research ethics topics also show a considerable variance within the regional medical schools. Approaches to teaching research ethics vary, even within the same country. The proposed model for education in this area is based on the United Nations Educational, Scientific and Cultural Organization Bioethics Core Curriculum. The model curriculum consists of topics in medical ethics, bioethics and research ethics, as a single course, over 30 teaching hours.


RepA protein- and oriR-dependent initiation of R1 plasmid replication: identification of a rho-dependent transcription terminator required for cis-action of repA protein.
Initiation of R1 plasmid replication is dependent on cis-acting repA protein and the 188 base-pair (bp) sequence, oriR. RepA protein synthesized in vitro preferentially activates oriR in cis, regardless of the orientation and location of oriR on the template DNA. RepA protein is not reusable after it activates oriR in the cis-position. Cis-action of repA protein is also dependent on the presence of CIS, a 170 bp sequence, between repA and oriR. CIS contains a rho dependent transcription terminator of the repA transcript, deletion of which results in decrease in transformation efficiency and rapid loss of plasmid in the absence of selection. The significance of transcription termination events in replication was indicated by decreased replication activity in vivo caused by premature termination of the repA transcript between repA and CIS. A model which may account for the role of CIS in mediating the cis-action of the repA protein is presented.


Dopaminergic influences on emotional decision making in euthymic bipolar patients.
We recently reported that the D2/D3 agonist pramipexole may have pro-cognitive effects in euthymic patients with bipolar disorder (BPD); however, the emergence of impulse-control disorders has been documented in Parkinson's disease (PD) after pramipexole treatment. Performance on reward-based tasks is altered in healthy subjects after a single dose of pramipexole, but its potential to induce abnormalities in BPD patients is unknown. We assessed reward-dependent decision making in euthymic BPD patients pre- and post 8 weeks of treatment with pramipexole or placebo by using the Iowa Gambling Task (IGT). The IGT requires subjects to choose among four card decks (two risky and two conservative) and is designed to promote learning to make advantageous (conservative) choices over time. Thirty-four BPD patients completed both assessments (18 placebo and 16 pramipexole). Baseline performance did not differ by treatment group (F=0.63; p=0.64); however, at week 8, BPD patients on pramipexole demonstrated a significantly greater tendency to make increasingly high-risk, high-reward choices across the five blocks, whereas the placebo group's pattern was similar to that reported in healthy individuals (treatment × time × block interaction, p<0.05). Analyses of choice strategy using the expectancy valence model revealed that after 8 weeks on pramipexole, BPD patients attended more readily to feedback related to gains than to losses, which could explain the impaired learning. There were no significant changes in mood symptoms over the 8 weeks, and no increased propensity toward manic-like behaviors were reported. Our results suggest that the enhancement of dopaminergic activity influences risk-associated decision-making performance in euthymic BPD. The clinical implications remain unknown.


Histological quantitation of aluminum in iliac bone from patients with renal failure.
Aluminum in undemineralized, methyl methacrylate-embedded iliac bone from 16 hemodialyzed renal patients and six normal controls was stained by a modification of the aluminon method and quantitated histomorphometrically, and the results were compared with the aluminum values measured by atomic absorption spectrophotometry. There was a high degree of correlation between the two techniques (r = 0.955, p less than 0.0001). The amount of stained aluminum also showed a statistically significant relationship with the amount of bone that is not mineralized (r = 0.840, p less than 0.001), with the TM (r = 0.841, p less than 0.001), and with osteoid width (r = 0.771, p less than 0.0001). The aluminum was found as bright red bands mainly in the junction of mineralized bone and osteoid but was also observed in cement lines of mature bone, on neutral surfaces, and surrounding the osteocytes. Analysis of serial sections for aluminum and tetracycline showed that a majority of the sites with aluminum did not take up tetracycline, implying defective mineralization. Occasionally tetracycline uptake was observed in such places, suggesting that the aluminum "block" might be overcome eventually. The presence of aluminum in cement lines of mature bone supports this hypothesis. The histochemical staining of aluminum provides a quick and easy method for identifying, localizing, and quantifying aluminum within the bone and may prove to be a useful tool in the study of the mechanisms of metabolic bone disease.