[Therapy of multiple myeloma. Results of sequential treatment with melphalan-prednisolone and the M2 protocol].
Melphalan and prednisolone were intermittently used for treatment of 101 patients with multiple myeloma. In case of progression, either initially or after remission or after a phase of stabilisation, treatment was changed to the M-2 protocol. By primary treatment, remission was obtained in 24 patients, a stable phase in 47 patients. The treatment of second choice which became necessary in a total of 46 patients led to remission in 5 patients (11%), 21 (46%) became stabilised and in 20 (43%) the disease progressed. The median survival time for all 101 patients was 40.5 months after commencement of treatment. In patients with remission it was clearly longer (67 months) than in those with stable disease (38.1 months) or in those with progression (16.8 months). The mean time of survival after commencement of treatment of second choice was 12.5 months. Compared to other studies the remission rates are slight, the time of survival, however, is very remarkable. The overall survival rates are equivalent to the results thus far obtained by more aggressive treatment schemes.


Clonidine, but not bradykinin or ANP, inhibits Na+ and water transport in Dahl SS rat CCD.
We examined the actions of potentially natriuretic autacoids in the isolated perfused cortical collecting duct (CCD) dissected from inbred Dahl (Rapp strain) salt-sensitive rats (SS). Atrial natriuretic peptide (ANP, 10 nM), bradykinin (BK, 10 nM), and clonidine (1 microM) were studied to determine their effects on the lumen-to-bath flux of 22Na+ (J1-->b, pmol min-1 mm-1), hydraulic conductivity (Pf, micron/s), and transepithelial voltage (VT, mV). ANP and BK have been shown by others to significantly reduce net Na+ reabsorption and hydraulic conductivity in the Sprague-Dawley (SD) rat CCD, but previous results from our laboratory showed no ANP or BK effect in the SD CCD. In the present study, we were also unable to observe any effect of either ANP or BK in the SS rat CCD. However, in the presence of AVP, clonidine (a partial alpha 2-adrenergic receptor agonist) significantly reduced J1-->b and Pf from 139 +/- 6 (SEM) to 88 +/- 7 and from 959 +/- 176 to 490 +/- 73, respectively. In addition, clonidine significantly depolarized VT from -14.5 +/- 2.8 to -11.2 +/- 1.8. However, unlike its effects in the SD rat CCD, yohimbine (300 nM, an alpha 2-adrenergic receptor antagonist) did not significantly reverse the effects of clonidine on J1-->b, Pf or VT in the SS rat CCD.


Visualization of thermally fluctuating surface structure in noncontact atomic-force microscopy and tip effects on fluctuation: theoretical study of Si(111)-(square root[3] x square root[3])-Ag surface.
We investigated noncontact atomic-force microscopy (NC-AFM) images of a thermally fluctuating surface structure together with tip effects based on the first-principles electronic state calculation. As an example the Si(111)-(square root[3] x square root[3])-Ag (square root[3]-Ag) surface is studied. We have succeeded in theoretically visualizing the thermal fluctuation of the square root[3]-Ag surface at room temperature, and in reproducing the observed NC-AFM image for the first time. Further, the pinning effect of the thermal fluctuation of the square root[3]-Ag surface by the tip is clarified, which shows a novel ability of NC-AFM to modify the surface structure.


The effects of intravenously administered methylxanthines on the proportion of goblet cells containing fucosylated glycoconjugates in rabbit tracheal epithelium.
The proportion of goblet cells containing various fucosylated glycoconjugates was evaluated with the use of lectin histochemistry in rabbit tracheal epithelium at 15 and 30 min after intravenous administration of either aminophylline (Syntophyllin) or a mixture of etophylline and theophylline (Oxantil). Methylxanthine derivatives are nonspecific inhibitors of phosphodiesterases that are used to treat bronchial asthma; the proportion of fucosylated glycoconjugates strongly affects rheologic properties of respiratory tract mucus. It is concluded that administration of Syntophyllin dramatically lowered the proportion of goblet cells containing fucosylated glycoconjugates in rabbit tracheal epithelium, especially at 30 min after exposure. This decrease was strongest in the levels of alpha(1-2)-fucosylated glycoconjugates. Therefore, Syntophyllin substantially altered the composition and viscoelastic properties of mucus of the upper respiratory tract. The vasodilator Oxantil exerted less pronounced changes in the proportion of goblet cells, but the strongest effect was again found in the levels of alpha(1-2)-fucosylated glycoconjugates.


Identification of volatile biomarkers of Giardia duodenalis infection in children with persistent diarrhoea.
Currently, chronic diarrhoea syndrome in children is a very common pathology whose aetiology is sometimes difficult to identify. Methodologies for the diagnosis of infections have diversified, and gas chromatography/mass spectrometry (GC/MS) is a very useful tool. The aim of this study was to identify volatile biomarkers of the presence of Giardia duodenalis in the faeces of patients with chronic diarrhoea (with and without giardiasis) using static headspace extraction followed by GC/MS. The analysis of the volatiles extracted from the headspace had enough sensitivity to detect differences in the volatile profiles in the faeces of the patients with and without Giardia duodenalis infection and discriminate between them. Dimethyl disulphide and trisulphide were found in the faeces of patients without giardiasis but not in the faeces of patients with G. duodenalis. Finally, three possible biomarkers, acetic acid, 1,4-dimethoxy-2,3-butanediol and 1,3-dimethoxy-2-propanol, were proposed to identify patients with giardiasis; these compounds were not present in the patients without the parasite. Multivariate analysis revealed that principal component 1 separated the stool samples according to the presence of infection by G. duodenalis despite the inter-individual variability in biological specimens such as faeces.


Lemniscular [16]Cycloparaphenylene: A Radially Conjugated Figure-Eight Aromatic Molecule.
A cycloparaphenylene-based molecular lemniscate (CPPL) was obtained in a short synthesis involving masked p-phenylene equivalents. The strained figure-eight geometry of CPPL is sustained by the incorporated 9,9'-bicarbazole subunit, which also acts as a stereogenic element. The shape of the distorted [16]cycloparaphenylene nanohoop embedded in CPPL is accurately approximated with a Booth lemniscate. The structure of CPPL, investigated using NMR and Raman spectroscopic methods, revealed strain-dependent features, consistent with the variable curvature of the ring. The electronic and optical properties of CPPL combine features more characteristic of smaller cycloparaphenylenes, such as a reduced optical bandgap and red-shifted fluorescence. CPPL was resolved into enantiomers, which are configurationally stable and provide strong chiroptical responses, including circularly polarized luminescence.


Endothelial heparanase regulates heart metabolism by stimulating lipoprotein lipase secretion from cardiomyocytes.
After diabetes mellitus, transfer of lipoprotein lipase (LPL) from cardiomyocytes to the coronary lumen increases, and this requires liberation of LPL from the myocyte surface heparan sulfate proteoglycans with subsequent replenishment of this reservoir. At the lumen, LPL breaks down triglyceride to meet the increased demand of the heart for fatty acid. Here, we examined the contribution of coronary endothelial cells (ECs) toward regulation of cardiomyocyte LPL secretion. Bovine coronary artery ECs were exposed to high glucose, and the conditioned medium was used to treat cardiomyocytes. EC-conditioned medium liberated LPL from the myocyte surface, in addition to facilitating its replenishment. This effect was attributed to the increased heparanase content in EC-conditioned medium. Of the 2 forms of heparanase secreted from EC in response to high glucose, active heparanase released LPL from the myocyte surface, whereas latent heparanase stimulated reloading of LPL from an intracellular pool via heparan sulfate proteoglycan-mediated RhoA activation. Endothelial heparanase is a participant in facilitating LPL increase at the coronary lumen. These observations provide an insight into the cross-talk between ECs and cardiomyocytes to regulate cardiac metabolism after diabetes mellitus.


Biventricular pacing decreases sympathetic activity compared with right ventricular pacing in patients with depressed ejection fraction.
Although there have been few studies in which the hemodynamic effects of right ventricular (RV) and left ventricular (LV) pacing were compared with those of biventricular (BV) pacing, the autonomic changes during these different pacing modes remain unknown. We hypothesized that BV pacing results in improved hemodynamics and a decrease in sympathetic nerve activity (SNA) compared with single-site pacing. Thirteen men with a mean ejection fraction of 0.28+/-0.7 were enrolled in the study. Arterial blood pressure (BP), central venous pressure (CVP), and SNA were recorded during 3 minutes of right atrial (RA)-RV, RA-LV, and RA-BV pacing at a rate 10 beats faster than sinus rhythm. BP was greater during LV (151+/-7/85+/-3 mm Hg) and BV (151+/-6/85+/-3 mm Hg) pacing than during RV pacing (146+/-7/82+/-3 mm Hg) (P:<0.05). There were no differences in CVP among all pacing modes (P:=0.27). SNA was significantly less (P:<0.02) during both LV (606+/-35 U) and BV (582+/-41 U) pacing compared with RV pacing (685+/-32 U) (P:<0.02). Although not statistically significant (P:=0. 08 to 0.14), there was a trend for patients with a narrow QRS to have a lower mean BP and higher SNA during LV pacing than during BV pacing (r=0.42 to 0.49). LV-based pacing results in improved hemodynamics and a decrease in SNA compared with RV pacing in patients with LV dysfunction regardless of the QRS duration.


Multidetector computed tomography myocardial perfusion imaging during adenosine stress.
The purpose of this study is to validate the accuracy of multidetector computed tomography (MDCT) to measure differences in regional myocardial perfusion during adenosine stress in a canine model of left anterior descending (LAD) artery stenosis, during first-pass, contrast-enhanced helical MDCT. Myocardial perfusion imaging by MDCT may have significant implications in the diagnosis and treatment of coronary artery disease. Eight dogs were prepared with a LAD stenosis, and contrast-enhanced MDCT imaging was performed 5 min into adenosine infusion (0.14 to 0.21 mg/kg/min). Images were analyzed using a semiautomated approach to define the regional signal density (SD) ratio (myocardial SD/left ventricular blood pool SD) in stenosed and remote territories, and then compared with microsphere myocardial blood flow (MBF) measurements. Mean MBF in stenosed versus remote territories was 1.37 +/- 0.46 ml/g/min and 1.29 +/- 0.48 ml/g/min at baseline (p = NS) and 2.54 +/- 0.93 ml/g/min and 8.94 +/- 5.74 ml/g/min during adenosine infusion, respectively (p < 0.05). Myocardial SD was 92.3 +/- 39.5 HU in stenosed versus 180.4 +/- 41.9 HU in remote territories (p < 0.001). There was a significant linear association of the SD ratio with MBF in the stenosed territory (R = 0.98, p = 0.001) and between regional myocardial SD ratio and MBF <8 ml/g/min, slope = 0.035, SE = 0.007, p < 0.0001. Overall, there was a significant non-linear relationship over the range of flows studied (LR chi-square [2 degrees of freedom] = 31.8, p < 0.0001). Adenosine-augmented MDCT myocardial perfusion imaging provides semiquantitative measurements of myocardial perfusion during first-pass MDCT imaging in a canine model of LAD stenosis.


Changing dietary patterns and body mass index over time in Canadian Inuit communities.
The International Polar Year (IPY) Inuit Health Survey provided an opportunity to compare dietary and body mass index (BMI) data with data collected a decade earlier for the same communities. A dietary survey included 1,929 randomly selected participants aged 15 years or older, selected from 18 Inuit communities in 1998-1999. The IPY survey included 2,595 randomly selected participants aged 18 years or older, selected from 36 Inuit communities in 2007-2008. Data from the same 18 communities included in both surveys were compared for adults 20 years and older. Twenty-four-hour dietary recall data were analysed to assess the percentage of energy from traditional and market foods by sex and age groups. Body mass index (BMI) was assessed to establish the prevalence of obesity by sex and age groups in both surveys. There was a significant decrease (p≤0.05) in energy contribution from traditional food and a significant increase in market food consumption over time. Sugar-sweetened beverages, chips and pasta all increased as percentages of energy. BMI increased overall for women and for each age stratum evaluated (p<0.05). The nutrition transition continues in the Canadian Arctic with a concurrent increase in BMI.


Transference interpretation in the psychotherapy of borderline patients: a high-risk, high-gain phenomenon.
The effectiveness of transference interpretation in the psychodynamic psychotherapy of patients with borderline personality disorder has been highly controversial. Both highly expressive approaches that stress the value of transference interpretation and supportive strategies that eschew transference work have been advocated in the literature. We review this literature and identify three emerging trends in thought: (1) Primarily interpretive approaches should be reserved for patients with greater levels of ego strength. (2) Whichever technique is used, a strong therapeutic alliance is the foundation of treatment. (3) Expressive and supportive techniques should not be juxtaposed as polarized opposites; supportive interventions often pave the way for transference interpretation. Our psychotherapy process study revealed that transference interpretations tended to have greater impact--both positive and negative--than other interventions made with patients with borderline personality disorder. We conclude that such factors as neuropsychologically based cognitive dysfunction, a history of early trauma, patterns of object relations involving interpersonal distance, masochistic tendencies, and anaclitic rather than introjective psychopathology are among the patient characteristics that influence the impact of transference interpretation on the therapeutic alliance. Bias toward expressive technique and countertransference issues appear to be relevant to the therapist's difficulty in shifting to a more supportive approach when indicated.


Elements of a general theory of joints. 7. Mechanical structures of the relative motion of adjacent vertebrae.
The mechanical structure of the relative motion of adjacent vertebrae is given by the interplay of the artt. zygapophysiales, the intervertebral disk and the ligaments, respectively. This structure will be described in the 3 main functional planes. Horizontal plane (parallel to the disk): The two artt. zygapophysiales structurally represent a force locked, kinematically contrained four-bar-chain (link quadrangle). It possesses but one degree of freedom. The intervertebral disk does not influence, therefore, the kinematic structure but the extent of the motion. Frontal-coronal plane: The four-bar-chain of the frontal plane seems to be inflexible. Lateral bending is only possible combined with a right or left handed axial rotation. Vertical-sagittal plane: The artt. zygapophysiales work as a single dimeric link chain. The momentary rotational center lies on the common normal of the two articular surfaces. Its position upon that is determined by the geometry of the applied force system according to the "reciprocal theorem of synarthroses". Solely one direction of translation is possible. A translation perpendicular to the intervertebral disk is impossible in the cervical and thoracic region.


Polymer-graphene oxide quadlayer thin-film assemblies with improved gas barrier.
Layer-by-layer assembly was used to create quadlayers (QLs) of chitosan (CH), poly(acrylic acid) (PAA), CH, and graphene oxide (GO). Electron microscopy confirmed GO coverage over the film and a highly ordered nanobrick wall structure. By varying pH deviation between CH and PAA, a thick and interdiffused polymer matrix was created because of the altered chain conformation. A 5 CH (pH 5.5)/PAA (pH 3)/CH (pH 5.5)/GO QL assembly (48 nm) exhibits very low oxygen permeability (3.9 × 10(-20) cm(3) cm cm(-2) Pa(-1) s(-1)) that matches SiOx barrier coatings. In an effort to maintain barrier performance under high humidity, GO was thermally reduced to increase hydrophobicity of the film. This reduction step increased H2/CO2 selectivity of a 5 QL film from 5 to 215, exceeding Robeson's upper bound limit. This unique water-based multilayer nanocoating is very promising for a variety of gas purification and packaging applications.


Blood phobics and nonphobics: psychological differences and affect during exposure.
This study compared psychological dimensions of blood phobics and nonphobic controls, examined affect in response to phobic and neutral stimuli, and investigated the relationship between reported feelings of faintness and blood pressure. Blood phobics (24 adults with extreme Mutilation Questionnaire scores) and 24 nonphobics completed several psychological measures and viewed one of two 60 sec surgery scenes and a 60 sec neutral scene in counterbalanced order. Subjective, psychophysiologic, and motoric measures of affect were assessed. On questionnaires, phobics reported greater anxiety sensitivity, empathic distress, fear and insecurity, and nightmares, but no difference in autonomic arousal, muscle tension, motion sickness, or other empathy domains. During surgery scenes, phobics had more negative affect than controls; however, phobics were more anxious during only one of the two surgeries, and often only when the surgery was presented prior to the neutral scene. Fainting did not occur, and self-reported feelings of faintness were unrelated to blood pressure changes. The findings highlight the lack of information on blood phobic stimulus properties, fainting's relationship to self-reports and blood pressure, and the specific emotion experienced in blood phobia.


Improved cardiovascular risk factors in pediatric renal transplant recipients on steroid avoidance immunosuppression: A study of the Midwest Pediatric Nephrology Consortium.
Several centers have examined the implementation of immunosuppression protocols that minimize steroid exposure. This study retrospectively examined cardiovascular risk factors in 70 pediatric renal transplant recipients on steroid avoidance-based immunosuppression over three yr compared to matched pediatric patients maintained on chronic corticosteroids. Although higher rates of acute rejection were noted in the steroid-avoidant group (22% vs. 16%, p = 0.034), graft function was similar (67 + 10 mL/min/1.73 m(2) vs. 72 + 12 mL/min/1.73 m(2)) (p = 0.053). The steroid-avoidant group demonstrated improved growth (height z-score -0.41 + 5.9 vs. -1.1 + 0.041) with a decrease in the prevalence of obesity (24% vs. 34%, p = 0.021). Indexed systolic blood pressures were lower beginning at six months post-transplant in the steroid-avoidant group (1.21 + 0.15 vs. 1.51 + 0.22, p = 0.020). Indexed diastolic blood pressures were lower beginning at 12 months post-transplant (0.91 + 0.11 vs. 1.12 + 0.18, p = 0.037). Differences in total serum cholesterol values and serum glucose values were not statistically significant. Beginning at 12 months, a statistically significant decrease in left ventricular mass index (39.2 + 11.3 vs. 49.4 + 14.5, p = 0.014) was noted in patients on steroid-avoidant immunosuppression, which corresponded to a significant decrease in the prevalence of left ventricular hypertrophy in these patients by two yr post-transplant (35% vs. 48%, p = 0.012). Systolic blood pressure and BMI were independent predictors of left ventricular hypertrophy.


Nanofocusing of optical energy in tapered plasmonic waveguides.
We predict theoretically that surface plasmon polaritons propagating toward the tip of a tapered plasmonic waveguide are slowed down and asymptotically stopped when they tend to the tip, never actually reaching it (the travel time to the tip is logarithmically divergent). This phenomenon causes accumulation of energy and giant local fields at the tip. There are various prospective applications in nano-optics and nanotechnology.


Physical fitness related to age and physical activity in older persons.
This study investigated physical fitness as a function of age and leisure time physical activity (LTPA) in a community-based sample of 624 persons aged 57 yr and older. LTPA during the last 12 months was assessed through personal interviews. A wide range of physical fitness components was measured using performance-based tests. Physical fitness was associated with the interaction age by LTPA in only a few components, in a gender-specific way, with generally larger differences in fitness between active and less active persons with increasing age. All LTPA, including low intensity LTPA, is positively and age-independent associated with most physical fitness components. The importance of LTPA typically participated in by the general population lies not so much in the delaying of the motor aging process but rather in a general, age-independent, positive effect.


Changes in stress and coping during late childhood and preadolescence.
Little is known about what produces stress in preadolescents. Secondary data analysis using hierarchical linear modeling was performed to determine how stressors and coping changed over time in 1,568 preadolescents (51% Hispanic) in 28 public schools. Over time, participants experienced most stressors less intensely, but intensity varied by gender. Children in lower grades and girls found that coping strategies used were more effective compared with those in upper grades or boys. There were several interactions of grade level, sex, and ethnicity. Changes in stress and coping among preadolescents have implications for nurses concerned with mental health.


[Morphological studies on estrogen-induced PRL cell hyperplasia in rat pituitary gland--immunocytochemical and scanning electron microscopic studies].
It is well known that the tumorous lesion in rat anterior pituitary gland occurs after continuous administration of estrogen, which is mainly composed of PRL secreting cells. In this study the changes in rat PRL cells by estrogen treatment were studied histologically and immunocytochemically. Furthermore, intracellular structures in PRL cells were observed by high resolution scanning electron microscopy using the aldehyde prefix osmium-DMSO-osmium method. These rat pituitary tumors were established in Fischer-344 female rats by weekly injections of 2.5 mg estradiol valerate. The estrogen-induced, autonomous and transplantable rat pituitary tumors (MtT/F84) were also studied in the same manner and the findings were compared with estrogen-induced tumors. The increase of weight of pituitary gland and of serum PRL level correlated significantly with the number of estrogen administration. On the basis of histological findings, estrogen-induced pituitary tumors can be classified into two different stages, that is, hypertrophic hyperplasia and adenomatous hyperplasia. The former corresponds to the period until 6 weeks after the beginning of estrogen treatment and the latter to the period thereafter. In the former, PRL cells were significantly increased in number and volume, and were diffusely immunoreactive for PRL in the cytoplasm. Cell atypism was rarely found. The extent of acidophil staining was well preserved in hematoxylin-eosin staining. Ultrastructurally, organelles were well developed, especially rough endoplasmic reticulum showed whorl formation. Immature, irregularly shaped secretory granules were frequently observed within the Golgi area. Mature granules were richly stored at the periphery of the cell.(ABSTRACT TRUNCATED AT 250 WORDS)


Psychological distress, somatization, and defense mechanisms associated with quality of life in inflammatory bowel disease patients.
Clinical parameters predict health-related quality of life (HRQOL) in inflammatory bowel disease (IBD), but some patients have impaired HRQOL despite being in clinical remission. To identify personality and psychological distress variables associated with HRQOL in IBD. In a cross-sectional study of 185 IBD patients, the General Health Questionnaire, the Hopkins' Symptoms Distress Checklist, the Defense Style Questionnaire and the Life Style Index were administered. The Inflammatory Bowel Disease Questionnaire was used for the assessment of HRQOL. Psychological distress was associated with impaired HRQOL in a dose-response fashion. Somatization mediated the relationships of anxiety and depression with HRQOL. Few years of education, more extensive use of the reaction-formation defense mechanism and higher rates of somatization were the variables most closely and independently associated with impaired HRQOL. Somatization and reaction-formation are independent correlates of disease-specific HRQOL in IBD patients, and this could be relevant to psychological interventions.


Desipramine rescues emotional memory deficit induced by low doses of reserpine.
Emotional memory deficit is a well-known complication in early Parkinson's disease. However, its molecular mechanism is still not well known. To address this issue, we examined the cue-related fear-conditioning task and long-term potentiation (LTP) of the thalamus to the lateral amygdala in rats treated with low doses of reserpine (Res). We found that low-dose Res treatment impaired emotional memory and LTP. We also found that exogenous upregulation of norepinephrine (NE) ameliorated the impairment of LTP by facilitating β-adrenergic receptors. Finally, acute treatment with NE or desipramine rescued the impaired emotional memory induced by a low-dose of Res. These results imply a pivotal role for NE in synaptic plasticity and associative fear memory in rats treated with low doses of Res and suggest that desipramine is a potential candidate for treating Parkinson's disease-related emotional memory deficit.


Glatiramer acetate antibodies, gene expression and disease activity in multiple sclerosis.
Glatiramer acetate (GA) treatment suppresses disease activity in multiple sclerosis (MS). The immunological response to treatment may differ in patients who are stable on GA therapy and patients with breakthrough disease activity, but the results of previous studies are inconsistent. We studied the immunological response to GA and its relationship with disease activity. Anti-GA antibodies in plasma and the expression of genes encoding cytokines and T-cell-polarizing transcription factors in blood cells were analysed by flow cytometric bead array and polymerase chain reaction (PCR) analysis in 39 untreated and 29 GA-treated relapsing-remitting MS patients. Definition of breakthrough disease was based on the occurrence of relapses, disability progression, or gadolinium (Gd)-enhanced MRI. The expression of T helper type 1 (Th1) and Th17 cytokines and transcription factors was reduced during long-term treatment, but there was no relationship between the expression of cytokines and transcription factors and anti-GA antibodies. High expression of mRNA encoding GATA3 and lymphotoxin-β (LT-β) was associated with low disease activity in Gd-enhanced MRI studies. None of the variables studied were associated with clinical disease activity. GA treatment resulted in the development of IgG and IgG4 anti-GA antibodies during the first months of treatment, persisting during long-term treatment. The observed relationship between the expression of mRNA encoding GATA3 and LT-β expression and MRI disease activity deserves further analysis in future studies. The development of anti-GA antibodies was observed in all patients treated with GA, but this was not related with measures of cellular immunity, clinical or MRI disease activity.


Empowerment through use of the Code of Professional Conduct.
The UKCC's Code of Professional Conduct is a source of empowerment. It presents a statement of the values of the nursing profession which may be used to promote the interests of patients and clients.


Knockout Corner: 5-HT(1A) receptor inactivation: anxiety or depression as a murine experience.
5-HT(1A) receptors play a critical role in the pathophysiology of anxiety and depression as well as in the mode of action of anxiolytic and antidepressant drugs. Mice with a targeted inactivation of the 5HT(1A) receptor show a phenotype that is associated with a gender-modulated and gene/dose-dependent increase of anxiety-related and antidepressant-like behaviours. Since this behavioural phenotype was observed in animals in which the mutation was bred into mice of different genetic backgrounds, 5-HT(1A) receptor knockout mice represent a useful model system for advanced investigations of 5-HT(1A) genotype/phenotype interaction.


Rapid detection and identification of beer-spoilage lactic acid bacteria by microcolony method.
We evaluated a microcolony method for the detection and identification of beer-spoilage lactic acid bacteria (LAB). In this approach, bacterial cells were trapped on a polycarbonate membrane filter and cultured on ABD medium, a medium that allows highly specific detection of beer-spoilage LAB strains. After short-time incubation, viable cells forming microcolonies were stained with carboxyfluorescein diacetate (CFDA) and counted with muFinder Inspection System. In our study, we first investigated the growth behavior of various beer-spoilage LAB by traditional culture method, and Lactobacillus lindneri and several L. paracollinoides strains were selected as slow growers on ABD medium. Then the detection speeds were evaluated by microcolony method, using these slowly growing strains. As a result, all of the slowly growing beer-spoilage LAB strains were detected within 3 days of incubation. The specificity of this method was found to be exceptionally high and even discriminated intra-species differences in beer-spoilage ability of LAB strains upon detection. These results indicate that our microcolony approach allows rapid and specific detection of beer-spoilage LAB strains with inexpensive CFDA staining. For further confirmation of species status of detected strains, subsequent treatment with species-specific fluorescence in situ hybridization (FISH) probes was shown as effective for identifying the CFDA-detected microcolonies to the species level. In addition, no false-positive results arising from noise signals were recognized for CFDA staining and FISH methods. Taken together, the developed microcolony method was demonstrated as a rapid and highly specific countermeasure against beer-spoilage LAB, and compared favorably with the conventional culture methods.


Problems associated with hemodialysis and travel.
While there is a recognized risk of hepatitis C acquisition associated with dialysis away from the "home" center, there is little documented data on the effect that dialysis while traveling has on the dialysis patient's health. This study was designed to examine the incidence of travel within a hemodialysis population and to ascertain whether travel was associated with morbidity for patients on hemodialysis. Travel data were collected prospectively over a 6-month period, from April 2009, for all patients receiving maintenance hemodialysis across our dialysis centers. Biochemical, microbiological, and hematological parameters as well as hepatitis serology and antibiotic starts were recorded for 12 weeks prior to and following dialysis away from center. A total of 172 individuals traveled on 200 occasions. The blood stream infection rate for travelers with a central venous catheter was 0.25 versus 0.83/1,000 access days (p = 0.038) in the 12 weeks pre-travel versus post-travel. Parenteral and oral antibiotic starts were both significantly elevated post-travel and were mainly instituted for either chest or urinary sepsis. There was evidence of raised inflammatory markers and anemia on return to center but no evidence of hepatitis B or hepatitis C seroconversion. Travel and dialysis away from a patient's usual hemodialysis unit is a common occurrence but is associated with an increased risk of bacterial infection, anemia, and inflammatory response. This study provides evidence for the concern that hemodialysis away from center is associated with increased morbidity.


Soft-tissue vibrations in the quadriceps measured with skin mounted transducers.
The purpose of this study was to develop a method to characterize the frequency and damping of vibrations in the soft tissues of the leg. Vibrations were measured from a surface-mounted accelerometer attached to the skin overlying the quadriceps muscles. The free vibrations in this soft tissue were recorded after impact whilst the muscle was performing isometric contractions at 0, 50, and 100% maximum voluntary force and with the knee held at 20, 40, and 60 degrees angles of flexion. The acceleration signals indicated that the soft tissue oscillated as under-damped vibrations. The frequency and damping coefficients for these vibrations were estimated from a model of sinusoidal oscillations with an exponential decay. This technique resolved the vibration coefficients to 2 and 7% of the mean values for frequency and damping, respectively.


Chemical synthesis of Nd2Fe14B/Fe3B nanocomposites.
High exchange-coupled Nd2Fe14B/Fe3B nanocomposites were synthesized by an integrative procedure of thermal decomposition and reductive annealing processes. The molar ratio of the resulting products of Nd/Fe/B can be tuned by adjusting the raw material proportion. The as-prepared nanocomposites exhibited an exchanged coupled effect with a large coercivity of 11 100 Gs, enhanced remanence Mr of 42.0 emu g(-1), and Mr/M3T of 0.59.


Cardiogenic shock due to tachycardiomyopathy after heart transplantation: successful treatment with ivabradine.
Tachycardia-induced cardiomyopathy refers to an impairment in left-ventricular function due to chronic or prolonged tachycardia. We describe a heart transplant patient who developed cardiogenic shock due to tachycardia-induced cardiomyopathy. Low-output failure was further aggravated by administration of a short-acting beta-blocker during invasive hemodynamic monitoring. In contrast, heart rate control by administration of increasing doses of ivabradine supported recovery from cardiogenic shock and led to an improvement in the patient's clinical condition as well as left-ventricular function during follow-up.


Cognitive and familial risk evidence converged: A data-driven identification of distinct and homogeneous subtypes within the heterogeneous sample of reading disabled children.
The evident degree of heterogeneity observed in reading disabled children has puzzled reading researchers for decades. Recent advances in the genetic underpinnings of reading disability have indicated that the heritable, familial risk for dyslexia is a major risk factor. The present data-driven, classification attempt aims to revisit the possibility of identifying distinct cognitive deficit profiles in a large sample of second to fourth grade reading disabled children. In this sample, we investigated whether genetic and environmental risk factors are able to distinguish between poor reader subtypes. In this profile, we included reading-related measures of phonemic awareness, letter-speech sound processing and rapid naming, known as candidate vulnerability markers associated with dyslexia and familial risk for dyslexia, as well as general cognitive abilities (non-verbal IQ and vocabulary). Clustering was based on a 200 multi-start K-means approach. Results revealed four emerging subtypes of which the first subtype showed no cognitive deficits underlying their poor reading skills (Reading-only impaired poor readers). The other three subtypes shared a core phonological deficit (PA) with a variable and discriminative expression across the other underlying vulnerability markers. More specific, type 2 showed low to poor performance across all reading-related and general cognitive abilities (general poor readers), type 3 showed a specific letter-speech sound mapping deficit next to a PA deficit (PA-LS specific poor readers) and type 4 showed a specific rapid naming deficit complementing their phonological weakness (PA-RAN specific poor readers). The first three poor reader profiles were more characterized by variable environmental risk factor, while the fourth, PA-RAN poor reader subtype showed a significantly strong familial risk for dyslexia. Overall, when we zoom in on the heterogeneous phenomenon of reading disability, unique and distinct cognitive subtypes can be identified, distinguishing between those poor readers more influences by the role of genes and those more influenced by environmental risk factors. Taking into account this diversity of distinct cognitive subtypes, instead of looking at the reading disabled sample as a whole, will help tailor future diagnostic and intervention efforts more specifically to the needs of children with such a specific deficit and risk pattern, as well as providing a more promising way forward for genetic studies of dyslexia.


Condylomata acuminata of the cervix: histopathology and association with cervical neoplasia.
Cervical specimens from 531 consecutive cases were reviewed to determine the frequency of cervical condylomata acuminata (CLA) and their association with cervical intraepithelial neoplasia (CIN). CLA and CIN were subclassified into three groups based on the criteria previously established in the literature. Our data indicate that CLA was present in 103 (19%) cases and that 44 (43%) of these had CIN as well. The total number of cases of CIN in this series was 146 (27%) with 44 of these (30%) having associated CLA. Of the eight CIN cases in women under 20 years of age, seven (99.6%) had associated CLA. The mean age of patients with CIN alone was 31 years; patients with CIN and CLA were younger, with a mean age of 24 years. These findings indicate that cervical CLA is a common lesion affecting young women and that there is a significant association of CLA with CIN. Women with CLA appear to be at significant risk for development of CIN.


Frida Kahlo: Portrait of Chronic Pain.
The Mexican artist Frida Kahlo (1907-1954) is one of the most celebrated artists of the 20th century. Although famous for her colorful self-portraits and associations with celebrities Diego Rivera and Leon Trotsky, less known is the fact that she had lifelong chronic pain. Frida Kahlo developed poliomyelitis at age 6 years, was in a horrific trolley car accident in her teens, and would eventually endure numerous failed spinal surgeries and, ultimately, limb amputation. She endured several physical, emotional, and psychological traumas in her lifetime, yet through her art, she was able to transcend a life of pain and disability. Of her work, her self-portraits are conspicuous in their capacity to convey her life experience, much of which was imbued with chronic pain. Signs and symptoms of chronic neuropathic pain and central sensitization of nociceptive pathways are evident when analyzing her paintings and medical history. This article uses a narrative approach to describe how events in the life of this artist contributed to her chronic pain. The purpose of this article is to discuss Frida Kahlo's medical history and her art from a modern pain sciences perspective, and perhaps to increase our understanding of the pain experience from the patient's perspective.


Physician assisted death and hard choices.
We argue that after the passage of a physician assisted death law some inequities in the health care system which prevent people from getting the medical care they need will become reasons for choosing assisted death. This raises the issue of whether there is compelling moral reason to change those inequities after the passage of an assisted death law. We argue that the passage of an assisted death law will not create additional moral reasons for eliminating inequities simply because they become motives for someone to opt for assisted death. We also argue that it is not feasible to eliminate these reasons for opting for assisted death by granting a right to health care because of an intractable scarcity of medical resources.


Implementation of advanced laparoscopy into daily gynecologic practice: difficulties and solutions.
The implementation of laparoscopy into Dutch gynecologic practice is slow. This study was conducted to assess the current state of laparoscopy, to identify factors influencing the implementation and to find solutions toward a better implementation. In 2003 a questionnaire was sent to all 151 gynecologists who finished residency within the previous 5 years. The questionnaire addressed practice demographics, performance of laparoscopy, factors influencing use of laparoscopy in practice and means of obtaining laparoscopic skills after residency. Of 151 gynecologists, 124 (82%) responded, 46 (37%) male and 78 (63%) female. Mean age was 39 years (range 32-47 years). Respondents (73%) believed they were adequately trained during residency for basic laparoscopic procedures, but not for the more advanced procedures (82%). Lack of caseload, lack of being a primary surgeon, and lack of simulator training caused the deficiency of laparoscopic skills at the end of the residency. Causes of the slow implementation were long operating time, lack of attention for laparoscopy during residency, and budgetary problems, but not the financial compensation for gynecologists. In current practice, only 9% believed they reached their preferred level of competence. Hiring an advanced laparoscopic gynecologist was believed to be the best opportunity to reach the preferred level of competence. A minority of respondents supported a referral system or fellowship program. Basic laparoscopy is sufficiently mastered during residency training; however, advanced laparoscopy is not. More emphasis should be placed on laparoscopic training of advanced procedures during residency and for gynecologists in practice. Hiring a gynecologist with advanced laparoscopic skills is expected to be the solution for this problem. However, a referral system or fellowship program is not.


Transcriptional regulation of the rat apelin receptor gene: promoter cloning and identification of an Sp1 site necessary for promoter activity.
The genomic structure and transcriptional regulation of the rat apelin receptor (APJR) were analysed by rapid amplification of 5' cDNA ends (5'-RACE), transient expression assays and DNA-protein interaction. Analysis of the 5'-flanking region of a rat genomic clone shows no TATA box, but a putative CAAT box and several putative binding sites for transcription factors are present. Two transcriptional start sites were identified by 5'-RACE, RNase protection and primer extension analyses. Promoter activity was exhibited in the APJR- expressing SH-SY5Y cell line as well as in COS-7 and Chinese hamster ovary (CHO-K1) cells. Consecutive 5'-deletion analysis revealed the highest promoter activity in a region between bp -966 and -165. DNaseI footprint analysis revealed seven protected regions and electrophoretic mobility shift, super-shift and competition assays identified individual DNA-protein complexes capable of binding Sp1, estrogen receptor (ER)alpha, glucocorticoid receptor and CCAAT enhancer binding protein (C/EBP)gamma transcription factors. Site-directed mutagenesis identified an individual Sp1 motif that plays a major role in activation of the APJR promoter and also demonstrated constitutive transcriptional regulation of the promoter by estrogen and glucocorticoid receptors. Promoter regulation by the cAMP-dependent signal cascade was also shown.


Development of Mycobacterium tuberculosis whole cell screening hits as potential antituberculosis agents.
The global pandemic of drug sensitive tuberculosis (TB) as well as the increasing threat from various multidrug resistant forms of TB drives the quest for newer, safer, more effective TB treatment options. The general lack of success in progressing novel chemical matter from high throughput screens of Mycobacterium tuberculosis (M.tb) biochemical targets has prompted resurgence in interest and efforts in prosecuting mycobacterial phenotypic screens. Whole cell active compounds identified from such screens offer significant intrinsic advantages over biochemical screening hits, and derivatives of many of these have proven invaluable in helping to fill the current TB drug development pipeline. Modern techniques for "de-orphaning" such screening hits (i.e., determining their specific biological mechanism of action) offer the possibility of ultimately identifying improved next-generation chemical series by screening these essential, pharmacologically validated biochemical targets as well.


Effects of cypermethrin on some biochemical parameters and its amelioration through dietary supplementation of ascorbic acid in freshwater catfish Heteropneustes fossilis.
Static bioassays were made to evaluate efficiency of dietary supplementation of ascorbic acid to counter stress exerted by the pyrethroid pesticide, cypermethrin on freshwater catfish Heteropneustes fossilis. Three levels, a control and two sub-lethal concentrations (0.3 and 0.5 microg L(-1)) of cypermethrin were tested. Stress on H. fossilis was evident from the rise of plasma glucose level and reduction in the level of liver glycogen after 4 h of cypermethrin treatment. Acid and alkaline phosphatase activities of liver and ascorbic acid level of blood, liver and kidney also decreased significantly in H. fossilis exposed to cypermethrin as compared with control. Fish previously fed for 60 d with a diet supplemented by high level of ascorbic acid (1.0 g kg(-1)) could significantly reverse these effects. Fish fed a low level of dietary ascorbic acid (0.5 g kg(-1)) failed to counter the stress of cypermethrin.


Vision testing and the elderly driver: is there a problem meriting policy change?
The number of elderly drivers is increasing. The primary purpose of public policy requiring vision screening for driver's license renewal is to identify, and, when necessary, restrict drivers with functional vision impairments. Because of age-related ocular conditions, elderly drivers, as a group, have a higher incidence and prevalence of functionally impaired vision. To date, there is no empirical evidence of a significant predictive relationship between changes in vision function and automobile crashes. Most states require vision screening for driver's license renewal, whereas some do not. Among those states requiring vision screening, there is considerable variation in the frequency and level of testing. Efforts to determine the role of vision in driving, while suggestive, have not been useful in identifying at-risk older drivers. Researchers have observed that older drivers are often aware of their decreased functional capacity and voluntarily adjust their driving patterns by driving less frequently, for shorter distances, during daylight hours, more slowly, and during non-rush hours. However, although not statistically significant, the decline in the mean annual traffic fatality rates with increased state vision screening requirements suggests a possible beneficial effect of vision screening. Because older drivers are at risk for sight-threatening conditions, they are most affected by vision screening requirements for driver's license renewal. Since a loss of driving privileges represents a major loss of mobility and independence, there is a pressing need to better understand the relationship between age-related vision changes and the frequency and severity of traffic crashes.


[Mechanism of benign, peripheral, paroxysmal positional vertigo (BPPV)].
Positioning nystagmus of the peripheral benign type (BPPV) has long been considered to be due to cupulolithiasis-i.e. attachment of inorganic material to the cupula-of the posterior vertical semicircular canal. Meanwhile it has generally been recognized that not all characteristics of this type of nystagmus/vertigo can be explained by assuming a gravity-dependent reaction of the posterior canal. Canalolithiasis-i.e. floating material within the canal, heavier than endolymphe-of the posterior semicircular canal is now widely regarded as the cause of BPPV. Again, however, this is a concept far too simple to explain most of the properties of BPPV. Clearly defined, carefully performed positioning and positional maneuvers were carried out on 79 patients suffering from typical BPPV in order to reveal those positioning movements and positions necessary to elicit BPPV. Detailed analysis of those positioning maneuvers necessary to elicit BPPV clearly reveals that canalolithiasis cannot be the cause of BBPV. BPPV is a combination of positioning, and positional-dependent reactions in which the otolithic organs, and particularly the sacculus, seem to be involved.


Resveratrol inhibits the growth and induces the apoptosis of both normal and leukemic hematopoietic cells.
It is often postulated that trans-3,4',5-trihydroxystilbene (resveratrol, RES) exhibits cell growth regulatory and chemopreventive activities. However, mechanisms by which this polyphenol inhibits tumor cell growth, and its therapeutic potential are poorly understood. Using various human leukemia cells, we have first defined the anti-tumoral doses of this compound. RES inhibited the proliferation and induced the apoptosis of all tested lymphoid and myeloid leukemia cells with IC(50) = 5-43 microM. Prior to apoptosis, RES-induced caspase activity in a dose-dependent manner and cell cycle arrest in G(2)/M-phase, correlating with a significant accumulation of cyclins A and B. Leukemia cell death with RES required both caspase-dependent and -independent proteases, as it was significantly inhibited by simultaneous addition of Z-VAD-FMK and leupeptin to these cultures. While RES did not affect non-activated normal lymphocytes, this agent decreased the growth and induced the apoptosis of cycling normal human peripheral blood lymphocytes at lower concentrations (IC(50) <8 microM) than those required for most leukemia cells. RES also induced the apoptosis of early normal human CD34(+) cells and decreased the number of colonies generated by these precursor cells in a dose-dependent manner (IC(50) = 60 microM). Together, the data point to the complexity of RES-mediated signaling pathways and revealed the high anti-proliferative and proapoptotic activities of RES in normal cycling hemopoietic cells.


[Hereditary long QT syndrome].
The Long QT Syndrome (LQTS) is a hereditary disease, characterized by a prolonged QT-interval on the electrocardiogram and a high risk of syncope and sudden death due to ventricular arrhythmias. LQTS must be suspected in apparently healthy children and young people with syncope after emotional or physical stress. Untreated symptomatic patients have a high mortality, which is markedly reduced by sympathetic block. The knowledge of the diagnostic criteria for the LQTS, a detailed history including a family history and an ECG-recording with measurement of the QT-interval in every patient with inexplicable syncope will advance the diagnosis of the LQTS and improve the survival of these patients after proper therapy. The current knowledge on the molecular genetics, epidemiology, mechanisms of arrhythmias and therapy are presented with special emphasis on the defects in the control of ionic transport over the cell membrane caused by mutations in ion channels.


Dose adaptation of antineoplastic drugs in patients with liver disease.
Dose adaptation for liver disease is important in patients treated with antineoplastic drugs because of the high prevalence of impaired liver function in this population and the dose-dependent, frequently serious adverse effects of these drugs. We classified the antineoplastic drugs marketed in Switzerland at the end of 2004 according to their bioavailability and/or hepatic extraction to predict their kinetic behaviour in patients with decreased liver function. This prediction was compared with kinetic studies carried out with these drugs in patients with liver disease. The studies were identified by a structured, computer-based literature search. Of the 69 drugs identified, 52 had a predominant extrarenal (in most cases hepatic) metabolism and/or excretion. For 49 drugs, hepatic extraction could be calculated and/or bioavailability data were available, allowing classification according to hepatic extraction. For 18 drugs, kinetic studies have been reported in patients with impaired liver function, with the findings generally resulting in quantitative recommendations for adaptation of the dosage. In particular, recommendations are precise for 16 drugs excreted by the bile (e.g. doxorubicin and derivatives and vinca alkaloids). Validation studies comparing such recommendations with kinetics and/or dynamics of antineoplastic drugs in patients with decreased liver function have not been published. We conclude that there are currently not enough data for safe use of cyctostatics in patients with liver disease. Pharmaceutical companies should be urged to provide kinetic data (especially hepatic extraction data) for the classification of such drugs and to conduct kinetic studies for drugs with primarily hepatic metabolism in patients with impaired liver function to allow quantitative advice to be given for dose adaptation.


Chronic Achilles tendon rupture associated with rupture of the peroneus brevis tendon. A clinical case.
The treatment of a chronic rupture of the Achilles tendon remains difficult. Different techniques such as augmentation or plasties are used for restoring the gap in these ruptures. The combination of Achilles tendon rupture with other tendon ruptures may increase the difficulty in their treatment. We present a rare combination of a delayed Achilles tendon rupture and a peroneus brevis tendon rupture and its management. The importance of the presented case is the combination of both ruptures that adds difficulty to the surgical technique. A thorough analysis should be made of the magnetic resonance findings when planning a tendon transposition. To our knowledge, there are no reports of a simultaneous combination of an Achilles tendon rupture and a peroneus brevis tendon rupture.


Positive smoker identity as a barrier to quitting smoking: findings from a national survey of smokers in England.
It has been proposed that positive smoker identity may be an important factor undermining smoking cessation but very little research exists on this. This study tested the hypothesis that a simple measure of positive smoker identity would predict quit attempts over and above other known predictors in a population sample. More tentatively it explored whether this measure would also predict quit success. A representative sample of adult smokers in England (n=9456) was included at baseline and 2099 were followed-up at six months. Demographic and smoking characteristics, a single item measure of positive smoker identity (endorsing the statement: 'I like being a smoker'), measures of smoking-related attitudes, quit attempts and quit success were included. A total of 18.3% (95% CI=17.5-19.2) of smokers reported a positive smoker identity. Adjusting for all other predictors, those with a positive smoker identity were more likely to be older (p<0.001), male (p=0.013), more nicotine dependent (p<0.001), have lower motivation to stop (p<0.001), have not made a quit attempt in the past year (p=0.025), enjoy smoking (p<0.001), and consider themselves to be addicted (p<0.001). Having a positive smoker identity independently predicted failure to make a quit attempt at six months (p=0.007). The independent association with quit success was similar in magnitude but did not reach statistical significance (p=0.053). Only a minority of smokers in England have a positive smoker identity. However, where it is present it may be an important barrier to quitting smoking and merits further study.


Optimization of in silico tools for predicting genetic variants: individualizing for genes with molecular sub-regional stratification.
Genes are unique in functional role and differ in their sensitivities to genetic defects, but with difficulties in pathogenicity prediction. This study attempted to improve the performance of existing in silico algorithms and find a common solution based on individualization strategy. We initiated the individualization with the epilepsy-related SCN1A variants by sub-regional stratification. SCN1A missense variants related to epilepsy were retrieved from mutation databases, and benign missense variants were collected from ExAC database. Predictions were performed by using 10 traditional tools with stepwise optimizations. Model predictive ability was evaluated using the five-fold cross-validations on variants of SCN1A, SCN2A, and KCNQ2. Additional validation was performed in SCN1A variants of damage-confirmed/familial epilepsy. The performance of commonly used predictors was less satisfactory for SCN1A with accuracy less than 80% and varied dramatically by functional domains of Nav1.1. Multistep individualized optimizations, including cutoff resetting, domain-based stratification, and combination of predicting algorithms, significantly increased predictive performance. Similar improvements were obtained for variants in SCN2A and KCNQ2. The predictive performance of the recently developed ensemble tools, such as Mendelian clinically applicable pathogenicity, combined annotation-dependent depletion and Eigen, was also improved dramatically by application of the strategy with molecular sub-regional stratification. The prediction scores of SCN1A variants showed linear correlations with the degree of functional defects and the severity of clinical phenotypes. This study highlights the need of individualized optimization with molecular sub-regional stratification for each gene in practice.


Inhibition of programmed cell death in mouse embryonic palate in vitro by cortisol and phenytoin: receptor involvement and requirement of protein synthesis.
In an in vitro model cortisol and phenytoin inhibit the precisely timed process of palatal development, the lysosomally mediated cell death of the medial edge palatal epithelium. This inhibition of programmed cell death of the palatal midline epithelium by each drug is virtually completely blocked by the antiglucocorticoid, cortexolone, whose blocking action results from competitive binding of the glucocorticoid receptor site. The inhibition produced by each of these drugs is prevented by the protein synthesis blocker, cycloheximide. Thus, blockade of programmed cell death by each of these drugs involves the glucocorticoid receptor site and requires protein synthesis.


Mutations in the X-linked form of Charcot-Marie-Tooth disease in the French population.
The present study reports eight additional mutations in the connexin32 gene associated with the X-linked form of Charcot-Marie-Tooth disease. One of these mutations was found twice in two apparently unrelated families. This form of the disease is demyelinating and dominant. However, patient selection for mutational screening should not be limited to these criteria since presentation can either be familial or sporadic, and some patients may be incorrectly classified as suffering from an 'axonal' form. These new mutations complete our previously published work on 12 other mutations and enable meaningful observation in a representative sample of the French population. Mutations are found in all regions of the gene. The most frequently observed mutations were those affecting arginines and mainly involved CpG sequences. Compared with other sources, some of the mutations were present at a higher frequency in the French population.


Lactate as an insidious metabolite due to the Warburg effect.
Although oncogenetics remains a critical component of cancer biology and therapeutic research, recent interest has been taken towards the non-genetic features of tumour development and progression, such as cancer metabolism. Specifically, it has been observed that tumour cells are inclined to preferentially undergo glycolysis despite presence of adequate oxygen. First reported by Otto Warburg in the 1920s, and now termed the 'Warburg effect', this aberrant metabolism has become of particular interest due to the prevalence of the fermentation phenotype in a variety of cancers studied. Consequently, this phenotype has proven to play a pivotal role in cancer proliferation. As such Warburg's observations are now being integrated within the modern paradigms of cancer and in this review we explore the role of lactate as an insidious metabolite due to the Warburg effect.


Recognition of antioxidants and photosensitizers in Dyssodia pinnata by EPR spectroscopy.
Previous studies report the isolation mainly of terthiophene derivatives and flavonoids from Dyssodia species. Terthiophenes are known as photosensitizers by their capacity to generate singlet oxygen (1 O2 ), and flavonoid antioxidant activity is recognized. These opposite properties could represent interesting options in photodynamic therapy. To determine the antioxidant and photosensitizer activities of extracts and isolates of Dyssodia pinnata by electron paramagnetic resonance (EPR). Extracts and isolates were evaluated as antioxidants by the interactions with copper ion (Cu2+ ) observed in EPR, and by the 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) and the thiobarbituric reactive substances (TBARS) methods. Their abilities as 1 O2 producers were also estimated by EPR. Terthiophenes were obtained from hexane (DPH) and acetone (DPA) extracts, and flavonoids from DPA and methanol (DPM) extracts. The interaction with Cu2+ of extracts and isolated compounds of Dyssodia pinnata showed two effects in EPR: reduction and chelation; flavonoids exhibited both effects, while terthiophenes showed only reduction. DPA, DPM, and flavonoids were active in DPPH and TBARS assays. Quercetagetin-7-O-β-glucoside showed the highest antioxidant and chelating activities, 3-glycosidated flavonoids were less active. Upon irradiation extracts and terthiophenes induced 1 O2 formation. Flavonoid reducing activity on Cu2+ and free radical scavenging capacity were related to the number of hydroxy groups and to the conjugation between the B and C rings. All tested flavonols showed a major complex with Cu2+ , with the most probable site of chelation between the 5-hydroxy and 4-oxo groups. Extracts and terthiophene derivatives showed photosensitizer activity. Thus, EPR is useful to evaluate free radical scavenging and pro-oxidant properties.


Assessment of the internal fit and marginal integrity of interim crowns made by different manufacturing methods.
The adaptation of interim crowns made by subtractive and additive manufacturing has not been well investigated. The purpose of this in vitro study was to evaluate the internal fit and marginal discrepancy of interim crowns made by different manufacturing methods. A dentoform mandibular left first molar was prepared for a ceramic crown and scanned for the fabrication of 48 stereolithical resin dies and interim crowns. Group CAM included 16 ZCAD interim crowns made by computer-aided design and computer-aided manufacturing (CAD-CAM) technology; group 3DP, 16 NextDent MFH interim crowns made by digital light processing technology; and group APP, 16 Jet interim crowns manually made by using autopolymerizing acrylic resin and used as controls. The silicone replica technique was used to determine the internal discrepancy volume before definitive cementation. All crowns were cemented with Temp-Bond NE under a 50-N load and bench set for 10 minutes before microcomputed tomographic (μCT) scan assessment. The volume of cement space was measured by using the μCT scan 3-dimensional (3D) images, and gap distance at assigned locations was recorded by using the μCT scan 2-dimensional (2D) images. The marginal discrepancy was measured by the polyvinyl siloxane (PVS) impression technique and using a stereomicroscope. Data were analyzed by ANOVA and the Tukey honestly significant difference tests (α=.05). The association between different measuring techniques was analyzed by the Pearson correlation test. The gap distance between interim crowns and dies from all 3 groups measured by using the μCT scan 2D images ranged from 0.13 mm to 0.55 mm, with the highest value found at the central occlusal location in group APP. The total average gap distance values recorded for group APP were significantly higher than those for group CAM and group 3DP (P<.05). No significant differences were found in internal discrepancy and cement space volume between group CAM and group 3DP (P>.05). The Pearson correlation test showed a moderate correlation (r=0.69) between the silicone replica technique and the μCT scan technique in determining cement space volume. When the PVS impression technique was used to measure the marginal discrepancy, the mean values obtained from group APP were statistically significantly higher than those from group CAM and group 3DP (P<.05). No statistically significant difference in marginal discrepancy was found between group CAM and group 3DP (P=.70). Digitally fabricated interim crowns had better internal fit and smaller marginal discrepancy than manually constructed interim crowns. The silicone replica technique and μCT scan technique measurements had a moderate correlation in assessing the adaptation of cemented interim crowns.


Variability of calcium absorption.
Variability in calcium absorption was estimated in three groups of normal subjects in whom Ca absorption was measured by standard isotopic-tracer methods at interstudy intervals ranging from 1 to 4 mo. Fifty absorption tests were performed in 22 subjects. Each was done in the morning after an overnight fast with an identical standard breakfast containing a Ca load of approximately 250 mg. Individual fractional absorption values were normalized to permit pooling of the data. The coefficient of variation (CVs) for absorption for the three groups ranged from 10.57 to 12.79% with the size of the CV increasing with interstudy duration. One other published study presenting replicate absorption values was analyzed in a similar fashion and was found to have a CV of absorption of 9.78%. From these data we estimate that when the standard double-isotope method is used to measure Ca absorption there is approximately 10% variability around any given absorption value within an individual human subject and that roughly two-thirds of this represents real biological variability in absorption.


High-harmonic spectroscopy: from femtosecond to attosecond molecular dynamics.
The principles of high-harmonic spectroscopy (HHS) are illustrated using two recent examples from the literature. The method can be applied as a probe to measure the evolution of the electronic structure of a molecule during a chemical reaction on the femtosecond time scale. Alternatively, it can be used as a comprehensive method unifying pump and probe steps into a single laser cycle to measure electronic dynamics on the attosecond time scale.


Antagonism of vasopressin V2 receptor improves albuminuria at the early stage of diabetic nephropathy in a mouse model of type 2 diabetes.
Vasopressin is increased in diabetes and was shown to contribute to development of diabetic nephropathy through V2 receptor (V2R) activation in an experimental model of type 1 diabetes. The role of V2R in type 2 diabetes remains undocumented. This study addresses the issue in a mouse model of type 2 diabetes. Male obese diabetic db/db mice were treated for 12weeks with a selective V2R antagonist (SR121463) and compared to non-treated db/db and non-diabetic db/m mice. All animals were previously uninephrectomized. The V2R antagonist did not alter glycemia or glycosuria in db/db mice. It induced a two-fold increase in urine output and a 52% decrease in urine osmolality compared to non-treated db/db mice. After four weeks of treatment urinary albumin to creatinine ratio was 50% lower in treated mice compared to non-treated mice, and remained significantly lower until end of experiment. Glomerular filtration rate increased significantly over time in non-treated db/db mice but remained stable in treated mice. This study shows that vasopressin contributes to albuminuria and glomerular hyperfiltration via V2R in a mouse model of type 2 diabetes. It documents causality behind the association of vasopressin with renal disease observed in diabetic patients.


[Inhibitory effect on proliferation of KG1a cell line by methyltransferase inhibitors].
To explore the possibility of a new therapeutic strategy for leukemia by intervening in the DNA methylation to re-express p15 suppressor gene, methylation inhibitors, 5-Aza-2'-deoxycytidine (5-Aza-CdR) and cell differentiation agent (CDAII) were used to treat myelogenous leukemia cell line KG1a in which p15 gene expression was suppressed due to DNA hypermethylation. The biological characteristics of KG1a cells untreated or treated with the agents were investigated and analyzed using morphology, methylation specific-PCR (MSP), (3)H-labeled microassay technique, restriction endonuclease reaction, flow cytometry and immunofluorescence methods. The results indicated that both agents showed concentration-dependent and time-dependent inhibition of cell proliferation. 5-Aza-CdR and CDAII induced apoptosis and cell differentiation with G(2) and G(0)/G(1) arrest respectively. Furthermore, DNA methyltransferase activity and level of methylation in genomic DNA were decreased and p15 protein was re-expressed partially. It is concluded that it is possible to treat leukemia by intervening in the DNA methylation using methyltransferase inhibitors and it is worth to make a thorough study on mechanism of the new strategy.


Computing the non-Markovian coarse-grained interactions derived from the Mori-Zwanzig formalism in molecular systems: Application to polymer melts.
Memory effects are often introduced during coarse-graining of a complex dynamical system. In particular, a generalized Langevin equation (GLE) for the coarse-grained (CG) system arises in the context of Mori-Zwanzig formalism. Upon a pairwise decomposition, GLE can be reformulated into its pairwise version, i.e., non-Markovian dissipative particle dynamics (DPD). GLE models the dynamics of a single coarse particle, while DPD considers the dynamics of many interacting CG particles, with both CG systems governed by non-Markovian interactions. We compare two different methods for the practical implementation of the non-Markovian interactions in GLE and DPD systems. More specifically, a direct evaluation of the non-Markovian (NM) terms is performed in LE-NM and DPD-NM models, which requires the storage of historical information that significantly increases computational complexity. Alternatively, we use a few auxiliary variables in LE-AUX and DPD-AUX models to replace the non-Markovian dynamics with a Markovian dynamics in a higher dimensional space, leading to a much reduced memory footprint and computational cost. In our numerical benchmarks, the GLE and non-Markovian DPD models are constructed from molecular dynamics (MD) simulations of star-polymer melts. Results show that a Markovian dynamics with auxiliary variables successfully generates equivalent non-Markovian dynamics consistent with the reference MD system, while maintaining a tractable computational cost. Also, transient subdiffusion of the star-polymers observed in the MD system can be reproduced by the coarse-grained models. The non-interacting particle models, LE-NM/AUX, are computationally much cheaper than the interacting particle models, DPD-NM/AUX. However, the pairwise models with momentum conservation are more appropriate for correctly reproducing the long-time hydrodynamics characterised by an algebraic decay in the velocity autocorrelation function.


Circulating immune complexes in patients with acquired immune deficiency syndrome contain the AIDS-associated retrovirus.
Using the polyethylene glycol (PEG) precipitation technique, circulating immune complexes were demonstrated in 88% of patients with acquired immune deficiency syndrome (AIDS) and AIDS-related disorders. The finding of immune complexes in these individuals was confirmed by using a second assay, the commercially available Clq-IgG enzyme-linked immunosorbent assay kit. The material precipitated by PEG contained elevated levels of IgG, IgM, Clq, and C3. There was no substantial correlation between the level of IgG in the complexes and the serum IgG concentration, although there was a significant association between complexed IgG and the titer of anti-AIDS-associated retrovirus (ARV) antibody in the serum. In some precipitates, ARV antigens were detected by immunoblot analysis. Coculture of other precipitates with normal lymphocytes revealed the presence of infectious virus. These data confirm the existence of immune complexes in the sera of AIDS patients and indicate that in some cases they may comprise ARV and anti-ARV antibodies.


Combined effect of reticular stimulation and physostigmine on hippocampal theta activity.
The effect of electrical stimulation of the mesencephalic reticular formation and of physostigmine on hippocampal theta activity was combined in rats with implanted electrodes. The effect was not additive. The threshold for evoking motor activity by means of reticular stimulation decreased after physostigmine administration.


Novel cysteine proteinase inhibitors homologous to the proregions of cysteine proteinases.
Propeptides of papain-like cysteine proteinases such as papain, cathepsins B, L and S are potent inhibitors of their cognate cysteine proteinases with Ki values in the nanomolar range, and they exhibit highest inhibition selectivity for enzymes from which they originate. Recent studies have identified novel inhibitor proteins that are homologous to the proregions of papain-like cysteine proteinases. Mouse activated T-lymphocytes express cytotoxic T-lymphocyte antigen (CTLA-2), which is homologous to the proregion of mouse cathepsin L. CTLA-2 exhibits inhibitory activities to several cysteine proteinases. We have also identified a similar propeptide-like cysteine proteinase inhibitor, Bombyx cysteine proteinase inhibitor (BCPI), in the silkmoth Bombyx mori. BCPI is a slow and tight binding inhibitor of cathepsin L-like cysteine proteinases with Ki values in picomolar range, and the inhibition is highly selective towards these proteinases just like the propeptides. Recent genome analyses have shown the expression of similar propeptide-like proteins in Drosophila and rat, suggesting the presence of a novel class of cysteine proteinase inhibitors in a variety of organisms. Studies of the gene structures and phylogenetic analysis have shown that genes of the propeptide-like cysteine proteinase inhibitors have emerged from ancestor genes of their parental enzymes.


Convening a conference--facilitating networking among delegates.
This article provides conference convenors and delegates with suggestions to enhance networking by delegates attending conferences. Our suggestions are based on a literature search (which revealed very little); what we have found to work well at the research conferences we have convened; conference evaluations; and our experiences at conferences we have attended.


E-Learning Training to Improve Pediatric Parenteral Nutrition Practice: A Pilot Study in Two University Hospitals.
Education and training may improve the prescription of pediatric parenteral nutrition. The aim was to evaluate the impact of an e-learning method on parenteral nutrition prescription skills among pediatric residents in 2 pediatric hospitals. A randomized double-blind control study was conducted over a 9-month period among pediatric residents in HOSP1, Geneva, Switzerland, where physicians prescribe parenteral nutrition directly, and in HOSP2, Montreal, Canada, where physicians prescribe only occasionally because clinical pharmacists are devoted to this activity. The intervention consisted of an e-learning session about key issues of parenteral nutrition. Physician parenteral nutrition knowledge was evaluated with a standardized questionnaire based on clinical cases before and after the e-learning in the intervention groups; in the control groups, only the 2 tests were conducted. In HOSP1, participants also underwent iterative tests every 2 months to measure the retention of knowledge. Sixty-five physicians participated. Initial knowledge scores were higher in HOSP1 (pretest scores 180 ± 29 vs 133 ± 24, p < 0.001). Overall, there was no significant difference in the impact of the e-learning intervention between the control and e-learning groups (p > 0.05). A significant knowledge improvement was observed in HOSP2 in the e-learning group (p = 0.033). Iterative tests in HOSP1 showed persistence of knowledge without significant differences between the groups. E-learning satisfaction among the participants was outstanding (100%). E-learning seems to be an effective method for teaching parenteral nutrition among pediatric residents and fellows at the beginning of the training. High satisfaction with this teaching method was observed in this study.


Topology specific stabilization of promoter over telomeric G-quadruplex DNAs by bisbenzimidazole carboxamide derivatives.
Various potential G-quadruplex forming sequences present in the genome offer a platform to modulate their function by means of stabilizing molecules. Though G-quadruplex structures exhibit diverse structural topologies, the presence of G-quartets as a common structural element makes the design of topology specific ligands a daunting task. To address this, the subtle structural variations of loops and grooves present in the quadruplex structures can be exploited. To this end, we report the design and synthesis of quadruplex stabilizing agents based on bisbenzimidazole carboxamide derivatives of pyridine, 1,8-naphthyridine, and 1,10-phenanthroline. The designed ligands specifically bind to and stabilize promoter quadruplexes having parallel topology over any of the human telomeric quadruplex topologies (parallel, hybrid, or antiparallel) and duplex DNAs. CD melting studies indicate that ligands could impart higher stabilization to c-MYC and c-KIT promoter quadruplexes (up to 21 °C increment in Tm) than telomeric and duplex DNAs (ΔTm ≤ 2.5 °C). Consistent with a CD melting study, ligands bind strongly (Kb = ∼10(4) to 10(5) M(-1)) to c-MYC quadruplex DNA. Molecular modeling and dynamics studies provide insight into how the specificity is achieved and underscore the importance of flexible N-alkyl side chains attached to the benzimidazole-scaffold in recognizing propeller loops of promoter quadruplexes. Overall, the results reported here demonstrate that the benzimidazole scaffold represents a potent and powerful side chain, which could judiciously be assembled with a suitable central core to achieve specific binding to a particular quadruplex topology.


Natural killer cell-mediated lysis of hepatoma cells via specific induction of NKG2D ligands by the histone deacetylase inhibitor sodium valproate.
Natural killer (NK) cells as components of the innate immunity substantially contribute to antitumor immune responses. However, the tumor-associated ligands engaging activating NK cell receptors are largely unknown. An exception are the MHC class I chain-related molecules MICA and MICB and the UL16-binding proteins (ULBP) which bind to the activating immunoreceptor NKG2D expressed on cytotoxic lymphocytes. A therapeutic induction of NKG2D ligands that primes cancer cells for NK cell lysis has not yet been achieved. By microarray studies, we found evidence that treatment of human hepatocellular carcinoma cells with the histone deacetylase inhibitor (HDAC-I) sodium valproate (VPA) mediates recognition of cancer cells by cytotoxic lymphocytes via NKG2D. VPA induced transcription of MICA and MICB in hepatocellular carcinoma cells, leading to increased cell surface, soluble and total MIC protein expression. No significant changes in the expression of the NKG2D ligands ULBP1-3 were observed. The induction of MIC molecules increased lysis of hepatocellular carcinoma cells by NK cells which was abolished by addition of a blocking NKG2D antibody. Importantly, in primary human hepatocytes, VPA treatment did not induce MIC protein expression. Taken together, our data show that the HDAC-I VPA mediates specific priming of malignant cells for innate immune effector mechanisms. These results suggest the clinical evaluation of HDAC-I in solid tumors such as hepatocellular carcinoma, especially in combination with immunotherapy approaches employing adoptive NK cell transfer.


Enhancing Children's Safety by Barcoding Implementation at Breast Milk Feeding.
When newborns remain hospitalized in a neonatal intensive care unit, they are often unable to feed themselves and receive human milk through enteral nutrition devices such as orogastric or nasogastric probes. Therefore, the Nursing staff is responsible for the fractionation, storage and administration of human milk. Breast milk has a great biological complexity being the optimal food for the baby to provide all the nutrients needed. At the same time, it is a bodily fluid that carries the risk of disease transmission if not administered properly. Patient safety should be a priority in healthcare, and health information technologies could be used to avoid preventable adverse events. Barcoding technology has the ability to accurately verify patient identity and prescription accuracy before milk administration. This paper describes the steps followed to implement breast milk barcoding technology in an academic tertiary hospital.


Hemorrhage risk after quinsy tonsillectomy.
The goal of the study was to evaluate the incidence and possible predictive factors of post-tonsillectomy hemorrhage (PTH) in patients with peritonsillar abscess, treated by acute abscess tonsillectomy. A retrospective cohort study was performed on 205 patients who underwent bilateral abscess tonsillectomy under general anesthesia. Age, sex, smoking habits, history of recurrent tonsillitis or prior peritonsillar abscess, current medical treatment, side of the peritonsillar abscess, initial treatment, surgeon's experience, procedure duration, intra- and postoperative anti-inflammatory medications, and side of bleeding were analyzed. Bleeding occurred in 27 patients (13%). Ipsilateral hemorrhage was observed in 8 patients (4%) and contralateral hemorrhage in 19 patients (9%). The higher incidence of PTH in the side contralateral to the abscess was found to be statistically significant (P = 0.02). Male gender (P = 0.042), smoking (P = 0.009), and aspirin intake (P = 0.008) were statistically significant factors associated with an increased PTH risk. The risk of bleeding following abscess tonsillectomy seems higher than reported in elective tonsillectomy. This high incidence is mainly due to patients with prior aspirin intake or to bleeding in the side contralateral to the abscess. Postoperative bleeding could be reduced by performing a unilateral acute abscess tonsillectomy in selected patients. An algorithm is proposed for the management of peritonsillar abscess based on age, prior history of pharyngo-tonsillar infections, aspirin intake, and clinical improvement after initial drainage and antibiotherapy. C.


Regional dependence of cerebral reperfusion after circulatory arrest in rats.
The severity of neurologic dysfunction after circulatory arrest depends on cerebral reperfusion during and after resuscitation. The objective of current study was to investigate the temporal and spatial patterns of the cerebral perfusion immediately after resuscitation. Precise control of circulatory arrest was achieved in rats by combination of asphyxia and transient blockage of cardiac-specific beta-adrenergic receptors with esmolol, an ultra-short-acting beta-blocker. Animals were randomized into 3 groups with resuscitation starting 0.5 (sham group, no asphyxia, n = 5), 4 (Group 2, n = 5), or 12 minutes (Group 3, n = 8) later by retrograde intraarterial infusion of donor blood along with a resuscitation mixture. Cerebral perfusion was measured by magnetic resonance imaging (MRI) using arterial spin labeling. The average perfusion before arrest was 163 +/- 27 mL 100 g(-1) min(-1) under isoflurane anesthesia. Resuscitation led to transient perfusion increase, which started from thalamus and hypothalamus and later shifted to the cortex. Severe hypoperfusion to as low as 6% to 20% of the normal level developed in the first 10 to 20 minutes of reperfusion and lasted for at least 2 hours. On the fifth day after circulatory arrest, all animals showed a normal level of perfusion (159 +/- 57 mL 100 g(-1) min(-1) ) and minimal neurologic deficit. Nevertheless, histologic examination revealed extensive changes in the CA1 region of the hippocampus consistent with global ischemia and reperfusion damage. The combination of an improved circulatory arrest model and noninvasive MRI cerebral perfusion measurements provides a powerful tool for investigations of circulatory arrest and resuscitation, allowing for evaluation of therapies aimed at modulating cerebral reperfusion.


Chronic cough--about a clinical case...
Gastro-oesophageal reflux is the third most frequent cause of chronic cough. The diagnosis is not easy, especially when gastro-oesophageal reflux disease (GORD) has extra-oesophageal manifestations. The treatment is based on behavioural, pharmacological or surgical measures. The patient, aged 20 years, was a chronic smoker and since 2004 had a chronic cough. Therefore she had repeatedly been treated by several physicians with different antibiotics for respiratory infections. The family doctor in 2008 decided to investigate this cough. After investigation of the cardiorespiratory tract revealed no changes, the family doctor investigated the upper digestive tract. An endoscopy diagnosed gastro-oesophageal reflux. The patient was treated with pantoprazole 40 mg, with symptomatic relief. One important issue is that the patient went to see her family doctor instead of resorting to complementary consultations. It is also necessary to emphasize the need to think of the extra-oesophageal manifestations of GORD.


Acellular human lung scaffolds to model lung disease and tissue regeneration.
Recent advances in whole lung bioengineering have opened new doors for studying lung repair and regeneration ex vivo using acellular human derived lung tissue scaffolds. Methods to decellularise whole human lungs, lobes or resected segments from normal and diseased human lungs have been developed using both perfusion and immersion based techniques. Immersion based techniques allow laboratories without access to intact lobes the ability to generate acellular human lung scaffolds. Acellular human lung scaffolds can be further processed into small segments, thin slices or extracellular matrix extracts, to study cell behaviour such as viability, proliferation, migration and differentiation. Recent studies have offered important proof of concept of generating sufficient primary endothelial and lung epithelial cells to recellularise whole lobes that can be maintained for several days ex vivo in a bioreactor to study regeneration. In parallel, acellular human lung scaffolds have been increasingly used for studying cell-extracellular environment interactions. These studies have helped provide new insights into the role of the matrix and the extracellular environment in chronic human lung diseases such as chronic obstructive pulmonary disease and idiopathic pulmonary fibrosis. Acellular human lung scaffolds are a versatile new tool for studying human lung repair and regeneration ex vivo.


Outbreaks of contagious ecthyma in camels (Camelus dromedarius) in the Turkana district of Kenya.
Mortality among camel calves (Camelus dromedarius) is one of the most serious problems faced by camel herdsmen and, although there are several reasons for this mortality, diseases play a major role. In an investigation of outbreaks of contagious ecthyma in camels in the Turkana district of Kenya, four outbreaks were detected involving only camel calves. The principal lesions were distinct or largely coalesced pustules on the mouth, nose and muzzle. Direct electron microscopy of infected scabs was used to confirm the presence of the virus. Secondary infection of the pustules was common in affected calves. Morbidity in affected herds was 100%, with no adult involvement. Affected calves were unable to suckle properly. In all outbreaks, there was a concurrent outbreak of contagious pustular dermatitis in goat kids in the same household. Contagious ecthyma is an important disease in camels, contributing to calf debility and eventually to high calf mortality.


Reduction of Pnn by RNAi induces loss of cell-cell adhesion between human corneal epithelial cells.
Pinin (Pnn/DRS/memA) plays an important role in regulating cell-cell adhesion of corneal epithelial cells. In the nucleus, Pnn interacts with both transcriptional repressor and pre-mRNA processing machinery. Here we investigated the consequences of "knocking down" Pnn expression with short hairpin RNAi (shRNAi) on the corneal epithelial cell phenotype. Cultured human corneal epithelial (HCE-T) cells were cotransfected with a shRNAi-expressing construct containing an inverted repeat of a Pnn specific 21 nucleotide sequence (Pnn shRNAi) and a GFP vector as a marker of transfected cells. After 24-48 h, cells were fixed and immunostained with antibodies against Pnn, keratin, desmoplakin, desmoglein, E-cadherin, ZO-1, SR-proteins, and SRm300. To demonstrate specificity of the Pnn knock down, a rescue vector was designed by incorporating three conservative nucleotide substitutions within the Pnn-shRNAi targeting sequences of the full length Pnn-GFP construct, thus generating a Pnn construct to produce mRNA that Pnn shRNAi could not target (Pnn-CS3-GFP). HCE-T cells were cotransfected with Pnn shRNAi and GFP vectors and after 24 and 48 h exhibited significantly reduced immunostaining for Pnn. Western blot analyses of Pnn and E-cadherin protein expression in cells transfected with Pnn-shRNAi and GFP vectors revealed marked reduction in levels of both proteins compared to those observed in cells transfected with GFP alone. The cells receiving Pnn-shRNAi appeared to be less adherent to neighboring nontransfected cells, often exhibited altered cell shape, downregulated cell adhesion and cell junction molecules, and escaped from the epithelium. The Pnn shRNAi transfected cells exhibited fewer keratin filaments anchored to desmosomes and a concurrent increase in the perinuclear bundling of filaments. SR proteins and SRm300 showed an altered distribution in the Pnn knock down cells. Cotransfection of Pnn-CS3-GFP with Pnn shRNAi demonstrated that the conservatively mutated Pnn could maintain cell-cell adhesion. Our results indicate that knocking down Pnn expression leads to a loss of epithelial cell-cell adhesion, changes in cell shape, and movement of Pnn shRNAi transfected cells out of the epithelium. We suggest that Pnn plays an integral role in the establishment and maintenance of epithelial cell-cell adhesion via its activity within nuclear multi-protein complexes.


One-Pot Acid-Catalyzed Ring-Opening/Cyclization/Oxidation of Aziridines with N-Tosylhydrazones: Access to 1,2,4-Triazines.
A new, three-step, telescoped reaction sequence for the regioselective conversion of N-tosyl hydrazones and aziridines to 3,6-disubstituted and 3,5,6-trisubstituted 1,2,4-triazines is described. The process involves an efficient nucleophilic ring opening of the aziridine, giving access to a wide range of aminohydrazones, isolated with excellent yields. A "one-pot" procedure, combining the ring opening with a cyclization and an oxidation step, allows the preparation of diversified triazines in good yields.


Effects of alanine on gluconeogenesis in isolated rat hepatocytes.
The effect of alanine on pyruvate kinase was investigated in isolated rat hepatocytes. Alanine at concentrations of 2, 5 or 10 mM increased glucose production by 73% during the first 30 minutes of incubation of hepatocytes with 9 mM lactate and 1 mM pyruvate. After 3 minutes, the rate was not affected by the addition of alanine. A dose-response study showed that maximal stimulation of gluconeogenesis was achieved with 0.5 mM alanine. Using a method that measured recycling of phosphoenolpyruvate to pyruvate, it was found that in either fed or starved rats, alanine significantly decreased the percentage of phosphoenolpyruvate recycling when lactate was the substrate. However, no significant change in recycling was noted when either pyruvate or lactate-pyruvate was the glucose precursor. This study suggests that in intact liver cells, alanine has an inhibitory effect on pyruvate kinase. However, the inhibition is not of sufficient magnitude to completely account for the increase in glucose production when lactate is the substrate. It is hypothesized that alanine may have other effects on gluconeogenesis in addition to that of inhibiting pyruvate kinase.


Long-range imino proton-13C J-couplings and the through-bond correlation of imino and non-exchangeable protons in unlabeled DNA.
Thanks to rather large (5-9 Hz) long-range imino proton-13C J-couplings, heteronuclear correlation experiments in H2O provide unambiguous assignment of imino protons by intranucleotide through-bond connectivities to guanosine H8 and thymidine CH3 protons, or sequence-specific assignment of non-exchangeable protons when the imino protons are identified independently. This method is demonstrated in the Dickerson dodecamer [d(CGCGAATTCGCG)]2 and in a human telomeric fragment of 22 nucleotides.


When should we measure vitamin D concentration in clinical practice?
The many recently published data on vitamin D have raised much interest in the medical community. One of the consequences has been a great increase in the prescription of vitamin D concentration measurements in clinical practice. It must be reminded that only the measurement of 25-hydroxyvitamin D (25(OH)D) concentration is indicated to evaluate vitamin D status. Furthermore, since vitamin D insufficiency is so common, since treatment is inexpensive and has a large safety margin, and since we already have much data suggesting that besides its classic effects on bone and mineral metabolism, vitamin D may potentially be helpful for the prevention/management of several diseases, perhaps should it be prescribed to everyone without prior testing? In our opinion, there are however groups of patients in whom estimation of vitamin D status is legitimate and may be recommended. This includes patients in whom a "reasonably" evidence-based target concentration (i.e., based on randomized clinical trials when possible) should be achieved and/or maintained such as patients with rickets/osteomalacia, osteoporosis, chronic kidney disease and kidney transplant recipients, malabsorption, primary hyperparathyroidism, granulomatous disease, and those receiving treatments potentially inducing bone loss. Other patients in whom vitamin D concentration may be measured are those with symptoms compatible with a severe vitamin D deficiency or excess persisting without explanation such as those with diffuse pain, or elderly individuals who fall, or those receiving treatments which modify vitamin D metabolism such as some anti-convulsants. Measurement of Vitamin D concentrations should also be part of any exploration of calcium/phosphorus metabolism which includes measurement of serum calcium, phosphate and PTH.


Scalar modifications to gravity from unparticle effects may be testable.
Interest has focused recently on low energy implications of a nontrivial scale invariant sector of an effective field theory with an IR fixed point, manifest in terms of "unparticles" with peculiar properties. If unparticle stuff exists it could couple to the stress tensor and mediate a new "fifth" force ("ungravity"). Under the assumption of strict conformal invariance in the hidden sector down to low energies, we compute the lowest order ungravity correction to the Newtonian gravitational potential and find scale invariant power law corrections of type (R_(G)/r)(2d)_(U)(-1), where d_(U) is an anomalous unparticle dimension and R_(G) is a characteristic length scale where the ungravity interactions become significant. It is shown that a discrimination between extra dimension models and ungravity is possible in future improved submillimeter tests of gravity.


Antibody-drug conjugates in triple negative breast cancer.
Triple negative breast cancer (TNBC) is a heterogeneous disease that comprises 15-20% of all breast cancers and is more frequently seen in younger women, African-Americans, and BRCA1 expression. Advanced TNBC carries aggressive features and is associated with overall poor outcomes. Unfortunately, there are no targeted therapies available for non-BRCA associated TNBC, which remains a high unmet therapeutic need. One emerging treatment modality includes antibody-drug conjugates which are highly selective monoclonal antibodies conjugated to cytotoxic agents, designed to deliver cytotoxic drugs to antigen-expressing tumor cells. This review will highlight three antibody-drug conjugates currently being evaluated in TNBC (CDX-011, SGN-LIV1a, IMMU-132), including one that has been given Breakthrough Therapy designation from the US FDA.