Population-based prevalence and mutational landscape of von Willebrand disease using large-scale genetic databases.
Von Willebrand disease (VWD) is a common bleeding disorder caused by mutations in the von Willebrand factor gene (VWF). The true global prevalence of VWD has not been accurately established. We estimated the worldwide and within-population prevalence of inherited VWD by analyzing exome and genome data of 141,456 individuals gathered by the genome Aggregation Database (gnomAD). We also extended our data deepening by mining the main databases containing VWF variants i.e., the Leiden Open Variation Database (LOVD) and the Human Gene Mutation Database (HGMD) with the goal to explore the global mutational spectrum of VWD. A total of 4,313 VWF variants were identified in the gnomAD population, of which 505 were predicted to be pathogenic or already reported to be associated with VWD. Among the 282,912 alleles analyzed, 31,785 were affected by the aforementioned variants. The global prevalence of dominant VWD in 1000 individuals was established to be 74 for type 1, 3 for 2A, 3 for 2B and 6 for 2M. The global prevalences for recessive VWD forms (type 2N and type 3) were 0.31 and 0.7 in 1000 individuals, respectively. This comprehensive analysis provided a global mutational landscape of VWF by means of 927 already reported variants in the HGMD and LOVD datasets and 287 novel pathogenic variants identified in the gnomAD. Our results reveal that there is a considerably higher than expected prevalence of putative disease alleles and variants associated with VWD and suggest that a large number of VWD patients are undiagnosed.


Influence of the eluent composition on the ionization efficiency for morphine of pneumatically assisted electrospray, atmospheric-pressure chemical ionization and sonic spray.
A comparative study of three atmospheric-pressure ionization (API) sources for liquid chromatography/mass spectrometry (LC/MS), namely pneumatically assisted electrospray or ionspray (IS), atmospheric-pressure chemical ionization (APCI), and sonic spray (SS), with respect to the influence of the eluent composition on the ionization of morphine, is presented. The effect of organic modifiers, volatile acids, and buffer systems (with and without pH adjustment) in the LC mobile phase on the ionization efficiency of each interface is described. We conclude that for all three ion sources, the composition of the liquid phase had a serious impact on the ionization of the target compound. For IS and SS, very similar behavior towards the LC eluent was observed. In both cases, an increase in organic modifier resulted in an increase in ionization, while an increasing amount of volatile acid or buffer caused signal suppression. APCI, on the other hand, proved to respond completely differently towards the changes in the eluent. Again, an increased ionization was observed with an increase in organic modifier content but this time also in the presence of mobile phase additives like acids or buffers. Finally, we concluded that APCI proved to be the preferred ion source for the test compound because of its robust character and its direct applicability in traditional LC analysis.


Modulatory action of endogenous and exogenous nitric oxide on survival of alveolar macrophages from normal and bleomycin-treated rats.
To investigate the modulatory action of endogenous and exogenous nitric oxide (NO) on survival of alveolar macrophages (AMs) in different cellular states, AMs from normal rats (normal AMs) and from bleomycin (BLM)-treated rats (BLM AMs) were incubated by sodium nitroprusside (SNP, NO donor) and L-arginin (L-Arg, NO precursor), respectively. The survival of AMs was evaluated by apoptosis and cell cycles. The molecular mechanisms were investigated by the contents of Bcl-2, Bax proteins in AMs. The results are as follows: (1) The degree of BLM AMs apoptosis was higher than that of normal AMs; the number of BLM AMs in G(0)/G(1) phases was less than that of normal AMs; there was no significant difference in S+G(2)M phases between the number of BLM AMs and that of normal AMs. (2) Down-regulation of Bcl-2 and up-regulation of Bax occurred in BLM AMs, compared to those in normal AMs. (3) Apoptosis of AMs, either normal AMs or BLM AMs, was induced by both SNP and L-Arg, when compared to their respective control; only the number of BLM AMs in S+G(2)M phases was increased by L-Arg. (4) SNP and L-Arg induced a down-regulation of Bcl-2 and an up-regulation of Bax proteins in normal AMs, but did not induce the same change pattern in BLM AMs. (5) The Bax in BLM AMs was down-regulated by L-Arg. It is concluded that NO can induce the apoptosis of BLM AMs and normal AMs; that Bcl-2 and Bax are implicated in NO-induced apoptosis of normal AMs, whereas they are not involved in that of BLM AMs, suggesting the differential molecular mechanisms underlying the NO-induced apoptosis of normal AMs and BLM AMs; and that endogenous NO promotes proliferation of BLM AMs, which might be associated with down-regulation of Bax.


DNMT3B deficiency presenting as severe combined immune deficiency: A case report.
Immunodeficiency, Centromeric instability and Facial anomalies (ICF) syndrome is a group of rare autosomal recessive disorders. The immune disease in the ICF syndrome consists mainly of humoral immunodeficiency. T-cell dysfunction has previously been suspected to be part of the syndrome's spectrum. However, patients with ICF display, at a young age, a normal number of T cells that tend to decline throughout disease progression due to apoptosis. Biallelic mutations in the DNMT3B gene account for around 50% of ICF cases (ICF type 1). The remaining half may be linked to ZBTB24, CDCA7 or HELLS. Here we report a novel homozygous DNMT3B mutation (NM_ 006892; p.R826H) in a Lebanese family presenting in early infancy with severe combined immune deficiency (SCID). This work expands the clinical spectrum of the ICF syndrome and confirms the importance of tailoring therapeutic approaches for each patient with ICF syndrome, according to the clinical manifestations of his disease.


On-demand deep brain stimulation for essential tremor: a report on four cases.
Deep brain stimulation (DBS) is an established therapy for essential tremor (ET), but loss of efficacy due to tolerance can occur. Our objective was to evaluate if it is feasible to use DBS only on-demand and if this would prevent tolerance. We report on the effects of left-side thalamic DBS in 4 ET patients who were instructed to switch on stimulation only when using their right hand for motor tasks and were followed-up to 30 months after surgery. The patients were capable of using DBS only on-demand (DBS use of 22.0+/-13.5%/day). DBS led to a stable suppression of right arm tremor throughout the follow-up. No problems associated with tolerance such as tremor rebound or late therapy failure occurred. In comparison to publications stating that ET patients had been using DBS continuously during the daytime, the use of on-demand DBS saves battery life, which delays surgical replacement of the stimulator. Thus, on-demand DBS saves money, may help to prevent tolerance, and should be adopted for the long-term treatment of ET patients.


The new paradigm of systemic therapies for metastatic melanoma.
New treatments for metastatic melanoma work through distinct mechanisms: enhancing the immune response and blocking cellular proliferation. Agents that enhance the immune response include ipilimumab, pembrolizumb, and nivolumab; agents that block cellular proliferation include vemurafenib, dabrafenib, trametinib, cobimetinib, binimetinib, and selumetinib. The translational impact of laboratory discoveries has revolutionized management of metastatic melanoma and enhanced the prognosis of affected patients.


Endothelial tyrosine kinase receptor B prevents VE-cadherin cleavage and protects against atherosclerotic lesion development in ApoE-/- mice.
Tyrosine kinase receptor B (TrkB) is a high-affinity receptor for brain-derived neurotrophic factor (BDNF). In addition to its nervous system functions, TrkB is also expressed in the aortic endothelium. However, the effects of endothelial TrkB signaling on atherosclerosis remained unknown. Immunofluorescence analysis revealed that TrkB expression is downregulated in the endothelium of atherosclerotic lesions from ApoE-/- mice compared with the atheroma-free aorta of WT mice. Endothelial TrkB knockdown led to increased lesion size, lipid deposition and inflammatory responses in the atherosclerotic lesions of the ApoE-/- mice compared with the control mice. Mechanistic studies showed that TrkB activation prevented VE-cadherin shedding by enhancing the interaction between vascular endothelial protein tyrosine phosphatase and VE-cadherin, maintaining VE-cadherin in a dephosphorylated state. Our data demonstrate that TrkB is an endothelial injury-response molecule in atherogenesis. Endothelial BDNF/TrkB signaling reduces VE-cadherin shedding and protects against atherosclerotic lesion development in ApoE-/- mice.


Exploration for the multi-effect of cardamom in's resistance to multiple myeloma.
This paper aimed to probe the cardamom in effect on the viability, proliferation, apoptosis and periodic function of the multiple myeloma, and explore its mechanism. We used CCK-8 method to evaluate the effect of cardamom in on the viability of PBMNCs (Persom Blood Mononuclear Normal Cells). EdU can test the influence of small cell proliferation. We used the method of PI single-staining flow cytometry, in order to test the influence of tumor cell cycle. AO (Acridine Orange), EB (Ethidium Bromide) double staining fluorescene microscope was applied to observe the influence of tumor apoptotic morphology. It can be concluded that cardamom in can inhibit the viability and proliferation of MM (Multiple Myeloma) cells and cardamom in is the anti-myeloma drug with strong viability.


Elemental content of the placenta: A comparison between two high-risk obstetrical populations, adult women carrying multiples and adolescents carrying singletons.
The placenta is responsible for the exchange of nutrients and for preventing harmful compounds from entering the fetal circulation. With increasing industrialization, exposures to commercial and toxic metals become a concern for both pregnant women and those planning a pregnancy. The understanding of transport mechanisms and pharmacokinetics for most inorganic elements is incomplete and limited to normal term deliveries. To obtain novel data on 46 inorganic elements in placentae from two high-risk obstetric populations, women carrying multiples and adolescents carrying singletons, evaluating differences, if present, and identifying predictors of placental content. Placental tissue was collected from adolescents carrying singletons and adults carrying multiples. Elemental content was analyzed using inductively coupled plasma-mass spectrometry (ICP-MS). Multivariate regression and factor analyses were used. With the exception of Au and Pt, almost all placentae contained quantifiable concentrations of each element analyzed. All placentae contained the essential elements Ca, Fe, Mg, Se and Zn, which clustered together onto the same factor. Most elements were higher in placentae from women carrying multiples. Differences in placental content disappeared after adjusting for maternal age. Rare earth elements (REEs) clustered together and remained higher in the multiples even after adjusting for maternal age. Human placentae contain a wide range of elements, including REEs. Ranges differed considerably between cohorts. Elements with similar chemical properties, like REEs or nutritionally essential elements, clustered together. Maternal age, and therefore longer environmental exposure, was significantly associated with elevated element concentrations in the placenta. Placental concentrations of several metals that are known to be nutritionally essential (e.g., Fe, Ca, Mg, and Zn) did not differ significantly between cohorts, suggesting tight regulation, whereas concentrations of environmental contaminants differed significantly between groups, even after adjusting for maternal age.


Correlations between Age, Pain Intensity, Disability, and Tactile Acuity in Patients with Chronic Low Back Pain.
Chronic low back pain is an overwhelming problem for a wide range of people and leads to tactile acuity deficits. We aimed to investigate the correlations among age, pain severity, disability, and tactile acuity in patients with chronic low back pain by using multiple tactile acuity tests. A total of 58 participants (36.40 ± 14.95 years) with chronic low back pain were recruited, and two-point discrimination, point-to-point test, and two-point estimation were performed on their painful low back areas. The correlations between age, pain intensity, disability, and tactile acuity were characterized with Pearson's correlation coefficients. Subgroup analyses according to the median values of age, pain intensity, and disability were used to compare the intergroup difference in tactile acuity. Results illustrated significant negative associations among age, pain intensity, disability, and tactile acuity. Subgroup analyses revealed that patients with below-the-median values of age, pain intensity, and disability had better performance in tactile acuity tests than those with above-the-median values. This study indicated that tactile acuity was negatively associated with age, pain intensity, and disability in young patients with chronic low back pain.


Microbial characteristics of post-traumatic infective keratitis.
To determine the demographics, risk factors, clinical and microbiological characteristics, and treatment outcome of post-traumatic infective keratitis. Consecutive patients with post-traumatic infective keratitis presenting to the Ophthalmology Department of a tertiary referral hospital in Singapore between March 2012 and March 2016 were prospectively identified. A standardized data collection form was used to document patient demographics, microbiological diagnosis, antibiotic sensitivity, and pretreatment and posttreatment ocular characteristics. Any contact lens-induced keratitis was excluded from the study. In total, 26 patients were included for analysis. The mean age was 40.0 years (SD ± 19.4) and 84.6% of the patients were male. The majority of the patients (69.2%, n = 18) had sustained work-related injury in their eyes. Gram-negative organisms were predominant isolates (75.0%, n = 12) in culture-positive corneal scrapings (n = 16). Pan-sensitive Pseudomonas aeruginosa was the commonest organism isolated among the culture-positive cases (56.2%, n = 9). Three patients (18.7%) had developed fungal keratitis and Acanthamoeba was isolated in 1 patient (6.2%) with polymicrobial keratitis. Infections resolved with medical treatment in 22 eyes (84.6%) and 4 eyes (15.3%) required therapeutic corneal transplantation. A shift of practice in post-traumatic infective keratitis should be considered in tropical countries to include Gram-negative cover. Work safety practices with vigilance in initiating treatment and education by front-line physicians such as ophthalmology and general practitioners should be reinforced.


[Molecular-cellular mechanisms of the biological action of low x-ray doses on isolated mammalian cells].
The colony-formation was stimulated by X-irradiation (0.03-0.30 Gy) of cultured Chinese hamster cells. The stimulation was only noted in the nonadhered cells which was perhaps associated with a change in their adhesive properties. It was shown that low-level radiation induced changes in the structural organization of the cell membrane.


Normal suppressive T cell function of Epstein-Barr virus-induced B cell activation in Graves' disease.
Several studies have demonstrated abnormalities of T cell regulation of Epstein-Barr virus (EBV)-induced B cell activation in systemic autoimmune diseases, such as rheumatoid arthritis, systemic lupus erythematosus, and systemic sclerosis. To investigate whether this abnormality is a common feature of other autoimmune diseases, we studied 10 EBV-immune normal subjects and 22 EBV-immune patients with Graves' disease (GD); 11 had newly diagnosed hyperthyroidism, and 11 had received carbimazole treatment for hyperthyroidism for at least 6 months. Peripheral B lymphocytes infected with EBV were cultured for 20 days in the presence or absence of autologous T cells at different ratios. Immunoglobulins M and G secretion into the supernatants was determined using an enzyme-linked immunosorbent assay. The extent of suppression when T cells were added, as measured by a suppression ratio, was not significantly different in normal subjects and newly diagnosed GD patients (0.65 vs. 0.63 on the 16th day and 0.77 vs. 0.72 on the 20th day of culture, respectively). In carbimazole-treated patients, the appearance of functional suppressor T cells was delayed slightly, but the overall suppression ratios on the 16th and 20th days were normal. Thus, a T cell regulation abnormality of EBV-induced B cell activation could not be demonstrated in patients with untreated hyperthyroid GD, suggesting that the autoimmune reactivity in such patients is probably dependent upon a specific thyroid suppression defect rather than a generalized suppression defect.


Molecular epidemiology of dengue viruses in three provinces of Lao PDR, 2006-2010.
Few data on dengue epidemiology are available for Lao PDR. Here, we provide information on the complexity of dengue epidemiology in the country, demonstrating dynamic circulation that varies over space and time, according to serotype. We recruited 1,912 consenting patients presenting with WHO dengue criteria at Mahosot Hospital, Vientiane (central Laos), between 2006 and 2010. Between 2008 and 2010, 1,413 patients with undifferentiated fever were also recruited at Luang Namtha (LNT) Provincial Hospital (northern Laos) and 555 at Salavan (SV) Provincial Hospital (southern Laos). We report significant variations in Dengue virus (DENV) circulation between the three sites. Peaks of DENV infection were observed in the rainy seasons, although 11% of confirmed cases in the provinces and 4.6% in the capital were detected during the dry and cool seasons (between December and February). Four DENV serotypes were detected among the 867 RT-PCR positive patients: 76.9% DENV-1, 9.6% DENV-2, 7.7% DENV-4 and 5.3% DENV-3. DENV-1 was the predominant serotype throughout the study except in LNT in 2008 and 2009 when it was DENV-2. Before July 2009, DENV-2 was not detected in SV and only rarely detected in Vientiane. DENV-3 and DENV-4 were commonly detected in Vientiane, before 2008 for DENV-4 and after 2009 for DENV-3. The phylogenetic analyses of DENV envelope sequences suggest concurrent multiple introductions of new strains as well as active DENV circulation throughout Laos and with neighboring countries. It is therefore of great importance to develop and strengthen a year-round nation-wide surveillance network in order to collect data that would allow anticipation of public health issues caused by the occurrence of large dengue outbreaks.


The crystal structure and oxygen reduction reaction of Ni(II)-complex templated borate-sulfate and borate.
Three nickel borate compounds, [Ni(1-MI)6]·[B(OH)3]4·SO4 (1-MI = 1-methylimidazole) (1), [Ni(H2O)3(1-MI)3]·[B5O6(OH)4]2 (2) and [Ni(DMA3)]·[B6O7(OH)6]·3.5H2O (DMA3 = N,N-dimethylethylenediamine) (3), have been synthesized. It is noteworthy that the structures of 1 and 2 can be adjusted by varying the ratio of amine. Compound 3 has shown an unexpected example of unique water clusters in its structure. The three frameworks exhibit different interlinkage modes, resulting in channels varying in their size and shape. These compounds have been characterized by FTIR, UV-vis and PXRD. In addition, 1, 2, and 3 exhibited different wide band gaps (4.4 eV for 1, 4.5 eV for 2 and 4.4 eV for 3), and ORR activities with a half-wave potential of 0.78 V for 1, 0.74 V for 2 and 0.79 V for 3.


[Treatment of pancreatic fistulas].
Persisting fistulae of the pancreas may develop after necrotomy or operation of abscesses or pseudocysts with drainage following pancreatitis. More rare causes are lesions of the glandula during operation in the upper abdomen or due to accidents. First of all conservative therapy is indicated. When spontaneous closure during two or three months will not take place, closure of the fistula by drainage of its origin into a jejunal loop (Roux) and for fistulas of the head of the pancreas the implantation in the duodenal stump after B II-resection is indicated.


Human malaria parasite adenosine deaminase. Characterization in host enzyme-deficient erythrocyte culture.
Human malaria infected erythrocytes show a dramatic increase in adenosine deaminase activity in vitro. Using recently developed culture techniques, adenosine deaminase-deficient human erythrocytes were infected in vitro with the major human pathogen Plasmodium falciparum. Adenosine deaminase activity was undetectable in the uninfected host red cells, but increased by 2-fold over normal levels in these cells with an 8% parasitemia. The enzyme in these cells appeared unique in that its activity was markedly elevated over that of other parasite purine enzymes, was not cross-reactive with antibody against human erythrocyte adenosine deaminase, and though inhibited competitively by deoxycoformycin was relatively insensitive to erythro-9-(2-hydroxy-3-nonyl) adenine. The use of adenosine deaminase-deficient erythrocytes for the in vitro cultivation of Plasmodium provides a unique system for the study of parasite enzyme and allows further insight into the purine metabolism of the intraerythrocytic malaria parasite.


Optimal Design of an Hourglass in-Fiber Air Fabry-Perot Microcavity-Towards Spectral Characteristics and Strain Sensing Technology.
An hourglass in-fiber air microcavity Fabry-Perot interferometer is proposed in this paper, and its second reflecting surface of in-fiber microcavity is designed to be a concave reflector with the best curvature radius in order to improve the spectral characteristics. Experimental results proved that the extinction ratio of Fabry-Perot interferometer with cavity length of 60 μm and concave reflector radius of 60 μm is higher than for a rectangular Fabry-Perot interferometer with cavity length of 60 μm (14 dB: 11 dB). Theory and numerical simulation results show that the strain sensitivity of sensor can be improved by reducing the microcavity wall thickness and microcavity diameter, and when the in-fiber microcavity length is 40 μm, the microcavity wall thickness is 10 μm, the microcavity diameter is 20 μm, and the curvature radius of reflective surface II is 50 μm, the interference fringe contrast of is greater than 0.97, an Axial-pull sensitivity of 20.46 nm/N and resolution of 1 mN can be achieved in the range of 0-1 N axial tension. The results show that the performance of hourglass in-fiber microcavity interferometer is far superior to that of the traditional Fabry-Perot interferometer.


Outdoor Uphill Exercise Testing for Trail Runners, a More Suitable Method?
There is extensive knowledge about uphill running on a treadmill, although paucity regarding the influence of uphill trail running on exercise capacity in runners. The purpose of this study was to compare an uphill field test with a treadmill test with 1% inclination especially considering cardiopulmonary exercise variables. The difference in those tests between a group of trail runners and a group of road runners was tested for establishing a test specifically for trail runners. Ten male endurance road runners and ten male trail runners performed one maximal incremental treadmill test with 1% inclination and a maximal incremental field test on a hill at 16% inclination which they were instructed to run up four times for three minutes, each time with a higher pace. A mobile cardiopulmonary exercise testing unit was used to measure ventilation. There were no significant differences between trail runners and road runners. The equivalence factor was comparable between both groups. V ˙ O 2 p e a k was comparable for both test protocols. However, there was a significant difference between the two test protocols regarding lactate concentration, the respiratory exchange ratio, running cost, heart rate, Breathing reserve, and O2pulse. The greater lactate concentration and running cost recorded uphill indicate a higher energy demand during trail running than level road running.


Placental gas exchange in the guinea-pig: fetal blood gas tensions following the reduction of maternal oxygen capacity with carbon monoxide.
The effect of CO hypoxia on the placental exchange of respiratory gases was studied in anaesthetized pregnant guinea-pigs near term. Fetal PO2 and PCO2 were measured by mass spectrometry from a blood gas catheter in the right atrium. Administration of 5 ml CO over 65 s reduced maternal oxygen capacity by 26%. There was a rapid fall in fetal arterial PO2 and a more gradual rise in fetal PCO2. It was shown in separate experiments that the carboxyhaemoglobin content of fetal blood did not alter greatly in the first few min. after CO administration, which is the interval within which fetal PO2 was seen to fall. The alteration in fetal gas tensions can therefore be ascribed to the increased oxygen affinity and reduced oxygen capacity occasioned by the presence of carboxyhaemoglobin in the maternal blood. The alteration in placental oxygen transfer was calculated from the experimental findings, using a mathematical model of placental gas exchange in the guinea-pig. The total reduction in the oxygen transfer was 32% of the initial value. It was calculated that the reduction in maternal oxygen capacity was responsible for about two-thirds of this decrease, the remainder being due to the increased oxygen affinity of maternal blood.


Smartphone App-Based Assessment of Gait During Normal and Dual-Task Walking: Demonstration of Validity and Reliability.
Walking is a complex cognitive motor task that is commonly completed while performing another task such as talking or making decisions. Gait assessments performed under normal and "dual-task" walking conditions thus provide important insights into health. Such assessments, however, are limited primarily to laboratory-based settings. The objective of our study was to create and test a smartphone-based assessment of normal and dual-task walking for use in nonlaboratory settings. We created an iPhone app that used the phone's motion sensors to record movements during walking under normal conditions and while performing a serial-subtraction dual task, with the phone placed in the user's pants pocket. The app provided the user with multimedia instructions before and during the assessment. Acquired data were automatically uploaded to a cloud-based server for offline analyses. A total of 14 healthy adults completed 2 laboratory visits separated by 1 week. On each visit, they used the app to complete three 45-second trials each of normal and dual-task walking. Kinematic data were collected with the app and a gold-standard-instrumented GAITRite mat. Participants also used the app to complete normal and dual-task walking trials within their homes on 3 separate days. Within laboratory-based trials, GAITRite-derived heel strikes and toe-offs of the phone-side leg aligned with smartphone acceleration extrema, following filtering and rotation to the earth coordinate system. We derived stride times-a clinically meaningful metric of locomotor control-from GAITRite and app data, for all strides occurring over the GAITRite mat. We calculated stride times and the dual-task cost to the average stride time (ie, percentage change from normal to dual-task conditions) from both measurement devices. We calculated similar metrics from home-based app data. For these trials, periods of potential turning were identified via custom-developed algorithms and omitted from stride-time analyses. Across all detected strides in the laboratory, stride times derived from the app and GAITRite mat were highly correlated (P<.001, r2=.98). These correlations were independent of walking condition and pocket tightness. App- and GAITRite-derived stride-time dual-task costs were also highly correlated (P<.001, r2=.95). The error of app-derived stride times (mean 16.9, SD 9.0 ms) was unaffected by the magnitude of stride time, walking condition, or pocket tightness. For both normal and dual-task trials, average stride times derived from app walking trials demonstrated excellent test-retest reliability within and between both laboratory and home-based assessments (intraclass correlation coefficient range .82-.94). The iPhone app we created enabled valid and reliable assessment of stride timing-with the smartphone in the pocket-during both normal and dual-task walking and within both laboratory and nonlaboratory environments. Additional work is warranted to expand the functionality of this tool to older adults and other patient populations.


The effects of arousal reappraisal on stress responses, performance and attention.
This study examined the effects of arousal reappraisal on cardiovascular responses, demand and resource evaluations, self-confidence, performance and attention under pressurized conditions. A recent study by Moore et al. [2015. Reappraising threat: How to optimize performance under pressure. Journal of Sport and Exercise Psychology, 37(3), 339-343. doi: 10.1123/jsep.2014-0186 ] suggested that arousal reappraisal is beneficial to the promotion of challenge states and leads to improvements in single-trial performance. This study aimed to further the work of Moore and colleagues (2015) by examining the effects of arousal reappraisal on cardiovascular responses, demand and resource evaluations, self-confidence, performance and attention in a multi-trial pressurized performance situation. Participants were randomly assigned to either an arousal reappraisal intervention or control condition, and completed a pressurized dart throwing task. The intervention encouraged participants to view their physiological arousal as facilitative rather than debilitative to performance. Measures of cardiovascular reactivity, demand and resource evaluations, self-confidence, task performance and attention were recorded. The reappraisal group displayed more favorable cardiovascular reactivity and reported higher resource evaluations and higher self-confidence than the control group but no task performance or attention effects were detected. These findings demonstrate the strength of arousal reappraisal in promoting adaptive stress responses, perceptions of resources and self-confidence.


A Comparison of Applanation and Rebound Tonometers in Young Chicks.
To assess the validity of and compare applanation and rebound tonometry readings of intraocular pressure in alert normal chicks from ages 3 to 45 days. Intraocular pressures (IOPs) were measured weekly in awake White Leghorn chicks, from ages 3-45 days (n = 22-30 per age group), with both applanation Tono-Pen and rebound TonoLab tonometers. Three repeated measurements on individual eyes were used to derive variance data for both instruments at each age. Calibration curves were also derived for each instrument and each age, weekly from ages 10-45 days (n = 3-4 per age group), from in situ manometry data collected over IOP settings of 0 to 100 mmHg in 5 mmHg steps in cannulated eyes. The TonoLab showed less within measurement variability, but more variability with age, than the Tono-Pen. The coefficient of variation ranged from 3.8-8.3% for the TonoLab, compared to 11.0-19.7% for the Tono-Pen across all ages. For the youngest, 3 day-old chicks, mean IOPs recorded with the Tono-Pen and TonoLab were not significantly different (17.0 ± 5.6 and 15.2 ± 3.7 mmHg, respectively, P = .27). However, with increasing age, IOP readings significantly increased for the TonoLab (P < .001), whereas Tono-Pen readings did not. Compared to manometry settings, the Tono-Pen tended to underestimate IOPs while the TonoLab overestimated IOPs over the range 20-60 mmHg, saturating thereafter; there were also age-dependent differences for the TonoLab. Both the Tono-Pen and TonoLab gave IOP readings that differed from manometry settings in normal young chicks over some or all of the ages tested. These results reinforce the importance of calibrating clinical tonometers in animal studies involving IOP as a key variable.


Resetting the detection level of cord blood thyroid stimulating hormone (TSH) for the diagnosis of congenital hypothyroidism.
An appraisal of a 17-year primary thyroid stimulating hormone (TSH) screening programme for the detection of congenital hypothyroidism was carried out to establish the reference interval of cord blood TSH in unaffected infants; the mean cord blood TSH concentration of affected infants and the incidence of congenital hypothyroidism in the Najran province of Saudi Arabia. Our findings show a reference interval of cord blood TSH of 2.0-16.8 mU/l in unaffected infants; a mean cord blood TSH concentration of 399 mU/l in affected infants; a false positive rate for the diagnosis of at-risk infants of 1.02% and a congenital hypothyroidism incidence rate of 34/100 000 (1 : 2931) live births. These findings suggest that there is a need to reset the cord blood TSH concentration for the detection of at-risk infants. We suggest that the detection level of cord blood TSH for the recognition of at-risk infants can be set at 90 mU/l rather than the recommended level of 30 mU/l. This should reduce the false positive rate for detection of infants at risk of congenital hypothyroidism.


Correction: Programmable RNA microstructures for coordinated delivery of siRNAs.
Correction for 'Programmable RNA microstructures for coordinated delivery of siRNAs' by Jaimie Marie Stewart et al., Nanoscale, 2016, 8, 17542-17550.


Involvement of a novel cAMP signaling mediator for beige adipogenesis.
Exposure to cold temperature stimulates the sympathetic nervous system that activates β-adrenergic receptor signals in brown and beige adipocytes, leading to the induction of adaptive thermogenesis in mammals. Prominin-1 (PROM1) is a pentaspan transmembrane protein that is widely identified as a marker for stem cells, although the role of this protein as a regulator of many intracellular signaling cascades has been recently delineated. The main focus of the current study is to identify the previously unknown role of PROM1 in beige adipogenesis and adaptive thermogenesis. Prom1 whole body knockout (Prom1 KO) mice, Prom1 adipogenic progenitor (AP) cell-specific knockout (Prom1 APKO) mice and Prom1 adipocyte-specific knockout (Prom1 AKO) mice were constructed and were subject for the induction of adaptive thermogenesis. The effect of systemic Prom1 depletion was evaluated by hematoxylin and eosin staining, immunostaining, and biochemical analysis in vivo. Flow cytometric analysis was performed to determine the identity of PROM1-expressing cell types, and the resultant cells were subject to beige adipogenesis in vitro. The potential role of PROM1 and ERM in cAMP signaling was also assessed in undifferentiated AP cells in vitro. Finally, the specific effect of Prom1 depletion on AP cell or mature adipocytes on adaptive thermogenesis was evaluated by hematoxylin and eosin staining, immunostaining, and biochemical analysis in vivo. Prom1 KO mice displayed an impairment in cold- or β3-adrenergic agonist-induced adaptive thermogenesis in subcutaneous adipose tissues (SAT) but not in brown adipose tissues (BAT). By fluorescence-activated cell sorting (FACS) analysis, we identified that PROM1 positive cells are enriched in PDGFRα+Sca1+ AP cells from SAT. Interestingly, Prom1 knockout stromal vascular fractions showed reduced PDGFRα expression, suggesting a role of PROM1 in beige adipogenic potential. Indeed, we found that Prom1-deficient AP cells from SAT showed reduced potential for beige adipogenesis. Furthermore, AP cell-specific depletion of Prom1, but not adipocyte-specific depletion of Prom1, displayed defects in adaptive thermogenesis as evidenced by resistance to cold-induced browning of SAT and dampened energy expenditure in mice. We found that PROM1 positive AP cells are essential for the adaptive thermogenesis by ensuing stress-induced beige adipogenesis. Identification of PROM1 ligand might be useful in the activation of thermogenesis that could be potentially beneficial in combating obesity.


Sonographic patterns of ribs with tumor involvement.
We analyzed the sonographic appearance of the ribs when there was tumor involvement in 16 patients (12 men and 4 women). The tumors included 11 carcinomas, 3 nonepithelial malignancies and 2 benign tumors. Three sonographic patterns on the cross section of abnormal ribs were identified. Pattern I was an eccentrical hyperechoic plate-like shadow inside a hypoechoic tumor. Pattern II was a round hyperechoic shadow or ring in the center of a hypoechoic tumor. Pattern III was a hypoechoic tumor only. The Pancoast tumor manifested pattern I. Metastatic cancer, mostly squamous cell carcinoma, manifested pattern II, and both malignant and benign tumors could manifest pattern III. In sonography, the margin and shape of the tumors, the change in the acoustic shadow of the rib and the pleural line did not differentiate the malignant tumors from benign tumors. When we routinely took a biopsy of these tumors under sonar guidance, the diagnostic yield was 100% without any complications. We conclude that the rib should be evaluated in chest sonography, and that pattern recognition and biopsy under sonar guidance are most useful.


Effect of gas disturbance on combustion characteristics of flammable refrigerants near LFLs.
Refrigerants with low global warming potential (GWP), will play an important role in the research of alternative refrigerants and the development of new refrigerants, but most of them are flammable. And the actual environment of the refrigeration system varies with different factors, especially gas disturbance. In this paper, the combustion characteristics of four refrigerants, i.e., isobutane (R600a), ethylene (R1150), propane (R290), and 2,3,3,3-Tetrafluoroprop-1-ene (R1234yf) under the conventional static and the gas disturbance were studied. Firstly, the flame characteristics and lower flammability limits (LFLs) of the refrigerants were investigated. Then the special combustion phenomena were observed and analyzed in the presence of the gas disturbance, and an improved method for better defining the LFLs of the flammable gases under the disturbance was proposed. Finally, the experimental results show that the LFLs of the three natural refrigerants under specific low disturbance are lower than that in normal conditions, but except for R1234yf. It was found that the specific disturbance may enhance the combustion intensity by comparing the burning time of visible flame under the two conditions. The results have guiding significance for the safe application of flammable refrigerants.


Semiotic aspects of cognitive development: illustrations from early mathematical cognition.
The premise of this article is that cognitive development involves both conceptual and semiotic achievements. From this perspective, the authors emphasize the distinctness of the semiotic issues and develop a differentiated appreciation of semiotic aspects of cognition, particularly in the field of elementary mathematical cognition. The authors provide semiotic analyses of the differences between counting, adding, and multiplying and of the conventional place-value sign system. The authors introduce the concept of the field of reference of a sign, the differentiation of the field into foreground and background, and the dynamics within the field of reference. Finally, the authors relate these ideas to the dynamics between two dimensions of semiotic relations: the sign-referent dimension and the sign-sign dimension.


Risk of pacemaker implantation after femur fracture in patients with and without a history syncope: a Danish nationwide registry-based follow-up study.
It has previously been described that fall-associated injuries including fractures are commonly observed among patients with bradyarrhythmia. However, knowledge on the risk of pacemaker implantation after admission due to femur fracture from large population-based epidemiologic data is lacking. Therefore, we investigated the risk of pacemaker implantation following femur fracture in patients with and without a history of previous syncope. All patients with femur fracture between 2005-2017 were identified using the Danish Nationwide Patient Registry. Among these, patients already having a pacemaker were excluded. Primary outcome was one-year risk of pacemaker implantation and secondary outcome was one-year all-cause mortality. Multivariable logistic regression was used to obtain absolute and relative risks of the study endpoint in relation to patients with versus without history of syncope and standardized to the age, sex, selected comorbidity and pharmacotherapy distribution of all patients. Of 93,093 patients with femur fracture, 5508 (5.9%) had a history of syncope within five years. Patients with prior syncope were slightly older (84 vs. 83 years), more often male (33.6% vs. 29.4%), and had more often comorbidities relative to those without history of syncope. All-cause mortality was significantly higher among those with previous history of syncope compared to those without previous syncope (29.9% vs. 28.6%, P = 0.021). The relative mortality risk was 1.05 (95% CI: 1.01-1.09, P = 0.021). A total of 695 (0.8%) patients underwent pacemaker implantation within 5 years following femur fracture, and a significantly higher proportion of patients with syncope had a pacemaker implanted within one year (1.6% vs. 0.7%, P < 0.001; relative risk, 2.01 [95% CI: 1.55-2.46]). In patients with femur fracture, a history of syncope was significantly associated with a higher one-year risk of pacemaker implantation.


[Changes in hemodynamics during isometric loading in ischemic heart disease].
The hemodynamics and contractility of the myocardium were investigated in 38 men with coronary heart disease using maximal leg isometric load. It was shown that the systemic response to the isometric load in CHD patients unlike in normal subjects was mediated through increased total peripheral resistance rather than through an increase in myocardial contractility and stroke volume. The degree of these changes corresponded to the exercise tolerance as determined by bicycle ergometry. Leg isometric load is an accessible method for evaluating myocardial contractility in CHD which makes it possible to detect its disorders at the earliest stages in functional class I patients.


Removal of the N-terminal methionine improves the sweetness of the recombinant expressed sweet-tasting protein brazzein and its mutants in Escherichia coli.
The sweet-tasting protein brazzein is the smallest sweet protein with high sweet potency. Overexpression of this protein in a heterogenous host is an essential way for its production in food industry. In this study, the gene of minor form brazzein was cloned into the pET-SUMO vector with optimized codon usage and expressed in E. coli BL21-CodonPlus (DE3)-RIL. The recombinant protein in absence of the N-terminal methionine displayed a sweetness threshold about 1.5 μg/ml, which is the sweetest brazzein protein reported up to now. The unexpected sweet potency of the protein was validated by a series of mutants (7Val → Arg, 9Glu → Lys and 9Glu → Asp), in which E9K exhibited about 50% enhancement of sweetness than the wild type. The superior sweetness of recombinant brazzein and its sweeter mutants suggest their potential applications in food and beverages. PRACTICAL APPLICATIONS: The sweet-tasting proteins are natural, low-, or non-caloric and nutritive, showing to be promising replacers of sugars and artificial sweeteners, and can be used as sweet additives in the fields of food, medicine, and biotechnology. In the present study, we report that the recombinant brazzein protein expressed in E. coli exhibits superior and improved sweetness than those previously reported. Furthermore, sweeter mutants were obtained with the expression procedure. The superior sweetness of recombinant brazzein and its sweeter mutants suggest their potential applications in food, beverages, and other fields.


Bodychecking rules and concussion in elite hockey.
Athletes participating in contact sports such as ice hockey are exposed to a high risk of suffering a concussion. We determined whether recent rule changes regulating contact to the head introduced in 2010-11 and 2011-12 have been effective in reducing the incidence of concussion in the National Hockey League (NHL). A league with a longstanding ban on hits contacting the head, the Ontario Hockey League (OHL), was also studied. A retrospective study of NHL and OHL games for the 2009-10 to 2011-12 seasons was performed using official game records and team injury reports in addition to other media sources. Concussion incidence over the 3 seasons analyzed was 5.23 per 100 NHL regular season games and 5.05 per 100 OHL regular season games (IRR 1.04; 95% CI 1.01, 1.50). When injuries described as concussion-like or suspicious of concussion were included, incidences rose to 8.8 and 7.1 per 100 games respectively (IRR 1.23; 95% CI 0.81, 1.32). The number of NHL concussions or suspected concussions was lower in 2009-10 than in 2010-11 (IRR 0.61; 95% CI 0.45, 0.83), but did not increase from 2010-11 to 2011-12 (IRR 1.05; 95% CI 0.80, 1.38). 64.2% of NHL concussions were caused by bodychecking, and only 28.4% of concussions and 36.8% of suspected concussions were caused by illegal incidents. We conclude that rules regulating bodychecking to the head did not reduce the number of players suffering concussions during NHL regular season play and that further changes or stricter enforcement of existing rules may be required to minimize the risk of players suffering these injuries.


Recovery of nuclei from glycol-methacrylate-embedded tissue.
The analysis of antigens, enzyme histochemical markers, and DNA has become an important part of the classification of some leukemias, lymphomas, and other neoplastic diseases. Many of the relevant antigens and most of the relevant enzyme histochemical activities are destroyed and others are less than optimally preserved in tissues embedded in hot paraffin. Most enzymatic activities and antigens are well preserved in tissues embedded at 4 degrees C in glycol methacrylate (GMA). The measurement of DNA content in neoplastic cells with the most commonly employed techniques depended on the availability of fresh suspensions of cells until the development by Hedley of methods that permit the recovery of nuclei from paraffin blocks for this purpose. In order to facilitate the analysis of antigens, enzymatic markers, and DNA from the same sample of tissue, we have developed a means of recovery of nuclei from GMA-embedded tissues. Twenty-microns-thick sections of GMA-embedded tonsil were either pretreated with an organic solvent (absolute ethanol or 2-ethoxyethanol) followed by rehydration in phosphate buffered saline (PBS) or directly rehydrated in PBS. The suspensions were formed mechanically by gentle sonication. The type of fixative and length of PBS rehydration were varied. Tissue fixed in 100% acetone, embedded in GMA, and rehydrated directly in PBS for six days gave the highest average yield of nuclei, 3.7 x 10(7) nuclei per gram tissue. In order to assess DNA binding of fluorescent dyes, 2-microns-thick GMA sections were stained with chromomycin, Hoechst 33342 (Sigma Chemical, St Louis, MO), and propidium iodide. Hoechst 33342 bound specifically to the nuclei with low background staining.


Training and Certification for Domestic Violence Service Providers: The Need for a National Standard Curriculum and Training Approach.
Domestic violence (DV) continues to constitute an enormous public health problem in the United States. Knowledge and understanding of the complexities involved in DV has grown significantly in recent years revealing a need for providers who have broad training in a variety of legal, safety, developmental, and clinical issues that face families impacted by DV. This paper reviews current approaches to training and the ability of such methods to adequately prepare providers. There are no national standards for providers at any level from DV advocates to batterer interventionists, to clinicians with the required hours of training in most states at an alarmingly low level. Few states require cross training for those working as victim advocates or batterer interventionists. The systems that currently provide segregated and limited training create silos of service that are less effective. A proposed set of standards and training guidelines are proposed for DV advocates, batterer interventionists, and clinicians along with a discussion of the implications of such standards for the field.


A novel method of caenophidian snake sex identification using molecular markers based on two gametologous genes.
Sex identification provides important information for ecological and evolutionary studies, as well as benefiting snake conservation management. Traditional methods such as cloacal probing or cloacal popping are counterproductive for sex identification concerning very small species, resulting in difficulties in the management of their breeding programs. In this study, the nucleotide sequences of gametologous genes (CTNNB1 and WAC genes) were used for the development of molecular sexing markers in caenophidian snakes. Two candidate markers were developed with the two primer sets, and successfully amplified by a single band on the agarose gel in male (ZZ) and two bands, differing in fragment sizes, in female (ZW) of 16 caenophidian snakes for CTNNB1 and 12 caenophidian snakes for WAC. Another candidate marker was developed with the primer set to amplify the specific sequence for CTNNB1W homolog, and the PCR products were successfully obtained in a female-specific 250-bp DNA bands. The three candidate PCR sexing markers provide a simple sex identification method based on the amplification of gametologous genes, and they can be used to facilitate effective caenophidian snake conservation and management programs.


Rapid Jumps and Bioluminescence Elicited by Controlled Hydrodynamic Stimuli in a Mesopelagic Copepod, Pleuromamma xiphias.
Actively vertically migrating mesopelagic copepods are preyed upon by a wide variety of fishes and invertebrates. Their responses to predatory attacks include vigorous escape jumps and discharge of bioluminescent material. Escape jumps and bioluminescent discharges in the calanoid copepod Pleuromamma xiphias were elicited by quantified hydrodynamic disturbances. Brief weak stimuli (peak water velocity 64 +/- 21 {mu}m s-1) elicited weak (peak force 6.5 dynes) propulsive responses ("jumps") and no bioluminescence. Moderate stimuli (1580 +/- 780 {mu}m s-1) produced strong propulsive responses consisting of long trains of coordinated power strokes by the four pairs of swimming legs ("kicks"). Peak forces averaged 42 dynes. Strong stimuli (5520 +/- 3420 {mu}m s-1) were required to elicit both a jump and a bioluminescent discharge. In several cases, multiple stimuli were needed to evoke bioluminescence, given the limits on stimulus magnitude imposed by the apparatus. Repeated bioluminescent discharges could be evoked, but this responsiveness waned rapidly. Latencies for the jump response (14 +/- 4 ms) were shorter than for the accompanying bioluminescent discharge (49 +/- 26 ms). The higher threshold for eliciting bioluminescent discharge compared to escape jumps suggests that the copepods save this defense mechanism for what is perceived to be a stronger threat.


Influence of calcium entry blockade on alpha 1- and alpha 2-adrenoceptor mediated vasoconstriction in the forearm of hypertensive patients.
The influence of treatment with the calcium entry blockers PY 108-068 (PY) and PN 200-110 (PN) on alpha 1- and alpha 2-adrenoceptor mediated vasoconstriction has been investigated in the forearms of hypertensive patients. Changes in forearm vascular resistance (FVR) in response to the intra-arterial infusion of drugs were determined at the end of a placebo period and after 2-4 weeks of treatment with PY or PN. The drugs used were the selective agonists methoxamine (alpha 1) and B-HT 933 (alpha 2). During placebo, basal FVR was dose-dependently increased by methoxamine and B-HT 933. Basal blood pressure was lowered during PN but not during PY. Treatment with the calcium entry blockers did not influence the effect of methoxamine, but the vasoconstriction induced by B-HT 933 was attenuated by both of the calcium entry blockers. These results confirm the findings in animal studies that calcium entry blockers preferentially inhibit the alpha 2-adrenoceptor mediated vasoconstriction induced by selective agonists.


The effect of NK cell therapy on sepsis secondary to lung cancer: A case report.
Patients with sepsis face high mortality rates and a bleak prognosis, prompting the need for advanced therapeutic interventions. A male patient diagnosed with moderately low-differentiated squamous cell carcinoma received diverse treatments, including radiotherapy, chemotherapy, immunotherapy, and targeted therapy to inhibit angiogenesis. Subsequently, he developed sepsis after comprehensive treatment, and conventional antibiotic combinations proved ineffective in combating the infection. As an experimental approach, allogeneic natural killer (NK) cell infusion was administered. Following the NK cell infusion, the patient regained consciousness, and laboratory analyses showed reduced infection-related markers, suppressed serum inflammatory cytokines, and elevated anti-tumor cytokines. However, the therapeutic effect only lasted 2-3 days. In vitro investigations demonstrated that the allogeneic NK cell product reduced interleukin-6 levels in the patient's serum. Moreover, subsequent co-cultivation of the NK cell product with the patient's serum resulted in a decrease in the proportion of cytotoxic subpopulations of NK cells and a downregulation of the expression of NK-mediated killing molecules. In conclusion, adoptive transfusion of allogeneic NK cells may improve sepsis symptoms in patients with tumor-related sepsis. In vitro co-culture tests hold promise in providing predictive biomarkers for treatment effectiveness.


A method for determination of human urinary inactive kallikrein (prekallikrein).
Inactive kallikrein (prekallikrein) in human urine was discovered recently. A more accurate method to measure this zymogen and total kallikrein is described. With trypsin pretreatment, it was found that 35.3 +/- 2.7% of total urinary kallikrein activity was prekallikrein. The results are supported by a new direct radioimmunoassay which measures predominantly active kallikrein.


[Aspects of the history of the deaconess movement].
Having existed for 60 years as a modern, respected and highly esteemed hospital the patients of Sankt Lukas Stiftelsen (St. Lukas Foundation) in Gentofte were taken over by the Public Health Service in 1992. In the article is given a short survey of the founding of modern deaconess work at Kaiserswerth (Germany) and how it spread to Denmark. A special description is given of the development of the St. Lukas Foundation Hospital and the changes which have followed since 1992. The deaconesses continue their work by taking up new aspects of social work e.g. the founding of the first hospice in Den mark.


Optic atrophy in prematurity: pathophysiology and clinical features.
Optic atrophy is an important cause of visual impairment in children, and the aetiological profile has changed over time. Technological advancements led by neuroimaging of the visual pathway and imaging of the optic nerve with optical coherence tomography have accelerated the understanding of this condition. In the new millennium, an increasing prevalence of prematurity as a cause of optic atrophy in children has been highlighted. This new shift has been linked with increasing rates of premature births and improved neonatal survival of preterm infants. The available literature is limited to hospital and registry-based cohorts with modest sample sizes, methodological heterogeneity and selection bias limitations. Larger studies that are better designed are required to better understand the contribution of prematurity to the disease burden. In addition to considering other life-threatening aetiologies, screening for premature birth should be covered as part of a comprehensive history when evaluating a child with paediatric optic atrophy.


A new fermentation process allows large-scale production of human milk oligosaccharides by metabolically engineered bacteria.
When fed to a beta-galactosidase-negative (lacZ(-)) Escherichia coli strain that was grown on an alternative carbon source (such as glycerol), lactose accumulated intracellularly on induction of the lactose permease. We showed that intracellular lactose was efficiently glycosylated when genes of glycosyltransferase that use lactose as acceptor were expressed. High-cell-density cultivation of lacZ(-) strains that overexpressed the beta 1,3 N acetyl glucosaminyltransferase lgtA gene of Neisseria meningitidis resulted in the synthesis of 6 g x L(-1) of the expected trisaccharide (GlcNAc beta 1-3Gal beta 1-4Glc). When the beta 1,4 galactosyltransferase lgtB gene of N. meningitidis was coexpressed with lgtA, the trisaccharide was further converted to lacto-N-neotetraose (Gal beta 1-4GlcNAc beta 1-3Gal beta 1-4Glc) and lacto-N-neoheaxose with a yield higher than 5 g x L(-1). In a similar way, the nanA(-) E. coli strain that was devoid of NeuAc aldolase activity accumulated NeuAc on induction of the NanT permease and the lacZ(-) nanA(-) strain that overexpressed the N. meningitidis genes of the alpha2,3 sialyltransferase and of the CMP-NeuAc synthase efficiently produced sialyllactose (NeuAc alpha 2-3Gal beta 1-4Glc) from exogenous NeuAc and lactose.


Randomized controlled trial of a self-efficacy enhancement program for the cardiac rehabilitation of Thai patients with myocardial infarction.
This study examined the effects of a self-efficacy enhancement program for the cardiac rehabilitation of Thai patients who had a myocardial infarction. Sixty-six hospitalized patients of various ages and both genders were randomly assigned to either an experimental or a control group. Participants in the experimental group took part in three individualized in-hospital education sessions and three weekly sessions of telephone counseling. The control group primarily engaged in a supervised exercise and activities of a daily living performance regimen, and received education in this regard. Self-efficacy and functional status were measured via questionnaire. Four weeks after discharge, the experimental group was found to have significantly higher total self-efficacy and functional status scores than the control group. In addition, the experimental group exhibited significantly higher subscale scores on social activity, household tasks, occupation, and exercise self-efficacy than the control group. These results indicate that the program is effective in improving the self-efficacy and functional status of Thai patients who have had a myocardial infarction.


[The influence of new hybrid antioxidants ichphans on the kinetics of ascorbate-induced reduction of spin probe radical centers in liposomes].
The influence of new hybrid antioxidants ichphans on the structure of liposome lipid bilayer was studied. The analysis of the kinetics of the ascorbate-induced reduction of spin probe radical centers incorporated in a membrane revealed that ichphans produce a modifying effect on the structure of liposome membrane. It was established that derivates with distinct hydrophobic properties transform different sites of the membrane that are the sites of localization of ichphans in the intramembraneous space. The data obtained suggest that lipid components play an important role in the effects of ichphans on biological membranes.


Parlodel treatment of uremic hypogonadism in men.
Parlodel, 2.5 mg daily, was given during 1-3 months to 14 uremic hypogonadal patients (placebo controlled in 6). Except for a decrease in serum prolactin levels on Parlodel (p less than 0.01), no significant changes in hormonal values (gonadotropins and testosterone) were observed either on Parlodel or on placebo. Parlodel treatment improved the potency of 10 patients, eliminated gynecomastia in 2 and normalized spermatogenesis in 6; the best results were obtained in patients on low-protein diet. On placebo, potency improved only in 1 patient; no improvement in spermatogenesis was registered. Thus, Parlodel can improve sexual function and spermatogenesis in hypogonadal uremic males.


The Aurora-Kinase A Phe31-Ile polymorphism as possible predictor of response to treatment in head and neck squamous cell carcinoma.
Recently the Aurora-Kinases (Aurk) moved into the focus as novel disease related biomarkers and therapeutic targets. Elevated Aurora-Kinase expression has been found in a number of malignancies, amongst them HNSCC. For esophageal cancer, the AurkA Phe31-Ile polymorphism has previously been associated with tumor progression. Here we evaluated the treatment efficiency of HNSCC cell radiation as a function of Aurora-Kinases in HNSCC cell lines. Moreover, we investigated a potential sensitization to radiation by a cell treatment with the inhibitors Alisertib, Barasertib, Docetaxel and VX-680. In parallel the radiation dependent expression and regulation of AurkA/B, p-Akt Ser 473 and Survivin and the AurkA polymorphism were investigated in primary tumor samples. We identified a high-risk collective with elevated AurkA and Survivin or AurkA and p-Akt Ser 473 expression. High AurkA, AurkB, and p-Akt Ser 473 expression was exclusively found in the heterozygous cell line. We found a polymorphism dependent sensitivity to treatments with different Aurk inhibitors: The homozygous cell line UD-SCC-5 could be sensitized to radiation with Docetaxel in combination with any of the Aurora-Kinase inhibitors. In contrast, treatment with Docetaxel or radiation did not enhance the inhibitory effect of Barasertib or VX-680 in the heterozygous SAS cell line. These findings indicate that the Aurora-Kinase A Phe31-Ile-polymorphism is a possibly predictive factor for response to radiation in combination with Docetaxel and Aurora-Kinase inhibitor treatments.


Characterization of the substrate specificity of the major cysteine protease (cruzipain) from Trypanosoma cruzi using a portion-mixing combinatorial library and fluorogenic peptides.
The substrate specificity of the major cysteinyl proteinase of the parasitic protozoan Trypanosoma cruzi (cruzipain) was investigated, by combinatorial replacement of amino acid residues at positions P5-P'5, using a fluorescent quenched solid-phase library assay. Positively charged residues appear to be a general preference in the P5-P3 and the P'5-P'3 positions, while a hydrophobic residue was always required at the P2 position. A broad range of amino acids could be accepted at the P'1 position. A clear difference in terms of specificity between cruzipain and human cathepsin L was observed for the accommodation of Pro at the P2 position. The P1 specificity was investigated by a more detailed enzyme kinetic analysis using peptidyl-MCA (where MCA is methylcoumarin amide) and Abz-peptidyl-EDDnp [where Abz is o-aminobenzoic acid and EDDnp is N-(2,4-dinitrophenyl)ethylenediamine] as substrates, and the results were compared with those obtained using human cathepsin L. Cruzipain showed a clear preference for benzyl-Cys or Arg at the P1 position. Human cathepsin L presented similar behaviour to that of cruzipain for the hydrolysis of the epsilon-NH2-Cap-Leu-Xaa-MCA (where Cap is epsilon-aminocaproyl) and Abz-Lys-Leu-Xaa-Phe-Ser-Lys-Gln-EDDnp series, whereas the mammalian enzyme was able to tolerate large P1 residues, such as phenylalanine, better than cruzipain in the latter series.


NOS1AP associates with Scribble and regulates dendritic spine development.
The formation and function of the neuronal synapse is dependent on the asymmetric distribution of proteins both presynaptically and postsynaptically. Recently, proteins important in establishing cellular polarity have been implicated in the synapse. We therefore performed a proteomic screen with known polarity proteins and identified novel complexes involved in synaptic function. Specifically, we show that the tumor suppressor protein, Scribble, associates with neuronal nitric oxide synthase (nNOS) adaptor protein (NOS1AP) [also known as C-terminal PDZ ligand of nNOS (CAPON)] and is found both presynaptically and postsynaptically. The Scribble-NOS1AP association is direct and is mediated through the phosphotyrosine-binding (PTB) domain of NOS1AP and the fourth PDZ domain of Scribble. Further, we show that Scribble bridges NOS1AP to a beta-Pix [beta-p21-activated kinase (PAK)-interacting exchange factor]/Git1 (G-protein-coupled receptor kinase-interacting protein)/PAK complex. The overexpression of NOS1AP leads to an increase in dendritic protrusions, in a fashion that depends on the NOS1AP PTB domain. Consistent with these observations, both full-length NOS1AP and the NOS1AP PTB domain influence Rac activity. Together these data suggest that NOS1AP plays an important role in the mammalian synapse.


Antibody-dependent cell-mediated cytotoxicity in rheumatoid arthritis. Effect of rheumatoid serum fractions on normal lymphocytes.
Reduced antibody-dependent cell-mediated cytotoxicity (ADCC) was demonstrated in lymphocytes of patients with rheumatoid arthritis (RA). Rheumatoid factor (RF) positive sera inhibited ADCC of normal lymphocytes by reacting both with effector and target cells (sensitized chicken red blood cells). These sera were fractionated by specific adsorption or gradient ultracentrifugation, and isolated RF or RF negative fractions were tested for their ability to inhibit ADCC by reacting with normal human lymphocytes or target cells. RF was ineffective on normal lymphocytes but it strongly inhibited the reaction by interaction with target cells. IgG RF negative fractions of certain sera were inhibitory by direct interaction with effector cells.


Physical interaction of the activator protein-1 factors c-Fos and c-Jun with Cbfa1 for collagenase-3 promoter activation.
Previously, we determined that the activator protein-1 (AP-1)-binding site and the runt domain (RD)-binding site and their binding proteins, c-Fos.c-Jun and Cbfa, regulate the collagenase-3 promoter in parathyroid hormone-treated and differentiating osteoblasts. Here we show that Cbfa1 and c-Fos.c-Jun appear to cooperatively bind the RD- and AP-1-binding sites and form ternary structures in vitro. Both in vitro and in vivo co-immunoprecipitation and yeast two-hybrid studies further demonstrate interaction between Cbfa1 with c-Fos and c-Jun in the absence of phosphorylation and without binding to DNA. Additionally, only the runt domain of Cbfa1 was required for interaction with c-Jun and c-Fos. In mammalian cells, overexpression of Cbfa1 enhanced c-Jun activation of AP-1-binding site promoter activity, demonstrating functional interaction. Finally, insertion of base pairs that disrupted the helical phasing between the AP-1- and RD-binding sites also inhibited collagenase-3 promoter activation. Thus, we provide direct evidence that Cbfa1 and c-Fos.c-Jun physically interact and cooperatively bind the AP-1- and RD-binding sites in the collagenase-3 promoter. Moreover, the AP-1- and RD-binding sites appear to be organized in a specific required helical arrangement that facilitates transcription factor interaction and enables promoter activation.


Demethylation of methyl-accepting chemotaxis proteins in Escherichia coli induced by the repellents glycerol and ethylene glycol.
The addition of glycerol or ethylene glycol caused not only severe tumbling but also a drastic decrease in the methylation level of methyl-accepting chemotaxis proteins (MCPs) in Escherichia coli. Experiments with various mutants having defects in their MCPs showed that the demethylation occurred in all three kinds of MCPs, MCPI, II, and III. The addition of an attractant to the glycerol- or ethylene glycol-treated cells resulted in a distinct increase in the methylation level of the relevant MCP, indicating that glycerol and ethylene glycol do not directly damage the methylation-demethylation system in the cell. The time courses of adaptation and MCP demethylation upon addition of these repellents were consistent with each other. Furthermore, both the response time and the extent of MCP demethylation were increased in parallel with increasing concentrations of glycerol or ethylene glycol. These results indicate that the adaptation to these repellents is performed by the demethylation of MCPs. Thus, glycerol and ethylene glycol are novel repellents, which utilize not just one but all three kinds of MCPs for both information processing and adaptation.


Tuning Radical Relay Residues by Proton Management Rescues Protein Electron Hopping.
Transient tyrosine and tryptophan radicals play key roles in the electron transfer (ET) reactions of photosystem (PS) II, ribonucleotide reductase (RNR), photolyase, and many other proteins. However, Tyr and Trp are not functionally interchangeable, and the factors controlling their reactivity are often unclear. Cytochrome c peroxidase (CcP) employs a Trp191•+ radical to oxidize reduced cytochrome c (Cc). Although a Tyr191 replacement also forms a stable radical, it does not support rapid ET from Cc. Here we probe the redox properties of CcP Y191 by non-natural amino acid substitution, altering the ET driving force and manipulating the protic environment of Y191. Higher potential fluorotyrosine residues increase ET rates marginally, but only addition of a hydrogen bond donor to Tyr191• (via Leu232His or Glu) substantially alters activity by increasing the ET rate by nearly 30-fold. ESR and ESEEM spectroscopies, crystallography, and pH-dependent ET kinetics provide strong evidence for hydrogen bond formation to Y191• by His232/Glu232. Rate measurements and rapid freeze quench ESR spectroscopy further reveal differences in radical propagation and Cc oxidation that support an increased Y191• formal potential of ∼200 mV in the presence of E232. Hence, Y191 inactivity results from a potential drop owing to Y191•+ deprotonation. Incorporation of a well-positioned base to accept and donate back a hydrogen bond upshifts the Tyr• potential into a range where it can effectively oxidize Cc. These findings have implications for the YZ/YD radicals of PS II, hole-hopping in RNR and cryptochrome, and engineering proteins for long-range ET reactions.


Patient-reported fatigue in breast cancer patients receiving radiation therapy.
Fatigue or tiredness is one of the most commonly reported symptoms in breast cancer patients treated with radiation therapy (RT). This study aimed to identify characteristics associated with fatigue in breast cancer patients receiving adjuvant RT. Patients with non-metastatic breast cancer receiving RT at the Odette Cancer Centre from 2011 to 2017 were included in our study if they completed at least one ESAS pre- and post-RT. Information regarding patient, disease and treatment characteristics was retrieved from chart review. To identify variables associated with fatigue scores pre-RT, post-RT and changes in fatigue scores, a univariate and multivariate general linear regression analysis was conducted; p < 0.05 was considered statistically significant. Our study included 1223 female patients (mean age 59 years old) who completed ESAS on average 28 days before, and 142 days after RT. In multivariate analysis, higher baseline fatigue scores were found in women with higher disease stages (p = 0.001), and those who receive locoregional radiation (p < 0.001). No variables were significantly associated with post-RT fatigue scores. While adjuvant chemotherapy and locoregional RT were associated with higher baseline scores in univariate analysis, in multivariate analysis, they were associated with significant reduction in fatigue post-RT (p = 0.01, p = 0.007 respectively). Fatigue is associated with higher disease stage and receipt of locoregional radiation. While the relationship between anxiety or depressive symptoms and fatigue is well-established, a major gap exists in our understanding of its etiology and treatment; further investigation to address this can better improve patient quality of life.


Glutamate as a putative neurotransmitter in the buccal central pattern generator of Helisoma trivolvis.
1. The effects of L-glutamate superfusion over identified neurons within the buccal ganglia of Helisoma trivolvis were examined. Glutamate mirrored the effect of activity of subunit 2 (S2) of the tripartite feeding central pattern generator (CPG) on S2 postsynaptic neurons. Neurons that are excited by S2 are depolarized by glutamate, whereas neurons that are inhibited by S2 are hyperpolarized by glutamate. Glutamate also stimulated rhythmic S2 activity. 2. Different glutamate agonists could mimic specific components of the effects of glutamate on buccal neurons. Kainate produced depolarizations in neurons that receive S2 excitatory postsynaptic potentials (EPSPs) and activated rhythmic S2 activity. Quisqualate produced hyperpolarizations in neurons that receive S2 inhibitory postsynaptic potentials (IPSPs). 3. The non-N-methyl-D-aspartate glutamate receptor antagonist cyano-7-nitroquinoxaline-2,3-dione (CNQX) blocked the effects of S2 EPSPs and depolarizations produced by application of glutamate and kainate, but was ineffective in blocking S2 IPSPs or hyperpolarizations produced by application of glutamate and quisqualate. 4. These data support the hypothesis that glutamate is the transmitter of S2 of the feeding CPG in Helisoma, acting at CNQX-sensitive kainate-like receptors at excitatory synapses and CNQX-insensitive quisqualate-like receptors at inhibitory synapses.


Patients choice for method of early abortion among comprehensive abortion care (CAC) clients at Kathmandu Medical College Teaching Hospital (KMCTH).
The over all objective of the study was to determine different methods of abortion opted by CAC clients at KMCTH. The specific objective of the study was to know the reasons for pregnancy termination and to know the reasons opted for either medical or surgical method of abortion. A hospital based prospective study was carried out for a period of six months at KMCTH from 1st January 2006 to 31st June 2006. All the patient undergoing CAC services were included for the study. Clients were provided with written and verbal information regarding the methods of terminating early abortion and its associated complications. After that they were asked to give their informed choice and decision. All the pertinent information was entered on pre-structured questionnaire. During the study period a total of 100 patients underwent CAC services. The commonest reason for termination pregnancy was no desire for additional children (60%) followed by youngest child too small or short spacing (21%). 74% of the patients opted for surgical abortion, 23% patient opted for medical abortion and 3% of the patient remain undecided. Reasons for favouring surgical method of abortion was that surgical abortion is complete (35), repeated visits are avoided (18), quick (10) would be with service provider and feel safe (5), lack of expectancy (2) side effect of medical treatment (1), twin pregnancy (1), easy (1), fear of pain (1). Medical method of abortion was favoured due to fear of surgery (9), easy and less painful (8) and maintains privacy (6). Factors affecting the choice of abortion method appear to be numerous and complex. Providers need to be sensitive to differences in women's values and life circumstances when counselling them about an abortion method. In particular, providers should incorporate into their counselling sessions what women need to know about the characteristics of abortion methods and help women to identify what is the best option for them. Key words: Early abortion medical methods, surgical methods, choice.


Antitumor potential of dark sweet cherry sweet (Prunus avium) phenolics in suppressing xenograft tumor growth of MDA-MB-453 breast cancer cells.
This study investigated in vivo the antitumor activity of dark sweet cherry (DSC) whole extracted phenolics (WE) and fractions enriched in anthocyanins (ACN) or proanthocyanidins (PCA) in athymic mice xenografted with MDA-MB-453 breast cancer cells. Mice were gavaged with WE, ACN or PCA extracts (150 mg/kg body weight/day) for 36 days. Results showed that tumor growth was suppressed at similar levels by WE, ACN and PCA compared to control group (C) without signs of toxicity or significant changes in mRNA oncogenic biomarkers in tumors or mRNA invasive biomarker in distant organs. Tumor protein analyses showed that WE, ACN and PCA induced at similar levels the stress-regulated ERK1/2 phosphorylation, known to be linked to apoptosis induction. However, ACN showed enhanced antitumor activity through down-regulation of total oncogenic and stress-related Akt, STAT3, p38, JNK and NF-kB proteins. In addition, immunohistochemistry analysis of Ki-67 revealed inhibition of tumor cell proliferation with potency WE ≥ ACN ≥ PCA. Differential quantitative proteomic high-resolution nano-HPLC tandem mass spectrometry analysis of tumors from ACN and C groups revealed the identity of 66 proteins associated with poor breast cancer prognosis that were expressed only in C group (61 proteins) or differentially up-regulated (P<.05) in C group (5 proteins). These findings revealed ACN-targeted proteins associated to tumor growth and invasion and the potential of DSC ACN for breast cancer treatment. Results lead to a follow-up study with highly immunodeficient mice/invasive cell line subtype and advanced tumor development to validate the anti-invasive activity of DSC anthocyanins.


Wood fibers are a crucial microhabitat for cellulose- and xylan- degrading bacteria in the hindgut of the wood-feeding beetle Odontotaenius disjunctus.
Wood digestion in insects relies on the maintenance of a mosaic of numerous microhabitats, each colonized by distinct microbiomes. Understanding the division of digestive labor between these microhabitats- is central to understanding the physiology and evolution of symbiotic wood digestion. A microhabitat that has emerged to be of direct relevance to the process of lignocellulose digestion is the surface of ingested plant material. Wood particles in the guts of some termites are colonized by a specialized bacterial fiber-digesting microbiome, but whether this represents a widespread strategy among insect lineages that have independently evolved wood-feeding remains an open question. In this study, we investigated the bacterial communities specifically associated with wood fibers in the gut of the passalid beetle Odontotaenius disjunctus. We developed a Percoll-based centrifugation method to isolate and enrich the wood particles from the anterior hindgut, allowing us to access the wood fibers and their associated microbiome. We then performed assays of enzyme activity and used short-read and long-read amplicon sequencing of the 16S rRNA gene to identify the composition of the fiber-associated microbiome. Our assays demonstrated that the anterior hindgut, which houses a majority of the bacterial load, is an important site for lignocellulose digestion. Wood particles enriched from the anterior hindgut contribute to a large proportion of the total enzyme activity. The sequencing revealed that O. disjunctus, like termites, harbors a distinct fiber-associated microbiome, but notably, its community is enriched in insect-specific groups of Lactococcus and Turicibacter. Our study underscores the importance of microhabitats in fostering the complex symbiotic relationships between wood-feeding insects and their microbiomes. The discovery of distinct fiber-digesting symbionts in O. disjunctus, compared to termites, highlights the diverse evolutionary paths insects have taken to adapt to a challenging diet.


Excretion of infectious hepatitis E virus into milk in cows imposes high risks of zoonosis.
Hepatitis E virus (HEV) represents the main cause of acute hepatitis worldwide. HEV infection in immunocompromised patients involves a high risk for the development of chronic hepatitis. Because HEV is recognized as a zoonotic pathogen, it is currently believed that swine is the primary reservoir. However, this is not sufficient to justify the strikingly high seroprevalence of HEV in both developing and Western countries. Thus, this study aimed to identify new zoonotic sources that bear a high risk of transmission to humans. We collected fecal, blood, and milk samples of cows in a typical rural region of Yunnan Province in southwest China, where mixed farming of domestic animals is a common practice. HEV RNA was quantified by quantitative real-time polymerase chain reaction, and the whole genome was sequenced. HEV infectivity was assessed in rhesus macaques. We found a high prevalence of active HEV infection in cows as determined by viral RNA positivity in fecal samples. Surprisingly, we discovered that HEV is excreted into milk that is produced by infected cows. Phylogenetic analysis revealed that all HEV isolates from cow/milk belong to genotype 4 and subtype 4h. Gavage with HEV-contaminated raw and even pasteurized milk resulted in active infection in rhesus macaques. Importantly, a short period of boiling, but not pasteurization, could completely inactivate HEV. Infectious HEV-contaminated cow milk is recognized as a new zoonotic source that bears a high risk of transmission to humans; these results call attention to understanding and establishing proper measurement and control of HEV zoonotic transmission, particularly in the setting of mixed farming of domestic animals. (Hepatology 2016;64:350-359).


Clinical and histopathological characterization and typing of the human papillomavirus in common warts of kidney transplant recipients.
The prevalence of skin lesions caused by the human papillomavirus (HPV) is high in kidney transplant patients. Twenty recipients of kidney transplants with a diagnosis of common warts were evaluated. HPV detection was performed by polymerase chain reaction (PCR) using the MY09/MY11 and RK91 primers. HPV typing was performed by restriction fragment length polymorphism analysis and direct sequencing. The presence of HPV was identified in 10 patients (50%) and the types identified were HPV-2, 27, 29, 34 and 57.


Does Otovestibular Loss in the Autosomal Dominant Disorder DFNA9 Have an Impact of on Cognition? A Systematic Review.
Background and Purpose: Cognitive impairment has been observed in patients with bilateral vestibular loss (BVL) and in patients with sensorineural hearing loss (SNHL). DFNA9 is an autosomal dominant disorder that causes a combination of both sensory deficits by the 3rd to 5th decade. We therefore hypothesize a combined detrimental effect on cognition. The aim of this systematic review was to identify studies related to DFNA9 in general and its relationship with cognitive impairment more specifically. Materials and Methods: Several databases including Medline, Cochrane Database of Systematic Reviews, Cochrane Central Register of Controlled Trials, ISI Web of Knowledge, and Web of Science were searched to accumulate information about DFNA9-mutations, including phenotype, genotype, pathophysiology, quality of life (QOL), and imaging in general and cognitive function more specifically. A qualitative analysis was performed on the 55 articles that qualified. Results: The clinical features of DFNA9 are different along the 24 COCH mutations, described up to now. Vestibular symptoms generally present themselves a few years after SNHL onset in mutations associated with the vWFA-domain although they can precede SNHL onset in other mutations associated with the LCCL-domain. QoL has not been studied extensively in DFNA9, although scarce work is available on the positive impact of cochlear implantation to rehabilitate hearing. No studies were found evaluating cognition in DFNA9 patients. Conclusion: Although cognitive impairment has been demonstrated in patients with hearing loss as well as in patients with BVL, no studies have been reported on the combination of both sensory deficits, such as in DFNA9. Further research is warranted to correlate otovestibular status with cognition.


Renal osteodystrophy.
Over the past decade important advances in our understanding of the pathophysiology and treatment of renal osteodystrophy have been made. In particular, the role of calcitriol deficiency in the genesis of hyperparathyroidism in early renal failure is now better understood. So too are the effects of aluminium on bone, and whereas the more florid aluminium related disease is now unusual the more subtle effects of aluminium are now being appreciated. There is still a major problem in the long-term treatment of hyperparathyroid bone disease. The reasons why parathyroid gland proliferation continues to occur on dialysis therapy require a better understanding of cellular events regulating hormone production and parathyroid cell replication. The case for early intervention with vitamin D is now strong but whether such an approach materially influences the long-term outcome is not yet established. Changes in the approach to treatment and in the modalities used for renal replacement therapy will continue to modify the nature of the bone disease.


Expression of growth differentiation factor-9 messenger ribonucleic acid in ovarian and nonovarian rodent and human tissues.
Growth differentiation factor-9 (GDF-9) is a member of the transforming growth factor-beta family that is reported to be expressed exclusively in the ovary, specifically in the oocyte. Female mice deficient in GDF-9 are infertile, suggesting that GDF-9 receptor agonists and antagonists may specifically modulate fertility. We now report that GDF-9 messenger RNA (mRNA) is expressed in nonovarian tissues in mice, rats, and humans. GDF-9 mRNA was detected in mouse and rat ovary, testis, and hypothalamus by Northern blot and RT-PCR analyses. The localization of GDF-9 mRNA specifically in oocytes of the mouse ovary was confirmed by in situ hybridization histochemistry. In mouse testis, although localization in Sertoli cell cytoplasm could not be ruled out, mRNA expression was observed in large pachytene spermatocytes and round spermatids. The expression of GDF-9 mRNA in human tissues was assessed by Northern blot and RT-PCR analyses. GDF-9 mRNA was observed in ovary and testis and, surprisingly, in diverse nongonadal tissues, including pituitary, uterus, and bone marrow. Therefore, GDF-9 mRNA expression in rodents is not exclusive to the ovary, but includes the testis and hypothalamus. Furthermore, human GDF-9 mRNA is expressed not only in the gonads, but also in several extragonadal tissues. The function and relevance of nongonadal GDF-9 mRNA are not known, but may affect strategies for contraception and fertility that are based on GDF-9 activity.


Effects of indirubin and isatin on cell viability, mutagenicity, genotoxicity and BAX/ERCC1 gene expression.
Indigofera suffruticosa Miller (Fabaceae) and I. truxillensis Kunth produce compounds, such as isatin (ISA) and indirubin (IRN), which possess antitumour properties. Their effects in mammalian cells are still not very well understood. We evaluated the activities of ISA and/or IRN on cell viability and apoptosis in vitro, their genotoxic potentials in vitro and in vivo, and the IRN- and ISA-induced expression of ERCC1 or BAX genes. HeLa and/or CHO-K1 cell lines were tested (3 or 24 h) in the MTT, Trypan blue exclusion, acridine orange/ethidium bromide, cytokinesis-blocked micronucleus (CBMN) and comet (36, 24 and 72 h) tests after treatment with IRN (0.1 to 200 μM) or ISA (0.5 to 50 μM). Gene expression was measured by RT-qPCR in HeLa cells. Swiss albino mice received IRN (3, 4 or 24 h) by gavage (50, 100 and 150 mg/kg determined from the LD50 - 1 g/kg b.w.) and submitted to comet assay in vivo. IRN reduced the viability of CHO-K1 (24 h; 5 to 200 μM) and HeLa cells (10 to 200 μM), and was antiproliferative in the CBMN test (CHO-K1: 0.5 to 10 μM; HeLa: 5 and 10 μM). The drug did not induce apoptosis, micronucleus neither altered gene expression. IRN and ISA were genotoxic for HeLa cells (3 and 24 h) at all doses tested. IRN (100 and 150 mg/kg) also induced genotoxicity in vivo (4 h). IRN and ISA have properties that make them candidates as chemotherapeutics for further pharmacological investigations.


[Uniform designed research on the active ingredients assembling of Chinese medicine prescription for anti-liver fibrosis].
To explore the method and significance for studying active anti-liver fibrosis ingredients consisted Chinese medicine compound prescription based on Chinese medicine theory. Optimized prescription was screened out, adopting uniform block design with 4-factor 8-level table and regression analysis, through applying the four known effective ingredients (cordyceps sinensis polysaccharide, salvianolic acid B, amygdaloside and gypenosides) of Fuzheng Huayu Capsule (FZHYC, a new Chinese medine anti-liver fibrosis drug) to two rat liver fibrosis models established separately by dimethylnitrosamine (DMN) and CCl4, and taking the liver content of hydroxyproline (Hyp) as the screen index. Then a further study for comparing and verifying the efficacy of the obtained optimized prescription was conducted on the two former models respectively by observing the changes of Hyp content in liver, serum ALT activity and fibrosis pathology after medication, controlled by the original FZHYC and the recipe assembled by all the four ingredients. Two optimized prescriptions (OPA and OPB) were screened out separately in the studies conducted on the two models. Both of them were consisted of cordyceps sinensis polysaccharide, Amygdaloside and Gypenosides, but different in constituent ratio, i.e., the ratio in OPA was 60 : 80: 50, and that in OPB, 20: 160: 50. Verifying study showed both OPA and OPA were significantly effective, with the efficacy equivalent to that of FZHYC (P>0.05). However, when they were used in combining with salvianolic acid B (the cutout ingredient in the screening), the efficacy lowered surely. Uniform design is a valuable method in the compatibility research of Chinese Medicine drugs' composition. To assemble a new compound recipe reasonably based on the prescription of traditional compound recipe could make its effect equivalent to that of the original prescription. Ingredients or constituents in a prescription, either presented synergistic or antagonistic effects, are not randomly stacked together, and they should be orderly assembled in intrinsic rules of qualitative and quantitative changing.


Functional identities of thoracic respiratory interneurones in the cat.
1. Spike-triggered averaging was used to reveal focal synaptic potentials (FSPs) in the thoracic ventral horn resulting from impulses in individual respiratory interneurones situated in the contralateral ventral horn of the same segment (T6 or T7) in anaesthetized paralysed cats. 2. FSPs were of two types, negative-going or positive-going, and were of the same sign at each of several sites for each unit. These two types were interpreted as indicating excitatory or inhibitory actions respectively. 3. The positive-going FSPs were almost all the result of activity in the most strongly modulated, phasic respiratory interneurones, whereas most of the negative-going ones were derived from neurones with a continuous tonic component in their firing patterns. 4. Positive-going FSPs (particularly those from inspiratory neurones) were generally found at more ventral sites in the ventral horn than negative-going ones. 5. It is suggested that an important role for the phasically active interneurones is to provide phasic inhibition, which is combined with largely tonic excitation to produce strong respiratory modulation in the activity of other neurones in the thoracic spinal cord, particularly in motoneurones. However, some groups of respiratory interneurones, such as a group of tonic inspiratory ones which are located more dorsally than the phasic inspiratory ones, may not receive such phasic inhibition.


Inhalation toxicity and lung toxicokinetics of C60 fullerene nanoparticles and microparticles.
While several recent reports have described the toxicity of water-soluble C60 fullerene nanoparticles, none have reported the toxicity resulting from the inhalation exposures to C60 fullerene nanoparticles or microparticles. To address this knowledge gap, we exposed male rats to C60 fullerene nanoparticles (2.22 mg/m3, 55 nm diameter) and microparticles (2.35 mg/m3, 0.93 microm diameter) for 3 h a day, for 10 consecutive days using a nose-only exposure system. Nanoparticles were created utilizing an aerosol vaporization and condensation process. Nanoparticles and microparticles were subjected to high-pressure liquid chromatography (HPLC), XRD, and scanning laser Raman spectroscopy, which cumulatively indicated no chemical modification of the C60 fullerenes occurred during the aerosol generation. At necropsy, no gross or microscopic lesions were observed in either group of C60 fullerene exposures rats. Hematology and serum chemistry results found statistically significant differences, although small in magnitude, in both exposure groups. Comparisons of bronchoalveolar (BAL) lavage fluid parameters identified a significant increase in protein concentration in rats exposed to C60 fullerene nanoparticles. BAL fluid macrophages from both exposure groups contained brown pigments, consistent with C60 fullerenes. C60 lung particle burdens were greater in nanoparticle-exposed rats than in microparticle-exposed rats. The calculated lung deposition rate and deposition fraction were 41 and 50% greater, respectively, in C60 fullerene nanoparticle-exposed group than the C60 fullerene microparticle-exposed group. Lung half-lives for C60 fullerene nanoparticles and microparticles were 26 and 29 days, respectively. In summary, this first in vivo assessment of the toxicity resulting from inhalation exposures to C60 fullerene nanoparticles and microparticles found minimal changes in the toxicological endpoints examined. Additional toxicological assessments involving longer duration inhalation exposures are needed to develop a better and more conclusive understanding of the potential toxicity of inhaled C60 fullerenes whether in nanoparticle or microparticle form.


Intestinal MUC2 and gastric M1/MUC5AC in preneoplastic lesions induced by 1,2-dimethylhydrazine in rat: a sequential analysis.
Our study was performed to sequentially analyze the expression of the intestinal mucin MUC2 and of the gastric mucin MUC5AC as indicators during progression of preneoplastic biomarkers in rat colon. F344 rats were sacrificed 2, 4, 8, 12, 24 and 36 weeks after injection of 1,2-dimethylhydrazine (DMH, 200 mg/kg, i.p.). The expression of MUC2 and of MUC5AC was studied by immunohistochemistry in preneoplastic lesions classified in two categories: histologically altered foci (HAF) and beta-catenin accumulated crypts (BCAC). HAF appeared 4 weeks after DMH injection. Their crypt multiplicity stagnated with time (3-4 crypts/foci) but gastric MUC5AC mucin was always observed in some goblet cells of the lesions of this category. In contrast, MUC2-immunostaining was not modified compared to the adjacent crypts. Double-immunofluorescence revealed that goblet cells which produced MUC5AC continued to express MUC2. In BCAC, crypt multiplicity and mucin expression strongly evolved with time. These lesions were observed only 8 weeks after DMH-injection. At this stage, 20% of BCAC showed a decreased MUC2 expression and 33% were MUC5AC immunopositive. At the 36-week point, 43% of BCAC had a reduced MUC2 staining and 90% were positive for MUC5AC. This immunopositivity was often observed in all the cells of these lesions. Seldom, some BCAC were depleted at the same time in MUC2 and in MUC5AC. Similar alterations in mucin expression were observed in human colonic pre-neoplastic lesions. These findings suggest that a decrease in MUC2 expression and staining of MUC5AC in non-goblet-like cells predicts histological progression of preneoplastic lesions.


Inhibition of hTERT Gene Expression by Silibinin-Loaded PLGA-PEG-Fe3O4 in T47D Breast Cancer Cell Line.
Introduction : Nowadays, using drug delivery is an essential method to improve cancer therapy through decreasing drug toxicity and increasing efficiency of treatment. Silibinin (C25H22O10), a polyphenolic flavonoid which is isolated from the milk thistle plant, has various applications in cancer therapy but it has hydrophobic structure with low water solubility and bioavailability. To increase the effect of silibinin, silibinin-loaded PLGA-PEG-Fe3O4 was prepared to determine the inhibitory effect of this nanodrug on Telomerase gene expression. Methods : The rate of silibinin loaded into PLGA-PEG-Fe3O4 was measured. Then, the cytotoxic effect of silibinin-loaded PLGA-PEG-Fe3O4 was determined by Methyl Thiazol Tetrazolium (MTT) assay. After that, inhibition of Telomerase gene expression was indicated through Real-time PCR. Results : Data analysis from MTT assay showed that silibinin-loaded PLGA-PEG-Fe3O4 had dose dependent cytotoxic effect on T47D cell line. MTT assay showed no cytotoxic effect of free PLGA-PEG-Fe3O4 on T47D breast cancer cell line. Real Time PCR analysis showed that the level of telomerase gene expression more efficiently decreased with silibinin-loaded PLGA-PEG-Fe3O4 than with free silibinin alone. Conclusion : The present study indicates that this nanodrug causes down-regulation of Telomerase gene expression in cancer cells. Therefore, PLGA-PEG-Fe3O4 could be an appropriate carrier for hydrophobic agents such as silibinin to improve their action in cancer therapy.