Effect of SC-435 on the gastrointestinal migrating myoelectric complex in guinea pigs.
To determine whether SC-435, a new ileal apical sodium-codependent bile acid transporter (IBAT) inhibitor, can alter the gastrointestinal motility in guinea pigs. Sixty guinea pigs received regular diet or IBAT inhibitor (SC-435) diet for 2, 4, and 8 weeks, respectively. At the end of the feeding period, the gallbladder motility was assessed and then four bipolar silver electrodes were implanted on the antrum, duodenum, jejunum, and ileum. Seven days later, migrating motor complex (MMC) was recorded and the total bile acid pool size was measured according to the isotope dilution principle in the meantime. After feeding SC435, the gallbladder motility was declined in the 4-week group and the 8-week group. The bile acid pool size decreased by 17.11% (P <0.05) in the 4-week group and 48.35% (P < 0.05) in the 8-week group. The places of origin of MMC were changed where antral origins (37%) and duodenal origins (46%) decreased whereas jejunal origins (17%) increased. The MMC cycle period was prolonged in the duodenum (1.16 times in the 4-week group, P < 0.05; 1.38 times in the 8-week group, P < 0.05) whereas MMC amplitude fell in the duodenum (10.58% in the 4-week group, P <0.05; 49.17% in the 8-week group, P <0.05). There were not significant differences in all parameters of MMC between the control group and the 2-week group in guinea pigs. The IBAT inhibitor (SC-435) reduces the bile acid pool size and inhibits the MMC cycle activity. MMC is related to the enterohepatic circulation of bile acids, which is consistent with the changes of the bile acid pool size in guinea pigs.


One Side of the Story; Clues to Etiology in Patients with Asymmetric Chorea.
Chorea can be due to a large number of etiologies. Unilateral chorea is classically related to a contralateral structural lesion, e.g. of the putamen or subthalamic nucleus, however, based upon personal impressions, we have observed that systemic disease, in particular metabolic or autoimmune conditions, can also lead to a unilateral or markedly asymmetric presentations. We sought to investigate this impression by reviewing the literature. A PubMed search was conducted using the terms asymmetric" AND "chorea" OR "hemichorea" OR "unilateral" AND "chorea" OR "monochorea" OR "right greater than left" AND "chorea" OR "left greater than right" AND "chorea" OR "right more than left" AND "chorea" OR "left more than right" AND "chorea" as well as "hemiballismus" NOT "stroke" NOT "infarct" NOT "dyskinesia. A total of 243 sources were felt to meet criteria and were reviewed. The most common etiology of reported hemi- or asymmetric chorea was diabetic non-ketotic hyperglycemic hemichorea/hemiballismus. Other common diagnoses were Sydenham's disease, antiphospholipid syndrome and drug-induced chorea. The vast majority of patients with hemi- or asymmetric chorea had acquired rather than genetic, degenerative or congenital causes. Despite the potential limitations of our literature review, the evidence presented here supports the observation that the vast majority of asymmetric or unilateral chorea presentations are due to acquired causes, and in this situation an exhaustive search for reversible etiology should be undertaken. However, presentation with symmetric, generalized chorea does not exclude reversible causes, and investigations should address these in addition to genetic and neurodegenerative etiologies.


The white nipple sign.
The white nipple sign is a term used to describe a fresh fibrin clot on a varix that has bled recently. The authors describe a case to highlight the importance of prompt endo-therapy when this sign is recognised as disturbing this clot can cause brisk bleeding.


Cerebral Venous Sinus Thrombosis From a Tertiary Care Teaching Hospital in India.
The prognosis of cerebral venous sinus thrombosis (CVST) may be dependent on underlying causes and magnetic resonance imaging findings. We report the clinical, laboratory, and radiologic spectrum, and outcome of consecutive patients with CVST. Consecutive patients with CVST over the last 15 years were subjected to detailed history, clinical examination, and laboratory tests as per fixed protocol. The diagnosis of CVST was based on MR venography. The magnetic resonance imaging findings and the sinuses involved were noted. The patients were treated with low-molecular-weight heparin or unfractionated heparin for 14 days, followed by oral anticoagulation. Hospital mortality and 3-month outcome (good or poor) were recorded and correlated with various clinical and laboratory parameters. There were 86 patients whose median age was 30 (6 to 76) years; 47 of them were females. Headache was present in 91.9%, vomiting in 68.6%, focal weakness in 53.5%, seizure in 51.2%, and altered sensorium in 53.5%. Female-specific etiology was present in 34%, prothrombotic condition in 62.8%, infections in 19.8%, and malignancy in 2.8%; no cause could be established in 25.6% of patients. Hyperhomocystinemia was present in 45.9% of patients. Five patients died in the unfractionated heparin group. At 3 months, 77 patients were followed up, of whom 68 had complete, 3 had partial, and 6 had poor recovery. The 3-month outcome was related to Glasgow Coma Scale score and type of heparin used. In our study multiple prothrombotic conditions were common, and low-molecular-weight heparin reduces mortality and results in good outcome.


Aggregation of embryonic stem cells induces Nanog repression and primitive endoderm differentiation.
When embryonic stem cells are allowed to aggregate, the outer layer of the aggregated spheres (referred to as embryoid bodies) differentiates into primitive endoderm. This initial specification of cell lineage facilitates further differentiation of the inner mass of the embryoid bodies. These processes are considered to recapitulate early embryonic development from the blastocyst stage to the egg-cylinder stage. Formation of the primitive endoderm layer in the embryoid bodies was induced solely by aggregation of embryonic stem cells, in the presence of leukemia inhibitory factor/STAT3 and serum/BMP4, which were considered to be sufficient for embryonic stem cell self-renewal. Interestingly, cell aggregation by itself induced Nanog repression at the outer layer, which was essential for aggregation-induced primitive endoderm formation. These data illustrate aggregation-based cell-fate specification during early embryonic development, when downregulation of Nanog plays a crucial role.


The effect of pharmacotherapy for attention deficit hyperactivity disorder on risk of seizures in pediatric patients as assessed in an insurance claims database.
To estimate the rate of new-onset seizure in ADHD patients in relation to ADHD pharmacotherapy. A retrospective cohort study of 34,727 patients, ages 6 to 17, with at least two insurance claims bearing ADHD diagnoses during 2003 in the UnitedHealthcare database. Incidence of seizure was calculated for observation time during treatment with atomoxetine and stimulants/bupropion. Seizure incidence among ADHD patients was 4.5/1,000 person-years (p-y; 95% confidence interval 3.7 - 5.5). ADHD patients who received any ADHD medication had an incidence of 3.8/1,000 p-y (3.0 - 4.8) compared to 8.7 (5.8 - 12.4) for patients who did not receive any ADHD medication. The relative risk (RR) for current vs non-use of atomoxetine was 1.1 (0.6 - 2.1). For stimulants and bupropion, the RR for current vs non-use was 0.8 (0.6 - 1.3). Elevated seizure risks were found in association with central nervous system (CNS) disease (OR 3.9, 1.2 - 10.9), CNS medications (OR 2.2, 1.3 - 3.6), metabolic disease (OR 2.9, 1.1 - 6.8), and psychiatric disease risk factors (OR 1.7, 1.1 - 2.6). In this study, there was no statistically significant association between use of atomoxetine or stimulants and seizure risk in children ages 6 to 17 years with ADHD and without prior seizure disorder.


Anomalous capacitive sheath with deep radio-frequency electric-field penetration.
A novel nonlinear effect of anomalously deep penetration of an external radio-frequency electric field into a plasma is described. A self-consistent kinetic treatment reveals a transition region between the sheath and the plasma. Because of the electron velocity modulation in the sheath, bunches in the energetic electron density are formed in the transition region adjacent to the sheath. The width of the region is of order V(T)/omega, where V(T) is the electron thermal velocity, and omega is the frequency of the electric field. The presence of the electric field in the transition region results in a collisionless cooling of the energetic electrons and an additional heating of the cold electrons.


Heart transplantation in children after mechanical circulatory support: comparison of heart transplantation with ventricular assist devices and elective heart transplantation.
Heart transplantation (HTx) is an ultimate treatment for children with end-stage heart failure or inoperable congenital heart disease. The supply of hearts is inadequate; therefore, different mechanical support systems must be used as bridge to HTx in pediatric patients with postoperative low output. The use of ventricular assist devices (VADs) as bridge to HTx in children is limited because of size differences. The purpose of this study was to evaluate the overall long-term outcome of pediatric circulatory support before pediatric HTx. From 1989 through 2004, 91 pediatric patients underwent isolated HTx. Seven of them required mechanical support before transplantation. We reviewed retrospectively the course of 91 children (mean age 14.7 years) who underwent HTx. Group A consisted of elective HTx patients who were treated as outpatients before HTx, whereas group B was the VAD-HTx bridging group (n=7; mean age 12.31 +/- 2.8 years). Mean duration of VAD support was 108 +/- 98 days (minimum 1 day, maximum 258 days). Overall survival rate after HTx was 80% at 1 year without significant differences between groups. Five of seven patients survived and could be discharged after successful HTx, for a survival rate of 77%. The mean follow-up period was 16.76 +/- 10.6 months. No differences in posttransplantation long-term survival and rejection episodes occurred between patients transplanted with or without VAD. VAD therapy can keep pediatric patients with end-stage heart failure alive until successful HTx, and bridge to HTx is a safe procedure in pediatric patients. After HTx, survival rates of these children are similar to those of patients awaiting elective HTx.


Seasonal variation in acceleration and delay of sexual maturation in female mice by urinary chemosignals.
Data were compiled and analysed from experiments done over a 10-year period on the acceleration and delay of sexual maturation of female house mice by urinary chemosignals from conspecific males and females. Distinct seasonal patterns emerged for control mice treated with water and for both the acceleration of maturation due to a chemosignal in male mouse urine and the delay of maturation which results from treatment with urine from grouped females. Urine from males accelerated puberty during the period March to September, but not during the months of October through February. Urine from grouped females delayed puberty for the months of September to April, but not for the period May through August.


Cryptococcal Osteomyelitis in an Immunocompetent Patient.
Cryptococcal osteomyelitis is extremely rare. When cryptococcal infections occur, they are usually caused by Cryptococcus neoformans and involve the lungs or central nervous system in an immunocompromised individual. However, we report a case of osteomyelitis of the right ankle and right elbow due to C. neoformans in a 29-year-old immunocompetent male.


[Evaluation of immunohistochemistry staining and cytologic diagnosis by using cell block sections prepared with effusion fluid cytology specimens].
To study the values of immunohistochemistry staining and cytological diagnosis by using cell block sections prepared with the effusion fluid cytology specimens. Ninety-nine effusion cytology specimens with the diagnoses of reactive mesothelial hyperplasia, atypical cells and metastatic carcinoma were enrolled into the study. The cytospin preparations/smears, cell block sections and immunohistochemical study were performed and correlated with the clinical findings and follow-up data. Amongst the 99 cases studied, the percentage with positive diagnosis using cytospin preparations/smears was 68.7% (68/99). The percentages with negative and equivocal diagnoses were 16.2% (16/99) and 15.1% (15/99), respectively. As for cell block sections, the percentages were 71.7% (71/99), 16.2% (16/99) and 12.1% (12/99), respectively. On the other hands, the percentages became 76.8% (76/99), 20.2% (20/99) and 3.0% (3/99), respectively, when coupled with immunohistochemical findings. The overall percentages of positive, negative and equivocal diagnoses were 77.8% (77/99), 17.2% (17/99) and 5.0% (5/99), respectively, upon clinicopathologic correlation. The difference between cytospin preparations/smears and cell block sections was not statistically significant (P > 0.05). When coupled with immunohistochemical findings or clinicopathologic correlation, the difference in rates of equivocal diagnosis however carried statistical significance (P < 0.05). The false-negative rate of immunohistochemical study applied on cell block sections was 1.0% (1/99). Immunohistochemistry, when applied on cell block sections, is useful in delineation of the primary origins of the tumor cells in effusion fluid cytology specimens. Combination of morphologic examination, immunohistochemical findings and clinicopathologic correlation can further improve the rate of positive diagnosis.


Progressive multifocal leukoencephalopathy in a patient with Good's syndrome.
Good's syndrome (GS) is an immunodeficiency characterised by thymoma, hypogammaglobulinemia and impaired T-cell function. The clinical symptoms are recurrent or chronic infections from common or opportunistic pathogens and diarrhoea. Encephalitis is rare, mostly associated to cytomegalovirus. We present a 65-year-old woman who developed blindness, motor deficits and cognitive changes over a 4-month period. MRI of the brain showed symmetric subcortical white matter changes in the occipital lobes, first thought to correspond to posterior reversible encephalopathy syndrome. A thymoma was found and operated. The patient had no B cells, low immunoglobulins and an inverted CD4/CD8 ratio. GS was diagnosed. In the cerbrospinal fluid >1 million JC virus copies/mL were found and a repeat MRI now showed a picture compatible with progressive multifocal leucoencephalopathy (PML). Her disease had a fatal outcome. The present case is the second reported association between GS and PML.


Experience with 123I-iomazenil SPECT in acute cerebral infarction.
Neuronal cells are susceptible to cerebral ischaemia. As gamma-aminobutyric acid(A) (GABA(A)) receptors are specific for neurones, functional receptor imaging using I-iomazenil (IMZ), a ligand to the GABA benzodiazepine receptor, has been proposed as an imaging modality for the assessment of neuronal integrity. However, there is only limited experience with IMZ in patients with acute cerebral infarction. Therefore, the aim of this study was to evaluate IMZ single photon emission computed tomography (SPECT) in patients with acute cerebral ischaemia. IMZ SPECT was performed in 21 patients with acute cerebral infarction 7-10 days after stroke onset. Eleven patients underwent systemic thrombolysis within 6 h after symptom onset (group 1), whereas 10 patients were treated conservatively (group 2). IMZ (150-200 MBq) was injected intravenously and imaging was performed using a dedicated four-head SPECT camera at 5 min (perfusion) and 90 min (receptor distribution) post-injection, with an acquisition time of 50 min each. Images were analysed by visual inspection. Four patients showed normal IMZ distribution, and 17 patients showed abnormalities of IMZ uptake on both early and late images. In six patients with regional uptake deficits, a crossed cerebellar diaschisis was observed on early images. Cerebellar inhomogeneity of tracer uptake was absent at the time of late images in all six patients. In eight patients, areas of hypoperfusion corresponded exactly to the regions of receptor deficiency (match). In five patients, preserved neuronal integrity was present in hypoperfused areas (mismatch). In four patients, normally or even hyperperfused areas exhibited regional receptor deficiency (inverse mismatch). In conclusion, IMZ SPECT demonstrated differences between regional perfusion and receptor distribution in about one-half of patients 7-10 days after acute cerebral ischaemia. Interesting patterns between the early phase (perfusion) and the late phase (receptor distribution) were found. These patterns are indicative of the heterogeneous development of cerebral ischaemia where, even days after stroke onset, areas of hypoperfusion but preserved neuronal integrity may be present. However, the evaluation of the potential clinical and therapeutic impact of individual IMZ distribution patterns requires further investigation.


Novel Biosynthetic Route to the Isoquinoline Scaffold.
Isoquinoline alkaloids are a large class of natural products with a broad range of biological activities, including antimicrobial, antitumor, antileukemic and anti-inflammatory properties. Although mostly found in plants, isoquinolines can also be found in the extracts of bacterial and fungal cultures. Regardless of the origin, most of the reported biosynthetic routes for isoquinolines use tyrosine as a main biosynthetic precursor. Here, we report the identification of a new biosynthetic pathway for production of isoquinolinequinone alkaloid mansouramycin D in Streptomyces albus Del14. Using feeding, mass spectrometry, and nuclear magnetic resonance spectroscopy, we demonstrate that tryptophan serves instead of tyrosine as a main mansouramycin biosynthetic precursor. The biosynthetic genes were identified in the chromosome of the strain by using gene inactivation and heterologous expression. Insights into the biosynthesis of mansouramycins are also presented.


Motor-mediated alignment of microtubules in semidilute mixtures.
We propose and study a model of molecular motor-induced ordering in a cytoskeletal filament solution for the semidilute case. Motors attach to a pair of filaments and walk along the pair bringing them into closer alignment. In the semidilute regime multiple motors can bind a filament to several others and, for a critical motor density, induce a transition to an ordered phase with a nonzero mean orientation. The motors, on the one hand, cause closer filament alignment, and, on the other hand, induce fluctuations that are dependent on the relative orientation of the filaments to which the motors are attached. We develop a spatially homogenous, mean-field theory that explicitly accounts for a force-dependent detachment rate of motors, which in turn affects the mean and the fluctuations of the net force acting on a filament. This model considers each filament to be in motor contact with all other filaments in the solution. We show that the transition to the oriented state changes from second order to first order when the force-dependent detachment becomes important.


Radiation-modifying effect of oxygen in synchronized cells pre-treated with acute or prolonged hypoxia.
The objective of the present study was to measure the sensitizing effect of oxygen following acute as well as protracted hypoxia in cell cultures of maximum homogeneity. Human cells of the established line NHIK 3025 were synchronized by the method of mitotic selection and irradiated while in late G1 (aerobic conditions) or in the oxygen-sensitive restriction point near the G1/S border. Four different conditions of oxygen treatment were used: (1) Cells were irradiated under aerobic conditions 5 h after mitotic selection (G1 lasts for 6.5 h in these cells). (2) Cells were irradiated under extremely hypoxic conditions (i.e. < 4 ppm O2) following an acute (30 min) deoxygenation starting 5 h after mitotic selection. (3) Cells were irradiated under aerobic conditions immediately following 20 h of extreme hypoxia which was started 5 h following mitotic selection. (4) Cells were irradiated under extremely hypoxic conditions after 20 h of extreme hypoxia starting 5 h after mitotic selection. In addition, asynchronous cells were irradiated, either without any pretreatment, or after reoxygenation, following 20 h of extreme hypoxia. The data indicate that the sensitizing effect of oxygen (after protracted hypoxia) is strictly dose-modifying for cells rendered hypoxic while in late G1 and arrested in the oxygen-sensitive restriction point in late G1. There is a non-dose-modifying sensitization when synchronized control cells, irradiated under aerobic conditions, are compared with synchronized cells irradiated during acute hypoxia. In this case, however, one is comparing cells in two different biological states, i.e. cells in an ordinary G1 are compared with cells arrested in the oxygen-sensitive restriction point in G1. The non-dose-modifying nature of the oxygen sensitization as observed in this case may, therefore, reflect biological differences between the cell cultures that are compared rather than differences with respect to the radiochemical sensitization as induced by oxygen at small and large radiation doses. The present data indicate, however, that if cancer cells in human tumours are best represented by a mixture of aerobic cells and chronic hypoxic cells, oxygen may appear to exert no effect, or perhaps even a net protective effect, at low radiation doses.


Neutron powder diffraction study of Pb(HfxTi1-x)O3 ceramics (0.10<or=x<or=0.50).
The crystal symmetries of lead hafnate titanate (Pb(HfxTi1-x)O3, PHT) powders with 0.10<or=x<or=0.50 were investigated by high-resolution neutron powder diffraction. Samples with x<or=0.40 were tetragonal (space group P4 mm), while the sample with x=0.50 contained both monoclinic Cm and rhombohedral (modeled using the R3c space group) phases. The role of the B cations (Hf and Ti) and the oxygen octahedra network, in addition to the displacement of Pb ions from their ideal sites, in promoting the phase transformation between the P4 mm and Cm phases was considered. Two types of structural disorder were identified. Diffuse scattering between Bragg reflection peaks was assigned to Pb ion displacement. A second type of structural disorder, revealed by the weak intensities of observed pseudo-cubic 00l reflections with l even and as 00l reflection peak widths significantly broader than the l00 reflection peaks, was observed. This behavior was attributed to disorder in the arrangement of the O-B-O rows parallel to the c axis. For small values of x, this shift was predominantly along the c axis, whereas shifts perpendicular to the c axis increased with increasing x. These features were modeled using an hkl-dependent line-broadening model. The origin of the hkl-dependent line broadening was assigned to the microstrain accompanying a spatial-composition variation. Structural models were tested by computing valence sums and spontaneous polarization values.


A biomechanical assessment of ergometer task specificity in elite flatwater kayakers.
The current study compared EMG, stroke force and 2D kinematics during on-ergometer and on-water kayaking. Male elite flatwater kayakers (n = 10) performed matched exercise protocols consisting of 3 min bouts at heart and stroke rates equivalent to 85% of VO2peak (assessed by prior graded incremental test). EMG data were recorded from Anterior Deltoid (AD), Triceps Brachii (TB), Latissimus Dorsi (LD) and Vastus Lateralis (VL) via wireless telemetry. Video data recorded at 50 Hz with audio triggers pre- and post-exercise facilitated synchronisation of EMG and kinematic variables. Force data were recorded via strain gauge arrays on paddle and ergometer shafts. EMG data were root mean squared (20ms window), temporally and amplitude normalised, and averaged over 10 consecutive cycles. In addition, overall muscle activity was quantified via iEMG and discrete stroke force and kinematic variables computed. Significantly greater TB and LD mean iEMG activity were recorded on-water (239 ± 15 vs. 179 ± 10 μV. s, p < 0.01 and 158 ± 12 vs. 137 ± 14 μV.s, p < 0.05, respectively), while significantly greater AD activity was recorded on-ergometer (494 ± 66 vs. 340 ± 35 μV.s, p < 0.01). Time to vertical shaft position occurred significantly earlier on-ergometer (p < 0.05). Analysis of stroke force data and EMG revealed that increased AD activity was concurrent with increased external forces applied to the paddle shaft at discrete phases of the on-ergometer stroke cycle. These external forces were associated with the ergometer loading mechanism and were not observed on- water. The current results contradict a previous published hypothesis on shoulder muscle recruitment during on-water kayaking. Key pointsWhen exercising at fixed heart and stroke rates, biomechanical differences exist between onergometer and on-water kayaking.Ergometer kayaking results in significantly greater Anterior Deltoid activity but significantly lower Triceps Brachii and Latissimus Dorsi activity, compared with on-water kayaking.The altered muscle recruitment patterns observed on-ergometer are most likely a result of additional forces associated with the ergometer loading mechanism, acting upon the paddle shaft.


Measuring information overload and message fatigue toward COVID-19 prevention messages in USA and China.
COVID-19 prevention messages are a crucial component of disease mitigation strategies and the primary driver of health decision-making during the global pandemic. However, the constant and repetitive nature of COVID-19 messaging may cause unintended consequences. Among the commonly observed phenomena are information overload and message fatigue, which might be experienced differently depending on cultural background. Using measurement invariance testing, this study compared how individuals from two countries-USA (n = 493) and China (n = 571)-experienced information overload and message fatigue toward COVID-19 prevention messages. Findings revealed that people in China showed significantly lower level of information overload and message fatigue than those in the USA. This study explores the extent of the unintended persuasive effects that people have experienced during the COVID-19 pandemic in different societies, a comparison which has never been studied before, even outside of the context of COVID-19. The study also provides much-needed practical insights to develop public health initiatives that improve COVID-19 prevention communication, which can further reduce these unintended effects in both countries, and has implications for other countries as well.


Human Trophoblast Progenitor Cells Express and Release Angiogenic Factors.
Trophoblast stem cells develop from polar trophectoderm and give rise to the cells that generate the placenta. Trophoblast cells are responsible for the uterine invasion and vascular remodeling during the implantation of the embryo. However this knowledge is not yet to be confirmed for trophoblast progenitor cells (TPCs). In this study, we aimed to demonstrate that human TPCs (hTPCs) express and release angiogenic factors for the first time. TPCs were isolated from the term placenta. Then immunophenotyping was performed by FACS method by analyzing caudal type homeobox 2 (CDX2) and eomesodermin (EOMES). Immunofluorescence staining of CDX2 and EOMES was performed on these cells. Lastly, angiogenesis-related proteins were detected by western blot and ELISA assays. The isolated cells were positive for trophoblast stem cell markers CDX2 and EOMES in 92.5% and 92.7% of cells, respectively showing the characteristics of TPCs. The investigation of vascular endothelial growth factor (VEGF), vascular endothelial growth factor receptor 1 (VEGFR1), and vascular endothelial growth factor receptor 2 (VEGFR2) at protein and mRNA level in comparison with human umbilical vein endothelial cells (HUVECs), revealed that human TPCs (hTPCs) have higher levels of VEGF and VEGFR1 transcripts. Additionally soluble forms of VEGF and VEGFR1 were detected in supernatants of hTPCs. With this information, TPCs seem to be promising for regenerative cell therapies by increasing angiogenesis.


Phage typing of Salmonella Typhimurium - is it still a useful tool for surveillance and outbreak investigation?
Phage typing has for decades been useful as a phenotypical, definitive method for epidemiological characterisation of Salmonella Typhimurium. The system recommended by the World Health Organization (WHO) Collaborative Centre for phage typing of Salmonella has, however, become rather complex, and the present study illustrates the challenges of sufficient standardisation of the interpretation of lysis results to make sure that the same strain is assigned to the same phage type in different laboratories. Even though molecular typing methods will replace phenotypic characterisation methods in the future, it is our opinion that phage typing will remain for some time a useful tool to strengthen global Salmonella surveillance. Therefore, improved standardisation and quality assurance is essential to obtain a robust and harmonized method that allows comparison of results between laboratories.


First Report of Rust Caused by Tranzschelia discolor on Peach Leaves in Shenzhou, China.
Peach (Prunus persica [L.] Batsch) as an economically important fruit tree is widely cultivated in Shenzhou, China. In September 2021, peach rust was observed in the peach tree in Shenzhou City, Hebei Province (lat. 38°02'56'' N, long. 115°54'57'' E, altitude 22 m). We investigated a peach orchard with a planting area of 1334 m2, where a total of 162 peach trees were planted, and found that about 10% of peach trees exhibited severe disease symptoms. The leaves of infected plant developed 100% disease symptoms, in which 50% of the infected leaves showed about 10 small pale-yellow spots on the front of leaves and reddish-brown pustules on the corresponding abaxial surface of leaves. Urediniospores varied from obovoid to clavate in shape, sometimes in irregular shape. They were orange-brown, echinulate near base with spines smaller towards apex and often smooth at apex, with germ pores 3-4 at equator, size ranging from 25.4 to 38.6 × 10.1 to 18.7 µm (n=100), and with wall 1 to 1.5μm thick at sides and 5-7 μm thick at apex. Golden capitate paraphyses were present, ranging from 25 to 40 µm in length, with a head in diameter of 12 to 14 µm and a tail in width of 5.2 to 6.5 µm. Based on the rust morphological characters, this pathogen was primarily identified as Tranzschelia discolor (Fuckel) Tranzschel & Litv. (Hiratsuka et al. 1992). For molecular identification, total DNA was extracted from 2 isolates, respectively, and the internal transcribed spacer (ITS) region was PCR-amplified using the primer set ITS5-u and ITS4-u (Pfunder et al. 2001). Obtained sequences were compared with sequences in the GenBank repository using BLAST algorithm. BLAST showed a 100% sequence identify to T. discolor (accession nos. AB097449、MT786217、KU712078、KY764179、MH599069). The sequence has been deposited in GenBank with (accession NO. ON950745 and ON950747). Thus, combining morphological observations and molecular identification, the isolate was identified as T. discolor. The pathogenicity was verified by inoculating the abaxial surface of peach leaves with a suspension of 1 × 106 urediniospores/ml. Peach leaves sprayed with sterile water were used as controls. The inoculated peach trees were placed in a greenhouse at 20°C under dark for 24 hours and maintained at 100% relative humidity to promote disease development. Next, the peach trees were grown in a greenhouse at 20°C with a 12 h day length and symptoms were observed on the leaves 14 days after inoculation. In contrast, the control leaves were asymptomatic. Previous studies reported that peach rust occurred in Oman, Korea and Brazil was caused by T. discolor. (Deadman M L, et al.2007, Shin, H D, et al. 2019, Vidal G S, et al. 2021). To our knowledge, this is the first report of T. discolor as a causal agent causing peach leaf rust in Northern China, which will enable us to rapidly diagnose this disease, identify the occurrence of this disease and develop adequate management strategies to control it in China.


Does clinical training period support patient-centeredness perceptions of medical students?
Learning environment influences students' professional formation and patient-centered attitudes and behaviors. The purpose of this study is to investigate how hidden curriculum of learning environment and the previous experience with chronically ill patients affect patient-centeredness perceptions of medical students. We followed 144 students and determined their opinions on 'ideal patient-centered practice and learning environment' via patient-centeredness questionnaire (PCQ) just before (third year) and at the end (sixth year) of clinical training years of medical school. At the end of each clinical training year (fourth, fifth, and sixth years), we determined experiences of the students about 'patient-centeredness of the learning environment' using a relevant survey called communication, curriculum, and culture (C3) instrument. We also compared PCQ and C3 instrument scores of the participants who had chronically ill patient in their families/friends and who do not. C3 scores worsened over the years, namely, students faced increasing number of examples against patient centeredness. Final PCQ scores were worse than initial ones. C3 and PCQ scores of the students who had previous experience with chronically ill patients were not different from the scores of the remaining students. Medical students, even those who have a chronically ill patient in their families or friends, lose their idealism about patient centeredness to some degree possibly due to hidden curriculum of the medical school.


Distinct gene expression patterns in chronic lymphocytic leukemia defined by usage of specific VH genes.
The mutation status and usage of specific VH genes such as V3-21 and V1-69 are potentially independent pathogenic and prognostic factors in chronic lymphocytic leukemia (CLL). To investigate the role of antigenic stimulation, we analyzed the expression of genes involved in B-cell receptor (BCR) signaling/activation, cell cycle, and apoptosis control in CLL using these specific VH genes compared to VH mutated (VH-MUT) and VH unmutated (VH-UM) CLL not using these VH genes. V3-21 cases showed characteristic expression differences compared to VH-MUT (up: ZAP70 [or ZAP-70]; down: CCND2, P27) and VH-UM (down: PI3K, CCND2, P27, CDK4, BAX) involving several BCR-related genes. Similarly, there was a marked difference between VH unmutated cases using the V1-69 gene and VH-UM (up: FOS; down: BLNK, SYK, CDK4, TP53). Therefore, usage of specific VH genes appears to have a strong influence on the gene expression pattern pointing to antigen recognition and ongoing BCR stimulation as a pathogenic factor in these CLL subgroups.


Glycyrrhizic acid improved lipoprotein lipase expression, insulin sensitivity, serum lipid and lipid deposition in high-fat diet-induced obese rats.
The metabolic syndrome, known also as the insulin resistance syndrome, refers to the clustering of several risk factors for atherosclerotic cardiovascular disease. Dyslipidaemia is a hallmark of the syndrome and is associated with a whole body reduction in the activity of lipoprotein lipase (LPL), an enzyme under the regulation of the class of nuclear receptors known as peroxisome proliferator-activated receptor (PPAR). Glycyrrhizic acid (GA), a triterpenoid saponin, is the primary bioactive constituent of the roots of the shrub Glycyrrhiza glabra. Studies have indicated that triterpenoids could act as PPAR agonists and GA is therefore postulated to restore LPL expression in the insulin resistant state. Oral administration of 100 mg/kg of GA to high-fat diet-induced obese rats for 28 days led to significant reduction in blood glucose concentration and improvement in insulin sensitivity as indicated by the homeostasis model assessment of insulin resistance (HOMA-IR) (p < 0.05). LPL expression was up-regulated in the kidney, heart, quadriceps femoris, abdominal muscle and the visceral and subcutaneous adipose tissues but down-regulated in the liver--a condition in reverse to that seen in high-fat diet-induced obese rats without GA. With regard to lipid metabolism, GA administration led to significant hypotriglyceridemic and HDL-raising effects (p < 0.05), with a consistent reduction in serum free fatty acid, total cholesterol and LDL cholesterol and significant decrease in tissue lipid deposition across all studied tissue (p < 0.01). In conclusion, GA may be a potential compound in improving dyslipidaemia by selectively inducing LPL expression in non-hepatic tissues. Such up-regulation was accompanied by a GA-mediated improvement in insulin sensitivity, which may be associated with a decrease in tissue lipid deposition. The HDL-raising effect of GA suggests the antiatherosclerotic properties of GA.


[Prostate carcinoma: vaccination as a new option for treatment].
Immune therapy and tumor cell vaccination is a challenging option in prostate cancer therapy, especially as side effects rarely occur. This review highlights recent developments in vaccination therapy of prostate cancer. The FDA approved antigen presenting cell vaccine Sipuleucel-T is described and new strategies of immune therapy like RNA and peptide vaccination are discussed in detail. Currently the effect of prostate cancer vaccination has still limitations, at least partially due to the immune suppressive effects of the tumor microenvironment and regulatory T cells, which suppress the immune effector function. To overcome these hurdles the concept of immune checkpoint modulation, which has the aim to break tolerance mechanisms, is discussed. Potential clinical therapies of checkpoint modulation are outlined.


Assessment of resectability of carcinoma of the pancreatic head by ultrasonography and computed tomography. A retrospective analysis.
In order to establish the reliability of the assessment of tumour stage and hence of resectability of carcinoma of the pancreatic head by ultrasonography (US) and computed tomography (CT) a retrospective analysis was performed on 41 patients. Both direct imaging techniques were very accurate in detecting non-resectable disease, 100 and 85% respectively, but were much less accurate in predicting resectable cancer, 18 and 15% respectively. False-resectable results were established in 58% of US examinations and 42% of CT examinations. As demonstrated in this study, predicting resectability of carcinoma of the pancreatic head with US and CT is an inadequate method of assessing tumour stage and should be complemented by other techniques to ensure a reliable result.


Antigen-induced changes in B cell subsets in lymph nodes: analysis by dual fluorescence flow cytofluorometry.
Changes in the representation and surface phenotype of defined B cell subsets in murine lymph nodes stimulated with keyhole limpet hemocyanin or sheep red blood cells have been analyzed by two-color immunofluorescence fluorocytometric analysis. Shortly after immunization with either antigen there is a dramatic increase in both the frequency and absolute number of IgM+, IgD+ B cells, which is followed by the formation of germinal centers. Germinal center cells, as soon as they appear on day 3 after primary immunization, bind high levels of peanut agglutinin, bear low levels of surface IgM but no detectable surface IgD, and are characterized by lack of staining with MEL-14, a monoclonal antibody which recognizes a lymphocyte surface receptor involved in lymphocyte homing. The level of I-A and H-2K region-encoded surface antigens on early germinal center cells is higher than on PNAlo B cells. During the first 7 days of the germinal centers there is a progressive decrease in the average level of H-2K but not of Ia antigens. A similar decrease was observed for ThB. It is confirmed that the germinal center cell population contains the majority of antigen-binding cells in the stimulated lymph node. These findings indicate that B cells are recruited nonspecifically to antigen-stimulated lymph nodes, and that the antigen-specific cells then selectively participate in the formation of germinal centers where they undergo specific differentiation events.


Activation of the NRF2-ARE signalling pathway by the Lentinula edodes polysaccharose LNT alleviates ROS-mediated cisplatin nephrotoxicity.
The nephrotoxicity of cisplatin (cis-DDP) limits its general clinical applications. Lentinan (LNT), a dextran extracted from the mushroom Lentinula edodes, has been shown to have multiple pharmacological activities. The primary objective of the current study was to determine whether and how LNT alleviates cis-DDP- induced cytotoxicity in HK-2 cells and nephrotoxicity in mice. LNT did not interfere with cisplatin's anti-tumour efficacy in vitro and functioned cooperatively with cis-DDP to inhibit activity in HeLa and A549 tumour cells. LNT alleviated the cis-DDP-induced decrease in HK-2 cell viability, caspase-3 activation and cleavage of the DNA repair enzyme PARP, decreased HK-2 cell apoptosis and inhibited reactive oxygen species (ROS) accumulation in HK-2 cells. The inhibitor of ROS (N-acetyl-L-cysteine, NAC) could decreased the apoptosis of HK-2 cell. In addition, LNT significantly prevented cis-DDP-induced kidney injury in vivo. LNT itself could not eliminate ROS levels in vitro. Further studies demonstrated that LNT induced NF-E2 p45-related factor 2 (Nrf2) protein and mRNA expression in a time- and dose-dependent manner. LNT promoted Nrf2 translocation to the nucleus and binding to the antioxidant-response element (ARE) sequence and induced the transcription and translation of heme oxygenase 1 (HO-1), aldo-keto reductases 1C1 and 1C2 (AKR1C), and NADP(H):quinone oxidoreductase 1 (NQO1). Finally, we used hNrf2 siRNA and an Nrf2 agonist (tBHQ) to inhibit or enhance Nrf2 expression. The results demonstrated that the LNT-mediated alleviation of cis-DDP-induced nephrotoxicity was achieved by preventing the accumulation of ROS in a manner that depended on the activation of the Nrf2-ARE signalling pathway.


Forced disappearance in an era of globalization: biopolitics, shadow networks, and imagined worlds.
In this article, I argue that the practice of forced disappearance of persons on the part of paramilitary groups has become linked to specific processes of globalization. Global flows related to biopolitics, global crime networks, and dehumanizing imaginations reproduced by mass media together constitute a driving force behind forced disappearances. Drawing on ethnographic fieldwork in the Colombian city of Medellín, I analyze how these global flows interact with local armed actors, helping create a climate conducive to forced disappearance. These mechanisms in Colombia show similarities to those in some African and Asian countries. Gaining insight into the mechanisms behind forced disappearance may help prevent it from occurring in the future. Enhancing social inclusion of residents, unraveling the transnational crime networks in which perpetrators are involved, and disseminating rehumanizing images of victims all contribute to curbing the practice of forced disappearance.


Polyploidy and Environment in Arctic Alaska.
The frequency of polyploidy in the angiosperm flora of the Ogotoruk Creek-Cape Thompson area of northwestern Alaska is correlated with edaphic environmental gradients in the area. These results, when placed in a historical and ecological context, help to clarify the problem of increasing frequency of polyploidy with latitude in the northern hemisphere.


Autonomic Dysfunction Correlates with Clinical and Inflammatory Activity in Patients with Crohn's Disease.
Autonomic dysfunction has been implicated in Crohn's disease (CD). We aimed to investigate heart rate variability (HRV) as a marker of possible autonomic imbalance in patients with CD. Thirty patients with CD and 30 age- and gender-matched healthy controls were enrolled in a prospective cohort study and underwent HRV study. Anxiety level was scored using the STAI questionnaire and CD clinical activity was assessed by Harvey-Bradshaw index. Blood tests including inflammatory markers were obtained for all participants. CD subjects had lower mean blood pressure (85.51 ± 11.07 mm Hg, 91.51 ± 6.99, P = 0.015) and albumin and significantly higher CRP and IL-6 compared with controls (P < 0.002 for all comparisons). Mean HRV values of very low-frequency power and low-frequency power components were significantly lower among CD subjects (P = 0.038 and 0.027, respectively), implying a predominant sympathetic tone. Anxiety level scores were significantly higher among patients with CD for both state anxiety (P = 0.001) and trait anxiety (P < 0.0001). However, patients with active disease had similar anxiety scores as patients in remission, yet had a significantly lower BMI, lower albumin level, and higher CRP and IL-6 levels (P < 0.05 for all comparisons). Moreover, despite similar anxiety scores, patients with active disease had higher pulse rate (P = 0.02) and lower HRV indexes, which correlated with albumin levels(r = 0.7, P = 0.001). Although patients with CD have higher anxiety levels compared with controls, they exhibit depressed HRV independent of this anxiety state and in direct correlation with disease activity and inflammatory markers. These observations suggest an inherent imbalance of autonomic function associated with active inflammation.


Drugs acting on the intracellular signalling system.
Cellular response to extracellular messages is a basic process to maintain and to support cell life. Several signalling molecules important as sites of therapeutic drug action are involved in the response. Recent studies on life sciences have elucidated molecular properties of intracellular signalling factors and mechanisms of cascading. Novel drugs acting on signalling molecules and possessing new sites and mechanisms of action have been found. This article summarizes the properties (subtypes, structures, functions) of signalling factors (receptors, ion channels, GTP binding proteins, second messenger-generating enzymes, second messenger-metabolizing enzymes, second messengers protein kinases, protein phosphatases) and lists in Tables A-H drugs that act on signalling molecules and which should find clinical use.


Asymmetric dimethylarginine, oxidative stress, and vascular nitric oxide synthase in essential hypertension.
We reported impaired endothelium-derived relaxation factor/nitric oxide (EDRF/NO) responses and constitutive nitric oxide synthase (cNOS) activity in subcutaneous vessels dissected from patients with essential hypertension (n = 9) compared with normal controls (n = 10). We now test the hypothesis that the patients in this study have increased circulating levels of the cNOS inhibitor, asymmetric dimethylarginine (ADMA), or the lipid peroxidation product of linoleic acid, 13-hydroxyoctadecadienoic acid (HODE), which is a marker of reactive oxygen species. Patients had significantly (P < 0.001) elevated (means +/- SD) plasma levels of ADMA (P(ADMA), 766 +/- 217 vs. 393 +/- 57 nmol/l) and symmetric dimethylarginine (P(SDMA): 644 +/- 140 vs. 399 +/- 70 nmol/l) but similar levels of L-arginine accompanied by significantly (P < 0.015) increased rates of renal ADMA excretion (21 +/- 9 vs. 14 +/- 5 nmol/mumol creatinine) and decreased rates of renal ADMA clearance (18 +/- 3 vs. 28 +/- 5 ml/min). They had significantly increased plasma levels of HODE (P(HODE): 309 +/- 30 vs. 226 +/- 24 nmol/l) and renal HODE excretion (433 +/- 93 vs. 299 +/- 67 nmol/micromol creatinine). For the combined group of normal and hypertensive subjects, the individual values for plasma levels of ADMA and HODE were both significantly (P < 0.001) and inversely correlated with microvascular EDRF/NO and positively correlated with mean blood pressure. In conclusion, elevated levels of ADMA and oxidative stress in a group of hypertensive patients could contribute to the associated microvascular endothelial dysfunction and elevated blood pressure.


Anomalous exit of the C-6 nerve root via the C-6, C-7 foramen.
We report a case of the anomalous exit of the C-6 nerve root at the C-6, C-7 foramen. This caused intense radicular and cervical pain in a 25-year-old woman. At myelography, the 5th and 6th cervical roots appeared conjoined, and at operation the C-6 root and ganglion were found to turn acutely downward, lateral to the pedicle of C-6, and then to exit with the C-7 nerve root through the C-6, C-7 foramen. The ganglion of C-6 was placed nearly parallel to the dural sac and tethered at its origin on the dural sac and at the exit foramen with the C-7 root. The pedicle of C-6 was interposed between the dural sac and the ganglion of C-6. Cervical hemilaminectomy and resection of the pedicle of C-6 defined the anatomy and resulted in a clinical cure. The radiological features are described and contrasted with those of other anomalies. To our knowledge, this is the first report of such an anomalous course of exit of a cervical root.


Ephrin-A5 Is Involved in Retinal Neovascularization in a Mouse Model of Oxygen-Induced Retinopathy.
Retinal neovascularization (RNV) is an important pathological feature of vitreoretinopathy that can lead to severe vision loss. The purpose of this study was to identify the role of ephrin-A5 (Efna5) in RNV and to explore its mechanism. The expression pattern and biological significance of Efna5 were investigated in a mouse model of oxygen-induced retinopathy (OIR). The expression of Efna5 and downstream signaling pathway members was determined by RT-PCR, immunofluorescence, immunohistochemistry, and western blot analyses. shRNA was used to knockdown Efna5 in the retina of the OIR mouse model. Retinal flat mounts were performed to evaluate the impact of Efna5 silencing on the RNV process. We found that the Efna5 was greatly upregulated in the retina of OIR mice. Elevated Efna5 mainly colocalized with the retinal vessels and endothelial cells. We then showed that knockdown of Efna5 in OIR mouse retinas using lentivirus-mediated shRNA markedly decreased the expression of Efna5 and reduced the retinal neovascularization and avascular retina area. We further showed hypoxia stimulation dramatically increased both total and phosphorylation levels of ERK1/2 and the phosphorylation levels of Akt in OIR mice. More importantly, knockdown of Efna5 could inhibit the p-Akt and p-ERK signaling pathways. Our results suggested that Efna5 may regulate the RNV. This study suggests that Efna5 was significantly upregulated in the retina of OIR mice and closely involved in the pathological retinal angiogenesis.


7. Mortality after the Chernobyl catastrophe.
A detailed study reveals that 3.8-4.0% of all deaths in the contaminated territories of Ukraine and Russia from 1990 to 2004 were caused by the Chernobyl catastrophe. The lack of evidence of increased mortality in other affected countries is not proof of the absence of effects from the radioactive fallout. Since 1990, mortality among liquidators has exceeded the mortality rate in corresponding population groups. From 112,000 to 125,000 liquidators died before 2005--that is, some 15% of the 830,000 members of the Chernobyl cleanup teams. The calculations suggest that the Chernobyl catastrophe has already killed several hundred thousand human beings in a population of several hundred million that was unfortunate enough to live in territories affected by the fallout. The number of Chernobyl victims will continue to grow over many future generations.


Current perspectives on recurrent HPV-mediated oropharyngeal cancer.
In the recent years, the prevalence of HPV-positive oropharyngeal squamous cell carcinoma (OPSCC) has increased significantly. Currently, nearly 80-90% of all oropharynx tumors are HPV-positive. In addition, it is now recognized that HPV-positive tumor status is associated with good prognosis and improved response to chemoradiation. However, within this setting, there are still patients with HPV-positive OPSCC who will experience recurrence. With the increasing incidence of HPV-mediated OPSCC, recurrent HPV disease is also becoming more prevalent and there is an increasing need to understand the unique presentation and treatment of recurrent HPV-mediated disease. In this review, we will discuss epidemiology of recurrent HPV-positive OPSCC, role of surgical salvage, re-irradiation, and the role of upcoming novel treatments and immunotherapy. Historically, recurrent oropharyngeal disease has been associated with poor prognosis and high morbidity. However, recent advances have transformed the landscape for salvage treatment of HPV-mediated OPSCC. Liquid biomarkers offer potential for early detection of recurrence, robotic techniques may reduce morbidity of surgical salvage, improvements in re-irradiation approaches reduce toxicities, and novel immune based therapies on the horizon are offering promising results. These advances combined with the improved prognosis of HPV-positive disease offer to transform our approach to recurrent disease of the oropharynx.


Crystal structure, electronic structure, and optical properties of the novel Li4CdGe2S7, a wide-bandgap quaternary sulfide with a polar structure derived from lonsdaleite.
The novel quaternary thiogermanate Li4CdGe2S7 (tetralithium cadmium digermanium heptasulfide) was discovered from a solid-state reaction at 750 °C. Single-crystal X-ray diffraction data were collected and used to solve and refine the structure. Li4CdGe2S7 is a member of the small, but growing, class of I4-II-IV2-VI7 diamond-like materials. The compound adopts the Cu5Si2S7 structure type, which is a derivative of lonsdaleite. Crystallizing in the polar space group Cc, Li4CdGe2S7 contains 14 crystallographically unique ions, all residing on general positions. Like all diamond-like structures, the compound is built of corner-sharing tetrahedral units that create a relatively dense three-dimensional assembly. The title compound is the major phase of the reaction product, as evidenced by powder X-ray diffraction and optical diffuse reflectance spectroscopy. While the compound exhibits a second-harmonic generation (SHG) response comparable to that of the AgGaS2 (AGS) reference material in the IR region, its laser-induced damage threshold (LIDT) is over an order of magnitude greater than AGS for λ = 1.064 µm and τ = 30 ps. Bond valence sums, global instability index, minimum bounding ellipsoid (MBE) analysis, and electronic structure calculations using density functional theory (DFT) were used to further evaluate the crystal structure and electronic structure of the compound and provide a comparison with the analogous I2-II-IV-VI4 diamond-like compound Li2CdGeS4. Li4CdGe2S7 appears to be a better IR nonlinear optical (NLO) candidate than Li2CdGeS4 and one of the most promising contenders to date. The exceptional LIDT is likely due, at least in part, to the wider optical bandgap of ∼3.6 eV.


Teaching hospice medicine to medical students, house staff, and other caregivers in the United Kingdom.
This paper outlines the development of a medical education programme within the existing programme of the Macmillan Education Centre of the Dorothy House Foundation, Bath, England, between 1989 and 1992. A review of some of the significant contributions to the literature in this area is followed by a description of the initial research that was instrumental in designing these courses. A review of activities of the Centre is reported. A more detailed description of a course specifically designed for family doctors is made. A natural development from that course was one for all disciplines involved in caring for the terminally ill--this is described--along with some of the difficulties of such a method of education. Reference is made to evaluative work undertaken with reporting of initial data and work still in progress. Conclusions are drawn about the nature of such workshops and comment made on the role of facilitation.


Determination of imazalil residues in lemons by gas chromatography with nitrogen-phosphorus detection.
A simple and reliable method was developed for determination of imazalil residues in whole lemons. Imazalil was extracted from lemon samples with ethyl acetate under basic conditions. The ethyl acetate in the extract was evaporated, and the residue was dissolved in dichloromethane. The solution was applied to a solid-phase extraction (diol-bonded silica) cartridge. Imazalil was eluted with methanol and determined by gas chromatography (GC) with nitrogen-phosphorus detection. Recoveries at 4 fortified levels (0.1, 1.0, 2.0, and 5.0 micrograms/g) ranged from 93.3 to 97.8%, with coefficients of variation ranging from 0.2 to 3.0%. The detection limit was 0.05 microgram/g. Fifteen commercial lemon samples were analyzed for Imazalil residues; 10 were positive. Concentrations of imazalil residues ranged from 0.22 to 4.44 micrograms/g. Imazalil-positive samples were confirmed by GC with mass-selective detection (m/z 173, 215, and 217).


Regulation of macrophage gene expression by pro- and anti-inflammatory cytokines.
The anti-inflammatory cytokines IL-4 and IL-10 are well recognized as important negative regulators of proinflammatory gene expression in mononuclear phagocytes. The intracellular mechanisms which mediate these responses appear to be multifactorial. IL-4 is able to markedly suppress transcriptional activation of IFNgamma-responsive genes and the promoter sequences required for both IFNgamma and IL-4 sensitivity are identical. IFNgamma-activated STAT1 and IL-4-activated STAT6 can both form complexes on the same regulatory sequence element; while STAT1 functions to promote transcription, STAT6 is inactive. STAT6 is, however, required for the suppressive activity of IL-4. In this model, IL-4 appears to suppress IFNgamma-inducible proinflammatory gene expression through the ability of STAT6 to compete with STAT1 for occupancy of promoter sites necessary for IFNgamma-induced transcriptional initiation. In a second model, IL-10 suppresses the expression of genes induced in LPS-stimulated macrophages through a pathway involving destabilization of specific mRNAs. We have demonstrated that nucleotide sequences in the 3'-untranslated region of an IL-10-sensitive gene can both destabilize a stable reporter mRNA (CAT) and confer sensitivity to IL-10-mediated destabilization. Deletion and site-specific mutagenesis have mapped this to an AU-rich sequence motif similar to that found in many cytokine and growth factor mRNAs. IL-10 is able to modulate the activity of proteins capable of binding to this sequence and one or more of these may regulate the rate of mRNA degradation. Thus mechanisms through which IL-10 and IL-4 act to dampen inflammatory responses are mechanistically distinct and involve diverse intracellular signaling pathways.


Bone morphogenetic protein-2 inhibits serum deprivation-induced apoptosis of neonatal cardiac myocytes through activation of the Smad1 pathway.
Bone morphogenetic protein (BMP)-2 has been shown to induce ectopic expression of cardiac transcription factors and beating cardiomyocytes in non-precardiac mesodermal cells, suggesting that BMP-2 is an inductive signaling molecule that participates in cardiac development. However, direct evidence of the effects of BMP-2 on cardiac myocytes has not been reported. To examine the role of BMP-2 and its receptors, we studied the ability of BMP-2 to promote survival of isolated neonatal rat cardiac myocytes. BMP receptors IA, IB, and II and activin receptor I were found to be expressed in myocytes, and BMP-2 phosphorylated Smad1 and p38 MAPK. Interestingly, BMP-2 promoted survival and inhibited apoptosis of serum-deprived myocytes, although it did not strongly induce hypertrophic growth. To explore the mechanisms for this protective effect, an adenovirus-based vector system was used. Similar to BMP-2, Smad1 promoted survival that was repressed by Smad6. Moreover, BMP-2 and Smad1 enhanced the expression of the anti-apoptotic molecule Bcl-x(L). Antisense oligonucleotides to bcl-x(L) attenuated the survival effected by BMP-2. Overall, our findings suggest that BMP-2 prevents apoptosis of myocytes by induction of Bcl-x(L) via a Smad1 pathway and might be a novel survival factor without any hypertrophic effect on myocytes.


Localization of a protease in protoplast preparations in infected cells of French bean nodules.
Protoplasts from infected and uninfected cells were isolated from the central nitrogen fixing tissue of French bean (Phaseolus vulgaris L. cv Contender) root nodules. Successive filtrations allowed the separation of the infected cells, whereas the small uninfected cells were isolated on a discontinuous Percoll gradient. Higher yields of intact protoplasts were obtained from young (4-week-old) nodules whereas no protoplasts could be isolated from the oldest nodules. When proteolysis was determined in the cytosolic fraction of both infected and uninfected cells, at pH 5.0 and 8.0, with leghemoglobin or azocasein as substrate, activity was present only in infected cell protoplasts and increased with nodule age. A protease with an acidic pH optimum, mainly responsible for this increasing activity, was highly purified from senescing nodules by electro-elution after nondenaturing polyacrylamide gel electrophoresis and used to produce polyclonal antibodies. Western blots of nodule protein separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and probed with purified anti-protease immunoglobulin G showed the molecular mass of the protease to be 58 kilodaltons. Blots also confirmed that protease protein was located in infected cell protoplasts only, regardless of nodule age.


Social media and communications: developing a policy to guide the flow of information.
The use of social media is ubiquitous in the daily lives of nearly three quarters of the population in the United States. This article addresses the process and results of a policy development project for a National Network of Libraries of Medicine Regional Medical Library. Content, scheduling, editing, and author responsibilities are addressed for each of the chosen media outlets, as well as the preparation of prefatory material using an online social media policy development tool.


Pretreatment prostate-specific antigen doubling times: clinical utility of this predictor of prostate cancer behavior.
The distribution of pretreatment and posttreatment prostate specific antigen (PSA) doubling times (PSADT) varies widely. This report examines the pretreatment PSADT as an independent predictor of biochemical freedom from disease (bNED) and describes the clinical utility of PSADT. Ninety-nine patients with T1-3 NX, M-0 prostate cancer treated between February 1989 and November 1993 have pretreatment PSADTs calculated from three or more PSA levels. Biochemical disease-free (bNED) survival (failure is PSA > or = 1.5 ngm/ml and rising) is evaluated by multivariate analysis of common prognostic indicators and PSADT. Prostate-specific antigen doubling time (PSADT) is a significant predictor of survival along with radiation dose. Patients with a pretreatment PSADT of < 12 months show 50% failure by 18 months, while those with a PSADT that is not increasing show only 3% failure at 3 years. Prostate-specific antigen doubling time (PSADT) is a predictor of bNED outcome in prostate cancer. Patients with PSADT < 12 months have aggressive disease, and should be considered for multimodal therapy. Slow PSADT (> or = 5 years) is observed in 57% of patients, and this end point may be considered in the decision to observe rather than to treat. After treatment failure, the PSADT may be used to determine which patients do not need immediate androgen deprivation.


Activation of factor XII and prekallikrein with polysaccharide sulfates and sulfatides: comparison with kaolin-mediated activation.
The activation of Factor XII and prekallikrein by polysaccharide sulfates and sulfatides in the presence of high-molecular-weight (HMW) kininogen was studied, and compared with the kaolin-mediated activation reaction. Among a variety of artificially-sulfated polysaccharides and native polysaccharide sulfates, amylose sulfate (M.W.= 380,000 and sulfur content, 19.1%) and sulfatide were found to have the most efficient ability to trigger the activation of prekallikrein by Factor XII. The effects of these two kinds of negatively-charged surfaces on the following three activation reactions were compared; the activation of prekallikrein by Factor XII (reaction 1), the activation of Factor XII by kallikrein (reaction 2) and the activation of prekallikrein by Factor XIIa (reaction 3). All three reactions mediated by the selected surfaces were strongly accelerated by HMW kininogen and its derivatives, kinin-free protein and fragment 1.2-linked light chain, like the kaolin-mediated activation. However, this accelerating effect of HMW kininogen on the amylose sulfate- and sulfatide-mediated activations (reaction 1) was diminished after treatment with fluorescein iso-thiocyanate, whereas the effect on the kaolin-mediated activation was not influenced by fluorescein-labeling. In addition, reaction 2 mediated by amylose sulfate and sulfatide was extremely slow even in the presence of HMW kininogen, and the results also differed from those with kaolin. The sulfatide-mediated activation of reaction 1 was not inhibited by fragment 1.2 (His-rich fragment), which is released from HMW kininogen by the action of kallikrein, and is known to be a potent inhibitor of the kaolin-dependent activation. These results indicate that the mechanisms responsible for surface activation triggered by soluble amylose sulfate, sulfatide micelles and kaolin differ from each other as regards the molecular interaction with the contact factors.


The effects of intranasal midazolam on urodynamic studies in children.
To determine the influence of midazolam on the function of the lower urinary tract when used to sedate the patient. Urodynamic studies were performed on 20 patients (seven boys and 13 girls, mean age 4.04 years, range 1 day to 10 years) before and after the intranasal administration of 0.5 mg/kg of midazolam. The patients' heart rate, respiratory rate and oxygen saturation were monitored and the level of sedation recorded by an anaesthesiologist and a surgeon during the study. Heart rate and respiratory rate did not change and the percentage oxygen saturation remained stable throughout the study period. The childrens' behaviour began to change 3-5 min after the intranasal administration of midazolam. The sedation lasted 2 h and the full co-operation of the patient was assured. The difference between the initial and post-midazolam evaluation of maximal cystometric capacity, contractility, compliance, intravesical pressure, maximum flow rate, intravesical pressure at maximum flow and the amount of residual urine were statistically insignificant (P > 0.05). Electromyographic characteristics showed no significant change with the use of midazolam (P > 0.05). Despite the suggestion that benzodiazepines, especially diazepam, are useful for the relaxation of pelvic floor striated musculature in voiding dysfunctions, there was no effect on urodynamic variables when midazolam was used in a single dose intranasally. The anxiolytic and sedative effects allowed children to undergo urodynamic studies in comfort.


Tricuspid valve disease: diagnosis, prognosis and management of a rapidly evolving field.
Tricuspid valve disease, and particularly tricuspid regurgitation, is a highly prevalent condition with a complex pathophysiology and long-term adverse consequences. Although historically neglected, tricuspid valve disease has gained increasing recognition, with important advances in the assessment and management of this disorder over the past 2 decades. Surgical treatment remains the standard of care, but it continues to have one of the the highest death rates among all cardiac valve-related procedures, and a broad range of patients still do not receive effective therapy for tricuspid valve disease in contemporary clinical practice. Therefore, several alternative, less-invasive technologies for treating patients with severe, native tricuspid valve disease at high surgical risk have been developed in the past decade, with promising early results. This Review summarizes key findings and highlights the latest developments in the diagnosis and management framework that are transforming clinical practice in the complex field of tricuspid valve disease.


Transport of chenodeoxycholic acid and its 3-alpha- and 7-alpha-sulfates by isolated perfused rat liver.
In patients with cholestasis, levels of sulfated bile acids rise. Sulfate esters of chenodeoxycholic acid are the most abundant of these bile acid sulfates. These compounds are taken up by the liver and excreted into bile, although their plasma clearance and biliary excretion are reduced compared with that of unsulfated bile acids. It is not clear whether this is due to differences in intrinsic hepatic uptake or biliary excretion. In the present study, single-pass transport kinetics of chenodeoxycholic acid 3-alpha-sulfate, chenodeoxycholic acid 7-alpha-sulfate and unsulfated chenodeoxycholic acid were quantified in isolated perfused rat liver. Influx of the 7-alpha- and 3-alpha-sulfated derivatives was 57% and 20% that of chenodeoxycholic acid, respectively. These three compounds bound to albumin equally well, indicating that this was not a factor in their differential uptake. Although single-pass extraction of material taken up by the liver was identical. There was no difference in bile flow or biliary excretion of material taken up by the liver was identical. There was no difference in bile flow or biliary excretion rate, regardless of which bile acid sulfate was tested. These results indicate that the low plasma elimination of sulfated bile acids previously observed by others can be explained by low hepatic influx. The diminished transport into liver resulting from sulfation could lead to enhanced elimination of bile acids by the kidney.


Quinine-induced tinnitus in rats.
Quinine ingestion reportedly induces tinnitus in humans. To expand our salicylate-based animal model of tinnitus, a series of conditioned suppression experiments was performed on 54 male-pigmented rats using quinine injections to induce tinnitus. Quinine induced changes in both the extent of suppression and recovery of licking, which followed a pattern that paralleled those produced after salicylate injections, and which may be interpreted as the result of tinnitus perception in animals. These changes depended on the dose and time schedule of quinine administration. Additionally, the calcium channel blocker, nimodipine, abolished the quinine-induced effect in a dose-dependent manner.


Long-term treatment of rheumatoid arthritis comparing nabumetone with aspirin.
This report summarizes the results of a 17-investigator multicenter six-month randomized double-blind parallel group study. The safety and efficacy of nabumetone 1,000 mg taken at bedtime was compared with that of aspirin 900 mg four times daily in the treatment of adult patients with active class II or III classical or definite rheumatoid arthritis. Two hundred sixty-four patients were entered into the study. Two hundred fifty-seven (126 nabumetone and 131 aspirin) patients were evaluable for safety. Two hundred thirty-four (113 nabumetone and 121 aspirin) patients were evaluable for efficacy. There was significant improvement in each of six clinical measurements of efficacy in both treatment groups and little difference between groups. The somewhat greater improvement in articular index and duration of morning stiffness in the nabumetone-treated group did not reach statistical significance. There was an equal percentage of patient withdrawal for lack of efficacy in each group. Overall, the rate of patient withdrawal due to adverse experiences was greater (p = 0.01) for aspirin-treated patients. These experiences were usually dispepsia, abdominal pain, and tinnitus. It was concluded that nabumetone was an effective anti-inflammatory drug in the treatment of rheumatoid arthritis with less toxicity than aspirin.


Tumours in calves: comparative aspects.
In this paper, calf neoplasia is discussed in relation to a series of cases comprising (1). spontaneous congenital bovine tumours of fetuses and newborn animals, (2). spontaneous juvenile-type tumours in calves aged 2-12 months, and (3). iatrogenic tumours of calves. The congenital cases (n=14) consisted of tumours of a predominantly mesenchymal and malignant nature (malignant lymphoma, mesothelioma and mixed mesodermal tumour). In the juvenile cases (n=11), malignant lymphoma and sarcoma were the commonest forms. In comparing tumour patterns in calves with those reported in adult cattle, it was apparent that tumours were less common in the former (6 versus 60 per 100000) and that, with the exception of malignant lymphoma, the types of tumour differed. Carcinomas, which were virtually absent in calves, predominated in adults, probably due to the longer exposure of older animals to carcinogenic factors. In comparing tumour patterns in calves with those reported in pigs and children, it was clear that calf cases were mainly sporadic, with the notable exception of malignant lymphoma in twins. In young pigs, however, several types of tumour (some hereditary) were reported on a single farm as multiple cases. In children, tumours occurred more frequently than in calves, and many neoplasms in both children and calves could be regarded as embryonic tumours or hamartomas. Little is known about the pathogenetic pathways of tumours in calves, with the exception of congenital neuro-fibromatosis (hereditary) and possibly of mesotheliomatosis (due to asbestos). Modern methods of analysing chromosomal and gene aberrations may be helpful in clarifying the pathogenesis of congenital tumours.


Correlative light and electron microscopy analysis of the centrosome: A step-by-step protocol.
Correlative light and electron microscopy harnesses the best from each of the two modalities of microscopy it utilizes; while light microscopy provides information about the dynamic properties of the cellular structure or fluorescently labeled protein, electron microscopy provides ultrastructural information in an unsurpassed resolution. However, tracing a particular cell and its rare and small structures such as centrosomes throughout numerous steps of the experiment is not a trivial task. In this chapter, we present the experimental workflow for combining live-cell fluorescence microscopy analysis with classical transmission electron microscopy, adapted for the studies of the centrosomes and basal bodies. We describe, in a step-by-step manner, an approach that can be affordably and successfully employed in any typical cell biology laboratory. The article details all key phases of the analysis starting from cell culture, live-cell microscopy, and sample fixation, through the steps of sample preparation for electron microscopy, to the identification of the target cell on the electron microscope.


Mortality in Rohingya refugee camps in Bangladesh: historical, social, and political context.
Fifty-two maternal deaths occurred between September 2017 and August 2018 in the Rohingya refugee camps in Ukhia and Teknaf Upazilas, Cox's Bazar District, Bangladesh. Behind every one of these lives lost is a complex narrative of historical, social, and political forces, which provide an important context for reproductive health programming in Rohingya camps. Rohingya women and girls have experienced human rights violations in Myanmar for decades, including government-sponsored sexual violence and population control efforts. An extension of nationalist, anti-Rohingya policies, the attacks of 2017 resulted in the rape and murder of an unknown number of women. The socio-cultural context among Rohingya and Bangladeshi host communities limits provision of reproductive health services in the refugee camps, as does a lack of legal status and continued restrictions on movement. In this review, the historical, political, and social contexts have been overlaid below on the Three Delays Model, a conceptual framework used to understand the determinants of maternal mortality. Attempts to improve maternal mortality among Rohingya women and girls in the refugee camps in Bangladesh should take into account these complex historical, social and political factors in order to reduce maternal mortality.


Effect of a static magnetic field on Escherichia coli adhesion and orientation.
This preliminary study focused on the effect of exposure to 0.5 T static magnetic fields on Escherichia coli adhesion and orientation. We investigated the difference in bacterial adhesion on the surface of glass and indium tin oxide-coated glass when exposed to a magnetic field either perpendicular or parallel to the adhesion surface (vectors of magnetic induction are perpendicular or parallel to the adhesion surface, respectively). Control cultures were simultaneously grown under identical conditions but without exposure to the magnetic field. We observed a decrease in cell adhesion after exposure to the magnetic field. Orientation of bacteria cells was affected after exposure to a parallel magnetic field. On the other hand, no effect on the orientation of bacteria cells was observed after exposure to a perpendicular magnetic field.


Organic phosphorus affects the retention of arsenite and arsenate by goethite.
The hazardous effects of arsenic are closely linked to its speciation and interaction with different soil minerals, which influence both As mobility and bioavailability. Adsorption onto iron (oxyhydr)oxides is one of the main processes controlling the partitioning of arsenite [As(III)] and arsenate [As(V)] between aqueous and solid phases. Arsenic retention can be affected by changes in soil pH and the presence of competing anions, like phosphate. Although competition with inorganic phosphorus (P) for sorption sites on mineral surfaces has been widely studied, little is known about the interactions with organic P (Po ) compounds, in particular inositol phosphates, even though they may represent a large fraction of total soil P. We quantified the effects of myo-inositol hexaphosphate (InsP6) on the adsorption and retention of As(III) and As(V) on goethite as influenced by pH, the order of anion addition, and residence time. The efficiency of InsP6 in displacing adsorbed As(III) decreased with increasing pH values and interaction time, which may be attributed to the increase in bonding strength of the As(III) complexes on the surface of goethite. Adsorption and retention of As(V) by goethite generally decreased with increasing pH, particularly in the presence of InsP6 due to the similar pKa values and the competition for the same binding sites. The addition of InsP6 before, together with, or after adsorption of As(III) and As(V) strongly reduced the amounts of sorbed As, suggesting that the addition of Po -rich matrices to As-contaminated soils may strongly enhance As mobility.


Unforeseen rupture of pseudoaneurysm of common carotid artery: An arduous anesthetic challenge.
Pseudoaneurysm of the common carotid artery (CCA) is exceptionally unstable and unpredictable; it mandates quick medical attention in order to circumvent neurologic sequelae or hemorrhage. Unanticipated rupture is extremely lethal and a potential provocation for the anesthesia caregiver. It is an arduous challenge for an anesthetist to establish emergency airway when a huge bleeding pseudoaneurysm is compressing and deviating the trachea, securing invasive lines in collapsing vessels, volume and vasopressor resuscitation in deteriorating hemodynamics in order to maintain cerebral perfusion without compromising other vital organs, arranging huge amount of blood and blood products in a short span of time, and transferring an exsanguinating patient for the rapid institution of cardiopulmonary bypass. Not only preoperatively it also necessitates appropriate neuromonitoring and neuroprotection during and after surgery. The association of unforeseen rupture of common carotid artery pseudoaneurysm secondary to the tubercular spine and lifesaving management by the rapid institution of cardiopulmonary bypass (CPB) is a rare occasion. To the best of the authors' knowledge, there is not any similar case in the peer-reviewed literature. Therefore, the authors enumerate the clinical experience of an unexpected rupture of CCA pseudoaneurysm requiring lifesaving CPB and emphasize the "Timely Teamed Effort Approach" that can sustain a life in such an inevitable situation.


Endoscopically assisted fasciotomy: description of technique and in vitro assessment of lower-leg compartment decompression.
We describe a reliable method of endoscopically assisted fasciotomy for treating chronic exertional compartment syndrome in the lower leg and for assessing compartment decompression in an in vitro model. Endoscopically assisted fasciotomy was performed in the anterior and lateral compartments of 14 matched, fresh-frozen, through-the-knee amputation specimens using a 30 degrees endoscope. A one-incision technique used in 4 specimens failed to provide complete visualization, and a two-incision technique was then performed in 10 specimens. After decompression, the skin and subcutaneous tissues were removed to assess adequacy of release, nerve decompression, anatomic course of the superficial peroneal nerve, and potential complications. Endoscopic visualization of the fascial layer and subcutaneous neurovascular structures permitted consistent compartment decompression. Fascial release, expressed as a percentage of length from the proximal origin of the fascia to the inferior retinaculum, was 99.8% (range, 98.4% to 100%) for the anterior compartment and 96.4% (range, 82% to 100%) for the lateral compartment. There were no retained fascial bands, unrecognized fascial defects, or neurovascular injuries. Optimal visualization with endoscopically assisted fasciotomy may improve clinical outcome through 1) reliable compartment decompression, 2) identification of fascial defects, 3) decompression of nerve branches at the fascial foramen, and 4) reduction of iatrogenic risk to neurovascular and muscular structures.


AMPAR-mediated synaptic transmission in the CA1 hippocampal region of neonatal rats: unexpected resistance to repeated ethanol exposure.
Alpha-amino-3-hydroxyl-5-methyl-4-isoxazole-propionate glutamatergic receptors (AMPAR) mediate most of the fast excitatory synaptic transmission in mature neurons. In contrast, a number of developing synapses do not express AMPARs; these are gradually acquired in an activity-driven manner during the first week of life in rats, which is equivalent to the third trimester of human pregnancy. Neuronal stimulation has been shown to drive high conductance Ca(2+)-permeable AMPARs into the synapse, strengthening glutamatergic synaptic transmission. Alterations in this process could induce premature stabilization or inappropriate elimination of newly formed synapses and contribute to the hippocampal abnormalities associated with fetal alcohol spectrum disorder. Previous studies from our laboratory performed with hippocampal slices from neonatal rats showed that acute ethanol exposure exerts potent stimulant effects on CA1 and CA3 neuronal networks. However, the impact of these in vitro actions of acute ethanol exposure is unknown. Here, we tested the hypothesis that repeated in vivo exposure to ethanol strengthens AMPAR-mediated neurotransmission in the CA1 region by means of an increase in synaptic expression of Ca(2+)-permeable AMPARs. We exposed rats to ethanol vapor (serum ethanol concentration approximately 40 mM) or air for 4h/day from postnatal day (P) 2-6. In brain slices prepared at P4-6, we found no significant effect of ethanol exposure on input-output curves for AMPAR-mediated field excitatory postsynaptic potentials (fEPSPs), the contribution of Ca(2+)-permeable AMPARs to these fEPSPs, or the acute effect of ethanol on fEPSP amplitude. These results suggest that homeostatic plasticity mechanisms act to maintain glutamatergic synaptic strength and ethanol sensitivity in response to repeated developmental ethanol exposure.


[Progress in molecular chorea diagnosis. McLeod syndrome and chorea acanthocytosis].
McLeod syndrome and chorea-acanthocytosis are classified with the so-called neuroacanthocytosis group of syndromes. Both lead to progressive basal ganglia degeneration and were not easily distinguished in the past. With the discovery of their molecular bases, mutations of the X-linked gene XK and autosomal recessive mutations of the gene coding for chorein, respectively, the two phenotypes can now be differentiated and extend the diagnostic spectrum in patients presenting with chorea. The present review compares the two conditions and proposes a practical approach to diagnosis and treatment. Better-defined disease concepts should eventually replace the umbrella term of "neuroacanthocytosis." Animal models are needed to understand the underlying mechanisms. A final common pathway is likely for the pathogenesis of these conditions and is most probably shared with Huntington's disease.


Effects of Vitamin D3 Supplementation on Body Composition in the VITamin D and OmegA-3 TriaL (VITAL).
Although observational studies show inverse associations between vitamin D status and body weight/adiposity, there are few large randomized controlled trials (RCTs) investigating this relationship. To determine whether vitamin D3 supplementation lowers weight or improves body composition. The VITamin D and OmegA-3 TriaL (VITAL) was a double-blinded, placebo-controlled RCT including 25 871 US adults. This ancillary study was completed in a sub-cohort that underwent body composition assessments at baseline and 2-year follow-up (89% retention). Harvard Clinical and Translational Science Center in Boston. 771 participants (men ≥ 50 and women ≥ 55 years). 2 × 2 factorial design of supplemental vitamin D3 (2000 IU/day) and/or omega-3 fatty acids (1 g/day). Endpoints were 2-year changes in weight, body mass index (BMI), waist circumference, and total and/or regional fat and lean tissue measures determined by dual-energy X-ray absorptiometry. Effect modification by clinical variables and total and free 25-hydroxyvitamin D (25[OH]D) levels was explored. There were no effects of supplemental vitamin D3vs placebo on weight, BMI, or measures of adiposity and lean tissue. Effects did not vary by sex, race/ethnicity, fat mass index, or baseline total or free 25(OH)D levels. Vitamin D3 supplementation did slightly improve body fat percentage in participants with normal BMI at baseline, but not in the overweight or obese (P for interaction = 0.04). Daily vitamin D3 supplementation vs placebo in the general older population did not improve weight or body composition. Whether supplemental vitamin D3 may benefit individuals with normal BMI warrants further study.


An immunological investigation of catalase in three mammalian species.
An immunological comparison of catalase has been conducted in the mouse, rat and guinea pig. In each of these species, enzyme activity from the peroxisomal compartment was indicated as immunologically distinct from the cytosolic activity, with catalase in the small granule compartment displaying intermediate characteristics. When related to the established data on the biogenetic and degradative changes of catalase in the mouse, the present results support the proposal that peroxisomal alterations, epigenetic modifications and degradative persistence are general characteristics of catalase biology in mammalian species.


Longitudinal displacement and intramural shear strain of the porcine carotid artery undergo profound changes in response to catecholamines.
The effects of catecholamines on longitudinal displacements and intramural shear strain of the arterial wall are unexplored. Therefore, the common carotid artery of five anaesthetized pigs was investigated using an in-house developed noninvasive ultrasonic technique. The study protocol included intravenous infusion of low-dose epinephrine (β-adrenoceptor activation), as well as intravenous boluses of norepinephrine (α-adrenoceptor activation). Further, the effects of β-blockade (metoprolol) were studied. There were significant positive correlations between pulse pressure and longitudinal displacement of the intima-media complex (r = 0.72; P < 0.001), as well as between pulse pressure and intramural shear strain (r = 0.48; P < 0.001). Following administration of norepinephrine, the longitudinal displacement of the intima-media complex and intramural shear strain profoundly increased (median 190%, range 102-296%, and median 141%, range 101-182%, respectively, compared with baseline), also when given during β-blockade (median 228%, range 133-266%, and median 158%, range 152-235%, respectively). During infusion of low-dose epinephrine, the longitudinal displacement of the intima-media complex and intramural shear strain decreased (median 88%, range 69-122%, and median 69%, range 47-117%, respectively, compared with baseline). In conclusion, the present study shows, for the first time, that the longitudinal displacement and intramural shear strain of the porcine carotid artery undergo profound changes in response to catecholamines. Increase in longitudinal displacements seems to be strongly related to α-adrenoceptor activation. Thus metoprolol is insufficient to counteract a profound increase in longitudinal displacement and intramural shear strain following a surge of norepinephrine.


Synthesis and biological evaluation of 1-(1,2-benzisothiazol-3-yl)- and (1,2-benzisoxazol-3-yl)piperazine derivatives as potential antipsychotic agents.
Members of the series of title compounds were tested for potential antipsychotic activity in relevant receptor binding assays and behavioral screens. Structure-activity relationships within the series are discussed. Compound 24 (BMY 13859-1), a (1,2-benzisothiazol-3-yl)piperazine derivative, was selected for further study because of its potent and selective profile in primary CNS tests. It was active in the Sidman avoidance paradigm and blocked amphetamine-induced stereotyped behavior in dogs for up to 7 h. The compound's lack of typical neuroleptic-like effects in the rat catalepsy test and its failure to produce dopamine receptor supersensitivity following chronic administration indicate that it should not cause the movement disorders commonly associated with antipsychotic therapy. Although 24 has potent affinity for dopaminergic binding sites, its even greater affinity for serotonin receptors suggests that a serotonergic component may be relevant to its atypical profile. Compound 24 is currently undergoing clinical evaluation in schizophrenic patients.


Tumor-targeted drug delivery and MRI contrast enhancement by chlorotoxin-conjugated iron oxide nanoparticles.
This study examines the capabilities of an actively targeting superparamagnetic nanoparticle to specifically deliver therapeutic and MRI contrast agents to cancer cells. Iron oxide nanoparticles were synthesized and conjugated to both a chemotherapeutic agent, methotrexate, and a targeting ligand, chlorotoxin, through a poly(ethylene glycol) linker. Cytotoxicity of this nanoparticle conjugate was evaluated by Alamar Blue cell viability assays, while tumor-cell specificity was examined in vitro and in vivo by MRI. Characterization of these multifunctional nanoparticles confirms the successful attachment of both drug and targeting ligands. The targeting nanoparticle demonstrated preferential accumulation and increased cytotoxicity in tumor cells. Furthermore, prolonged retention of these nanoparticles was observed within tumors in vivo. The improved specificity, extended particle retention and increased cytotoxicity toward tumor cells demonstrated by this multifunctional nanoparticle system suggest that it possesses potential for applications in cancer diagnosis and treatment.


Current Development, Obstacle and Futural Direction of Induced Pluripotent Stem Cell and Mesenchymal Stem Cell Treatment in Degenerative Retinal Disease.
Degenerative retinal disease is one of the major causes of vision loss around the world. The past several decades have witnessed emerging development of stem cell treatment for retinal disease. Nevertheless, sourcing stem cells remains controversial due to ethical concerns and their rarity. Furthermore, induced pluripotent stem cells (iPSCs) and mesenchymal stem cells (MSCs) are both isolated from patients' mature tissues; thus, issues such as avoiding moral controversy and adverse events related to immunosuppression and obtaining a large number of cells have opened a new era in regenerative medicine. This review focuses on the current application and development, clinical trials, and latest research of stem cell therapy, as well as its limitations and future directions.


Sites of electron transfer to membrane-bound copper and hydroperoxide-induced damage in the respiratory chain of Escherichia coli.
Previous studies in Escherichia coli as a model system for peroxide toxicity (L. Rodríguez-Montelongo, L. C. De la Cruz-Rodríguez, R. N. Farías, and E. M. Massa, 1993, Biochim. Biophys. Acta 1144, 77-84) have shown that electron flow through the respiratory chain supports a membrane-associated Cu(II)/Cu(I) redox cycle involved in irreversible impairment of the respiratory system by tert-butyl hydroperoxide (t-BOOH). In this paper, E. coli mutants deficient in specific respiratory chain components have been used to determine the sites of copper reduction and the targets inactivated by t-BOOH. Two sites of electron transfer to membrane-bound copper were identified: one in the region between NADH and ubiquinone supported by NADH as electron donor and another localized between ubiquinone and the cytochromes supported by electrons coming from NADH, succinate, or D-lactate. Electron flow through the former site in the presence of t-BOOH led to inactivation of NADH dehydrogenase II, whereas electron flow through the latter site in the presence of the hydroperoxide led to damage of ubiquinone. In agreement with the above in vitro results with isolated membranes, copper-dependent inactivation of NADH dehydrogenase and ubiquinone was demonstrated in E. coli cells exposed to t-BOOH. It is proposed that the t-BOOH-induced damage is a consequence of t-butylalkoxy radical generation through a Fenton-type reaction mediated by redox cycling of membrane-bound copper at those two loci of the respiratory chain.


Aquaporin-1 expressed in cultured human trabecular meshwork cells.
To determine if aquaporin-1 could be detected in cultures of human trabecular meshwork cells. Using primers specific for aquaporin-1, reverse transcription combined with polymerase chain reaction (RT-PCR) yielded a product and its size with total RNA prepared from the human trabecular meshwork cells. SDS-PAGE and immunoblotting were also used in this study to detect the specific water channel. The presence of this product and its size (298 base pairs) are consistent with that of an aquaporin-1 message in these cells. A band of 28 kD in agreement with the molecular size of aquaporin-1 was showed in a film by immunoblotting. The presence of aquaporin-1 in human trabecular meshwork cells, the predominant cell-type of the primary outflow region of the human eye, suggests that water channels may be involved in the movement of aqueous fluid out of the eye. In addition, the existence of aquaporin-1 on cultures of human trabecular meshwork cells provides an in vitro model to study the endogenous expression of aquaporin-1 and its possible role in the regulation of aqueous outflow.


Dickkopf-1 is regulated by the mevalonate pathway in breast cancer.
Amino-bisphosphonates and statins inhibit the mevalonate pathway, and may exert anti-tumor effects. The Wnt inhibitor dickkopf-1 (DKK-1) promotes osteolytic bone lesions by inhibiting osteoblast functions and has been implicated as an adverse marker in multiple cancers. We assessed the effects of mevalonate pathway inhibition on DKK-1 expression in osteotropic breast cancer. Regulation of DKK-1 by bisphosphonates and statins was assessed in human breast cancer cell lines, and the role of the mevalonate pathway and downstream targets was analyzed. Moreover, the potential of breast cancer cells to modulate osteoblastogenesis via DKK-1 was studied in mC2C12 cells. Clinical relevance was validated by analyzing DKK-1 expression in the tissue and serum of women with breast cancer exposed to bisphosphonates. DKK-1 was highly expressed in receptor-negative breast cancer cell lines. Patients with receptor-negative tumors displayed elevated levels of DKK-1 at the tissue and serum level compared to healthy controls. Zoledronic acid and atorvastatin potently suppressed DKK-1 in vitro by inhibiting geranylgeranylation of CDC42 and Rho. Regulation of DKK-1 was strongest in osteolytic breast cancer cell lines with abundant DKK-1 expression. Suppression of DKK-1 inhibited the ability of breast cancer cells to block WNT3A-induced production of alkaline phosphates and bone-protective osteoprotegerin in preosteoblastic C2C12 cells. In line with the in vitro data, treatment of breast cancer patients with zoledronic acid decreased DKK-1 levels by a mean of 60% after 12 months of treatment. DKK-1 is a novel target of the mevalonate pathway that is suppressed by zoledronic acid and atorvastatin in breast cancer.