Influence of Buffers of Different pH and Composition on the Murine Skin Barrier, Epidermal Proliferation, Differentiation, and Inflammation.
The pH of the skin is tightly regulated by endogenous buffering systems. We examined the influence of buffers of different pH and composition on skin barrier repair, pH, inflammation, and epidermal thickness/proliferation/differentiation. After tape-stripping in hairless mice buffers with pH 4-7 were applied in patch test chambers. After removal of the chambers, skin pH and transepidermal water loss (TEWL) were monitored for 24 h, and biopsies were taken for histology/immunohistology. Hairless mice showed a basal skin pH of about 5.8. Following barrier disruption and application of water, the pH increased by 0.6 units; increase in pH was reduced by the pH 4 glycolate buffer, unchanged by pH 4 citrate and pH 5.5 buffers, and even increased by the pH 7 buffer. pH 5.5, pH 4 citrate, and pH 4 glycolate buffers led to a slight, while the pH 7 buffer led to a significant increase in TEWL after barrier disruption compared to water. The pH 7 buffers led to a significant increase in epidermal thickness/proliferation/differentiation and inflammation after barrier disruption, whereas buffers with pH 4 and 5.5 caused a slight increase. In conclusion, only the pH 4 glycolate buffer significantly reduced the skin barrier disruption-related increase in skin pH. This was accompanied by only slight increase in epidermal thickness and inflammation compared to water. Application of the pH 7 buffer led to a significant increase in the skin pH, TEWL, epidermal thickness, and inflammation. The results are important for the formulation of topical products for effective acidification in pathological skin conditions.


[Rationale for the use of bioactive cements by in vitro simulation of accidental pulp opening].
To study the features of reparative dentinogenesis when using biomaterials during accidental opening of the pulp in an experiment. In an experiment on 3 dwarf pigs of the Svetlogorsk breed a traumatic opening of the tooth cavity was carried out in 18 teeth (6 molars, 6 premolars, 6 incisors), then Biodentin (9 teeth) and Aurosil (9 teeth) were applied on the opened pulp horn. After 14, 30 and 60 days, dwarf pigs were euthanized, and jaw blocks taken. Histological examination of the teeth was carried out using the method of embedding in methyl methacrylate (OsteoBead, Sigma, USA). Thin sections of 35-50 microns were made from the samples and stained using the «Heavenly Trichrome» method. Documentation was carried out using a Leica DM100 microscope and an EC3 digital camera in reflected and transmitted light. Histological examination of the thin sections of teeth stained according to the «Heavenly Trichrome» method revealed that 2 weeks after the pulp opening and applying of Aurosil and Biodentin, comparable processes occurred in the tooth cavity. Thus, the proliferation of odontoblasts was noted on the lateral surfaces of the chamber. New dentin layers and odontoblasts spread over the lower surface of the material with the formation of dentinal bridges by 4 weeks of observation in both groups. By week 12, there was a decrease in the proliferative activity of odontoblasts and dentin maturation in all cases. At the initial stages of the experiment, some pulp edema and moderate plethora were noted, which was no longer observed at 4 weeks. By 12 weeks, because of the proliferation of odontoblasts, the volume of the tooth cavity was reduced by a third. By the 6th month in the experiment, no significant changes in the structure of dentin and pulp were revealed. According to morphological data the studied biomaterials (Aurosil and Biodentin) contribute to dentin regeneration equally.


Interleukin-2 and interferon-gamma production in follicular lymphomas.
In situ hybridization with specific RNA radiolabeled probes was used to analyze the production of interleukin-2 (IL-2) and interferon-gamma (IFN-gamma) by 21 low-grade follicular lymphomas (FLs). All of the 21 FLs tested contained lymphokine-synthesizing cells in the interfollicular and follicular areas. Enumeration of lymphokine synthesizing cells indicated heterogeneous IL-2 production among lymph nodes tested; 2 of the 21 had much higher densities of IL-2-producing cells (855 and 570/cm2) than did the remaining 19 (mean, 92 +/- 15/cm2). IFN-gamma-producing cells displayed no such variation (mean, 77 +/- 8 IFN-gamma-producing cells/cm2). The mean IL-2/IFN-gamma-producing cell ratio was 2.69 +/- 0.84, indicating preferential induction of IL-2. The detailed distribution of lymphokine-producing cells showed that IL-2 and IFN-gamma-producing cells were located mainly in the follicular areas. The mean follicular/interfollicular ratio was 1.82 +/- 0.16 for IL-2 and 1.92 +/- 0.19 for IFN-gamma-producing cells. The results show that T-cell activation, defined by lymphokine production, occurs in FL lymph nodes in direct contact with malignant B cells. Thus, lymphokine production may play an important role in the control of tumor growth, which is the result of interaction between tumor cells and host-derived immune reaction.


The adductor flap: a new method for transferring posterior and medial thigh skin.
Skin flaps from the medial aspect of the thigh have traditionally been based on the gracilis musculocutaneous unit. This article presents anatomic studies and clinical experience with a new flap from the medial and posterior aspects of the thigh based on the proximal musculocutaneous perforator of the adductor magnus muscle and its venae comitantes. This cutaneous artery represents the termination of the first medial branch of the profunda femoris artery and is consistently large enough in caliber to support much larger skin flaps than the gracilis musculocutaneous flap. In all 20 cadaver dissections, the proximal cutaneous perforator of the adductor magnus muscle was present and measured between 0.8 and 1.1 mm in diameter, making it one of the largest skin perforators in the entire body. Based on this anatomic observation, skin flaps as large as 30 x 23 cm from the medial and posterior aspects of the thigh were successfully transferred. Adductor flaps were used in 25 patients. On one patient the flap was lost, in one the flap demonstrated partial survival, and in 23 patients the flaps survived completely. The flap was designed as a pedicle island flap in 14 patients and as a free flap in 11. When isolating the vascular pedicle for free tissue transfer, the cutaneous artery is dissected from the surrounding adductor magnus muscle and no muscle is included in the flap. Using this maneuver, a pedicle length of approximately 8 cm is isolated. In addition to ample length, the artery has a diameter of approximately 2 mm at its origin from the profunda femoris artery. The adductor flap provides an alternative method for flap design in the posteromedial thigh. Because of the large pedicle and the vast cutaneous territory that it reliably supplies, the authors believe that the adductor flap is the most versatile and dependable method for transferring flaps from the posteromedial thigh region.


Central venous pressure measurements improve the accuracy of leg raising-induced change in pulse pressure to predict fluid responsiveness.
Passive leg raising (PLR) is a maneuver performed to test the cardiac Frank-Starling mechanism. We assessed the influence of PLR-induced changes in preload on the performance of PLR-induced change in pulse pressure (Delta(PLR)PP) and cardiac output (Delta(PLR)CO) for fluid responsiveness prediction. Sedated, nonarrhythmic patients with persistent shock were included in this prospective multicenter study. Cardiac output and pulse pressure were measured at baseline (patient supine), during PLR (lower limbs lifted to 45 degrees) and after 500-ml volume expansion. Patients were classified as responders or not. In the whole population (n = 102), the area under the receiver-operating characteristic curve (AUC) was 0.76 for Delta(PLR)PP and was higher for Delta(PLR)CO (0.89)(p < 0.05), but likelihood ratios were close to 1. In patients with a PLR-induced increase in central venous pressure (CVP) of at least 2 mmHg (n = 49), Delta(PLR)PP and Delta(PLR)CO disclosed higher AUCs than in the rest of the population (0.91 vs. 0.66 and 0.98 vs. 0.83; p < 0.05); positive/negative likelihood ratios were 9.3/0.14 (8% cutoff level) and 30/0.07 (7% cutoff level), respectively. A PLR-induced change in CVP > or =2 mmHg was required to allow clinical usefulness of PLR-derived indices. In this situation, Delta(PLR)PP performed well for predicting fluid responsiveness in deeply sedated patients.


Polyethylene glycol-stabilized sulphur nanoparticles: an effective antimicrobial agent against multidrug-resistant bacteria.
To elucidate the antibacterial efficacy of chemically synthesized and custom-made sulphur nanoparticles (SNPs) of two different sizes and surface modifications against a number of multidrug-resistant Gram-negative bacilli (GNB) harbouring the New Delhi metallo-β-lactamase 1 enzyme (NDM-1). Antimicrobial susceptibility of the isolates was determined. The strains were evaluated for the presence of carbapenemases, β-lactamases, 16S rRNA methylases and integrons. Chemically synthesized, polyethylene-glycol (PEG)-stabilized SNPs of 10 nm and custom-made non-capped SNPs of 60 nm were physicochemically characterized and evaluated for their antibacterial efficacy against multidrug-resistant GNB using the agar dilution method (ADM) and the broth microdilution method (BMD). The cytotoxicity of the chemically synthesized SNPs was evaluated with a human-derived hepatoma (HepG2) cell line using a WST-1 assay kit. All isolates were multidrug-resistant and possessed NDM-1 along with other β-lactamases, 16S rRNA methylases and integron 1. Chemically synthesized PEGylated SNPs showed a bactericidal effect against all tested strains at a concentration between 9.41 and 18.82 mg/L using BMD. The ADM data revealed that SNPs had uniform MICs (18.82 mg/L) for all tested strains. On the other hand, custom-made SNPs failed to impart any antibacterial effect at the equivalent concentrations of chemically synthesized SNPs. The WST-1 assay revealed no significant cytotoxicity of the PEGylated SNPs even at the highest concentration (94.08 mg/L). To the best of our knowledge, this is the first attempted study to show the effectiveness of nanoparticles against multidrug-resistant GNB harbouring NDM-1.


Localization of cortical dysfunction based on auditory and visual naming performance.
Naming is generally considered a left-hemisphere function without precise localization. However, recent cortical stimulation studies demonstrate a modality-related anatomical dissociation, in that anterior temporal stimulation disrupts auditory description naming ("auditory naming") but not visual object naming ("visual naming"), whereas posterior temporal stimulation disrupts naming on both tasks. We hypothesized that patients with anterior temporal abnormalities would exhibit impaired auditory naming, yet normal range visual naming, whereas patients with posterior temporal abnormalities would exhibit impaired performance on both tasks. Thirty-four patients with documented anterior temporal abnormalities and 14 patients with documented posterior temporal abnormalities received both naming tests. As hypothesized, patients with anterior temporal abnormalities demonstrated impaired auditory naming, yet normal range visual naming performance. Patients with posterior temporal abnormalities were impaired in visual naming; however, auditory naming scores were intact. Although these group patterns were statistically significant, on an individual basis, auditory-visual naming asymmetries better predicted whether individual patients had anterior or posterior temporal abnormalities. These behavioral findings are generally consistent with stimulation results, suggesting that modality specificity is inherent in the organization of language, with predictable neuroanatomical correlates. Results also carry clinical implications regarding localizing dysfunction, identifying and characterizing naming deficits, and potentially, in treating neurologically based language disorders.


Different subcellular localizations of discoidin I monomer and tetramer in Dictyostelium discoideum cells: using conformation-specific monoclonal antibodies.
We characterized four monoclonal antibodies that recognize monomeric and/or tetrameric forms of Dictyostelium lectin (discoidin I) to study intracellular localization of this lectin in early development. Three different PAGEs (native-, urea-, and SDS-PAGE) following immunoblot showed that three of the four mAbs exhibit preference for the tetrameric form while mAb DC2 reacts only with unreduced monomer. By immunofluorescence studies of Dictyostelium NC-4, we found that the anti-tetramer antibodies mainly stain multilamellar bodies, which are food vacuoles to be externalized from cells. In contrast, mAb DC2 stains the cytosol weakly but not the multilamellar bodies. The same pattern of distribution was confirmed by immunoelectron microscopy. These results clearly indicate that, upon packaging discoidin I into multilamellar bodies, the tetrameric form is selectively packaged from the cytosolic pool. In addition, to clarify the relation between the multilamellar bodies and the tetramer, we examined the localization of tetramer using both the cells fed by Escherichia coli and the A3 cells axenically grown. When the cells were fed by E. coli, the cells made multilamellar bodies that contained no lectin; instead the tetramer was found in the cytoplasm and the cells still exclude the lectin around the cells. In contrast, A3 cells grown axenically without bacteria do not make multilamellar bodies but the tetramer also was found in the cytoplasm evenly (also, see W.F. Loomis, Dictyostelium Discoideum, pp. 160-161, Academic Press, New York, 1975). The data suggest that, under these conditions, the tetramer can be excluded from cells via some organelle such as normal secretory vacuole but not multilamellar bodies. Moreover this represents the first example of differential localization of tetramer and monomer of discoidin I in the cellular slime mold cells.


[Duffy antigen receptor for chemokines attenuates breast cancer growth and metastasis: an experiment with nude mice].
To study the effects of Duffy antigen receptor for chemokines (DARC) on the tumorigenesis and metastasis of breast cancer. Human breast cancer cells of the line MDA-MB-435HM with a great potentiality of metastasis were cultured and then divided into 3 groups: MDA-MB-435HM-DARC cell group, to be transfected with pcDNA3/DARC FyB containing human DARC cDNA; MDA-MB-435HM-vect cell group, to be transfected with blank plasmid pcDNA3; and MDA-MB-435HM cell group without transfection. Female BALB/c nude mice were implanted with MDA-MB-435HM cells into the nipple. The size of the implanted cancer was measured once a week. The mice were killed 4, 6, and 8 weeks after the implantation respectively and their lungs were taken out to observe the number of metastatic tumor. Immunohistochemical staining was performed to calculate the microvascular density (MVD). Western blotting was used to detect the matrix metalloproteinase (MMP)-9 protein expression of the lung tissues. ELISA was used to detect the concentrations of the 2 DARC ligands: CNCL8 and CCL2 in the supernatant of culture fluid of the 3 kinds of cells and the metastatic tumors. The expression levels of DARC mRNA and protein of the MDA-MB-435HM cells were 38% those of the MDA-MB-435 cells. The expression levels of CXCL8 and CCL2 of the MDA-MB-435HM-DARC cells were significantly lower than those of the MDA-MB-435HM-vect cells and MDA-MB-435HM cells (both P < 0.05). All mice developed tumor with a tumorigenesis rate of 100%. However, the tumor occurred 17 days after implantation in those mice implanted with MDA-MB-435HM-DARC cells, significantly later then in those implanted with MDA-MB-435HM-vect cells and MDA-MB-435HM cells (9 and 10 days after implantation) the size of tumor in the former group being significantly smaller than those of the 2 latter groups (both P < 0.01). The number of metastatic foci in the lung of the MDA-MB-435HM-DARC group was significantly less than those of the other 2 groups (both P < 0.01). The level of CCI2 of the implanted tumor in the MDA-MB-435HM-DARC group was significantly lower than those of the other 2 groups (both P < 0.01). The MVD of the MDA-MB-435HM-DARC was significantly less than those of the other 2 groups (both P < 0.01) and most of the vessels of the MDA-MB-435HM-DARC group were not newly formed vessels with a diameter < 8 red blood cells. The MMP-9 protein expression levels of the MDA-MB-435HM-vect cells and MDA-MB-435HM-cells were 3.1 and 3.4 times that of the MDA-MB-435HM-DARC group (both P < 0.05). DARC, a chemokine decoy receptor, inhibits the chemokine-induced angiogenesis, thus inhibiting the growth and lung metastasis of breast cancer.


Phenotyping transgenic wheat for drought resistance.
Realistic experimental protocols to screen for drought adaptation in controlled conditions are crucial if high throughput phenotyping is to be used for the identification of high performance lines, and is especially important in the evaluation of transgenes where stringent biosecurity measures restrict the frequency of open field trials. Transgenic DREB1A-wheat events were selected under greenhouse conditions by evaluating survival and recovery under severe drought (SURV) as well as for water use efficiency (WUE). Greenhouse experiments confirmed the advantages of transgenic events in recovery after severe water stress. Under field conditions, the group of transgenic lines did not generally outperform the controls in terms of grain yield under water deficit. However, the events selected for WUE were identified as lines that combine an acceptable yield-even higher yield (WUE-11) under well irrigated conditions-and stable performance across the different environments generated by the experimental treatments.


Endothelin signaling via guanine exchange factor C3G in renal glomerular mesangial cells.
The guanine nucleotide exchange factor C3G is one of the mediators of endothelin-1 (ET-1) intracellular signaling cascades and is vital for kidney development and homeostasis. The aim of the current study was to analyze the specificity of ET-1-induced signaling via C3G in rat glomerular mesangial cells (GMC) and to investigate the biological significance of C3G during mesangioproliferative glomerulonephritis. In GMC, C3G expression was increased (1) in vivo after induction of the anti-Thy1 model of glomerulonephritis and (2) in cell culture experiments after fetal bovine serum incubation. To examine the consequences of C3G up-regulation, adenovirus-mediated gene transfer of C3G into cultured glomerular cells was done, and the GTP loading of the small G proteins Rap1 and R-Ras was analyzed. Overexpression of C3G in mesangial cells resulted in enhanced activation of Rap1, but failed to affect the GTP-bound status of R-Ras in ET-1-stimulated cells. C3G overexpression led to significant changes in GMC spreading and migration patterns in response to ET-1 stimulation and increased stress fiber formation, which was mimicked by Rap1A overexpression. Together, these findings suggest (1) the existence of regulatory mechanisms resulting in disease-related up-regulation of C3G in GMC and (2) that an increase in the C3G protein level may contribute to the resolution stage of mesangioproliferative glomerulonephritis by reducing GMC sensitivity to ET-1, modulating cellular motility, and actin dynamics.


Culture-independent characterization of the microbiota of the ant lion Myrmeleon mobilis (Neuroptera: Myrmeleontidae).
Ant lions are insect larvae that feed on the liquefied internal components of insect prey. Prey capture is assisted by the injection of toxins that are reportedly derived from both the insect and bacterial symbionts. These larvae display interesting gut physiology where the midgut is not connected to the hindgut, preventing elimination of solid waste until adulthood. The presence of a discontinuous gut and the potential involvement of bacteria in prey paralyzation suggest an interesting microbial role in ant lion biology; however, the ant lion microbiota has not been described in detail. We therefore performed culture-independent 16S rRNA gene sequence analysis of the bacteria associated with tissues of an ant lion, Myrmeleon mobilis. All 222 sequences were identified as Proteobacteria and could be subdivided into two main groups, the alpha-Proteobacteria with similarity to Wolbachia spp. (75 clones) and the gamma-Proteobacteria with similarity to the family Enterobacteriaceae (144 clones). The Enterobacteriaceae-like 16S rRNA gene sequences were most commonly isolated from gut tissue, and Wolbachia-like sequences were predominant in the head and body tissue. Fluorescence in situ hybridization analyses supported the localization of enterics to gut tissue and Wolbachia to nongut tissue. The diversity of sequences isolated from freshly caught, laboratory-fed, and laboratory-starved ant lions were qualitatively similar, although the libraries from each treatment were significantly different (P = 0.05). These results represent the first culture-independent analysis of the microbiota associated with a discontinuous insect gut and suggest that the ant lion microbial community is relatively simple, which may be a reflection of the diet and gut physiology of these insects.


Superdiffusion of quantized vortices uncovering scaling laws in quantum turbulence.
Generic scaling laws, such as Kolmogorov's 5/3 law, are milestone achievements of turbulence research in classical fluids. For quantum fluids such as atomic Bose-Einstein condensates, superfluid helium, and superfluid neutron stars, turbulence can also exist in the presence of a chaotic tangle of evolving quantized vortex lines. However, due to the lack of suitable experimental tools to directly probe the vortex-tangle motion, so far little is known about possible scaling laws that characterize the velocity correlations and trajectory statistics of the vortices in quantum-fluid turbulence, i.e., quantum turbulence (QT). Acquiring such knowledge could greatly benefit the development of advanced statistical models of QT. Here we report an experiment where a tangle of vortices in superfluid 4He are decorated with solidified deuterium tracer particles. Under experimental conditions where these tracers follow the motion of the vortices, we observed an apparent superdiffusion of the vortices. Our analysis shows that this superdiffusion is not due to Lévy flights, i.e., long-distance hops that are known to be responsible for superdiffusion of random walkers. Instead, a previously unknown power-law scaling of the vortex-velocity temporal correlation is uncovered as the cause. This finding may motivate future research on hidden scaling laws in QT.


Identification of a murine monoclonal antibody specific for an allotypic determinant on mouse CD3.
A murine monoclonal antibody (mAb; 7D6) that was mitogenic for T cells was derived from 129/Sv animals immunized with a T helper clone from C57BL/6 origin. Fluoresceinated 7D6 labeled T cells from most common mouse strains but not from 129/Sv and LP/J animals, and this labeling was inhibited by the anti-CD3 epsilon mAb 145-2C11. The mitogenicity of 7D6 for T cells had a similar strain specificity. The antibody immunoprecipitated the T cell receptor (TcR) complex from a T cell hybridoma. After dissociation of this immunoprecipitate with detergents, the CD3 gamma and epsilon chains were retained by the 7D6 antibody. Immunoprecipitation data were also obtained with COS cells transfected with the CD3 gamma, delta or epsilon chains alone, in pairs or together. They confirmed that 7D6 bound the CD3 gamma epsilon pair, suggesting that the antibody recognizes a conformational epitope formed by gamma epsilon pairing, whereas 145-2C11 bound both gamma epsilon and delta epsilon pairs. These results, therefore, add to current information about TcR structure and subunit stoichiometry. We have demonstrated that the 7D6 mAb specifically binds to a CD3 dimer comprised of gamma and epsilon chains. We thus provide additional evidence that indicates that two CD3 epsilon chains are found within the receptor, one linked to CD3 gamma and the other to CD3 delta.


Does a critical rotator cuff tear stage exist?: a biomechanical study of rotator cuff tear progression in human cadaver shoulders.
It is unknown at which stage of rotator cuff tear the biomechanical environment is altered. The purpose of this study was to determine if a critical rotator cuff tear stage exists that alters glenohumeral joint biomechanics throughout the rotational range of shoulder motion, and to evaluate the biomechanical effect of parascapular muscle-loading. Eight cadaver shoulders were used with a custom testing system. Four progressive rotator cuff tear stages were investigated on the basis of footprint anatomy. Three muscle-loading conditions were examined: rotator cuff only; rotator cuff with deltoid muscle; and rotator cuff, deltoid, pectoralis major, and latissimus dorsi muscles. Testing was performed in the scapular plane with 0°, 30°, and 60° of shoulder abduction. The maximum internal and external rotations were measured with 3.4 Nm of torque. The position of the humeral head apex with respect to the glenoid was calculated with use of a MicroScribe 3DLX digitizing system throughout the rotational range of motion. The abduction capability was determined as the abduction angle achieved with increasing deltoid load. Tear of the entire supraspinatus tendon significantly increased maximum external rotation and significantly decreased abduction capability with higher deltoid loads (p < 0.05). Tear of the entire supraspinatus tendon and half of the infraspinatus tendon significantly shifted the humeral head apex posteriorly at the midrange of rotation and superiorly at maximum internal rotation (p < 0.05). Loading the pectoralis major and latissimus dorsi muscles decreased the amount of humeral head elevation due to deltoid loading. Tear of the entire supraspinatus tendon was the critical stage for increasing rotational range of shoulder motion and for decreased abduction capability. Further tear progression to the infraspinatus muscle was the critical stage for significant changes in humeral head kinematics. The pectoralis major and latissimus dorsi muscles played an important role in stabilizing the humeral head as the rotator cuff tear progressed.


[Abnormal head position caused by incorrect prescription for astigmatism: case report].
Abnormal head position is a compensatory condition which improves patients' vision. It can be caused by ophthalmological problems such as oculomotor imbalances (strabismus, nystagmus) and high astigmatism. However, it results in esthetic impairment, orthopedic trouble and facial asymmetries. We describe a case of a girl, JL, 8 years, with abnormal head position tilted to the left since the last glasses were prescribed. The correction used by the patient was: right eye = +2.00 sph diopter -5.5 cyl 180 degrees and left eye = +2.25 sph diopter -5.75 cyl 180 degrees. In tilted position, the correct visual acuity was: right eye 6/12 and left eye 6/9. No deviations were noted by the cover test and the remaining ophthalmological examination was completely normal. Retinoscopy under cycloplegia and subjective test showed right eye = +3.50 sph diopter -6.00 cyl 10 degrees; and left eye = +3.50 sph diopter -6.00 cyl 170 degrees, with visual acuity 6/6 in both eyes. With adequate prescription, the head position was normalized. Wrong cylindrical positions for correction of high astigmatism may cause abnormal head position. Retinoscopy under cycloplegia and subjective test are essential for precise diagnoses and prescriptions.


Restriction of Human Cytomegalovirus Infection by Galectin-9.
Human cytomegalovirus (HCMV) is a ubiquitous human herpesvirus. While HCMV infection is generally asymptomatic in the immunocompetent, it can have devastating consequences in those with compromised or underdeveloped immune systems, including transplant recipients and neonates. Galectins are a widely expressed protein family that have been demonstrated to modulate both antiviral immunity and regulate direct host-virus interactions. The potential for galectins to directly modulate HCMV infection has not previously been studied, and our results reveal that galectin-9 (Gal-9) can potently inhibit HCMV infection. Gal-9-mediated inhibition of HCMV was dependent upon its carbohydrate recognition domains and thus dependent on glycan interactions. Temperature shift studies revealed that Gal-9 specific inhibition was mediated primarily at the level of virus-cell fusion and not binding. Additionally, we found that during reactivation of HCMV in hematopoietic stem cell transplant (HSCT) patients soluble Gal-9 is upregulated. This study provides the first evidence for Gal-9 functioning as a potent antiviral defense effector molecule against HCMV infection and identifies it as a potential clinical candidate to restrict HCMV infections.IMPORTANCE Human cytomegalovirus (HCMV) continues to cause serious and often life-threatening disease in those with impaired or underdeveloped immune systems. This virus is able to infect and replicate in a wide range of human cell types, which enables the virus to spread to other individuals in a number of settings. Current antiviral drugs are associated with a significant toxicity profile, and there is no vaccine; these factors highlight a need to identify additional targets for the development of anti-HCMV therapies. We demonstrate for the first time that secretion of a member of the galectin family of proteins, galectin-9 (Gal-9), is upregulated during natural HCMV-reactivated infection and that this soluble cellular protein possesses a potent capacity to block HCMV infection by inhibiting virus entry into the host cell. Our findings support the possibility of harnessing the antiviral properties of Gal-9 to prevent HCMV infection and disease.


Bone structure in two adult subjects with impaired minor spliceosome function resulting from RNU4ATAC mutations causing microcephalic osteodysplastic primordial dwarfism type 1 (MOPD1).
Microcephalic osteodysplastic primordial dwarfism type 1 (MOPD1), or Taybi-Linder syndrome is characterized by distinctive skeletal dysplasia, severe intrauterine and postnatal growth retardation, microcephaly, dysmorphic features, and neurological malformations. It is an autosomal recessive disorder caused by homozygous or compound heterozygous mutations in the RNU4ATAC gene resulting in impaired function of the minor spliceosome. Here, we present the first report on bone morphology, bone density and bone microstructure in two adult MOPD1 patients and applied radiographs, dual energy X-ray absorptiometry, high-resolution peripheral quantitative computed tomography and biochemical evaluation. The MOPD1 patients presented with short stature, low BMI but normal macroscopic bone configuration. Bone mineral density was low. Compared to Danish reference data, total bone area, cortical bone area, cortical thickness, total bone density, cortical bone density, trabecular bone density and trabecular bone volume per tissue volume (BV/TV) were all low. These findings may correlate to the short stature and low body weight of the MOPD1 patients. Our findings suggest that minor spliceosome malfunction may be associated with altered bone modelling.


Monoclonal antibodies produced against lipopolysaccharide from fimA Type II Porphyromonas gingivalis.
An important periodontal pathogen, Porphyromans gingivalis strains are classified into six genotypes (types I-V and Ib), based on the genotype of the fimbriae A (fimA). Among the genotypes, fimA type II strains are thought to be most strongly related to advanced periodontitis. To develop passive immunotherapy, over 300 hybridoma clones were constructed through immunization of cell extracts of fimA type II strain P. gingivalis TDC60 using hybridoma technology. Among these clones, 15 MAbs recognized TDC60 lipopolysaccharide (LPS) with an individual ladder-like structure by Western blot analysis. Further Western blotting of the 15 MAbs against LPS from TDC60, FDC381 (fimA type I), and W83 (fimA type IV) of P. gingivalis and Escherichia coli was carried out. None of these MAbs recognized E. coli LPS, and divided into at least three different Western blot patterns. To confirm the specificity to LPS, three clones were selected and competition assays were carried out using TDC60 LPS. All three MAbs reduced the reactivity against TDC60 LPS after absorption of the LPS in a dose-dependent manner. These findings suggest that MAbs recognizing different epitopes of P. gingivalis LPS were successfully constructed, and these MAbs may be useful in neutralizing P. gingivalis infection.


Patient-controlled analgesia. Its South African debut in a provincial hospital.
Patient-controlled analgesia (PCA) is a well-established technique for the relief of acute and chronic pain. It is widely used in Western hospitals. Patient and staff education is required to provide successful analgesia. This paper reports the successful introduction of PCA on a large scale in a provincial hospital. The considerable potential difficulties in communication and education appear to have been overcome. The widespread introduction of PCA in all our hospitals would appear to be feasible.


Defective adhesion in tumor infiltrating CD8+ T cells.
CD8(+) tumor-infiltrating lymphocytes (TIL) are defective in cytolysis due to tumor-induced inhibition of proximal TCR-mediated signaling, a defect that is relieved upon purification and brief culture. We show in this study that frequency of conjugation in vitro of nonlytic TIL with tumor cells is low in comparison with their lytic counterparts, and the strength of interaction and duration of conjugation are also reduced. Previous reports show that p56(lck) activation is required for TCR-initiated LFA-1 avidity up-regulation, raising the question: is low LFA-1 avidity the basis of reduced TIL conjugation frequency? When stimulated with phorbol ester, nonlytic TIL bind purified ICAM-1 equivalently as lytic TIL, suggesting that LFA-1 can be activated if proximal TCR signaling is bypassed. However, when treated with phorbol ester, the conjugation frequency of nonlytic TIL does not increase. CD2 and CD8 also mediate T cell adhesion to cognate target cells and are both expressed at lower levels in nonlytic TIL in addition to being excluded from the immune synapse formed upon conjugation. Collectively, these results imply that adhesion defects in nonlytic TIL result from a combination of decreased cell surface levels of adhesion molecules, deficient LFA-1 activation, and the failure to recruit essential adhesion receptors to the membrane contact site formed with cognate target cells.


ATP and other nucleotides stabilize the Rho-mRNA complex.
Transcription termination factor Rho from Escherichia coli is a protein that consists of a single 47 kDa protomeric unit that can form a hexameric structure. To determine whether active hexamers can form on an RNA by assembly of subunits, we measured the dependence of complex formation on the concentration of Rho protein in the presence and absence of various nucleotides and related the binding properties to association states determined from sedimentation properties. The results show that the presence of adenine nucleotides converts RNA binding from a multimeric process to a largely monomeric process and that the change correlates with the stabilization of multimers of Rho by the nucleotides. The experimental evidence also indicates that the hexameric form of Rho is stabilized slightly by binding to a transcript but that the protein on RNA is in equilibrium with nonhexameric forms. These results suggest that a Rho hexamer can form on a transcript by addition of subunits to a partial assembly, which means that the complex can consist of six subunits surrounding an RNA transcript as proposed in recent models for Rho action.


Rapid development of vitamin K deficiency in an adolescent boy receiving total parenteral nutrition following bone marrow transplantation.
In this case report the development of a vitamin K deficiency in a neutropenic adolescent receiving parenteral nutrition following bone marrow transplantation is described. The parenteral nutrition solution did not provide vitamin K and the patient had been receiving oral antibiotics prior to and during use of the intravenous feeding. Oral intake was minimal. Standard adult oral and intravenous multivitamin preparations for use in individuals older than 11 years do not routinely contain this vitamin. The function of vitamin K and causes for development of a deficiency are reviewed. Recommended intakes and guidelines for supplementation are also discussed.


Patient-perceived outcomes in thigh pain after primary arthroplasty of the hip.
Thigh pain after arthroplasty first was identified in the joint replacement literature in 1988; little information has been published about the functional status of patients who develop this complication. Eleven patients who reported thigh pain at 2 years after cementless primary total hip arthroplasty were matched with patients who did not report thigh pain on the following characteristics: age, gender, diagnosis, bone type, and surgical procedure (ie, unilateral or bilateral). Preoperative WOMAC function scores were different in these two groups (53.5 versus 39.7). There were no differences in SF-36 and Quality of Well Being scores in the two patient groups. There were also no differences in the WOMAC, SF-36, and Quality of Well Being scores at the 2-year followup. However, item analysis of the WOMAC indicated that patients with thigh pain were more likely to report at least some functional deficit relative to patients without thigh pain at the 2-year followup. Thigh pain does not seem to affect 2-year postoperative quality of life severely in patients who have had cementless hip arthroplasties with a tapered stem design. Selected functional activities will be impacted by the presence of thigh pain. Prognostic study, Level III (case-control study). See the Guidelines for Authors for a complete description of levels of evidence.


[Clinical characteristics of viral pneumonia in patients with chronic obstructive pulmonary disease].
Objective: To study the clinical and etiological characteristics of viral pneumonia in patients with chronic obstructive pulmonary disease(VP-COPD), and to identify the risk factors associated with poor prognosis. Methods: From August 1, 2017 to August 1, 2019, totally 235 patients in a general hospital in Beijing were prospectively enrolled in this research, and all patients were diagnosed with viral pneumonia by imaging and etiology. The patients were divided into VP-COPD group(n=60) and VP-nCOPD(viral pneumonia in non-COPD patients) group(n=175). Pathogen detection and clinical characteristics were compared between the two groups.Finally, the binomial logistic regression was used to explore the risk factors associated with severe VP-COPD. Results: Compared with the VP-nCOPD group, the VP-COPD group was older(76.5 vs 66.0 years, P=0.001), and the CURB-65 score(2 vs 1, P= 0.001) and the PSI score(111 vs 85, P<0.001) were higher at admission. Pseudomonas aeruginosa(χ²= 10.308, P= 0.001) and Staphylococcus aureus(χ²= 5.953, P=0.028) were the most common co-infection bacteria. In the VP-COPD group type Ⅱ respiratory failure was more common(23.3% vs 6.8%, P<0.001), the number of severely ill patients was larger(48.3% vs 30.3%, P=0.011), the length of hospital stay was longer(13 vs 8, P<0.001), and the mortality rate during hospitalization was higher(18.3% vs 7.4%, P=0.016) in the VP-nCOPD group. Multivariate analysis showed that the level of blood glucose(OR: 1.73, 95%CI: 1.22-2.44, P= 0.002) and pleural effusion(OR: 133.12, 95%CI: 7.57-2 340.36, P=0.001) were risk factors for severe VP-COPD patients. Conclusion: Viral pneumonia in patients with COPD tended to develop into severe cases and had a poor prognosis.


Kinetics of surface enrichment: A molecular dynamics study.
We use molecular dynamics to study the kinetics of surface enrichment (SE) in a stable homogeneous mixture (AB), placed in contact with a surface which preferentially attracts A. The SE profiles show a characteristic double-exponential behavior with two length scales: ξ(-), which rapidly saturates to its equilibrium value, and ξ(+), which diverges as a power-law with time (ξ(+)∼t(θ)). We find that hydrodynamic effects result in a crossover of the growth exponent from θ≃0.5 to θ≃1.0. There is also a corresponding crossover in the growth dynamics of the SE layer thickness.


FOXD3 regulates the lineage switch between neural crest-derived glial cells and pigment cells by repressing MITF through a non-canonical mechanism.
The first neural crest cells to emigrate from the neural tube are specified as neurons and glial cells and are subsequently followed by melanocytes of the skin. We wished to understand how this fate switch is controlled. The transcriptional repressor FOXD3 is expressed exclusively in the neural/glial precursors and MITF is expressed only in melanoblasts. Moreover, FOXD3 represses melanogenesis. Here we show that avian MITF expression begins very early during melanoblast migration and that loss of MITF in melanoblasts causes them to transdifferentiate to a glial phenotype. Ectopic expression of FOXD3 represses MITF in cultured neural crest cells and in B16-F10 melanoma cells. We also show that FOXD3 does not bind directly to the MITF promoter, but instead interacts with the transcriptional activator PAX3 to prevent the binding of PAX3 to the MITF promoter. Overexpression of PAX3 is sufficient to rescue MITF expression from FOXD3-mediated repression. We conclude that FOXD3 controls the lineage choice between neural/glial and pigment cells by repressing MITF during the early phase of neural crest migration.


Immunostimulatory oligonucleotides inhibit colonic proinflammatory cytokine production in ulcerative colitis.
We previously showed that Toll-like receptor-9 (TLR-9) ligands ameliorate experimental colitis. In this study, we evaluated the effect of TLR-9 ligands on the generation of proinflammatory cytokines by human colonic mucosa. Colonoscopic biopsies were obtained from patients with active ulcerative colitis (UC) and from normal subjects. The tissue was organ cultured for 24 hours in the presence or absence of different types of immunostimulatory (ISS) (CpG)-oligonucleotides (ODNs). Tumor necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta) levels in the medium were determined by enzyme-linked immunosorbent assay. In active UC, hTNF-alpha and hIL-lbeta generation by inflamed colonic mucosa is 7- and 3-fold higher, respectively, than their generation by normal mucosa. Class B CpG ODNs inhibited colonic TNF-alpha and IL-1beta generation by 50%, whereas class A or C ODNs had a partial or no effect, respectively. A novel class of ODNs that is based on multiple TCG repeats was as effective as class B ODNs. This inhibition resulted from the transcriptional suppression of IL-1beta that occurred within the first 2 hours after ISS-ODN incubation. The addition of chloroquine abolished the inhibitory effects of ISS-ODNs on colonic TNF-alpha and IL-1beta generation. Only certain classes of ISS-ODNs inhibit the enhanced TNF-alpha and IL-1beta generated ex vivo by inflamed colonic mucosa of patients with UC. The effect of ISS-ODNs is mediated by triggering of TLR-9. These results suggest a potential therapeutic value for ISS-ODNs in UC.


Effect of pacifier use on exclusive and any breastfeeding: a meta-analysis.
The objective of this meta-analysis was to determine with cross-sectional and cohort trails whether the use of pacifier increases the risk of early weaning from exclusive breastfeeding before six months of age or cessation of breastfeeding from any breastfeeding before 24 months of age. Additionally, the effect of the age for starting pacifier use on breastfeeding duration was analyzed in the cohort trails. The Medline database was searched (1980 to 2006) with "breastfeed, breastfeeding, or breast feed" and "pacifier, dummy, or soother" as individual keywords. Only human studies published in English were included. Unpublished data were not sought. Twelve trials with weaning from exclusive breastfeeding and 19 trials with cessation of any breastfeeding were included in the meta-analysis. The meta-analysis was performed with Stata 6.0 statistical package. Summary risk ratio for early weaning before six months of age in exclusive breastfeeding trails was 2.016 (95% CI: 1.619-2.511) for pacifier users compared with nonusers in studies with univariate analysis and 1.792 (95% CI: 1.452-2.212) in studies with multivariate analysis. Similarly, pacifier usage compared with nonusers reduced the duration of any breastfeeding in both univariate (2.760, 95% CI: 2.083-3.657) and multivariate trials (1.952, 95% CI: 1.662-2.293). The use of pacifiers was associated with shortened duration of exclusive and of any breastfeeding. Given the increase in the benefits with duration of breastfeeding, parents should be informed of the link between pacifier use and shortened breastfeeding duration in order to help them make informed decisions about their children's care.


Stable Low-Grade Degenerative Spondylolisthesis Does Not Compromise Clinical Outcome of Minimally Invasive Tubular Decompression in Patients with Spinal Stenosis.
Background and Objectives: In recent literature, the routine addition of arthrodesis to decompression for lumbar spinal stenosis (LSS) with concomitant stable low-grade degenerative spondylolisthesis remains controversial. The purpose of this study is to compare the clinical outcome, complication and re-operation rates following minimally invasive (MIS) tubular decompression without arthrodesis in patients suffering from LSS with or without concomitant stable low-grade degenerative spondylolisthesis. Materials and Methods: This study is a retrospective review of prospectively collected data. Ninety-six consecutive patients who underwent elective MIS lumbar decompression with a mean follow-up of 27.5 months were included in the study. The spondylolisthesis (S) group comprised 53 patients who suffered from LSS with stable degenerative spondylolisthesis, and the control (N) group included 43 patients suffering from LSS without spondylolisthesis. Outcome measures included complications and revision surgery rates. Pre- and post-operative visual analog scale (VAS) for both back and leg pain was analyzed, and the Oswestry Disability Index (ODI) was used to evaluate functional outcome. Results: The two groups were comparable in most demographic and preoperative variables. VAS for back and leg pain improved significantly following surgery in both groups. Both groups showed significant improvement in their ODI scores, at one and two years postoperatively. The average length of hospital stay was significantly higher in patients with spondylolisthesis (p-value< 0.01). There was no significant difference between the groups in terms of post-operative complications rates or re-operation rates. Conclusions: Our results indicate that MIS tubular decompression may be an effective and safe procedure for patients suffering from LSS, with or without degenerative stable spondylolisthesis.


Shortened dental arch therapy: views of consultants in restorative dentistry in the United Kingdom.
This paper reports the results of a study on the attitudes of the consultants in restorative dentistry (CRD) in the UK towards shortened dental arch therapy (SDAT). Data were collected by means of questionnaire, with a response of 67% achieved. The results indicate that 95% (n = 87) of the 91 participants in the study were of the opinion that SDA has a place in contemporary clinical practice. Eighty (88%) of the participants reported having prescribed SDA therapy during the last 5 years, the number of these patients requiring > 5 units of crown and bridgework being typically less than 25%. Forty-one (45%) of the participants reported patients being considered for SDAT expressing reservations about the outcome of their treatment, although around 75 (82%) of the participants indicated that SDAT was satisfactory in terms of oral function, comfort and well being. Experience of having to extend shortened dental arches following SDAT was limited to 34 (37%) of the participants. It is concluded that SDA is widely accepted but not widely practiced by CRDs in the UK and that, in patients satisfying existing criteria, the outcome of SDAT may be found to be acceptable in approximately 82% of cases.


Effects of the Children's Health Insurance Reauthorization Act on immigrant children's healthcare access.
To estimate the effects of Children's Health Insurance Reauthorization Act (CHIPRA), a policy that provided states the option to extend Medicaid/CHIP eligibility to immigrant children who have not been legal residents for five years or more, on insurance coverage, access, utilization, and health outcomes among immigrant children. Restricted use 2000-2016 National Health Interview Survey (NHIS). We used a difference-in-differences design that compared changes in CHIPRA expansion states to changes in non-expansion states. Our sample included immigrant children who were born outside the US, aged 0-18 with family income below 300% of the Federal Poverty Level (FPL). Subgroup analyses were conducted across states that did and did not have a similar state-funded option prior to CHIPRA (state-funded vs. not state-funded), by the length of time in the US (5 years vs. 5-14 years), and global region of birth (Latin American vs. Asian countries). We found that CHIPRA was associated with a significant 6.35 percentage point decrease in uninsured rates (95% CI: -11.25, -1.45) and an 8.1 percentage point increase in public insurance enrollment for immigrant children (95% CI: 1.26, 14.98). However, the effects of CHIPRA became small and statistically not significant 3 years after adoption. Effects on public insurance coverage were significant in states without state-funded programs prior to CHIPRA (15.50 percentage points; 95% CI:8.05, 22.95) and for children born in Asian countries (12.80 percentage points; 95% CI: 1.04, 24.56). We found no significant changes in health care access and utilization, and health outcomes, overall and across subgroups due to CHIPRA. CHIPRA's eligibility expansion was associated with increases in public insurance coverage for low-income children, especially in states where CHIPRA represented a new source of coverage versus a substitute for state-funded coverage. However, we found evidence of crowd-out in certain subgroups and no effect of CHIPRA on access to care and health. Our results suggest that public coverage may be an important tool for promoting the well-being of immigrant children but other investments are still needed.


Evidence supporting a circadian control of natural killer cell function.
Natural killer (NK) cells participate in the immune response against infection and cancer. An emerging body of epidemiological data supports that circadian homeostasis may constitute a factor risk for cancer development. Physiological rhythms under circadian control persist in the absence of light entrainment and ultimately rely on a molecular clock. We have previously shown that NK cell cytolytic activity follows a daily rhythm and that NK cells enriched from light-entrained rats present 24-h oscillations of clock genes, cytolytic factors, and cytokines. To investigate whether these oscillations are under a genuine circadian control, we assessed the daily expression of clock genes (Per1, Per2, Clock, and Bmal1), a clock-controlled gene (Dbp), cytolytic factors (granzyme B and perforin), and cytokines (IFN-gamma and TNF-alpha) in NK cells enriched from rats maintained in constant darkness (DD). In addition, we investigated whether the disruption of the NK cell clock by RNA interference (RNAi) affects the expression of cytolytic factors and cytokines. Persistent 24-h oscillations were found in the expression levels of clock genes, cytolytic factors, and cytokines in NK cells enriched from DD rats. In addition, RNAi-mediated Per2 knockdown caused a significant decrease of granzyme B and perforin levels in the rat derived NK cell line RNK16. Taken together, these results provide evidence supporting that NK cell function is under circadian regulation.


Identification of human genetic variants controlling circular RNA expression.
Circular RNAs (circRNAs) are abundant in eukaryotic transcriptomes and have been linked to various human disorders. However, understanding genetic control of circular RNA expression is in the early stages. Here we present the first integrated analysis of circRNAs and genome sequence variation from lymphoblastoid cell lines of the 1000 Genomes Project. We identified thousands of circRNAs in the RNA-seq data and show their association with local single-nucleotide polymorphic sites, referred to as circQTLs, which influence the circRNA transcript abundance. Strikingly, we found that circQTLs exist independently of eQTLs with most circQTLs having no effect on mRNA expression. Only a fraction of the polymorphic sites are shared and linked to both circRNA and mRNA expression with mostly similar effects on circular and linear RNA. A shared intronic QTL, rs55928920, of HMSD gene drives the circular and linear expression in opposite directions, potentially modulating circRNA levels at the expense of mRNA. Finally, circQTLs and eQTLs are largely independent and exist in separate linkage disequilibrium (LD) blocks with circQTLs highly enriched for functional genomic elements and regulatory regions. This study reveals a previously uncharacterized role of DNA sequence variation in human circular RNA regulation.


Single-walled carbon nanotubes exhibit limited transport in soil columns.
The increased production and commercial use of nanomaterials combined with a lack of regulation to govern their disposal may result in their introduction to soils and ultimately into groundwater systems. In this study, we investigated the transport behavior of carboxyl-functionalized single-walled carbon nanotubes (SWNTs) in columns packed with a natural soil. In general, SWNT deposition (filtration) rate increased with increasing solution ionic strength, with divalent cations (Ca(2+)) being more effective in increasing SWNT retention than monovalent cations (K(+)). However, SWNT deposition rate over a very wide range of monovalent and divalent cation concentrations (0.03 to 100 mM) was relatively high and changed only slightly above 0.3 mM KCl or 0.1 mM CaCl(2). In contrast, filtration of another type of engineered carbon-based nanomaterial, namely aqueous fullerene (C(60)) nanoparticles (radius of 51 nm), was more sensitive to solution ionic strength, displaying lower deposition rate and more effective transport in soil than SWNTs. These observations indicate that physical straining governs SWNT filtration and transport under all the solution chemistries investigated in the present study. It is proposed that SWNT shape and structure, particularly the very large aspect ratio and its highly bundled (aggregated) state in aqueous solutions, as well as the heterogeneity in soil particle size, porosity, and permeability, collectively contribute to straining in flow through soil media. Our results suggest that SWNTs of comparable properties to those used in the present study will not exhibit substantial transport and infiltration in soils because of effective retention by the soil matrix.


When catchers meet - a computational study on the dimerization of the Buckycatcher.
We study the dimerization of the buckycatcher in gas phase and in toluene. We created an extensive library of 36 different complexes, which were characterized at semi-empirical and DFT levels. Semi-empirical geometries and dimerization energies compare well against reference data or Density Functional Theory calculations we performed. Born-Oppenheimer molecular dynamics was used to understand what happens when two molecules of the buckycatcher meet, allowing us to infer on the lack of kinetic barriers when dimers form. Thermodynamically, it is possible that room temperature solutions contain dimerized buckycatcher. Using a very simple exchange model, it is shown, however, that dimerization cannot compete thermodynamically against complexation with fullerenes, which accounts for experimental observations.


Simulation study of amplitude-modulated (AM) harmonic motion imaging (HMI) for stiffness contrast quantification with experimental validation.
The objective of this study is to show that Harmonic Motion Imaging (HMI) can be used as a reliable tumor-mapping technique based on the tumor's distinct stiffness at the early onset of disease. HMI is a radiation-force-based imaging method that generates a localized vibration deep inside the tissue to estimate the relative tissue stiffness based on the resulting displacement amplitude. In this paper, a finite-element model (FEM) study is presented, followed by an experimental validation in tissue-mimicking polyacrylamide gels and excised human breast tumors ex vivo. This study compares the resulting tissue motion in simulations and experiments at four different gel stiffnesses and three distinct spherical inclusion diameters. The elastic moduli of the gels were separately measured using mechanical testing. Identical transducer parameters were used in both the FEM and experimental studies, i.e., a 4.5-MHz single-element focused ultrasound (FUS) and a 7.5-MHz diagnostic (pulse-echo) transducer. In the simulation, an acoustic pressure field was used as the input stimulus to generate a localized vibration inside the target. Radiofrequency (rf) signals were then simulated using a 2D convolution model. A one-dimensional cross-correlation technique was performed on the simulated and experimental rf signals to estimate the axial displacement resulting from the harmonic radiation force. In order to measure the reliability of the displacement profiles in estimating the tissue stiffness distribution, the contrast-transfer efficiency (CTE) was calculated. For tumor mapping ex vivo, a harmonic radiation force was applied using a 2D raster-scan technique. The 2D HMI images of the breast tumor ex vivo could detect a malignant tumor (20 x 10 mm2) surrounded by glandular and fat tissues. The FEM and experimental results from both gels and breast tumors ex vivo demonstrated that HMI was capable of detecting and mapping the tumor or stiff inclusion with various diameters or stiffnesses. HMI may thus constitute a promising technique in tumor detection (>3 mm in diameter) and mapping based on its distinct stiffness.


Drug resistance towards etoposide and cisplatin in human melanoma cells is associated with drug-dependent apoptosis deficiency.
Anticancer drugs kill susceptible cells through induction of apoptosis. Alterations of apoptotic pathways in drug-resistant tumor cells leading to apoptosis deficiency might represent a potent mechanism conferring drug resistance. We have assessed the effect of etoposide and cisplatin on the apoptotic pathways of the drug-sensitive human melanoma cell line MeWo as well as its etoposide- and cisplatin-resistant sublines (MeWo(Eto01), MeWo(Eto1), (and) MeWoCis01, MeWo(Cis1)). Etoposide and cisplatin induced apoptosis in drug-sensitive MeWo cells as indicated by dose-dependent (i) cytochrome c release, (ii) caspase activation, (iii) DNA fragmentation, and (iv) cleavage of poly(ADP-ribose)polymerase. In contrast, whereas low etoposide-resistant cells (MeWo(Eto01)) demonstrated reduced but detectable apoptotic activities, highly etoposide-resistant cells (MeWo(Eto1)) did not exhibit any of the apoptotic events observed in etoposide-induced cell death downstream of a strongly reduced cytochrome c release. Highly cisplatin-resistant cells (MeWo(Cis1)), however, demonstrated a reduced caspase 9 activity and cytochrome c release but the extent of effector caspase activation as well as DNA fragmentation was comparable to that of sensitive MeWo cells at equitoxic concentrations. In addition, poly(ADP-ribose)polymerase cleavage was strongly reduced in highly cisplatin-resistant sublines. Taken together, sensitive and drug-resistant MeWo cells utilized different apoptotic pathways upon drug exposure in a drug-dependent fashion and apoptosis deficiency was strongly associated with the drug-resistant phenotype.


Long-term expression of the lamin A mutant associated with dilated cardiomyopathy induces senescence.
Mutation of the lamin A gene (LMNA) causes a diverse range of diseases referred to as laminopathies. Because most laminopathies have a dominant inheritance pattern and progress gradually, cultured cells stably expressing mutant lamin A at the same level as endogenous wild-type cells are required for chronological analysis. In this study, we showed that an expression system involving a lentiviral vector that carries the human metallothionein gene basal promoter ensures stable and basal-level expression of proteins and is thus suitable for investigating the properties of lamin A mutants. The small ubiquitin-related modifier (SUMO) modification (SUMOylation)-defective E203G mutant that is associated with familial dilated cardiomyopathy exhibited abnormal subnuclear distribution and inhibited normal localization of WT lamin A in a dominant-negative manner. Low-level and long-term expression of the E203G mutant resulted in multinucleated giant cells, aberrant lipid droplet accumulation in the cytoplasm and premature senescence. Expression of another SUMOylation-defective mutant (K201R) did not induce any phenotypes observed in cells expressing E203G. These results indicate that the E203G mutant may inhibit the normal functions of wild-type lamin A in a dominant-negative manner, but a defect in SUMOylation itself may not be involved in disease pathogenesis.


Assessing alexithymia in French-speaking samples: psychometric properties of the Observer Alexithymia Scale-French translation.
In the present study, we evaluated the psychometric properties of the Observer Alexithymia Scale-French translation (OAS-F), a 33-item, observer-rated alexithymia measure. The scale, accessible to lay and professional raters, taps everyday expressions of alexithymia. French university students (N = 159) were asked to rate a person they knew well or ask an acquaintance to rate them. Those being rated (N = 159) were parents, siblings, children, and friends. OAS-F total and subscale scores were comparable to those in the English normative samples. Moreover, OAS scores were reliable, and the scale's five-factor structure (distant, uninsightful, somatizing, humorless, and rigid) was confirmed. Importantly, too, OAS total scores correlated 0.31 with (self-report) 20-item Toronto Alexithymia Scale (TAS) scores. The OAS-F appears to be a psychometrically sound observer-rated alexithymia measure.


Detailed evaluation of pyruvate dehydrogenase complex inhibition in simulated exercise conditions.
The mammalian pyruvate dehydrogenase complex (PDC) is a mitochondrial multienzyme complex that connects glycolysis to the tricarboxylic acid cycle by catalyzing pyruvate oxidation to produce acetyl-CoA, NADH, and CO2. This reaction is required to aerobically utilize glucose, a preferred metabolic fuel, and is composed of three core enzymes: pyruvate dehydrogenase (E1), dihydrolipoyl transacetylase (E2), and dihydrolipoyl dehydrogenase (E3). The pyruvate-dehydrogenase-specific kinase (PDK) and pyruvate-dehydrogenase-specific phosphatase (PDP) are considered the main control mechanism of mammalian PDC activity. However, PDK and PDP activity are allosterically regulated by several effectors fully overlapping PDC substrates and products. This collection of positive and negative feedback mechanisms confounds simple predictions of relative PDC flux, especially when all effectors are dynamically modulated during metabolic states that exist in physiologically realistic conditions, such as exercise. Here, we provide, to our knowledge, the first globally fitted, pH-dependent kinetic model of the PDC accounting for the PDC core reaction because it is regulated by PDK, PDP, metal binding equilibria, and numerous allosteric effectors. The model was used to compute PDH regulatory complex flux as a function of previously determined metabolic conditions used to simulate exercise and demonstrates increased flux with exercise. Our model reveals that PDC flux in physiological conditions is primarily inhibited by product inhibition (∼60%), mostly NADH inhibition (∼30-50%), rather than phosphorylation cycle inhibition (∼40%), but the degree to which depends on the metabolic state and PDC tissue source.


The neurology clerkship core curriculum.
Neurologic symptoms are common in all practice settings, and neurologic diseases comprise a large and increasing proportion of health care expenditures and global disease burden. Consequently, the training of all physicians should prepare them to recognize patients who may have neurologic disease, and to take the initial steps in evaluating and managing those patients. We present a core curriculum outlining the clinical neurology skills and knowledge necessary to achieve that degree of preparation. The curriculum emphasizes general principles and a systematic approach to patients with neurologic symptoms and signs. The ability to perform and interpret the neurologic examination is fundamental to that approach, so the curriculum delineates the essential components of the examination in three different clinical settings. The focus of the curriculum is on symptom-based rather than disease-based learning. The only specific diseases selected for inclusion are conditions that are common or require urgent management. This curriculum has been approved by the national organization of neurology clerkship directors and endorsed by the major national professional organizations of neurologists. It is intended as a template for planning a neurology clerkship and as a benchmark for evaluating existing clerkships. It should be especially helpful to clerkship directors, neurology chairs, deans of medical education, and members of external accreditation groups.


Platelet CD40L mediates thrombotic and inflammatory processes in atherosclerosis.
CD40 ligand (CD40L), identified as a costimulatory molecule expressed on T cells, is also expressed and functional on platelets. We investigated the thrombotic and inflammatory contributions of platelet CD40L in atherosclerosis. Although CD40L-deficient (Cd40l(-/-)) platelets exhibited impaired platelet aggregation and thrombus stability, the effects of platelet CD40L on inflammatory processes in atherosclerosis were more remarkable. Repeated injections of activated Cd40l(-/-) platelets into Apoe(-/-) mice strongly decreased both platelet and leukocyte adhesion to the endothelium and decreased plasma CCL2 levels compared with wild-type platelets. Moreover, Cd40l(-/-) platelets failed to form proinflammatory platelet-leukocyte aggregates. Expression of CD40L on platelets was required for platelet-induced atherosclerosis as injection of Cd40l(-/-) platelets in contrast to Cd40l(+/+) platelets did not promote lesion formation. Remarkably, injection of Cd40l(+/+), but not Cd40l(-/-), platelets transiently decreased the amount of regulatory T cells (Tregs) in blood and spleen. Depletion of Tregs in mice injected with activated Cd40l(-/-) platelets abrogated the athero-protective effect, indicating that CD40L on platelets mediates the reduction of Tregs leading to accelerated atherosclerosis. We conclude that platelet CD40L plays a pivotal role in atherosclerosis, not only by affecting platelet-platelet interactions but especially by activating leukocytes, thereby increasing platelet-leukocyte and leukocyte-endothelium interactions.


Regression of the Left Ventricular Hypertrophy in Patients with Essential Hypertension on Standard Drug Therapy.
PURPOSE: The American College of Cardiology/ American Heart Association 2017 and European Society of Cardiology/European Society of Hypertension 2018 guidelines were a paradigm shift in hypertension management in contemporary medicine. Lowering of blood pressure to less than 130 (systolic) and 80 (diastolic) mm of Hg irrespective of cardiovascular risk is recommended. While intensive blood pressure control is commonly achievable with rational pharmacotherapy, the magnitude of left ventricular hypertrophy regression is an independent factor in improvement in cardiovascular health. The regression of left ventricular hypertrophy has been adjudged as a clinically useful surrogate marker that reflects the efficacy of hypertension treatment. Though angiotensin converting enzyme inhibitors/ angiotensin receptor blockers (ACEI/ARB) are the preferred initial drug for greater regression of left ventricular mass, the choice of add-on therapy, if required, is still debatable. Therefore, in our observational study, we sought to compare the reduction in left ventricular mass index in hypertensives with left ventricular hypertrophy on standard ACEI/ARB based drug therapy. MATERIALS AND METHODS: The cohort (n=217) comprised of patients with uncontrolled hypertension (blood pressure>140/90 mm of Hg) and left ventricular hypertrophy (left ventricular mass index>115 and 95 gram/square meter in males and females respectively). The add-on drug in ACEI/ARB therapy was either thiazide diuretics (TD) or calcium channel blockers (CCB). Four sub-cohorts were constituted: mono-therapy - group A (n=70, ACEI/ARB), dual-therapy - group B (n=48, ACEI/ARB+TD) and group C (n=51, ACEI/ ARB+CCB), triple therapy - group D (n=48, ACEI/ ARB+TD+CCB). Left ventricular mass index was determined using echocardiography at baseline and after 24 weeks of therapy. RESULTS: There was no significant difference in baseline clinical or demographic variables between group B and group C. Baseline blood pressure and duration of hypertension was greater in group D compared to group A (P<0.001). The reduction in left ventricular mass index (mean ±SD) in the four groups (A to D) was 16.7±18.7, 21.0±20.8, 20.5±15.5 and 29.1±21.5 g/m2 respectively (D>A, P=0.011, B versus C, P=1.00). The corresponding change in blood pressure (systolic/diastolic) was 18.5±13.6/8.9±11.2, 27.5±19.2/12.2±9.3, 23.4±16.7/ 5.4±10.1, 26.6±19.5/10.7±12.8 mm of Hg respectively (systolic, B>A, P=0.027, D>A, P=0.048) (diastolic, B>C, P=0.013). CONCLUSION: Anti-hypertensive treatment with angiotensin converting enzyme inhibitors/angiotensin receptor blockers-based therapy produced graded regression of left ventricular hypertrophy with monotherapy, dual therapy and triple therapy. In dual therapy, add-on of either thiazide diuretics or calcium channel blockers to angiotensin converting enzyme inhibitors/angiotensin receptor blockers showed equal efficacy in regression of left ventricular hypertrophy independent of blood pressure reduction.


On the N2O correction used for mass spectrometric analysis of atmospheric CO2.
To obtain accurate values of delta(13)C(CO(2)) and delta(18)O(CO(2)) on environmental CO(2) by mass spectrometry, the raw isotope data must be corrected for the isobaric N(2)O contribution. This is one of the analytical problems limiting inter-laboratory delta(13)C(CO(2)) data consistency. The key parameter, the N(2)O relative ionisation efficiency (E(N2O)), cannot be determined with sufficient accuracy by direct measurements of pure N(2)O. The determination of (E(N2O)) by analyses on N(2)O--CO(2) mixtures of known isotope composition and mixing proportions has been recently suggested. In this work we propose a new method of N(2)O correction which uses the m/z 30 signal as a measure of the N(2)O/CO(2) ratio, so that determinations of (E(N2O)) and N(2)O content are not required. The method uses the fact that fragment-ion spectra of N(2)O and CO(2) are very specific. The formalism of the correction is considered. Various tests demonstrate that the new method is robust, stable and easy to implement in practice. The effective value (E(N2O)) (the key parameter for the new correction) has to be calibrated on known N(2)O--CO(2) mixtures by measuring (30)R signals only. The method accuracy we presently achieved is around 2.5% and any error which appears to come mostly from our N(2)O--CO(2) mixture preparation. Based on our tests and error considerations, the error of the proposed method that may be achieved is as low as +/-1.5% (relative to the correction magnitude). For tropospheric CO(2) this means +/-0.003 per thousand and +/-0.005 per thousand for delta(13)C(CO(2)) and delta(18)O(CO(2)), respectively. The proposed method may be valuable for small samples where no separate N(2)O determinations are available (e.g. ice core samples and CF-IRMS measurements) as well as for determination of (E(N2O)) and testing the 'traditional' N(2)O correction based on mass balance calculations.


Xenopus oocytes express a unitary glutamate receptor endogenously.
The results described here demonstrate that Xenopus oocytes endogenously express a unitary glutamate receptor subunit XenU1. The level of XenU1 mRNA expression reaches approximately 1/300 of that in the adult Xenopus brain. The endogenous expression of XenU1, which can functionally interact with N-methyl-D-aspartate receptor subunit NR1, explains the differences in NR1 subunit expression in mammalian cell lines (no functional expression without partner subunits) and in the Xenopus oocytes (NR1 forms functional receptors when expressed singly).


Polar Positioning of Phase-Separated Liquid Compartments in Cells Regulated by an mRNA Competition Mechanism.
P granules are non-membrane-bound RNA-protein compartments that are involved in germline development in C. elegans. They are liquids that condense at one end of the embryo by localized phase separation, driven by gradients of polarity proteins such as the mRNA-binding protein MEX-5. To probe how polarity proteins regulate phase separation, we combined biochemistry and theoretical modeling. We reconstitute P granule-like droplets in vitro using a single protein PGL-3. By combining in vitro reconstitution with measurements of intracellular concentrations, we show that competition between PGL-3 and MEX-5 for mRNA can regulate the formation of PGL-3 droplets. Using theory, we show that, in a MEX-5 gradient, this mRNA competition mechanism can drive a gradient of P granule assembly with similar spatial and temporal characteristics to P granule assembly in vivo. We conclude that gradients of polarity proteins can position RNP granules during development by using RNA competition to regulate local phase separation.


Glucosinolate-related glucosides in Alliaria petiolata: sources of variation in the plant and different metabolism in an adapted specialist herbivore, Pieris rapae.
Specialized metabolites in plants influence their interactions with other species, including herbivorous insects, which may adapt to tolerate defensive phytochemicals. The chemical arsenal of Alliaria petiolata (garlic mustard, Brassicaceae) includes the glucosinolate sinigrin and alliarinoside, a hydroxynitrile glucoside with defensive properties to glucosinolate-adapted specialists. To further our understanding of the chemical ecology of A. petiolata, which is spreading invasively in North America, we investigated the metabolite profile and here report a novel natural product, petiolatamide, which is structurally related to sinigrin. In an extensive study of North American populations of A. petiolata, we demonstrate that genetic population differences as well as developmental regulation contribute to variation in the leaf content of petiolatamide, alliarinoside, sinigrin, and a related glycoside. We furthermore demonstrate widely different metabolic fates of these metabolites after ingestion in the glucosinolate-adapted herbivore Pieris rapae, ranging from simple passage over metabolic conversion to sequestration. The differences in metabolic fate were influenced by plant β-glucosidases, insect-mediated degradation, and the specificity of the larval gut transport system mediating sequestration.


Evaluation of serum sialic acid and carcinoembryonic antigen for the detection of early-stage colorectal cancer.
Various expressions of elevated serum sialic acid (total sialic acid, TSA: lipid-associated sialic acid, LASA; LASA/TSA; TSA normalized to total protein, TSA/TP) have been evaluated and compared with increased serum carcinoembryonic antigen (CEA) levels for the detection of early-stage colorectal cancer. This evaluation was done blindly on a coded panel of 320 sera from staged colorectal cancer patients and controls provided by the Mayo Clinic--National Cancer Institute Diagnostic Bank. Unlike the findings of a previous preliminary study (Tautu et al., JNCI 80:1333-1337, 1988), the ratio of LASA/TSA was not useful for detecting early-stage (Dukes A and B) colorectal cancer. However, TSA and TSA/TP values were significantly elevated in each colorectal cancer subgroup compared with normal controls. TSA and TSA/TP values displayed a marginally better discriminatory power than CEA values in the case of Dukes A subgroup with respect to normal controls. CEA still appears to be the best single overall marker for discriminating between colorectal cancers and controls. However, multiple marker analysis using CEA and TSA (and related markers) appears to be more sensitive than CEA alone for detecting colorectal cancer.


Prescribing exercise for health: a simple framework for primary care.
Physicians need to become committed to promoting the preventive benefits of regular physical activity. Practical strategies can help physicians efficiently and effectively incorporate exercise counseling into their practices. These strategies include regularly asking about leisure activity and linking the agenda of an office visit to the benefits of exercise. The physician and the patient should work together to formulate a mutually acceptable plan for the patient to adopt and maintain a healthful exercise lifestyle. In the same way that prescriptions are written for medications, a prescription can be given for an exercise plan suited to the individual patient.


Molecular genetics of medullary thyroid carcinoma: the quest for novel therapeutic targets.
Medullary thyroid carcinoma (MTC) is a rare tumour arising from neural crest-derived parafollicular C-cells. Metastatic MTC patients are incurable because the cancer does not respond to radiotherapy or chemotherapy. The REarranged during Transfection (RET) proto-oncogene plays a key role in the development of MTC. However, one-half of the sporadic MTC do not carry RET mutations. Mice models and early evidence obtained in human samples suggest that other genes, including those encoding components of the RB1 (retinoblastoma) and TP53 tumour-suppressor pathways, may be involved in MTC formation. Here, we review the data on the involvement of genes acting in the RET and RB1/TP53 pathways in MTC. Understanding genetic lesions that occur in MTC is a prerequisite to identifying molecular therapeutic targets in MTC and in improving the efficacy of RET-targeted therapies.


Morphological study of cationic polymer-anionic surfactant complex precipitated in solution during the dilution process.
We investigated the phase diagrams and the morphology of the complexes that were formed by cationic polymers, cationic cellulose (CC) and cationic dextran (CD), and by anionic surfactant-based sodium poly(oxyethylene) lauryl ether sulfate (LES). The anionic charge of the LES-based surfactants was changed by adding an amphoteric surfactant, lauryl amidopropyl betaine acetate (LPB), or a nonionic surfactant, polyoxyethylene stearyl ether (C18EO25). We discuss the relationship between the complex aggregation process and the morphology of the precipitated complexes. The morphologies of CC complex aggregates, which precipitated during the dilution process in a model shampoo solution, changed from membranous forms to mesh-like forms by decreasing the charges of both the CC and the surfactant. Their touch on hair in the rinsing process changed from sticky to smooth and velvety, corresponding to their rheological properties. In contrast, CD complex aggregates had a membranous form and a smooth touch independently of the charges on the polymer and surfactant. These results suggested that the control of the charges of both the polymer and surfactant and the choice of polymer structure are important for excellent conditioning effects upon rinsing with shampoo.


Prospective study of external cephalic version in Glasgow: patient selection, outcome and factors associated with outcome.
Data were collected prospectively on all 67 women who underwent an attempt at external cephalic version (ECV) over 1 year in the four Glasgow maternity hospitals. Ultrasonography was used in all women. However, tocolytics were used in only two (6%) nulliparous women despite published evidence of their efficacy. Only 25 (37%) women undergoing ECV had a free presenting part which is known to be associated with success. Seventeen (25%) women were less than 37 weeks pregnant despite spontaneous version being common at this stage. ECV was successful in only 26 (39%) women and only 18 (27%) had a vaginal cephalic delivery. These results compare unfavourably with published results of around two-thirds for both end-points. Although publication bias is likely, patient selection, under-usage of tocolytics and lack of experience may also be factors. Consideration should be given to a reduced number of operators who can maximise their throughput and expertise.


Flow-rate fluctuations in the outpouring of grains from a two-dimensional silo.
We present experimental results obtained with a two-dimensional silo discharging under gravity through an orifice at the flat bottom. High-speed measurements provide enough time resolution to detect every single bead that goes out and this allows the measurement of the flow rate in short-time windows. Two different regimes are clearly distinguished: one for large orifices, which can be described by Gaussian fluctuations, and another for small orifices, in which extreme events appear. The frontier between those two regimes coincides with the outlet size below which jamming events are frequent. Moreover, it is shown that the power spectrum of the flow-rate oscillations is not dominated by any particular frequency.


Biochemical Plant Responses to Ozone : II. Induction of Stilbene Biosynthesis in Scots Pine (Pinus sylvestris L.) Seedlings.
Formation of the stilbenes pinosylvin and pinosylvin 3-methyl ether, as well as the activity of the biosynthetic enzyme stilbene synthase (pinosylvin-forming), were induced several hundred- to thousandfold in primary needles of 6-week-old pine (Pinus sylvestris L.) seedlings upon exposure to a single pulse of ozone of at least 0.15 microliters per liter. The seedlings required 4 hours of exposure as a minimum for the induction of stilbene biosynthesis when exposed to 0.2 microliters per liter ozone. Both stilbene synthase activity and stilbene accumulation increased with the duration of ozone treatment. The activity of phenylalanine ammonia-lyase and the activity of chalcone synthase, a key enzyme of the flavonoid pathway that uses the same substrates as stilbene synthase, were also stimulated about twofold by ozone. Stilbene biosynthesis appears to represent the first example of a dose-dependent biochemical response to ozone in a conifer species and may serve as a useful biomarker to study stress impacts on pine trees.


Prognostic significance of α5β1-integrin expression in cervical cancer.
The purpose of this study was to evaluate the association of expression of α5β1-integrin with clinicopathologic features and prognosis in cervical cancer. Levels of α5β1-integrin in normal cervical mucosa and cervical cancer tissue were detected with immunohistochemistry. Survival analysis by the Kaplan-Meier method was performed to assess prognostic significance. α5β1-integrin expression was detected in 84.6% (143/169) cervical cancer samples, significantly different from that in normal cervical mucosa (P < 0.05). Positive expression rates of α5β1-integrin in patients with poor histologic differentiation, lymph node metastasis, and recurrence were elevated. Using Kaplan-Meier analysis, a comparison of survival curves of low versus high expression of α5β1-integrin revealed a highly significant difference in human cervical cancer cases (P < 0.05), suggesting that overexpression of α5β1-integrin is associated with a worse prognosis.The α5β1-integrin promotes angiogenesis and associates with lymph node metastasis, vascular invasion and poor prognosis of cervical cancer. The current study indicated that α5β1-integrin may be an independent prognostic factor for cervical cancer patients.


A Scoping Review of Climate Change, Climate-Related Disasters, and Mental Disorders among Children in Low- and Middle-Income Countries.
Children, particularly those living in low- and middle-income countries (LMICs), are highly vulnerable to climate change and its impacts. Our main objective was to conduct a scoping literature review to determine how exposure to climate change and climate-related disasters influences the presence of mental disorders among children in LMICs. We also aimed to identify gaps in this area of scholarship. We included studies of children in LMICs that had a climate change or climate-related disaster exposure and mental disorder outcome. Twenty-three studies were included in the final synthesis. Fourteen studies were conducted in China, three in India, two each in Pakistan and the Philippines, and one each in Namibia and Dominica. All studies assessed the association between a climate-related disaster exposure and a mental disorder outcome, while none explored broader climate change-related exposures. Post-traumatic stress disorder (n = 21 studies) and depression (n = 8 studies) were the most common mental disorder outcomes. There was considerable between-study heterogeneity in terms of sample size, follow-up length, and outcome measurement. Overall, the literature in this area was sparse. Additional high-quality research is required to better understand the impacts of climate-related disasters and climate change on mental disorders within this population to ultimately inform future policies and interventions.


Chromium (III) thiamin diphosphate: a new probe of enzyme coenzyme--metal ion binding. Interaction with wheat germ pyruvate decarboxylase.
Chromium (III) thiamin diphosphate (CrTDP) is a substitutionally inert complex which is a physical and kinetic probe of the binding mode of metal ion and coenzyme in thiamin diphosphate (TDP) dependent enzymes. CrTDP is prepared by reaction of aquated Cr (III) and TDP and purifed by ion-exchange chromatography. CrTDP binds to the apoenzyme of wheat germ pyruvate decarboxylase, giving an inactive holoenzyme. Chromium (III) ion binds to the apoenzyme in a manner that suggests, in light of the results with CrTDP, that there are two metal ion binding sites. Extension of the use of CrTDP to other enzymes can give specific information about binding and can introduce an active site reporter group


Cardiorespiratory physical assessment for the acutely ill: 1.
This article, the first of two parts, aims to develop core cardiovascular and respiratory knowledge and to enhance practice for the practitioner caring for an acutely ill patient on a general ward setting. The second part will focus on advanced cardiorespiratory assessment. With increasing acuity of illness in general ward settings, patients' needs are becoming more complex in terms of their nursing and medical management. Nurses must be able to respond to subtle clinical signs and symptoms and act appropriately to ensure that a safe, patient-focused environment is maintained and that the care delivered is evidence-based. By increasing knowledge of the theory and practice of nursing physical assessment, and the ensuing management, acutely ill patients nursed in general ward areas may be provided with timely and appropriate care and may avoid further deterioration and an intensive care admission. This article describes core physical assessment skills, the normal physiology of the cardiovascular system, and the management of cardiovascular and respiratory dysfunction.


Genome-wide identification of chemosensitive single nucleotide polymorphism markers in gastric cancer.
A chemosensitive single nucleotide polymorphism (SNP) discovery schema is presented that utilizes (i) genome-wide SNP screening, with a human SNP array and an in vitro chemosensitivity assay, in 93 patients with gastric cancer (GC), and (ii) biological utility assessment using cell viability assays of transfected GC cells. Cytotoxicity analysis showed that most of the MKN1 and SNU638 clones transfected with the G allele of Deoxyribonuclease II beta (DNASE2B) rs3738573 were more sensitive to docetaxel than those with the C allele (p≤0.001-0.029) and most of the AGS and SNU638 clones transfected with the T allele of 5-hydroxytryptamine receptor IE (HTRIE) rs3828741 were more sensitive to paclitaxel than those with the C allele (p≤0.001-0.019). Our findings show that the two novel markers, DNASE2B rs3738573 and HTR1E rs3828741, have potential for improving the prediction of chemosensitivity of GC patients.


A reciprocal regulatory circuit between CD44 and FGFR2 via c-myc controls gastric cancer cell growth.
Despite their suggested importance, the mechanistic roles of FGFR2 and gastric cancer stem cell (GCSC) marker CD44 remain unclear. We investigated cross talk between CD44 and FGFR2. FGFR2 and CD44 positively regulate each other's expression. While FGFR2 suppresses c-Myc transcription, CD44 activates it. c-Myc in turn augments FGFR2 transcription. CD44 knockdown (KD) depleted FGFR2 and other GCSC markers, decreased c-Myc and Sox2 expression, and suppressed tumor growth, whereas CD44 activation led to FGFR2 induction. FGFR2 KD decreased most GCSC marker expression, including CD44, but increased c-Myc and Sox2 expression and attenuated tumor growth. FGFR2 kinase inhibitor and FGFR2 neutralizing antibody decreased the CD44+/hi GCSC fraction. Conversely, FGFR2 overexpression increased CD44 and accelerated tumor growth in mice. FGFR2 was co-expressed and colocalized diffusively with CD44, EpCAM, and LGR5. In contrast, phospho-FGFR2 colocalized densely with CD44, forming an aggregated signaling complex that was prevented by FGFR2 inhibition. The c-Myc KD depleted FGFR2 but not CD44. Similarly to CD44+/hi phenotypes, sorted FGFR+/hi cells had larger volumes, formed more tumor spheres, grew faster in vivo with bigger tumor mass, and expressed more CD44, EpCAM, and HER2. These findings suggest that FGFR2+/hi cells have stemness properties. Moreover, in situ FGFR2 expression in patient-derived gastric cancer tissue correlated with tumorigenic potential in a xenograft model. In conclusion, CD44 and FGFR2 maintain stemness in gastric cancer by differentially regulating c-Myc transcription.


Transient monocular visual loss from narrow-angle glaucoma.
Three patients had transient monocular visual loss from subacute attacks of narrow-angle glaucoma. In all three cases the visual loss had been previously diagnosed as being due to other neurologic causes.


Detection of parvalbumin, a common fish allergen gene in food, by real-time polymerase chain reaction.
Fish, as one of the most common causes of IgE-mediated food hypersensitivity, has recently received increasing attention from the food industry and legislative and regulatory agencies. A real-time polymerase chain reaction assay based on TaqMan-MGB probe technology was developed for the detection of parvalbumin, a major fish allergen gene. The assay had a sensitivity up to 5 pg purified fish DNA and had no cross-reaction with other species, such as cattle, sheep, swine, chicken, shrimp, lobster, crab, squid, clam, rice, soybean, maize, and potato. The coefficient of variation for both intra- and interexperimental variability demonstrated high reproducibility and accuracy. The assay proved to be a potential tool for the detection and label management of fish allergens in food.


The epidemiology of preterm birth.
Secular trends in the prevalence of preterm birth and international comparisons of the rates of preterm birth are difficult to interpret because of differences, both formal and informal, in the registration of extremely preterm births. Accurate estimation of gestational age is another problem in the measurement of preterm birth. Preterm birth is heterogeneous in several ways. It is heterogeneous in terms of the extent to which the birth is preterm (20-27 weeks, 28-31 weeks or 32-36 weeks of gestation); in whether the birth was elective or spontaneous; and among spontaneous idiopathic preterm births, in whether there was preterm labour or premature rupture of the membranes. Case-control study designs taking account of these subgroups have been a recent feature of epidemiologic approaches. The classic social associations of preterm birth--low socioeconomic status, extremes of maternal age, primiparity, being unmarried--apply to extremely preterm and moderately preterm births as well as to the mildly preterm group. The strength of these associations is small compared with factors in the prior reproductive history and with medical and obstetric complications of the current pregnancy. Recent epidemiological research activities have focused on the ways in which risk factors such as physical workload, drugs and alcohol, lack of social support and infection might be mediating factors between sociodemographic status and preterm birth. As Eastman (1947) pointed out almost 50 years ago, 'only when the factors causing prematurity are clearly understood can any intelligent attempt at prevention be made'.


Personifications of personal and typical death as related to death attitudes.
The present article examined differences in personifications of personal and typical death as a function of attitudes about death. Ninety-eight students enrolled in psychology classes were randomly assigned to personify death as a character in a movie depicting either their own deathbed scene or the deathbed scene of the typical person prior to completing the Death Attitude Profile-Revised. The results supported the conceptual distinction between attitudes about personal death and death in general. Participants in the personal death condition personified death more frequently as a gentle-comforting image and less frequently as a cold-remote image than did participants in the typical death condition. The results also further validated the relation between personifications of death and death attitudes. Across both conditions, participants who selected the grim-terrifying image reported more fear of death and death avoidance; whereas, participants who selected the cold-remote or robot-like images reported more neutral acceptance.


Multipotent progenitor cells are present in human peripheral blood.
To determine whether the peripheral blood in humans contains a population of multipotent progenitor cells (MPCs), products of leukapheresis were obtained from healthy donor volunteers following the administration of granulocyte colony-stimulating factor. Small clusters of adherent proliferating cells were collected, and these cells continued to divide up to 40 population doublings without reaching replicative senescence and growth arrest. MPCs were positive for the transcription factors Nanog, Oct3/4, Sox2, c-Myc, and Klf4 and expressed several antigens characteristic of mesenchymal stem cells. However, they were negative for markers of hematopoietic stem/progenitor cells and bone marrow cell lineages. MPCs had a cloning efficiency of approximately 3%, and following their expansion, retained a highly immature phenotype. Under permissive culture conditions, MPCs differentiated into neurons, glial cells, hepatocytes, cardiomyocytes, endothelial cells, and osteoblasts. Moreover, the gene expression profile of MPCs partially overlapped with that of neural and embryonic stem cells, further demonstrating their primitive, uncommitted phenotype. Following subcutaneous transplantation in nonimmunosuppressed mice, MPCs migrated to distant organs and integrated structurally and functionally within the new tissue, acquiring the identity of resident parenchymal cells. In conclusion, undifferentiated cells with properties of embryonic stem cells can be isolated and expanded from human peripheral blood after granulocyte colony-stimulating factor administration. This cell pool may constitute a unique source of autologous cells with critical clinical import.


Removal of total nitrogen from wastewater by a combination of Chlorella sp. and audible sound.
In developing countries, nitrogen in the traditional market wastewater is a critical environmental problem. In this study, the microalga Chlorella sp., which was isolated from wastewater, was used to remove the total nitrogen (TN) from conventional market wastewater in combination with audible sound (Vietnamese classical music). In addition, effects of sound exposure on removal efficiency at different initial cell densities were analyzed. Results revealed that music sound control demonstrates potential to improve the removal efficiency. TN removal efficiencies of 96%, 69.5%, and 4.3% were observed for treatments with Chlorella sp./audible sound, Chlorella sp., and without Chlorella sp., respectively. The significance of probability value (p-value) (<0.05) on the paired sample t-test confirmed the critical role of audible sound and Chlorella sp. density on the TN removal in screening experiments. The predicted optimal conditions for TN removal were as follows: a Chlorella sp. density of 4%, an audible sound of 52.5 dB, and a cultivation time of 4.6 days. Results based on statistical analysis revealed that the quadratic models for TN removal are significant at a low p-value (<0.05) and a high predicted coefficient of determination (R2 = 0.9452) value. The obtained statistical results also indicated that most of the variables are significant for the abatement of TN from market wastewater using Chlorella sp.