MinDE-dependent pole-to-pole oscillation of division inhibitor MinC in Escherichia coli.
By inhibiting FtsZ ring formation near the cell ends, the MinC protein plays a critical role in proper positioning of the division apparatus in Escherichia coli. MinC activity requires that of MinD, and the MinE peptide provides topological specificity by suppressing MinC-MinD-mediated division inhibition specifically at the middle of the cell. We recently presented evidence that MinE not only accumulates in an FtsZ-independent ring structure at the cell's middle but also imposes a unique dynamic localization pattern upon MinD in which the latter accumulates alternately in either one of the cell halves in what appears to be a rapidly oscillating membrane association-dissociation cycle. Here we show that functional green fluorescent protein-MinC displays a very similar oscillatory behavior which is dependent on both MinD and MinE and independent of FtsZ. The results support a model in which MinD recruits MinC to its site of action and in which FtsZ ring assembly at each of the cell ends is blocked in an intermittent and alternate fashion.


Mixed gonadal dysgenesis: whole life follow-up of a rare case.
There are two forms of gonadal dysgenesis - mixed and pure. In the mixed form, some differentiated gonads as well as some either ovarian or testicular rudiments are present. This form results in a number of phenotypes with a possibility of malignant transformation. In the pure form occurring in female gender, also some rudimental gonads are bilaterally present. In the case of simultaneous presence of Y chromosome, also some malignant transformation may appear (Siklar et al. 2007). Chromosomal aberrations are present in 2-7 % adult pairs with fertility disorders and in 0.6 % of newborns. However, only few cases with similar chromosomal aberrations were described so far (Roubin et al. 1977; Alexander et al. 1978; Teyssier et al. 1982; Caglayan et al. 2009). Mixed gonadal dysgenesis presents as a unilateral testis, usually intraabdominal, also with a streak gonad on contralateral side, and persistent mullerian structures. 45X/45XY karyotype is the most frequent in such cases with predominance of 45X cells in both peripheral lymphocytes and gonads. We present a rare case of a left undescended testis, normally descended right testis, with penoscrotal hypospadias, who had a normal karyotype and whose histopathological findings were endometrial tissue and fallopian tube in left testicular biopsy. Gonadal dysgenesis should always be kept in mind because of a possibility of undescended testis and proximal hypospadias. If karyotype reveals a 46XY gonadal dysgenesis, these patients need the careful follow-up to screen for gonadoblastoma in remaining normal testis. Subjecting the patients to prophylactic orchidectomy with hormone replacement can be an additional option in such patients.


Not all spondylolisthesis grading instruments are reliable.
Spondylolisthesis is the slippage of one vertebral body on an adjacent level, and occurs commonly at the lumbosacral junction in children. Many radiographic measurement instruments have been described to predict progression and need for intervention. We evaluated the reliability of eight common grading instruments. Four raters reviewed 30 lateral radiographs of the lumbar spine in patients with spondylolisthesis. Each rater measured each film twice, and had mean individual correlation coefficients of at least 0.76 (range, 0.76-0.91). Only three measurements had interobserver correlations greater than 0.75 (slip percentage, Meyerding's grade, and sacral inclination), which corresponded to excellent reliability. For intraobserver reliability, six measurements had correlations greater than 0.75 (all except kyphosis angle and lumbar index), indicating excellent agreement. Slip percent, Meyerding's grade, and sacral inclination had excellent interobserver agreement and intraobserver agreement.


Reproductive and hormonal factors, and ovarian cancer risk for BRCA1 and BRCA2 mutation carriers: results from the International BRCA1/2 Carrier Cohort Study.
Several reproductive and hormonal factors are known to be associated with ovarian cancer risk in the general population, including parity and oral contraceptive (OC) use. However, their effect on ovarian cancer risk for BRCA1 and BRCA2 mutation carriers has only been investigated in a small number of studies. We used data on 2,281 BRCA1 carriers and 1,038 BRCA2 carriers from the International BRCA1/2 Carrier Cohort Study to evaluate the effect of reproductive and hormonal factors on ovarian cancer risk for mutation carriers. Data were analyzed within a weighted Cox proportional hazards framework. There were no significant differences in the risk of ovarian cancer between parous and nulliparous carriers. For parous BRCA1 mutation carriers, the risk of ovarian cancer was reduced with each additional full-term pregnancy (P trend = 0.002). BRCA1 carriers who had ever used OC were at a significantly reduced risk of developing ovarian cancer (hazard ratio, 0.52; 95% confidence intervals, 0.37-0.73; P = 0.0002) and increasing duration of OC use was associated with a reduced ovarian cancer risk (P trend = 0.0004). The protective effect of OC use for BRCA1 mutation carriers seemed to be greater among more recent users. Tubal ligation was associated with a reduced risk of ovarian cancer for BRCA1 carriers (hazard ratio, 0.42; 95% confidence intervals, 0.22-0.80; P = 0.008). The number of ovarian cancer cases in BRCA2 mutation carriers was too small to draw definitive conclusions. The results provide further confirmation that OC use, number of full-term pregnancies, and tubal ligation are associated with ovarian cancer risk in BRCA1 carriers to a similar relative extent as in the general population.


Appendicular Mucocele: Possibilities and Limits of Laparoscopy. Brief Series and Review of the Literature.
Appendicular mucocele, a cystic dilatation of the appendix, is a rare disease, but unfortunately about 1/10 of cases evolves into pseudomyxoma peritonei. We performed a prospective study between 1 January 2010 to 31 December 2014 in order to track the incidence, symptoms, and circumstances of diagnosis, treatment and evolution of these rare tumors. A total of seven patients underwent curative surgery for a mucocele of the appendix: one woman and six men with an average age of 59.71 years. Clinical signs, present in two cases, were uncharacteristic. Ultrasound performed in all cases, could guide diagnosis in 5 cases. CT performed in 5 cases diagnosed only two cases. All cases were operated on: the open approach was used in four cases and a minimally invasive in three cases. We performed two right colectomies, an open appendectomy associated to anterior resection of the rectum, two laparoscopic appendectomies and two appendectomies and cecum resection with stapler, one by open approach and one by a minimally invasive approach. Intraoperative spillage of mucinous tumor did not occur in any case. The mean hospital stay was 5.7 days. Postoperative complications were present in 1 case (14.2%): wound infection. The average follow-up period was 40.28 months. (Range 6 to 48 months). No tumor recurrence or readmission, such as pseudomyxoma peritonei, has occurred. Appendicular mucocele is a rare entity; it can be found incidentally and it can mimic acute appendicitis, appendicular plastron or cecum tumor. Once diagnosed, surgical treatment is required for fear of perforation, tumor evolution and the emergence of the rule of complications. Laparoscopic approach in selected cases can be used, accompanied by safety measures to avoid iatrogenic perforation and peritoneal and parietal seeding.


Electrodeposition of V-VI Nanowires and Their Thermoelectric Properties.
Nanostructuration is an intensive field of research due to the appearance of interesting properties at the nanoscale. For instance, in thermoelectricity the most outstanding improvements obtained lately are related to phenomena that appear as a result of nano-engineering different materials. The thermoelectric effect is the direct conversion from temperature gradients into electricity and vice versa. When going to low dimensions, for example in the particular case of thermoelectric nanowires, the transport properties of phonons are modified with respect to those found in bulk leading to a higher thermoelectric figure of merit z. In more detail, this review tries to compile some of the landmarks in the electrodeposition of Bi2Te3-based nanowires. We will focus on the achievements using different templates, electrolytes and deposition modes. We will also summarize the measurements performed in those nanowires and the main conclusions that can be extracted from the published works. Finally, an update of nanowire-based thermoelectric generators is also included.


Comparison of hand hygiene antimicrobial efficacy: Melaleuca alternifolia essential oil versus triclosan.
this study aimed to evaluate the efficacy of hand hygiene performed with two different soap formulations: 0.3% Melaleuca alternifolia essential oil versus 0.5% triclosan, and to compare them with two reference hygiene procedures: the official methodology procedure (soft soap) versus the draft version of the procedure (soft soap + propan-2-ol). using the European EN 1499 method, logarithmic reduction factors were determined for the number of colony forming units of Escherichia coli K12 before and after hand hygiene of 15 volunteer subjects, and compared using the one-tailed Wilcoxon test. referring to the soft soap, there was no difference between the performance of soap with 0.3% M. alternifolia and soap containing 0.5% triclosan. The soft soap + propan-2-ol proved to be more effective than the other hand hygiene procedures. studies to verify the therapeutic efficacy of essential oil in hand hygiene can improve adherence to this practice.


Molecularly resolved, label-free nucleic acid sensing at solid-liquid interface using non-ionic DNA analogues.
Nucleic acid-based biosensors, where the capture probe is a nucleic acid, e.g., DNA or its synthetic analogue xeno nucleic acid (XNA), offer interesting ways of eliciting clinically relevant information from hybridization/dehybridization signals. In this respect, the application of XNA probes is attractive since the drawbacks of DNA probes might be overcome. Within the XNA probe repertoire, peptide nucleic acid (PNA) and morpholino (MO) are promising since their backbones are non-ionic. Therefore, in the absence of electrostatic charge repulsion between the capture probe and the target nucleic acid, a stable duplex can be formed. In addition, these are nuclease-resistant probes. Herein, we have tested the molecularly resolved nucleic acid sensing capacity of PNA and MO capture probes using a fluorescent label-free single molecule force spectroscopy approach. As far as single nucleobase mismatch discrimination is concerned, both PNA and MO performed better than DNA, while the performance of the MO probe was the best. We propose that the conformationally more rigid backbone of MO, compared to the conformationally flexible PNA, is an advantage for MO, since the probe orientation can be made more upright on the surface and therefore MO can be more effectively accessed by the target sequences. The performance of the XNA probes has been compared to that of the DNA probe, using fixed nucleobase sequences, so that the effect of backbone variation could be investigated. To our knowledge, this is the first report on molecularly resolved nucleic acid sensing by non-ionic capture probes, here, MO and PNA.


CD4 positive T helper cells contribute to retinal ganglion cell death in mouse model of ischemia reperfusion injury.
Neuron degeneration is a common pathological process associated with many disease conditions in the central nervous system including retina. Although immune responses have been proposed as one potential element in triggering neural damage, the mechanism of action of specific immune components underlying the pathogenesis is unclear. In this study we focus on adaptive immune activities to evaluate CD4 positive helper cells in the retinal ganglion cell (RGC) degeneration in response to transient retinal ischemic/reperfusion (I/R) injury. Transient retinal ischemia was induced in four mouse strains with different immune backgrounds, including wild type mice from C57BL/6 and BABL/c strains, severe combined immunodeficient (SCID) mice lacking T and B lymphocytes, SCID mice with transferred wild type CD4+ T cells, and the STAT6 deficient mice without T helper 2 (TH2) cells. In SCID mice RGCs showed a strong resistance to cell death in response to I/R injury (89% ± 3% of the survival cells in contralateral eye) compared with C57BL/6 (p = 0.018) and BALB/C (p = 0.038) wild types. By transferring the mature CD4+ T cells from matched wild type into SCID mice, the resistance of RGCs to injury was significantly compromised (p < 0.05). Furthermore a significant resistance of RGCs to cell death (p < 0.05) accompanied with an overexpression of STAT1 and STAT3 was confirmed in STAT6 deficient mice in response to I/R injury compared with the wild type controls, indicating that TH2 cells maturation might be involved in RGC damage. Adaptive immunity carried by CD4 T cells plays an essential role in RGC degeneration.


Diagonal Trajectory Posterior Screw Instrumentation for Compromised Bone Quality Spine: Groove-Entry Technique/Hooking Screw Hybrid.
In an attempt to increase anchoring strength of posterior instrumentation in spine with compromised bone quality, we introduced diagonal trajectory pedicle screwing (hooking screws) that do not rely on screw thread purchase in bone but rather hook onto the strong posterior elements of vertebrae from inside the bone. Between November 2016 and July 2017 we treated eight patients, mean age 80 years old (75-86 years old) with compromised bone quality for spinal instability. The diagnosis was osteoporotic fracture nonunion in three, ankylosed spine fracture in three, pyogenic spondylitis in two cases. All spines were percutaneously instrumented. Groove-entry technique was used for down-going thoracic screws. No additional hooks, cables, or any other augmentation was used. All patients were mobilized on post-operative day 1. 84 screws were inserted overall. Groove-entry technique was used for 42 screws insertion. On average, 5.3 spinal segments were fixed per case. Mean operation time was 252 min (46 min per one spinal segment). Mean intraoperative bleeding was 112 ml per case (21 ml per one fixed spinal segment). All cases achieved bony union of the fracture site or across the destroyed intervertebral disk. Mean time to union was 4 months postop (3-7 months). All patients were ambulatory at the time of discharge. No nerve injury, no skin irritation caused by implants, no screw loosening, no screw pullout, no loss of correction, and no junctional kyphosis were noted in this series. Diagonal screw instrumentation (our hooking screws and groove-entry technique) appears to provide sufficient anchoring strength while being minimally invasive and possibly helpful in prevention of junctional kyphosis.


Effective depth of spermatogonia in man II. Calculations for external high-energy beta rays.
An important criterion for dosemeter design for male gonad dosimetry in mixed beta-gamma fields is the effective spermatogonial depth. A detector set at this depth will register mean dose to the spermatogonia: this is the quantity of interest for genetic risk, which is proportional to the mean dose to the spermatogonia, averaged over the whole testicular volume. Factors affecting mean spermatogonial dose and effective depth of the spermatogonia were examined by computer using a simple mathematical model. Depth dose data were input from tables of beta dose. Mean spermatogonial dose shows a moderate dependence upon beta maximum energy, and a moderate inverse dependence upon scrotal thickness and source distance in air. Effective spermatogonial depth, while largely independent of source distance, shows strong dependence upon beta maximum energy: hence dosemeters designed for a given beta maximum energy will over-respond when exposed to betas of higher energy. Values of effective spermatogonial depth are given for two isotopes. In fields of mixed isotopes, weighting of the depths is essential for averaging.


Immune regulatory molecules as modifiers of semen and fertility: A review.
Declining fertility rates in both human and animals is a cause for concern. While many of the infertility cases are due to known causes, idiopathic infertility is reported in 30% of the infertile couples. In such cases, 18% of the infertile males carry antisperm antibodies (ASAs). Such data are lacking in livestock, wherein 20-30% of the animals are being culled due to low fertility. In males, the blood-testis barrier (BTB) and biomolecules in the semen provide an immuno-tolerant microenvironment for spermatozoa as they traverse the immunologic milieu of both the male and female reproductive tracts. For example, insults from environmental contaminants, infections and inflammatory conditions are likely to impact the immune privilege state of the testis and fertility. The female mucosal immune system can recognize allogenic spermatozoa-specific proteins affecting sperm kinematics and sperm-zona binding leading to immune infertility. Elucidating the functions and pathways of the immune regulatory molecules associated with fertilization are prerequisites for understanding their impact on fertility. An insight into biomolecules associated with spermatozoal immune tolerance may generate inputs to develop diagnostic tools and modulate fertility. High-throughput sequencing technologies coupled with bioinformatics analyses provides a path forward to define the array of molecules influencing pregnancy outcome. This review discusses the seminal immune regulatory molecules from their origin in the testis until they traverse the uterine environment enabling fertilization and embryonic development. Well-designed experiments and the identification of biomarkers may provide a pathway to understand the finer details of reproductive immunology that will afford personalized therapies.


The Effect of a Personalized Newsletter to Physical Therapists on Patient Recruitment: A Cluster Randomized Trial in Primary Physiotherapy Care.
To assess the effect of a personalized newsletter compared with a standard newsletter on patient recruitment in physiotherapy research. We performed a cluster-randomized trial including 120 physiotherapists who recruited patients for a prospective cohort and were randomly assigned to either receiving personalized feedback in a newsletter (intervention group) or a standard newsletter (control group). We calculated the difference in the number of patients included in the study corrected for inclusion time between both groups. The physiotherapists in the control group (n = 59) included 110 patients (35.4% of the total number of patients included) compared with an inclusion of 200 patients (64.6% of the total number of patients included) by the physiotherapists in the intervention group (n = 61), a difference of 90 patients in favor of the intervention group. However, when corrected for inclusion time and a cluster effect, we found no statistically significant difference between both groups. In addition, therapists who did not include a single patient (inactive therapists) were evenly divided between the 2 groups (n = 29 [49%] in the control group; n = 30 [49%] in the intervention group). A personalized newsletter does not significantly increase the number of recruited patients by physiotherapists. However, therapists receiving personalized feedback recruited nearly double the number of patients compared with the ones receiving standard feedback.


Structure of glycerol-3-phosphate dehydrogenase, an essential monotopic membrane enzyme involved in respiration and metabolism.
Sn-glycerol-3-phosphate dehydrogenase (GlpD) is an essential membrane enzyme, functioning at the central junction of respiration, glycolysis, and phospholipid biosynthesis. Its critical role is indicated by the multitiered regulatory mechanisms that stringently controls its expression and function. Once expressed, GlpD activity is regulated through lipid-enzyme interactions in Escherichia coli. Here, we report seven previously undescribed structures of the fully active E. coli GlpD, up to 1.75 A resolution. In addition to elucidating the structure of the native enzyme, we have determined the structures of GlpD complexed with substrate analogues phosphoenolpyruvate, glyceric acid 2-phosphate, glyceraldehyde-3-phosphate, and product, dihydroxyacetone phosphate. These structural results reveal conformational states of the enzyme, delineating the residues involved in substrate binding and catalysis at the glycerol-3-phosphate site. Two probable mechanisms for catalyzing the dehydrogenation of glycerol-3-phosphate are envisioned, based on the conformational states of the complexes. To further correlate catalytic dehydrogenation to respiration, we have additionally determined the structures of GlpD bound with ubiquinone analogues menadione and 2-n-heptyl-4-hydroxyquinoline N-oxide, identifying a hydrophobic plateau that is likely the ubiquinone-binding site. These structures illuminate probable mechanisms of catalysis and suggest how GlpD shuttles electrons into the respiratory pathway. Glycerol metabolism has been implicated in insulin signaling and perturbations in glycerol uptake and catabolism are linked to obesity in humans. Homologs of GlpD are found in practically all organisms, from prokaryotes to humans, with >45% consensus protein sequences, signifying that these structural results on the prokaryotic enzyme may be readily applied to the eukaryotic GlpD enzymes.


Deregulated expression of the Per1 and Per2 in human gliomas.
Growing evidence shows that the deregulation of the circadian clock plays an important role in the development of malignant tumors, including gliomas. However, the molecular mechanisms of genes controlling circadian rhythm in glioma cells have not been explored. Using reverse transcription polymerase chain reaction and immunohistochemistry techniques, we examined the expression of two important clock genes, Per1 and Per2, in 33 gliomas. In this study, out of 33 gliomas, 28 were Per1-positive, and 23 were Per2-positive. The expression levels of Per1 and Per2 in glioma cells were significantly different from the surrounding non-glioma cells (P<0.01). The difference in the expression rate of Per1 and Per2 in high-grade (grade III and IV) and low-grade (grade 1 and II) gliomas was insignificant (P>0.05). While there was no difference in the intensity of immunoactivity for Per2 between high-grade gliomas and low-grade gliomas (r=-0.330, P=0.061), the expression level of Per1 in high-grade gliomas was significantly lower than that in low-grade gliomas(r=-0.433, P=0.012). In this study, we found that the expression of Per1 and Per2 in glioma cells was much lower than in the surrounding non-glioma cells. Therefore, we suggest that disturbances in Per1 and Per2 expression may result in the disruption of the control of normal circadian rhythm, thus benefiting the survival of glioma cells. Differential expression of circadian clock genes in glioma and non-glioma cells may provide a molecular basis for the chemotherapy of gliomas.


[Complications and after care in aphakic and pseudophakic patients].
The aim of every cataract operation is optical rehabilitation. With the greater social demands made today by aphakic and pseudophakic patients, the recognition and treatment of diseases with early or late onset after cataract extraction assumes greater importance. The early aims are rapid mobilization, complete absence of irritation, and normalization of pressure. Regular administration of corticosteroids, essential in the post-operative treatment of pseudophakia, may mask a bacterial or mycotic infection. The sole indication for removal of an implant is the uveitis-glaucoma-hyphema syndrome. The prevention and treatment of cystoid macular edema is discussed: whether or not the posterior lens capsule is intact seems to play a major role in the pathogenesis of this early or late complication in both aphakia and pseudophakia.


Polyethylenimine-Mediated Electrostatic Assembly of MnO2 Nanorods on Graphene Oxides for Use as Anodes in Lithium-Ion Batteries.
In recent years, the development of electrochemically active materials with excellent lithium storage capacity has attracted tremendous attention for application in high-performance lithium-ion batteries. MnO2-based composite materials have been recognized as one of promising candidates owing to their high theoretical capacity and cost-effectiveness. In this study, a previously unrecognized chemical method is proposed to induce intra-stacked assembly from MnO2 nanorods and graphene oxide (GO), which is incorporated as an electrically conductive medium and a structural template, through polyethylenimine (PEI)-derived electrostatic modulation between both constituent materials. It is revealed that PEI, a cationic polyelectrolyte, is capable of effectively forming hierarchical, two-dimensional MnO2-RGO composites, enabling highly reversible capacities of 880, 770, 630, and 460 mA·h/g at current densities of 0.1, 1, 3, and 5 A/g, respectively. The role of PEI in electrostatically assembled composite materials is clarified through electrochemical impedance spectroscopy-based comparative analysis.


Cohesin-independent segregation of sister chromatids in budding yeast.
Cohesin generates cohesion between sister chromatids, which enables chromosomes to form bipolar attachments to the mitotic spindle and segregate. Cohesin also functions in chromosome condensation, transcriptional regulation, and DNA damage repair. Here we analyze the role of acetylation in modulating cohesin functions and how it affects budding yeast viability. Previous studies show that cohesion establishment requires Eco1p-mediated acetylation of the cohesin subunit Smc3p at residue K113. Smc3p acetylation was proposed to promote establishment by merely relieving Wpl1p inhibition because deletion of WPL1 bypasses the lethality of an ECO1 deletion (eco1Δ wpl1Δ). We find that little, if any, cohesion is established in eco1Δ wpl1Δ cells, indicating that Eco1p performs a function beyond antagonizing Wpl1p. Cohesion also fails to be established when SMC3 acetyl-mimics (K113Q or K112R,K113Q) are the sole functional SMC3s in cells. These results suggest that Smc3p acetylation levels affect establishment. It is remarkable that, despite their severe cohesion defect, eco1Δ wpl1Δ and smc3-K112R,K113Q strains are viable because a cohesin-independent mechanism enables bipolar attachment and segregation. This alternative mechanism is insufficient for smc3-K113Q strain viability. Smc3-K113Q is defective for condensation, whereas eco1Δ wpl1Δ and smc3-K112R,K113Q strains are competent for condensation. We suggest that Smc3p acetylation and Wpl1p antagonistically regulate cohesin's essential role in condensation.


Xenopus tropicalis egg extracts provide insight into scaling of the mitotic spindle.
The African clawed frog Xenopus laevis has been instrumental to investigations of both development and cell biology, but the utility of this model organism for genetic and proteomic studies is limited by its long generation time and unsequenced pseudotetraploid genome. Xenopus tropicalis, which is a small, faster-breeding relative of X. laevis, has recently been adopted for research in developmental genetics and functional genomics, and has been chosen for genome sequencing. We show that X. tropicalis egg extracts reconstitute the fundamental cell cycle events of nuclear formation and bipolar spindle assembly around exogenously added sperm nuclei. Interestingly, X. tropicalis spindles were approximately 30% shorter than X. laevis spindles, and mixing experiments revealed a dynamic, dose-dependent regulation of spindle size by cytoplasmic factors. Measurements of microtubule dynamics revealed that microtubules polymerized slower in X. tropicalis extracts compared to X. laevis, but that this difference is unlikely to account for differences in spindle size. Thus, in addition to expanding the range of developmental and cell biological experiments, the use of X. tropicalis provides novel insight into the complex mechanisms that govern spindle morphogenesis.


Excitation and scattering of guided waves: relationships between solutions for plates and pipes.
The detection of localized defects such as cracks and corrosion in pipes using guided elastic waves is now an established non-destructive testing procedure. However, the prediction of guided wave excitation and scattering in pipes is a complex three-dimensional (3D) problem with many parameters that can generally only be solved using numerical methods. In many important industrial applications, the diameter of a pipe is much larger than wall thickness. In this case an approximate theory is applicable, when a pipe is considered as an unwrapped isotropic plate. In this paper, a technique for obtaining pipe mode amplitudes in terms of the solution to the forced 3D problem on a plate is presented. The same principle is extended to relate guided wave scattering from defects in plates to scattered circumferential modal amplitudes from defects in pipe. This is of practical benefit as the scattering of guided waves by defects in a plate is a much simpler problem than that in a pipe, and one that, in some cases, can be solved using analytical methods. Results are shown that illustrate the application of the method to reflection from through-thickness circumferential cracks in pipes.


Severe anemia in young children after high and low malaria transmission seasons in the Kassena-Nankana district of northern Ghana.
Malaria and anemia accounted for 41% and 18% respectively of hospital deaths in the Kassena-Nankana district of northern Ghana during 1996. We measured hemoglobin (Hb), malaria prevalence, and anthropometric indices of 6--24-month-old infants and young children randomly selected from this community at the end of the high (May-October, n = 347) and low (November-April, n = 286) malaria transmission seasons. High transmission season is characterized by rainfall (the equivalent of 800-900 mm/yr.), while the remaining months receive less than 50 mm/yr. Severe anemia, defined as Hb < 6.0 g/dL, was 22.1% at the end of the high transmission season compared to 1.4% at the end of the low transmission season (Odds Ratio [OR] = 20.1; 95% CI: 7.1-55.3). Parasitemia was 71% and 54.3% at these time points (OR = 2.1; 95% CI: 1.5-2.9). Nutritional anemia appeared to have little impact upon this seasonal difference since anthropometric indices were comparable. Although the relative contributions of other causes of severe anemia were not assessed, repeated malaria infections may be a primary determinant of severe anemia among infants and young children during the high transmission season.


Successful treatment of a massive atenolol and nifedipine overdose with CVVHDF.
The aim of this paper was to describe a case of massive atenol and nifedipine poisoning, complicated by the co-existence of liver cirrhosis, where standard therapies (fluid replacement, vasopressors and inotropic agents, insulin, glucagon, calcium and bowel decontamination) were ineffective in restoring an adequate heart rate, blood pressure, renal and intestinal blood flow. This led to consequent anuric renal insufficiency and incipient multiple organ failure syndrome (MOFS). The patient recovered completely after Continuous Veno-Venous Hemo-Dia-Filtration (CVVHDF); this treatment removed atenolol from blood, with predicted clearance levels. The patient was a 45-year old female with a history of hypertension, liver cirrhosis, neurological and psychiatric disorders, with a massive atenolol (69.6 microg/mL) and nifedipine (63 ng/mL) overdose. CVVHDF at an ultrafiltration rate of 1 500 mL/h was started on day 1. From day 2 onwards, as the plasma atenolol concentration decreased, the blood pressure rose at a slow but constant rate. On day 5, there was restoration of an adequate blood pressure, which restored both renal and intestinal function, and also improved MOFS. The standard therapeutic approach was ineffective at eliminating both substances from the blood, and the clinical picture became worse due to incipient MOFS. CVVHDF was used in order to maintain the fluid and electrolyte balance and also to clear the beta blocker from the blood. The clearance kinetics of atenolol were consistent with the expected clearance values, on the basis of a CVVHDF ultrafiltration flow of 1 500 mL/h, which corresponds to a creatinine clearance of about 25 mL/min.


Precision of the XCT 3000 and comparison of densitometric measurements in distal radius scans between XCT 3000 and XCT 2000 peripheral quantitative computed tomography scanners.
Researchers conducting longitudinal studies face challenges as technology evolves and data collection equipment is upgraded from older to newer models. In this study, we report precision error and associations from correlation, regression equations, and Bland-Altman plots that compare results of distal radius images from the XCT 2000 and XCT 3000 peripheral quantitative computed tomography densitometers at the 4% and 20% distal radius site. The mean difference expressed as a percent of the measurement's mean was approximately 5% for cortical thickness, periosteal and endosteal circumferences, and total bone size at the 20% site, and less than 2% for all other measures. There were strong associations between bone parameter measures from the XCT 3000 and XCT 2000 with correlation coefficients ranging from 0.92 to 1.0. Strong associations between the 2 machines allow for the comparison of results with minor adjustments.


Local Parallel Cross Pattern: A Color Texture Descriptor for Image Retrieval.
Riding the wave of visual sensor equipment (e.g., personal smartphones, home security cameras, vehicle cameras, and camcorders), image retrieval (IR) technology has received increasing attention due to its potential applications in e-commerce, visual surveillance, and intelligent traffic. However, determining how to design an effective feature descriptor has been proven to be the main bottleneck for retrieving a set of images of interest. In this paper, we first construct a six-layer color quantizer to extract a color map. Then, motivated by the human visual system, we design a local parallel cross pattern (LPCP) in which the local binary pattern (LBP) map is amalgamated with the color map in "parallel" and "cross" manners. Finally, to reduce the computational complexity and improve the robustness to image rotation, the LPCP is extended to the uniform local parallel cross pattern (ULPCP) and the rotation-invariant local parallel cross pattern (RILPCP), respectively. Extensive experiments are performed on eight benchmark datasets. The experimental results validate the effectiveness, efficiency, robustness, and computational complexity of the proposed descriptors against eight state-of-the-art color texture descriptors to produce an in-depth comparison. Additionally, compared with a series of Convolutional Neural Network (CNN)-based models, the proposed descriptors still achieve competitive results.


Carumonam enhances reactivity of Escherichia coli with mono- and polyclonal antisera to rough Escherichia coli J5.
Escherichia coli O111 reacts only slightly with antiserum to its rough mutant E. coli J5 in an enzyme-linked immunosorbent assay. When E. coli O111 was grown in the presence of sub-MICs of the monocyclic beta-lactam antibiotic carumonam, however, the enzyme-linked immunosorbent assay titer increased from 1,280 to 81,920. When the bacteria were grown in the presence of carumonam, the titer that was obtained with antiserum against E. coli O111 was not affected. This reaction was abolished after this antiserum was absorbed with E. coli J5 in the case of the carumonam-treated strain, whereas this absorption did not affect the reaction with E. coli O111. Thus, the O-antigenic side chain of E. coli O111 seems to be affected if this strain is cultured in the presence of carumonam. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed a relative loss of the O polysaccharide in E. coli O111 when this strain was grown in the presence of carumonam. Also, a much stronger reaction of the antibiotic-affected lipopolysaccharide with a monoclonal antibody against E. coli J5 lipopolysaccharide was shown in immunoblots. The results of this study indicate that there is a synergism between certain antibiotics and monoclonal antibodies, something that could have clinical implications.


The mouse p (pink-eyed dilution) and human P genes, oculocutaneous albinism type 2 (OCA2), and melanosomal pH.
Recessive mutations of the mouse p (pink-eyed dilution) gene lead to hypopigmentation of the eyes, skin, and fur. Mice lacking a functional p protein have pink eyes and light gray fur (if non-agouti) or cream-colored fur (if agouti). The human orthologue is the P protein. Humans lacking a functional P protein have oculocutaneous albinism type 2 (OCA2). Melanocytes from p-deficient mice or OCA2 individuals contain small, minimally pigmented melanosomes. The mouse and human proteins are predicted to have 12 membrane spanning domains and possess significant sequence homology to a number of membrane transport proteins, some of which are involved in the transport of anions. The p protein has been localized to the melanosome membrane. Recently, it has been shown that melanosomes from p protein-deficient melanocytes have an abnormal pH. Melanosomes in cultured melanocytes derived from wild-type mice are typically acidic, whereas melanosomes from p protein-deficient mice are non-acidic. Melanosomes and related endosome-derived organelles (i.e., lysosomes) are thought to have an adenosine triphosphate (ATP)-driven proton pump that helps to generate an acidic lumen. To compensate for the charge of these protons, anions must also be transported to the lumen of the melanosome. In light of these observations, a model of p protein function is presented in which the p protein, together with the ATP-driven proton pump, regulates the pH of the melanosome.


Antiaging Potential of Fucoxanthin Concentrate Derived from Phaeodactylum tricornutum.
The aim of the study was to investigate the potential of a fucoxanthin concentrate prepared from Phaeodactylum tricornutum as a wrinkle care cosmetic agent. The concentrate (up to 25 µg/ml) did not affect the proliferation of human fibroblasts. In addition, the concentrate significantly increased procollagen synthesis in the fibroblasts at 12.5 and 25 µg/ml; however, it significantly decreased the expression of matrix metalloproteinase (MMP)-1, MMP-2, and MMP-9 at 25 µg/ml. In a follow-up study, a wrinkle care cream containing 0.03% of fucoxanthin concentrate was prepared and tested in women (aged 35-50 years, n = 21) for 8 weeks. The cream was applied twice daily. Safety assessment of the cream was carried out visually. In addition, interviews were conducted to investigate if adverse events such as erythema, edema, scaling, itching, stinging, burning, tightness, or prickling had occurred. No symptoms that threaten skin safety were reported. Evaluation of wrinkles around the eyes using the replica method showed a statistically significant decrease in wrinkles at week 8. Moreover, skin moisture and elasticity increased significantly from week 4. These results suggest that the fucoxanthin concentrate has no adverse effects on the skin and can be used as an active ingredient in wrinkle care cosmetics.


[Influenza in Poland in 2006].
A total number of 251,815 cases of influenza and influenza-like illness registered in Poland in 2006 (incidence 660.4 per 100,000 population). Regionally the incidence varied from 75.6 per 100,000 in swietokrzyskie voivodeship to 1,690.4 in mazowieckie. The highest reported incidence was observed in age group 0-4 years (1,543.1 per 100,000). Children and adolescents under 15 years of age accounted for 34.3% of all cases (age specific incidence 1,434.8 per 100,000). In this age group the incidence varied regionally from 190.2 in lubuskie voivodeship to 3,582.5 in lódzkie. 284 patients (0.11% of all cases) required hospital admission. There were 14 deaths due to influenza, in 71.4% these were persons over 70 years of age. In the epidemic season 2005/06 infections with influenza virus registered in Poland were mainly caused by type B similarly to other parts of Europe. Thirty five influenza strains were isolated, including 6 strains of subtype A/H1, 2 strains of subtype A/H3 and 27 strains of type B. All of these isolates with the exception of one strain of influenza B were antigenically similar to the vaccine strains recommended for the epidemic season 2005/06.


Effect of functional lateral shift of the mandible on hyaluronic acid metabolism related to lubrication of temporomandibular joint in growing rats.
Hyaluronic acid (HA) is a major molecular component of the articular cartilage of the temporomandibular joint (TMJ) influencing joint lubrication. Functional lateral shift of the mandible (FLSM) can lead to malocclusion. This study investigated the effects of FLSM on HA metabolism and lubrication of the TMJ in growing rats. Thirty 5-week-old male Wistar rats were divided into shift, recovery, and control groups. Rats in the shift and recovery groups were fitted with guiding plates to produce a 2-mm FLSM which were removed from the rats in the recovery group 14 days later. Animals were sacrificed at 14 and 28 days after the appliances were attached. Immunohistochemistry of HA-binding protein (HABP), hyaluronan synthase (HAS), and hyaluronoglucosaminidases (HYALs) was examined. The thickness of HABP-positively stained areas in the lateral regions in the bilateral condyle was reduced during the experimental period in the shift group compared with that in the control group. The proportion of HAS2-stained areas was bilaterally decreased in different regions of condylar cartilage during the experimental period in the shift group. The reduction of the HYAL2-stained area proportion in the condylar cartilage was more significant than that of HYAL1 at 14 days after appliance attachment in the shift group. HAS2 staining was not recovered in the recovery group. This research was based on animal experiments with a limited experimental period. FLSM altered lubrication related HA metabolism in the articular cartilage of the TMJ in growing rats.


Renal diseases in the elderly underwent to percutaneous biopsy of native kidneys.
The elderly population is growing and aging in better clinical conditions than in the past. However, the distribution and course of kidney diseases in elderly patients are not well known partially due to reluctance to indicate renal biopsies in those patients. To evaluate the distribution, clinical features, and outcomes of nephropathies diagnosed by biopsy in the elderly. Seventy-one patients (47 males, 24 females) aged 60 years or older (67.3 ± 6.5 years), undergoing biopsy from January 1990 to December 2006, were evaluated. They were grouped according to their clinical syndromes. Nephrotic syndrome was observed in 35 patients (49.3%), mainly associated with membranous nephropathy (17 patients), followed by amyloidosis and focal segmental glomerulosclerosis (seven patients each). Acute kidney injury (AKI) was diagnosed in 19 patients, and the main histopathological diagnoses were acute tubular necrosis (six patients) and cast nephropathy (three patients). Of those 19 patients, only two had a favorable course, while the others died early or progressed toward advanced chronic kidney disease. Twelve patients undergoing biopsy because of asymptomatic hematuria or proteinuria had different diagnoses, but most of them already had significant chronic nephropathy. In five patients with nephritic syndrome, the biopsies also showed several diagnoses. Nephrotic syndrome was the major indication for renal biopsy, and membranous nephropathy was the most frequent diagnosis. Among patients with AKI and asymptomatic hematuria or proteinuria, different diagnoses were found with high levels of advanced chronic nephropathy.


The influence of anxiety on action-specific perception.
Recent empirical research suggests that performance modulates perception. For example, baseball players judge the ball to be bigger when performing well as compared to when performing less successfully. Yet, the underlying processes that mediate the emergence of action-specific effects on perception are still largely unknown. To this end, we aimed to examine the impact of anxiety on the action-specific influence on perception as anxiety has been shown to affect both performance and perception. Thirty participants threw darts at a circle-shaped target and were then asked to judge the size of the target. The task was performed under conditions of low and high anxiety. Results replicated previously reported action-specific effects on perception under levels of low anxiety. However, these effects vanished in the high anxiety condition. Results seem to suggest that anxiety has a direct influence on the relation between performance and perception. Attentional control theory is discussed to explain the current findings.


Changes in pulmonary function after surgical treatment of congenital heart disease with pulmonary hyperflow.
Analysis of pulmonary status of pediatric patients in the postoperative phase of cardiac surgery. To assess pulmonary compliance and airway resistance in patients with congenital heart disease and pulmonary hyperflow submitted to surgical treatment with the use of extracorporeal circulation. Thirty-five patients were evaluated during surgery with measurements of static compliance and airway resistance at four different timepoints. Pulmonary measurements were performed non-invasively using end-inspiratory airway occlusion and specific mathematical formulas. The variables examined and related to pulmonary changes were: preoperative - age, weight, and relationship between systemic and pulmonary blood flow; intraoperative - perfusion times, anoxia times and minimum temperature; postoperative - time on mechanical ventilation and length of stay in the intensive care unit. Pulmonary compliance in all patients had an immediate and significant increase (P<0.001) at the end of surgery. Patients older than 30 months experienced a greater increase (P=0.0004). Those with more than 10 kg also had a greater increase (P=0.0006). In patients on extracorporeal circulation for more than 50 minutes, the increase in pulmonary compliance took longer to occur (P=0.04). Airway resistance was not significantly changed at the end of surgery (P=0.393). All patients experienced improved pulmonary compliance at the end of surgery, and this was significantly influenced by age, weight and time on extracorporeal circulation. Airway resistance, however, was not changed.


Ethnic and cultural determinants influence risk assessment for hepatitis C acquisition.
In the developed world hepatitis C virus (HCV) infection is predominantly associated with sharing contaminated equipment between injecting drug users (IDU). In developing countries inadequately sterilized medical equipment, transmission of infected blood and cultural practices have been implicated. Accurate risk factor assessment is essential for education targeted at risk reduction in culturally diverse populations. Ninety Australian-born Caucasians and 72 South-east Asian (SEA) HCV patients attending a Melbourne hospital liver clinic completed a questionnaire which assessed risk factor profile, perceived risk factors, knowledge of risk factors and methods to minimize transmission. Medical records were audited to identify doctor assessment of risk factors. Risk factors in Caucasians were IDU, body piercing and tattooing (89%, 47% and 32%, respectively). Risk factors in SEA patients were injection therapy, dental therapy and surgery (89%, 70% and 38%, respectively). Most Caucasian patients (94%) correctly identified their mode of acquisition compared with 33% of SEA patients (P < 0.0001). Accurate risk factor documentation in medical records was more common in Caucasians (96 vs 32%; P < 0.0001). The majority of patients identified blood-to-blood and sexual/vertical transmission as important modes of acquisition. However, 33% of SEA patients believed transmission occurred through food, water and poor hygiene and 80% did not identify therapeutic injection or traditional medical practices as risk factors. Education provided to SEA patients did not address less well established routes of transmission. Ethnicity influences perception and knowledge of risk factors. Improved assessment of risk factors in high-risk ethnic groups is needed. Education should be culturally appropriate and address the concerns of all populations with HCV.


Development of an ideal starch saccharification process using amylolytic enzymes from thermophiles.
The extensive efforts to screen thermophilic fungi and bacteria, isolated from various environmental samples, have resulted in the selection of Thermomucor indicae-seudaticae, Geobacillus thermoleovorans NP33 and G. thermoleovorans NP54 for the production of glucoamylase, amylopullulanase and alpha-amylase, respectively. Submerged and solid-state fermentation processes were optimized for maximizing the secretion of glucoamylase by T. indicae-seudaticae. The production of amylopullulanase and alpha-amylase by NP33 and NP54 in submerged fermentation was also optimized. Glucoamylase was optimally active at pH 7.0 and 60 degrees C and was shown to saccharify soluble as well as raw starches. Amylopullulanase and alpha-amylase exhibited optima at pH 7.0 and 100 degrees C and saccharified starch efficiently. Differential inhibition and action on mixed substrates clearly suggested that there are two separate active sites for alpha-amylase and pullulanase activities of amylopullulanase. Both alpha-amylase and amylopullulanase are high maltose-forming and Ca(2+)-independent. These amylolytic enzymes have been shown to be useful in starch saccharification alone and in combination.


Yerba mate aqueous extract improves the oxidative and inflammatory states of rats with adjuvant-induced arthritis.
Healthy and adjuvant-induced arthritic rats were treated for 23 days with daily doses of 400 and 800 mg kg-1Ilex paraguariensis extract. This treatment (a) diminished the ROS levels in the liver and brain, (b) decreased oxidative protein and lipid damage in liver and brain, (c) increased the plasma antioxidant capacity, (d) increased the GSH levels and the GSH/GSSH ratio in both the liver and the brain, (e) almost restored the enzymatic activities linked to the metabolism of GSH-GSSG, and (f) reversed the modified activities of xanthine oxidase, superoxide dismutase and catalase. The anti-inflammatory actions (firstly) and the antioxidant actions (in the second place) of the yerba mate constituents (e.g., chlorogenic acid derivatives) are the causes of these beneficial effects. Daily ingestion of traditional yerba mate beverages may be effective in attenuating the symptoms of inflammatory diseases, especially in older adults.


Plant-made vaccines against West Nile virus are potent, safe, and economically feasible.
The threat of West Nile virus (WNV) epidemics with increasingly severe neuroinvasive infections demands the development and licensing of effective vaccines. To date, vaccine candidates based on inactivated, live-attenuated, or chimeric virus, and viral DNA and WNV protein subunits have been developed. Some have been approved for veterinary use or are under clinical investigation, yet no vaccine has been licensed for human use. Reaching the milestone of a commercialized human vaccine, however, may largely depend on the economics of vaccine production. Analysis suggests that currently only novel low-cost production technologies would allow vaccination to outcompete the cost of surveillance and clinical treatment. Here, we review progress using plants to address the economic challenges of WNV vaccine production. The advantages of plants as hosts for vaccine production in cost, speed and scalability, especially those of viral vector-based transient expression systems, are discussed. The progress in developing WNV subunit vaccines in plants is reviewed within the context of their expression, characterization, downstream processing, and immunogenicity in animal models. The development of vaccines based on enveloped and non-enveloped virus-like particles is also discussed. These advancements suggest that plants may provide a production platform that offers potent, safe and affordable human vaccines against WNV.


The transition experience of Enrolled Nurses to a Bachelor of Nursing at an Australian university.
Endorsed Enrolled Nurses (EENs) articulating from diploma level to Bachelor of Nursing (BN) studies at university experience many transitional barriers. Flexible credit arrangements can create further difficulties because students may enter directly into the second year of a degree program, thus foregoing supportive interventions targeting first year students. This qualitative study explored the transitional barriers faced by EENs articulating to the second year of a BN program and the processes employed to adapt to the university learning environment. Lizzio's (2006) Five Senses of Success Model provided a framework for data analysis. Lizzio's model highlights how students' success at university depends on their sense of purpose, capability, resourcefulness, connectedness and academic culture. This study revealed EENs grapple with their dual identity, have difficulty reconciling their academic and clinical competence, and struggle to assimilate to the academic learning environment. Findings illuminate the importance of tailoring orientation and engagement activities to the specific transitional needs of articulating students.


Anal findings in sexual abuse of children (a descriptive study).
The aim of this study was to assess anal physical findings in children whose abuse was admitted by the perpetrator. Fifty children were studied in whom medical assessment took place remote in time from anal abuse which was admitted by the perpetrator. Medical assessment included examination to detect the presence of anal physical signs. Most frequent signs were anal scars and tags (either single or multiple) present, respectively, in 84 and 32% of cases. In some cases scars extended to the perianal region. Other signs included reflex anal dilatation (RAD) and venous congestion (VC) found, separately or associated with other signs, in over 33% of the cases. In 6% of the cases there were no abnormal anal findings. The results confirm earlier reports that physical signs, including scars, tags, RAD, funneled anus and extensive venous congestion, are often present in abused children, singly or in combination, and that anal examination should be undertaken even months after a known or suspected sexual assault. In the legal setting these physical signs are seen in association with anal abuse and support the child's statement. They do not per se provide proof of abuse.


The yellow gene influences Drosophila male mating success through sex comb melanization.
Drosophila melanogaster males perform a series of courtship behaviors that, when successful, result in copulation with a female. For over a century, mutations in the yellow gene, named for its effects on pigmentation, have been known to reduce male mating success. Prior work has suggested that yellow influences mating behavior through effects on wing extension, song, and/or courtship vigor. Here, we rule out these explanations, as well as effects on the nervous system more generally, and find instead that the effects of yellow on male mating success are mediated by its effects on pigmentation of male-specific leg structures called sex combs. Loss of yellow expression in these modified bristles reduces their melanization, which changes their structure and causes difficulty grasping females prior to copulation. These data illustrate why the mechanical properties of anatomy, not just neural circuitry, must be considered to fully understand the development and evolution of behavior.


Comparison of type I and II papillary renal cell carcinoma (RCC) and clear cell RCC.
To compare the pathological features of clear cell renal cell carcinoma (ccRCC) with papillary RCC (pRCC) and further differentiate type I and II pRCC as independent prognosticators for survival. From September 1994 to February 2007 557 RCCs were treated and reviewed. All patients underwent radical nephrectomy or nephron-sparing surgery. We reviewed patient data and correlated RCC subtypes to tumour size, pathological stage, nuclear grade, and 5-year cancer-specific survival (CSS). pRCC was re-evaluated in to type I and II. The 2002 Tumour-Node-Metastasis and Fuhrman classifications were used. In all, 391 (70%) patients had ccRCC, 96 (17%) had pRCC, 34 (6%) had chromophobe RCC, seven (1%) had ductus Bellini RCC and 29 (5%) had unclassified RCC. Upon re-evaluation 34 patients had type I pRCC and 62 had type II. The pRCCs were significantly smaller than the ccRCCs, at a mean (sd) of 4.5 (2.5) cm vs 5 (2.9) cm (P = 0.013), and multifocal (25% vs 12%, P = 0.001). Whereas patients with ccRCC had significantly more primary metastases (12% vs 3%, P = 0.014). The mean (sd) follow-up was 42.3 (41.4) months. The 5-year CSS for M0 patients was 84% for ccRCC and 90% for pRCC (P = 0.573). At multivariate analyses predictors for 5-year CSS were only tumour size (hazard ratio, HR 2.6, P < 0.001), pathological stage (HR 3.9, P < 0.001) and nuclear grade (HR 2.7, P < 0.001). The type I and II pRCCs had significantly different lymphovascular invasion (LVI) and 5-year CSS rates (94% vs 74%, P = 0.03). The ccRCCs were significantly larger at diagnosis than the pRCCs. The histological subtype (pRCC vs ccRCC) had no impact on the 5-year CSS in multivariate analyses. The type I and II pRCCs had similar histopathological features except for a significant difference in LVI. However, the 5-year CSS was significantly different in type I and II pRCC.


Nanowire array composites.
Long, nanometer-size metallic wires can be synthesized by injection of the conducting melt into nanochannel insulating plates. Large-area arrays of parallel wires 200 nanometers in diameter and 50 micrometers long with a packing density of 5 x 10(8) per square centimeter have been fabricated in this way. When charged, the ends of the wires generate strong, short-range electric fields. The nanowire electric fields have been imaged at high spatial resolution with a scanning force microscope.


Atopy patch testing in children with asthma and rhinitis symptoms allergic to house dust mite.
The atopy patch test (APT) is generally used to assess immunoglobulin E (IgE) mediated sensitization to allergens in patients with atopic dermatitis, but its diagnostic role in children with respiratory allergy is still controversial. The aim of the study was to evaluate APT with house dust mite (HDM) in children with asthma and rhinitis symptoms allergic to HDM and its relevance to skin prick test (SPT) diameters and specific IgE levels. The study population consisted of 33 children, aged 8-16 yr (median: 12 yr) with asthma and 30 children with allergic rhinitis in the same age range (median: 11 yr). All patients had positive SPT results and high serum specific IgE levels for Dermatophagoides pteronyssinus APT was performed on back skin of all patients with 200 index of reactivity (IR)/ml of D. pteronyssinus allergen extracts in petrolatum (Stallerpatch) and evaluated at 72 h. Of 63 patients, 16 (25%) showed a positive patch test result. APT with HDM showed 30% (10/33) positivity among the patients with asthma and 20% (6/30) positivity among the patients with allergic rhinitis. APT presented no significant correlation with age, SPT diameter, serum total and specific IgE levels for D. pteronyssinus, nasal provocation test or pulmonary function test results. Patch testing with HDM may partly identify mite sensitive children with respiratory allergy. Positive APT results may imply that delayed hypersensitivity reactions play a role in children with asthma and rhinitis allergic to HDM.


Transforming growth factor-beta induces nuclear import of Smad3 in an importin-beta1 and Ran-dependent manner.
Smad proteins are cytoplasmic signaling effectors of transforming growth factor-beta (TGF-beta) family cytokines and regulate gene transcription in the nucleus. Receptor-activated Smads (R-Smads) become phosphorylated by the TGF-beta type I receptor. Rapid and precise transport of R-Smads to the nucleus is of crucial importance for signal transduction. By focusing on the R-Smad Smad3 we demonstrate that 1) only activated Smad3 efficiently enters the nucleus of permeabilized cells in an energy- and cytosol-dependent manner. 2) Smad3, via its N-terminal domain, interacts specifically with importin-beta1 and only after activation by receptor. In contrast, the unique insert of exon3 in the N-terminal domain of Smad2 prevents its association with importin-beta1. 3) Nuclear import of Smad3 in vivo requires the action of the Ran GTPase, which mediates release of Smad3 from the complex with importin-beta1. 4) Importin-beta1, Ran, and p10/NTF2 are sufficient to mediate import of activated Smad3. The data describe a pathway whereby Smad3 phosphorylation by the TGF-beta receptor leads to enhanced interaction with importin-beta1 and Ran-dependent import and release into the nucleus. The import mechanism of Smad3 shows distinct features from that of the related Smad2 and the structural basis for this difference maps to the divergent sequences of their N-terminal domains.


Effects of vitamin B-6 deficiency on morphological changes in dendritic trees of Purkinje cells in developing cerebellum of rats.
The effect of maternal vitamin B-6 deficiency on the morphological changes in the dendritic growth of Purkinje cells of progeny was studied at four different ages (12, 15, 21, and 50 days) of postnatal development. By day 10, rat pups in the deficient groups (0.6 or 1.0 mg pyridoxine . HCl/kg diet) began to show gross neurological symptoms of the deficiency and before weaning approximately 12% of the pups in the 0.6 group had died. At each age studied, body weights were less and brain-body weight ratios were greater in the deficient groups compared to control (7.0 mg pyridoxine . HCl/kg diet) groups. Morphological changes of the Purkinje cell dendritic field, as determined from camera lucida drawings of Golgi-Cox impregnated cells of the mid-sagittal vermis, indicated significantly smaller width, height and area in the deficient groups compared to the control. Stereological measurements showed that dendritic branching density was significantly less in the 0.6 group compared with the 1.0 and 7.0 groups. The total length of Purkinje cell dendrites per cell was significantly reduced in the deficient groups compared to the 7.0 group. Qualitative analyses of photomicrographs of Purkinje cells indicated more dendritic and somatic aberrations in the deficient groups than in the control. Dendritic trees were larger in size and more complex in the 7.0 group compared to the deficient groups. The findings indicate that maternal vitamin B-6 deficiency interferes with normal development of the Purkinje cell dendrites.


Detection of anticonductive tissue autoantibodies in a patient with chronic intestinal pseudo-obstruction and sick sinus syndrome.
A 26-year-old patient was diagnosed as having chronic intestinal pseudo-obstruction with manometric and histopathologic features suggestive of an intestinal myopathy. Histology was characterized by smooth muscle degeneration without inflammatory or immune cells. The severe gut dysfunction required full parenteral nutritional support. After a few months, the patient developed symptomatic tachy-brady arrhythmia episodes with syncopes. A thorough diagnostic work-up led to a diagnosis of sick sinus syndrome, which was managed by pacemaker implantation and administration of β-blockers. This led to a partial improvement in tachy-brady arrhythmia episodes. Nonetheless, the patient continued to experience sustained supraventricular tachyarrhythmia runs, poorly responsive to increasing β-blocker doses. To investigate the origin of the cardiologic impairment, the patient was tested for anticonductive tissue autoantibodies, which were positive, thus supporting a possible autoimmune origin of the dysrhythmia. Other autoantibodies were negative. On the basis of these findings, the patient was treated with high-dose steroids, which were then tapered. The patient responded to the steroid treatment and did not experience further episodes of syncope and tachyarrhythmias. The severe gut dysfunction remained unchanged. This case highlights an association between severe gut dysfunction and cardiac conductive tissue abnormalities, with autoantibodies to conductive tissue possibly causing the dysrhythmia. The severe gut and heart (likely autoimmune-mediated) dysfunction presented in this case provides a basis to further assess a link between intestinal and cardiac abnormal rhythmicity.


Control of axon guidance and neurotransmitter phenotype of dB1 hindbrain interneurons by Lim-HD code.
Hindbrain dorsal interneurons (HDIs) are implicated in receiving, processing, integrating, and transmitting sensory inputs from the periphery and spinal cord, including the vestibular, auditory, and proprioceptive systems. During development, multiple molecularly defined HDI types are set in columns along the dorsoventral axis, before migrating along well-defined trajectories to generate various brainstem nuclei. Major brainstem functions rely on the precise assembly of different interneuron groups and higher brain domains into common circuitries. Yet, knowledge regarding interneuron axonal patterns, synaptic targets, and the transcriptional control that govern their connectivity is sparse. The dB1 class of HDIs is formed in a district dorsomedial position along the hindbrain and gives rise to the inferior olive nuclei, dorsal cochlear nuclei, and vestibular nuclei. dB1 interneurons express various transcription factors (TFs): the pancreatic transcription factor 1a (Ptf1a), the homeobox TF-Lbx1 and the Lim-homeodomain (Lim-HD), and TF Lhx1 and Lhx5. To decipher the axonal and synaptic connectivity of dB1 cells, we have used advanced enhancer tools combined with conditional expression systems and the PiggyBac-mediated DNA transposition system in avian embryos. Multiple ipsilateral and contralateral axonal projections were identified ascending toward higher brain centers, where they formed synapses in the Purkinje cerebellar layer as well as at discrete midbrain auditory and vestibular centers. Decoding the mechanisms that instruct dB1 circuit formation revealed a fundamental role for Lim-HD proteins in regulating their axonal patterns, synaptic targets, and neurotransmitter choice. Together, this study provides new insights into the assembly and heterogeneity of HDIs connectivity and its establishment through the central action of Lim-HD governed programs.


Construction of Sierpiński triangles with the coexistence of C60 or MnPc molecules.
Sierpiński triangles up to the fourth order were successfully prepared using 4,4''-dicyano-1,1':3',1''-terphenyl molecules and Fe or Co atoms with the coexistence of the third molecules (C60 or MnPc) on a reconstructed Au(111) substrate. Sierpiński triangles could be disturbed and restored by C60 molecules after annealing at 400 and 360 K, respectively. The dispersed MnPc molecules did not affect the formation of Sierpiński triangles when annealed at high temperatures. The selective adsorption of MnPc molecules at certain holes of Sierpiński triangles was observed.


Parametric embedding for class visualization.
We propose a new method, parametric embedding (PE), that embeds objects with the class structure into a low-dimensional visualization space. PE takes as input a set of class conditional probabilities for given data points and tries to preserve the structure in an embedding space by minimizing a sum of Kullback-Leibler divergences, under the assumption that samples are generated by a gaussian mixture with equal covariances in the embedding space. PE has many potential uses depending on the source of the input data, providing insight into the classifier's behavior in supervised, semisupervised, and unsupervised settings. The PE algorithm has a computational advantage over conventional embedding methods based on pairwise object relations since its complexity scales with the product of the number of objects and the number of classes. We demonstrate PE by visualizing supervised categorization of Web pages, semisupervised categorization of digits, and the relations of words and latent topics found by an unsupervised algorithm, latent Dirichlet allocation.


[Minor surgery in laryngeal cancer].
The article presents the results of conservative surgery in 102 patients with laryngeal cancer T2 and T3. Frontolateral laryngeal resections in different variants (endoprosthesis, extended, an inferior variant) were conducted in 85 (83.3%) patients, horizontal laryngeal resection - in 17 (16.7%) patients (a combined variant -in 2 of them). The treatment resulted in primary healing of a postoperative wound in 92 (90.2%) patients. Complications consisted in development of pharyngeal fistula, suppuration of the postoperative wound, first of all in combined and extended resections, after a total dose of radiotherapy. Minor operations on the larynx enabled decannulation of 98 (96.1%) patients. Four (3.9%) patients continue to carry cannula because of marked scarry alterations.


Silver- and sulfadiazine-loaded nanostructured silica materials as potential replacement of silver sulfadiazine.
Silver sulfadiazine (AgSD) is the leading topical antibacterial agent for the treatment of burn wound infections. Antibacterial effect of AgSD is limited by its poor aqueous solubility, and antibacterial activity develops only by decomposition of AgSD to silver ions and sulfadiazine. In this study, it is for the first time that application of silver-modified nanoporous silica carriers (MCM-41 or SBA-15) loaded with sulfadiazine (SD), instead of silver sulfadiazine, overcoming the abovementioned disadvantages has been demonstrated. By direct or post synthesis methods, 5-15 nm sized silver nanoparticles can be stabilized in the channels or on the outer surface of nanoporous silica supports; moreover, the empty channels can be loaded by SD molecules. The SD-loaded, silver-modified materials show sustained release properties and similar or even better antimicrobial properties than AgSD. Adsorption of AgSD on nanoporous silica particles significantly improves its water solubility.


A small molecule discrimination map of the antibiotic resistance kinome.
Kinase-mediated resistance to antibiotics is a significant clinical challenge. These enzymes share a common protein fold characteristic of Ser/Thr/Tyr protein kinases. We screened 14 antibiotic resistance kinases against 80 chemically diverse protein kinase inhibitors to map resistance kinase chemical space. The screens identified molecules with both broad and narrow inhibition profiles, proving that protein kinase inhibitors offer privileged chemical matter with the potential to block antibiotic resistance. One example is the flavonol quercetin, which inhibited a number of resistance kinases in vitro and in vivo. This activity was rationalized by determination of the crystal structure of the aminoglycoside kinase APH(2″)-IVa in complex with quercetin and its antibiotic substrate kanamycin. Our data demonstrate that protein kinase inhibitors offer chemical scaffolds that can block antibiotic resistance, providing leads for co-drug design.


What determines severity among patients with painful functional bowel disorders?
For patients with painful functional bowel disorders (FBD), physicians frequently make diagnostic and treatment decisions based on the severity of the pain reported; patients with severe painful complaints may receive extensive diagnostic tests and treatments. Therefore, it would be important to determine what clinical factors contribute to the judgment of severity among patients with FBD. The aim of this study was to identify the psychosocial, behavioral, and physiological (visceral sensitivity) factors that predicted severity in patients with moderate to severe FBD. Two hundred eleven female patients with moderate or severe FBD, as determined by the Functional Bowel Disorder Severity Index, entered a multicenter treatment trial at the University of North Carolina and the University of Toronto. Patients filled out diary cards and were given questionnaires and physiological testing (rectal sensitivity using barostat). Analysis of covariance and logistic regression adjusting for demographic factors were performed to determine which factors distinguished patients at study entry with moderate from those with severe FBD. Patients with severe FBD were characterized by greater depression and psychological distress, poorer physical functioning and health-related quality of life, more maladaptive coping strategies, and greater health care utilization. There was a trend for patients with severe FBD to have lower rectal sensation thresholds. Regression analysis indicated that severity was best predicted by behavioral features: poorer daily physical function, difficulties related to eating, more phone calls to the physician, and more days in bed for GI symptoms. We conclude that patient illness behaviors are best correlated with severity in FBD. The use of psychopharmacological agents (e.g., antidepressants) and psychological treatments to treat psychiatric comorbidity and to improve behavioral coping styles is recommended. Training to help medical physicians identify and respond to psychosocial and behavioral features of these conditions is likely to improve patient satisfaction with their care and the clinical outcome.


Provision of paediatric intensive care services.
Paediatric intensive care has developed into a highly specialized and labour-intensive clinical activity. The provision of adequate numbers of properly staffed children's intensive care beds within the UK, and the lack of national planning and coordination are both problematic. This article examines these problems and discusses current thinking on the provision of paediatric intensive care services.


Observation of orbital currents in CuO.
Orbital currents are proposed to be the order parameter of the pseudo-gap phase of cuprate high-temperature superconductors. We used resonant x-ray diffraction to observe orbital currents in a copper-oxygen plaquette, the basic building block of cuprate superconductors. The confirmation of the existence of orbital currents is an important step toward the understanding of the cuprates as well as materials lacking inversion symmetry, such as magnetically induced multiferroics. Although observed in the antiferromagnetic state of cupric oxide, we show that orbital currents can occur even in the absence of long-range magnetic moment ordering.


Periodontitis and the incidence of metabolic syndrome: An 8-year longitudinal study of an adult Japanese cohort.
To examine the effect of periodontitis on the development of metabolic syndrome (MetS) and MetS components. This study included 4761 participants aged 30-74 years who underwent health examinations at both baseline and 8-year follow-up. The Japanese MetS criteria were used for diagnosis. The Community Periodontal Index was used to assess periodontal status. The association between periodontal status and MetS incidence was examined by Poisson regression analysis. Multivariate analysis revealed that individuals with a ≥6 mm periodontal pocket had a significantly higher relative risk (RR) for MetS onset, as compared to individuals without deep periodontal pockets (adjusted RR 1.30, 95% confidence interval [CI]: 1.01-1.67). Compared to individuals without a deep periodontal pocket, individuals with a ≥6 mm periodontal pocket had significantly higher RRs for developing two components of MetS; the RRs were 1.25 (95% CI: 1.01-1.56) for abdominal obesity and 1.39 (95% CI: 1.03-1.86) for hyperglycaemia. Individuals with periodontitis had a significantly higher risk of MetS onset, possibly due to the influence of periodontitis on abdominal obesity and hyperglycaemia.


Lubricating properties of single metal ions at interfaces.
The behaviour of ionic solutions confined in nanoscale gaps is central to countless processes, from biomolecular function to electrochemistry, energy storage and lubrication. However, no clear link exists between the molecular-level behaviour of the liquid and macroscopic observations. The problem mainly comes from the difficulty to interrogate a small number of liquid molecules. Here, we use atomic force microscopy to investigate the viscoelastic behaviour of pure water and ionic solutions down to the single ion level. The results show a glassy-like behaviour for pure water, with single metal ions acting as lubricants by reducing the elasticity of the nano-confined solution and the magnitude of the hydrodynamic friction. At small ionic concentration (<20 mM) the results can be quantitatively explained by the ions moving via a thermally-activated process resisted by the ion's hydration water (Prandtl-Tomlinson model). The model breaks down at higher salt concentrations due to ion-ion interaction effects that can no longer be neglected. The correlations are confirmed by direct sub-nanometre imaging of the interface at equilibrium. The results provide a molecular-level basis for explaining the tribological properties of aqueous solutions and suggest that ion-ion interactions create mesoscale effects that prevent a direct link between nanoscale and macroscopic measurements.


Successful Long-Term Multimodality Management of Facial Lesions in Tuberous Sclerosis Complex in an Adult Patient.
Angiofibroma and shagreen patches are common cutaneous manifestations of tuberous sclerosis complex (TSC) and have significant physical and psychological repercussions for patients. Several treatments have been proposed to improve lesions; however, clear treatment guidelines have not yet been presented. Thus, we introduce a case of angiofibroma and shagreen patch improved by application of pulsed dye laser, ablative fractional CO2 laser, and topical rapamycin, and present clinical implications for the treatment of angiofibroma and shagreen patch in TSC.


Urinary adenosine excretion in patients receiving amphotericin B.
Intravenous amphotericin B (AMB) administration in animals causes renal vasoconstriction, ischemia, and oliguria that may result in irreversible renal injury; the mechanism of AMB nephrotoxicity may be similar in human beings. Adenosine is excreted in urine by the ischemic kidney. We hypothesized that adenosine excretion and oliguria would be a marker for patients who later would manifest AMB-associated renal insufficiency and that pre-AMB saline administration (which ameliorates AMB nephrotoxicity) would negate the change in adenosine excretion and urine output. Twenty hospitalized patients being treated at the direction of their attending physician and who were receiving AMB (15 to 75 mg intravenously) had urine collected for 1 hour before and for 2 hours during AMB infusion. Eleven patients received normal saline solution (500 ml intravenously) before the AMB infusion; the other nine formed the comparator group. An aliquot of each urine collection was precipitated with perchloric acid to remove protein and cellular elements and centrifuged, and the supernatant was assayed for adenosine by using high-pressure liquid chromatography. Infusion of AMB was associated with a decrease in mean urine output both in patients who received saline solution (245 before versus 149 ml/hr during AMB infusion, p = 0.04) and in patients in comparator group (139 versus 89 ml/hr, p = 0.027). The mean urinary adenosine excretion was unchanged in the saline-loaded group (0.1354 before versus 0.1255 mmol/hr during drug infusion, p = 0.25) and was decreased in the comparator group (0.2276 versus 0.1127 mmol/hr, p = 0.01). Development of renal insufficiency did not correlate with the change in urine output or adenosine excretion. AMB infusion in human beings results in decreased urine output and decreased adenosine excretion. The latter effect is prevented by a pre-AMB saline load. The changes in urine output and adenosine excretion are not predictive of the development of renal insufficiency.


A large-scale evaluation of computational protein function prediction.
Automated annotation of protein function is challenging. As the number of sequenced genomes rapidly grows, the overwhelming majority of protein products can only be annotated computationally. If computational predictions are to be relied upon, it is crucial that the accuracy of these methods be high. Here we report the results from the first large-scale community-based critical assessment of protein function annotation (CAFA) experiment. Fifty-four methods representing the state of the art for protein function prediction were evaluated on a target set of 866 proteins from 11 organisms. Two findings stand out: (i) today's best protein function prediction algorithms substantially outperform widely used first-generation methods, with large gains on all types of targets; and (ii) although the top methods perform well enough to guide experiments, there is considerable need for improvement of currently available tools.


Proline is biosynthesized from arginine in Staphylococcus aureus.
Staphylococcus aureus NCTC 8325 exhibited a long lag phase (11 h) when inoculated into defined medium lacking proline, that could be shortened by increasing the concentration of arginine in the medium, or by supplying ornithine. Radioactivity from L-[14C]arginine, but not L-[14C]glutamate was incorporated into a spot with the chromatographic mobility of [14C]proline in the pool metabolites fraction. Selection for transposon Tn917-lacZ mutants impaired in arginine catabolism yielded four proline auxotrophs. Enzyme assays and precursor feeding experiments suggested that the major pathway for proline biosynthesis in S. aureus was from arginine via ornithine and delta'-pyrroline 5-carboxylate, rather than from glutamate. Strain 8325 Pro+, a proline prototrophic variant obtained by cultivation of 8325 in the absence of proline, accumulated L-[14C]arginine from the medium at about eight times the rate of strain 8325, suggesting its response to proline starvation was to increase arginine uptake.


Congenital Factor VII Deficiency in Association With Bicuspid Aortic Valve and Multicystic Dysplastic Kidney Disease in a Child.
Congenital factor VII deficiency is a rare bleeding disorder, with an incidence of 1:500,000. This case report describes an exceptionally unusual combination of congenital factor VII deficiency, multicystic dysplastic kidney disease and bicuspid aortic valve disease, in the same patient.


Identification of three novel mutations in the major human skeletal muscle chloride channel gene (CLCN1), causing myotonia congenita.
Myotonia congenita (MC) is a genetic disease characterized by mutations in the CLCN1 gene (OMIM*118425) encoding the skeletal muscle voltage-gated chloride channel (ClC-1). Autosomal dominant and recessive forms are observed, characterized by impaired muscle relaxation after forceful contraction (myotonia), which is more pronounced after inactivity and improves with exercise. We report three novel and one known mutations of the CLCN1 gene in four unrelated MC families. In two families the mutations were missense: 803C>T (T268M) and 1272C>G (I424M) in exons 7 and 12, respectively. The third was a splice mutation in intron 5 (696+2T>A), which induced a frame shift with a stop codon in exon 6 (fs213X). In the fourth family the previously-reported missense mutation 689G>A (G230E) was found. We also report two known polymorphisms: 261C>T (T87T) and 2154T>C (D718D) in exons 2 and 17 of two MC families; also found in 14 (33%) and 28 (67%) of 42 healthy controls, respectively. These findings expand our knowledge of mutations responsible for myotonia congenita, reducing the proportion of MC patients in whom genetic alterations have not been found.


Health benefits of isoflavones in functional foods? Proteomic and metabonomic advances.
There is currently considerable interest in the potential health benefits of isoflavones in functional foods and in the future prospects for the development of new products of benefit to the consumer. The potential health benefits of isoflavones may include protection against age-related diseases including cardiovascular disease, osteoporosis, hormone-dependent cancer and loss of cognitive function. The mechanisms involved may include weak oestrogenic action and antioxidant activity. Our proteomic investigations of changes in the human serum profile in response to the consumption of isoflavones in soya functional foods suggest potentially beneficial modulation of the levels of a number of serum proteins, including increased apolipoprotein E (involved in lipid metabolism) and caeruloplasmin (antioxidant and copper regulatory properties) levels and decreased alpha-1-acid glycoprotein (involved in immunomodulation) levels that may contribute to vascular protection. Furthermore, preliminary metabonomic data indicates an alteration in the urinary metabolite profile after isoflavone consumption, which may be of significance.


Does physical activity in adolescence have site-specific and sex-specific benefits on young adult bone size, content, and estimated strength?
The long-term benefits of habitual physical activity during adolescence on adult bone structure and strength are poorly understood. We investigated whether physically active adolescents had greater bone size, density, content, and estimated bone strength in young adulthood when compared to their peers who were inactive during adolescence. Peripheral quantitative computed tomography (pQCT) was used to measure the tibia and radius of 122 (73 females) participants (age mean ± SD, 29.3 ± 2.3 years) of the Saskatchewan Pediatric Bone Mineral Accrual Study (PBMAS). Total bone area (ToA), cortical density (CoD), cortical area (CoA), cortical content (CoC), and estimated bone strength in torsion (SSIp ) and muscle area (MuA) were measured at the diaphyses (66% tibia and 65% radius). Total density (ToD), trabecular density (TrD), trabecular content (TrC), and estimated bone strength in compression (BSIc ) were measured at the distal ends (4%). Participants were grouped by their adolescent physical activity (PA) levels (inactive, average, and active) based on mean PA Z-scores obtained from serial questionnaire assessments completed during adolescence. We compared adult bone outcomes across adolescent PA groups in each sex using analysis of covariance followed by post hoc pairwise comparisons with Bonferroni adjustments. When adjusted for adult height, MuA, and PA, adult males who were more physically active than their peers in adolescence had 13% greater adjusted torsional bone strength (SSIp , p < 0.05) and 10% greater adjusted ToA (p < 0.05) at the tibia diaphysis. Females who were more active in adolescence had 10% larger adjusted CoA (p < 0.05), 12% greater adjusted CoC (p < 0.05) at the tibia diaphysis, and 3% greater adjusted TrC (p < 0.05) at the distal tibia when compared to their inactive peers. Benefits to tibia bone size, content, and strength in those who were more active during adolescence seemed to persist into young adulthood, with greater ToA and SSIp in males, and greater CoA, CoC, and TrC in females.


Capsaicin-induced vasoconstriction in the mouse knee joint: a study using TRPV1 knockout mice.
Capsaicin is the pungent component of chilli peppers that concomitantly activates and desensitizes C-fibre and Adelta sensory nerve fibres. Stimulation causes an acute neurogenic response including vasodilation, plasma extravasation and hypersensitivity. However, in the present study we have shown that capsaicin produces a dose-dependent vasoconstrictor effect in the mouse knee joint via Transient Receptor Potential Vanilloid 1 (TRPV1) receptor activation. A (125)I-albumin accumulation technique showed that the intravascular volume of capsaicin-treated joints in wild type (WT) mice was significantly reduced compared to TRPV1 knockout mice (p<0.01). Similarly, a laser Doppler technique showed significantly reduced blood flow in the capsaicin-treated joints of WT compared to TRPV1 knockout mice (p<0.001). Pretreatment with guanethinidine (50 mg kg(-1), i.p.) had no effect on the vasoconstriction. These data are important considering the involvement of TRPV1 receptors in joint disease. The mechanisms underlying the vasoconstriction therefore require further investigation.


Distribution of valproate to subdural cerebrospinal fluid, subcutaneous extracellular fluid, and plasma in humans: a microdialysis study.
We sought to study the time course of the distribution of valproate (VPA) to subdural cerebrospinal fluid (CSF) in relation to subcutaneous extracellular fluid (ECF) and plasma after a single oral dose and to study the distribution to these three compartments under steady-state conditions. Microdialysis was used to estimate unbound VPA concentrations in subdural CSF and subcutaneous ECF, and blood samples were drawn for estimation of total and unbound VPA plasma concentrations in four patients with drug-resistant partial epilepsy undergoing presurgical evaluation with subdural EEG monitoring. Three patients were given a single oral dose of VPA, and one patient was receiving regular VPA treatment. VPA was analyzed by gas chromatography with flame ionization detection. The distribution of VPA to subdural CSF was rapid (Tmax, 3.5 h in two patients and 5.5 h in one patient) and subject to a minor delay in all three patients compared with that in the subcutaneous tissue ECF (Tmax, 2.5 h in all three patients), which in turn exhibited no evidence of a distribution delay compared with plasma. Subdural CSF levels of VPA were slightly lower than subcutaneous ECF levels (mean ratio, 0.78) and unbound plasma levels (mean ratio, 0.91). VPA rapidly enters the subdural CSF in unbound concentrations marginally lower than those obtained in subcutaneous ECF and plasma. These findings provide a pharmacokinetic rationale for acute administration of VPA. The good correlation between VPA concentrations in subcutaneous ECF and subdural CSF indicates that estimation of unbound VPA concentrations in subcutaneous tissue using microdialysis sampling has the potential to be useful for monitoring purposes.


Acromiothoracic Venous Supercharge for Abdominal-Based Free Flaps in Breast and Chest Wall Reconstruction.
Supercharging the venous drainage of free abdominal flaps in breast reconstruction has been well described in the literature, with diverse options used to augment venous drainage. In this study, we present our experience in using the acromiothoracic vein (ATV)/thoracoacromial vein (TAV) as a secondary recipient vein for the superficial inferior epigastric vein (SIEV) of free, muscle-sparing transverse rectus abdominis myocutaneous flaps in breast and chest wall reconstruction. We retrospectively reviewed 523 free, muscle-sparing transverse rectus abdominis myocutaneous flaps the senior author (H.H.K.) performed between 2009 and 2022 for breast and chest wall reconstruction; 46 cases required venous super drainage. Seventeen patients had ipsilateral SIEV anastomosed into the second internal mammary vein, 5 had ipsilateral SIEV anastomosed into flap second deep inferior epigastric vein, and 24 required the use of the (ATV)/(TAV), which will be the focus of this study. The study included 24 female (20 breast and 4 chest wall reconstruction) patients ranging in ages between 39 and 72 years. They had a median follow-up of 26 months. Combined muscle splitting and cutting techniques were used to expose the ATV/TAV. Increase in operative time ranged between 10 and 20 minutes (median, 12 minutes). Vein coupler sizes were 1.5 to 3 mm. The mean weight of the flap was 740 g (range, 460-1300 g). There was 1 flap failure (salvage with latissimus dorsi flap performed), whereas 23 flaps wholly survived. The ATV/TAV is a suitable recipient for venous supercharging free flaps used to reconstruct breast and chest wall defects.


A Multipurpose Toolkit to Enable Advanced Genome Engineering in Plants.
We report a comprehensive toolkit that enables targeted, specific modification of monocot and dicot genomes using a variety of genome engineering approaches. Our reagents, based on transcription activator-like effector nucleases (TALENs) and the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 system, are systematized for fast, modular cloning and accommodate diverse regulatory sequences to drive reagent expression. Vectors are optimized to create either single or multiple gene knockouts and large chromosomal deletions. Moreover, integration of geminivirus-based vectors enables precise gene editing through homologous recombination. Regulation of transcription is also possible. A Web-based tool streamlines vector selection and construction. One advantage of our platform is the use of the Csy-type (CRISPR system yersinia) ribonuclease 4 (Csy4) and tRNA processing enzymes to simultaneously express multiple guide RNAs (gRNAs). For example, we demonstrate targeted deletions in up to six genes by expressing 12 gRNAs from a single transcript. Csy4 and tRNA expression systems are almost twice as effective in inducing mutations as gRNAs expressed from individual RNA polymerase III promoters. Mutagenesis can be further enhanced 2.5-fold by incorporating the Trex2 exonuclease. Finally, we demonstrate that Cas9 nickases induce gene targeting at frequencies comparable to native Cas9 when they are delivered on geminivirus replicons. The reagents have been successfully validated in tomato (Solanum lycopersicum), tobacco (Nicotiana tabacum), Medicago truncatula, wheat (Triticum aestivum), and barley (Hordeum vulgare).


Developmental changes in cognitive and behavioural functioning of adolescents with fragile-X syndrome.
Individuals with fragile-X syndrome exhibit developmental delay, hyperexcitation and social anxiety; they also show lack of attention and hyperactivity. Few studies have investigated whether levels of functioning change with increasing age. Here, we explored developmental changes across adolescence in the cognitive and behavioural profile of individuals with fragile-X syndrome. To this scope, we assessed intellectual functioning, adaptive behaviour, autistic symptomatology, behavioural problems (e.g. hyperactivity/lack of attention) and strengths (prosocial behaviours). Thirty-six participants underwent standardised outcome measures (i.e. the Wechsler Intelligence Scales-Revised, the Childhood Autism Rating Scale, the Vineland Adaptive Behavior Scales, and the Strengths and Difficulty Questionnaire) in three time points (Time 1: 9-11; Time 2: 11-13, and Time 3: 13-15 years). Verbal IQ improved across time, whereas Nonverbal IQ declined and Full Scale IQ was quite unchanged. Autism ratings decreased; communication and social aspects of adaptive behaviour also enhanced. Finally, elevated levels of hyperactivity/lack of attention at Time 1 significantly improved across the three time points, whereas emotional symptoms, behavioural difficulties, problems with peers and prosocial behaviours remained stable over time. These findings revealed specific developmental changes in cognitive and behavioural functioning of individuals with fragile-X syndrome, likely related to a progressive maturation of brain systems devoted to attentional control.


Vasopressin and atrial natriuretic peptide release in cardiopulmonary denervated dogs.
To assess the effect of cardiopulmonary denervation on the release of arginine vasopressin (AVP) and atrial natriuretic peptide (ANP), either hemorrhage (1.0 ml.kg-1.min-1, for 40 min) or an infusion of 3% dextran in lactated Ringer solution (RL) (3% dextran in RL, 1.0 ml.kg-1.min-1, for 40 min) was carried out in chronic cardiopulmonary denervated (CPD, n = 7) and sham-operated (SO, n = 7) dogs under anesthesia. Plasma AVP increased significantly in both groups during hemorrhage, but its rise was significantly attenuated in CPD dogs. Plasma ANP decreased similarly during hemorrhage in both groups. An infusion of 3% dextran in RL decreased significantly plasma AVP during its whole period in SO dogs, but AVP release was only suppressed in the late period in SO dogs. Plasma ANP increased significantly during its infusion in both groups. These results indicate that cardiopulmonary denervation may attenuate the release of AVP in response to either hemorrhage or blood volume expansion but may not affect the release of ANP in response to the blood volume change. Moreover, a large increase in plasma ANP may attenuate the release of AVP.


Hepatic acetyl CoA links adipose tissue inflammation to hepatic insulin resistance and type 2 diabetes.
Impaired insulin-mediated suppression of hepatic glucose production (HGP) plays a major role in the pathogenesis of type 2 diabetes (T2D), yet the molecular mechanism by which this occurs remains unknown. Using a novel in vivo metabolomics approach, we show that the major mechanism by which insulin suppresses HGP is through reductions in hepatic acetyl CoA by suppression of lipolysis in white adipose tissue (WAT) leading to reductions in pyruvate carboxylase flux. This mechanism was confirmed in mice and rats with genetic ablation of insulin signaling and mice lacking adipose triglyceride lipase. Insulin's ability to suppress hepatic acetyl CoA, PC activity, and lipolysis was lost in high-fat-fed rats, a phenomenon reversible by IL-6 neutralization and inducible by IL-6 infusion. Taken together, these data identify WAT-derived hepatic acetyl CoA as the main regulator of HGP by insulin and link it to inflammation-induced hepatic insulin resistance associated with obesity and T2D.