Synergistic anti-liver fibrosis actions of total astragalus saponins and glycyrrhizic acid via TGF-β1/Smads signaling pathway modulation.
Huangqi decoction (HQD) is a well-known traditional Chinese herbal formulation, It is an effective treatment for consumptive disease and chronic liver diseases. It consists of Radix Astragali (Astragalus membranceus(Fisch.) Bge. Root, Huangqi) and Radix Glycyrrhizae (Glycyrrhiza uralensis Fisch., root and rhizome, Gancao). Total astragalus saponins (AST) is a main component of Radix Astragali and glycyrrhizic acid(GA) is a main component of Radix Glycyrrhizae. Our primary results showed that the combination of AST and GA had an obvious synergistic effect in reducing liver collagen deposition and decreasing serum alanine aminotransferase (ALT) activity in dimethylnitrosamine (DMN)-induced liver fibrosis. Through in vivo and in vitro experiments, we aimed at investigating the key anti-fibrosis signal pathway TGF-β1/Smads to further explore the synergistic mechanism of AST and GA. Two hepatic fibrosis animal models, bile duct ligation-induced (BDL) and DMN-induced, were utilized. Rats were treated orally with AST, GA or AST/GA, with the effects evaluated via liver histopathology, hydroxyproline (Hyp) levels, and α-SMA expression. In the hepatic stellate cell line JS-1, cells were treated with AST/GA for 24h, followed by a cell viability assessment using Cell Counting Kit-8(CCK-8) and Real-time PCR and Western blot analysis of α-SMA, ColⅠ and TGF-β1/Smads signaling pathway related components. The AST/GA combination attenuated liver tissue inflammation, collagen deposition, Hyp levels, and α-SMA expression in both BDL-and DMN-stimulated hepatic fibrosis rats. In vitro results showed that the AST/GA combination significantly inhibited JS-1 cell viability, significantly suppressed α-SMA, ColⅠ, TGF-β1, Smad2 and Smad3 mRNA and protein expression, as well reduced p-Smad2/3. Compared with AST or GA treatment alone, the AST/GA combination significantly reduced Smad3 mRNA expression levels and TGF-β1, Smad3, and p-Smad2/3 protein levels. AST and GA synergistically alleviated both BDL-and DMN-induced hepatic fibrosis via TGF-β1/Smads signaling pathway inhibition in hepatic stellate cells.


Ih current stabilizes excitability in rodent DRG neurons and reverses hyperexcitability in a nociceptive neuron model of inherited neuropathic pain.
We show here that hyperpolarization-activated current (Ih ) unexpectedly acts to inhibit the activity of dorsal root ganglion (DRG) neurons expressing WT Nav1.7, the largest inward current and primary driver of DRG neuronal firing, and hyperexcitable DRG neurons expressing a gain-of-function Nav1.7 mutation that causes inherited erythromelalgia (IEM), a human genetic model of neuropathic pain. In this study we created a kinetic model of Ih and used it, in combination with dynamic-clamp, to study Ih function in DRG neurons. We show, for the first time, that Ih increases rheobase and reduces the firing probability in small DRG neurons, and demonstrate that the amplitude of subthreshold oscillations is reduced by Ih . Our results show that Ih , due to slow gating, is not deactivated during action potentials (APs) and has a striking damping action, which reverses from depolarizing to hyperpolarizing, close to the threshold for AP generation. Moreover, we show that Ih reverses the hyperexcitability of DRG neurons expressing a gain-of-function Nav1.7 mutation that causes IEM. In the aggregate, our results show that Ih unexpectedly has strikingly different effects in DRG neurons as compared to previously- and well-studied cardiac cells. Within DRG neurons where Nav1.7 is present, Ih reduces depolarizing sodium current inflow due to enhancement of Nav1.7 channel fast inactivation and creates additional damping action by reversal of Ih direction from depolarizing to hyperpolarizing close to the threshold for AP generation. These actions of Ih limit the firing of DRG neurons expressing WT Nav1.7 and reverse the hyperexcitability of DRG neurons expressing a gain-of-function Nav1.7 mutation that causes IEM. KEY POINTS: Hyperpolarization-activated cyclic nucleotide-gated (HCN) channels, the molecular determinants of hyperpolarization-activated current (Ih ) have been characterized as a 'pain pacemaker', and thus considered to be a potential molecular target for pain therapeutics. Dorsal root ganglion (DRG) neurons express Nav1.7, a channel that is not present in central neurons or cardiac tissue. Gain-of-function mutations (GOF) of Nav1.7 identified in inherited erythromelalgia (IEM), a human genetic model of neuropathic pain, produce DRG neuron hyperexcitability, which in turn produces severe pain. We found that Ih increases rheobase and reduces firing probability in small DRG neurons expressing WT Nav1.7, and demonstrate that the amplitude of subthreshold oscillations is reduced by Ih . We also demonstrate that Ih reverses the hyperexcitability of DRG neurons expressing a GOF Nav1.7 mutation (L858H) that causes IEM. Our results show that, in contrast to cardiac cells and CNS neurons, Ih acts to stabilize DRG neuron excitability and prevents excessive firing.


A preliminary report: the changes in the neuropeptide containing epidermal innervation in response to inflammatory reactions elicited in human breast skin.
A pilot study was performed to demonstrate the involvement of the epidermal innervation in cutaneous neurogenic inflammation following injury or skin incision. Scratching human skin is known to evoke an inflammatory reaction dependent in part on sensory nerves (neurogenic inflammation) in the dermis. Human breast skin from four patients was scratched using a blunt metal ruler to elicit an inflammatory reaction. Samples of skin along the scratch line were then removed before and up to 5 min after scratching. Tissue samples were then stained using immunocytochemical techniques for the neuropeptides PGP 9.5, SP, CGRP and VIP. Quantitative assessment using image analysis demonstrated a marked variation between patients in the levels of epidermal PGP 9.5 innervation prior to scratching of the skin. In addition both increases and decreases in the epidermal innervation of PGP 9.5 were observed in different patients immediately after scratching. It is considered that epidermal innervation may have an important role in the inflammatory reaction to trauma and surgical operations and that a more extensive study is merited.


A hermeneutic phenomenological analysis of midwives' ways of knowing during childbirth.
to explore the ways of knowing used by the midwife while attending women during childbirth through textual analysis of poems written by American midwives. a hermeneutic phenomenology and human science research method inspired by van Manen was used. Midwifery ways of knowing during childbirth were thematically derived from 10 poems written by midwives about attending childbirth or the experience of being a midwife. Textual analysis included examination of the poems as a whole, via verse and metaphor, and via individual lines of prose. 10 American midwives wrote the poems used in this study. The poems were discovered through online searches of many databases using the key words 'poetry, poems, midwifery and childbirth' and through a national call for poetry by the researcher over a period of 4 years (1996-2000), undertaken in order to publish an anthology of poetry written by midwives. three authoritative ways of knowing that guided the care given by the midwife to women during childbirth were discovered. They were self-knowledge from the belief system of the individual midwife, grounded knowledge from the midwife's personal lived experience with childbirth, and informed knowledge from objective and scholarly sources. midwives must continue to develop their own body of knowledge in order to move the profession forward. Multiple ways of knowing including the use of experiential/contextual and intuitive knowledge is legitimate and humane, if provision of care is holistic. Care of women during childbirth can be enhanced with the use of multiple knowers and multiple ways of knowing. This study captured a unique and fresh interpretation of the lived experience of midwifery knowledge. Midwifery educational programmes should offer opportunities for students to explore the artistry as well as the science of midwifery practice.


Increased brain levels of F2-isoprostane are an early marker of behavioral sequels in a rat model of global perinatal asphyxia.
Perinatal asphyxia is a major cause of immediate and postponed brain damage in the newborn. It may be responsible for several delayed neurologic disorders and, in this respect, early markers of brain injury would be relevant for therapeutic intervention as well as for identification of infants at high risk for developmental disabilities. Biochemical measurements (brain F2-isoprostane levels) and behavioral tests (ultrasonic vocalization pattern on postnatal days (pnd) 5, 8, and 11, spontaneous motor behaviors on pnd 7 and 12, and homing response on pnd 10) were performed in a rat model of global perinatal asphyxia in the immature neonate. Caesarean section was performed in rats and the pups, still in uterus horns, were placed into a water bath at 37 degrees C for either 10 or 20 min. Caesarean delivered pups were used as controls. Pups experiencing severe (20 min), in contrast to those undergoing the 10 min, asphyctic insult presented with detectable abnormalities including early (two hours after the insult) increase in brain F2-isoprostane (a direct marker of oxidative injury) without detectable changes in PGE2, COX-2 and iNOS levels, and delayed physical (reduced weight gain on pnd 5 and thereafter) and behavioral disturbances (alterations in ultrasound emission on pnd 11 and spontaneous motricity levels mainly). These findings suggest that increased brain F2-isoprostane levels shortly after the asphyctic insult are predictive of delayed behavioral disturbances in the newborn rat. The present 20-min asphyxia model might serve for the assessment of preventive and curative strategies to treat neurologic/behavioral disturbances associated with perinatal asphyxia.


Comparison of endoscopic ultrasound, computed tomography and magnetic resonance imaging in assessment of detailed structures of pancreatic cystic neoplasms.
To evaluate the advantages of endoscopic ultrasound (EUS) in the assessment of detailed structures of pancreatic cystic neoplasms (PCNs) compared to computed tomography (CT) and magnetic resonance imaging (MRI). All patients with indeterminate PCNs underwent CT, MRI, and EUS. The detailed information, including size, number, the presence of a papilla/nodule, the presence of a septum, and the morphology of the pancreatic duct of PCNs were compared among the three imaging modalities. The size of each PCN was determined using the largest diameter measured. A cyst consisting of several small cysts was referred to as a mother-daughter cyst. Disagreement among the three imaging modalities regarding the total number of mother cysts resulted in the assumption that the correct number was the one in which the majority of imaging modalities indicated. A total of 52 females and 16 males were evaluated. The median size of the cysts was 42.5 mm by EUS, 42.0 mm by CT and 38.0 mm by MRI; there was no significant difference in size as assessed among the three imaging techniques. The diagnostic sensitivity and ability of EUS to classify PCNs were 98.5% (67/68) and 92.6% (63/68), respectively. These percentages were higher than those of CT (73.1%, P < 0.001; 17.1%, P < 0.001) and MRI (81.3%, P = 0.001; 20.3%, P < 0.001). EUS was also able to better assess the number of daughter cysts in mother cysts than CT (P = 0.003); however, there was no significant difference between EUS and MRI in assessing mother-daughter cysts (P = 0.254). The papilla/nodule detection rate by EUS was 35.3% (24/68), much higher than those by CT (5.8%, 3/52) and MRI (6.3%, 4/64). The detection rate of the septum by EUS was 60.3% (41/68), which was higher than those by CT (34.6%, 18/52) and by MRI (46.9%, 30/64); the difference between EUS and CT was significant (P = 0.02). The rate of visualizing the pancreatic duct using EUS was 100%, whereas using CT and MRI it was less than 10%. EUS helps visualize the detailed structures of PCNs and has many advantages over CT and MRI. EUS is valuable in the diagnosis and assessment of PCNs.


Design and implementation of a CORBA-based genome mapping system prototype.
CORBA (Common Object Request Broker Architecture), as an open standard, is considered to be a good solution for the development and deployment of applications in distributed heterogeneous environments. This technology can be applied in the bioinformatics area to enhance utilization, management and interoperation between biological resources. This paper investigates issues in developing CORBA applications for genome mapping information systems in the Internet environment with emphasis on database connectivity and graphical user interfaces. The design and implementation of a CORBA prototype for an animal genome mapping database are described. The prototype demonstration is available via: http://www.ri.bbsrc.ac.uk/ark_corba/. email@example.com


An unusual duplication of the inferior vena cava in a patient with endovascular repair for abdominal aortic aneurysm.
A 66-year-old Caucasian male, with sensation of abdominal pulsation was admitted to our hospital. In multidetector 64-row computed tomography (CT) angiography, an abdominal aortic aneurysm was observed. Endovascular aortic repair was performed. Control CT confirmed prosperity with stent graft fixation and absence of any vascular complications. Investigation also showed asymmetrical duplication of the inferior vena cava (IVC). Right (RIVC) and left (LIVC) inferior vena cava arose from the confluence of the right and left iliac veins. The LIVC continued as left renal vein.


On the physical nature of mesophases of guanosine gels.
The unusual columnar aqueous mesophases of self-assembled guanosine stacks, such as 2'-deoxyguanosine 5'-monophosphate and 2'-deoxyguanosine 3'-monophosphate, are analyzed in terms of a general theory of azimuthal correlations between the charged helices. This theory considers forces, specific to the helical structure of each macromolecule, which depend on the azimuthal orientations of the molecules about their long axes. More specifically, in determining the magnitudes and decay lengths of these helix specific forces we utilize the Kornyshev-Leikin theory of electrostatic interaction between helical macromolecules and quantitatively fit experimental data. Together with explaining a number of the observed features of these mesophases, several new effects are predicted. Possible limitations and developments of our theoretical model are discussed, as well as new experiments to test the implications of the theory.


Endowments and parental investments in infancy and early childhood.
This article tests whether parents reinforce or compensate for child endowments. We estimate how the difference in birth weight across siblings impacts specific parental investments: breast-feeding, well-baby visits, immunizations, and preschool attendance. Our results indicate that normal-birthweight children are 5%-11% more likely to receive early childhood parental investments than their low-birth-weight siblings. Moreover, the presence of additional low-birth-weight siblings in the household increases the likelihood of investments such as well-baby visits and immunizations for normal-birth-weight children. These results suggest that parental investments in early childhood tend to reinforce endowment differences.


Effects of the high-affinity Peptide reversin 121 on multidrug resistance proteins in experimental pancreatic cancer.
Multidrug resistance (MDR) is the major obstacle in the treatment of pancreatic carcinoma. Recent studies indicate resistance-modulating effects of high-affinity peptides. Pancreatic cancer was induced in an ultrasound-guided orthotopic mouse model. Expression and function of MDR proteins (MRPs) P-glycoprotein MRP1 and MRP3 were analyzed in vitroandin vivo by multicolor flow-cytometric analysis. Proliferation of tumor cells was evaluated with a nonradioactive proliferation assay. Animals and cells received standard chemotherapy or chemotherapy plus the high-affinity peptide reversin 121 (R121). The proportions of P-glycoprotein-, MRP1- and MRP3-positive tumor cells increased significantly after chemotherapy in vitro and in vivo. Drug efflux activity increased after chemotherapy. Addition of R121 to chemotherapeutic regimens significantly reduced the proportions of tumor cells positive for MRPs in vitroandin vivo. Tumor size and prevalence of metastases decreased significantly compared to untreated controls. Beneficial effects of high-affinity peptide R121 on reversing chemotherapy-induced MDR were demonstrated in pancreatic cancer.


Chiropractic spinal manipulative therapy for cervicogenic headache: a single-blinded, placebo, randomized controlled trial.
Cervicogenic headache is a disabling headache where pharmacological management have limited effect. Thus, non-pharmacological management is warranted. Our objective was therefore to investigate the efficacy of chiropractic spinal manipulative therapy versus placebo (sham manipulation) and control (continued usual but non-manual management) for cervicogenic headache in a prospective 3-armed single-blinded, placebo, randomized controlled trial of 17 months' duration. Nineteen participants were equally randomized into the three groups, and 12 participants completed the randomized controlled trial. Headache frequency improved at all time points in the chiropractic spinal manipulative therapy and the placebo group. Headache index improved in the chiropractic spinal manipulative therapy group at all time points, while it improved at 6 and 12 months' follow-up in the placebo group. The control group remained unchanged during the whole study period. Adverse events were few, mild and transient. Blinding was concealed throughout the RCT. Thus, our results suggest that manual-therapy might be a safe treatment option for participants with cervicogenic headache, but data need to be confirmed in a randomized controlled trial with sufficient sample size and statistical power. Trial registration ClinicalTrials.gov identifier: NCT01687881, 11 September 2012.


Natural and manufactured objects activate the fusiform face area.
Recent fMRI studies indicate that the anterior fusiform gyrus (the fusiform face area, FFA) is specialized for face recognition. However, the analyses used to determine face selectivity have not ruled out the possibility that other object categories produce significant activation in the FFA, relative to baseline. In the current fMRI study, we use a conservative hypothesis testing approach to show that FFA activation is not selective for faces. Rather, the FFA response is almost completely explained by a graded response in which faces produce more activation than either manufactured or natural objects, but those categories produce a statistically greater response than the baseline task. These findings question whether the FFA can be interpreted as a specialized module for face recognition.


IL-12 plasmid delivery by in vivo electroporation for the successful treatment of established subcutaneous B16.F10 melanoma.
Interleukin-12 (IL-12) has been used in numerous immunotherapy protocols against melanoma. However, delivery of IL-12 in the form of recombinant protein can result in severe toxicity, and gene therapy has had limited success against B16.F10 murine melanoma. The purpose of this study was to examine the effectiveness of in vivo electroporation for the delivery of plasmid DNA encoding IL-12 as an antitumor agent against B16.F10 melanoma. We treated mice bearing established B16.F10 melanoma tumors with intratumoral (i.t.) or intramuscular (i.m.) injections of a plasmid encoding IL-12, followed by in vivo electroporation. For i.t. treatments, we used an applicator containing six penetrating electrodes to deliver 1500-V/cm, 100-micros pulses. We administered i.m. pulses with an applicator containing four penetrating electrodes delivering 100-V/cm, 20-ms pulses. The i.t. treatment resulted in the cure of 47% of tumor-bearing mice, and 70% of cured mice were resistant to challenge with B16.F10 cells. The i.m. treatment did not result in tumor regression. We found that i.t. treatment resulted in increased levels of IL-12 and interferon-gamma (IFN-gamma) within the tumors, the influx of lymphocytes into the tumors, and reduction in vascularity. Neither i.m. nor i.t. treatment was successful against B16.F10 tumors in a nude mouse model, supporting a role for T cells in regression of this tumor model.


Analytical study on the kinematic and dynamic behaviors of a knee joint.
A knee model in the sagittal plane is established in this study. Specifically, the model is used to study the effects of inertia, articular surfaces of the knee joint, and patella on the behaviors of a knee joint. These behaviors include the joint surface contact point, ligament forces, instantaneous center and slide/roll ratio between the femur and tibia. Model results are compared to experimental cadaver studies available in the literature, as well as between the quasistatic and dynamic models. We found that inertia increases the sliding tendency in the latter part of flexion, and lengthens the cruciate ligaments. Decreasing the curvature of the femur surface geometry tends to reduce the ligament forces and moves the contact points towards the anterior positions. The introduction of the patellar ligament in the model seems to stabilize the behaviors of the knee joint as reflected by the behavior of the instant centers and the contact point pattern on the tibia surface. Furthermore, we found that different magnitudes of the external load applied to the tibia do not alter the qualitative behaviors of the knee joint.


Drug effectiveness explained: the mathematics of antiviral agents for HIV.
Antiretroviral therapy has improved the quality and length of life of millions of individuals affected by human immunodeficiency virus type 1 (HIV-1). The capacity of these drugs to indefinitely suppress HIV-which has a well-known capacity for escaping antiviral pressures-is surprising. In this issue of Science Translational Medicine, Shen et al. use a combination of mathematical modeling and experiment to examine the potency of anti-HIV drugs. They show that non-nucleoside reverse transcriptase inhibitors and protease inhibitors exhibit cooperative dose-response curves-a finding that has implications for the treatment of HIV as well as other viral infections, such as hepatitis C.


Malignant obstructive jaundice - brachytherapy as a tool for palliation.
Malignant obstructive jaundice (MOJ) is relieved by stenting via endoscopic retrograde cholangiopancreatography (ERCP) or percutaneous transhepatic cholangiography and biliary drainage (PTCD). Stent occlusion rates of 30-45% have been reported in literature due to tumor ingrowth or overgrowth. We prospectively evaluated the feasibility and the role of intraluminal brachytherapy (ILBT) in preventing stent blockage in patients with MOJ after PTCD and stenting. Twelve patients with MOJ who underwent PTCD followed by self expanding metallic stent (SEMS) placement were prospectively enrolled in this study. Written informed consent was obtained. Intraluminal brachytherapy was done once patient was stable and serum bilirubin was less than 2 mg% or 50% of baseline value. On the day of ILBT, 6 French brachytherapy catheters were placed across malignant stricture under fluoroscopic guidance with placement of the tip 1 cm distal to stricture. A dose of 10 to 14 Gy was delivered at 1 cm from central axis of the source. Suitable patients also received external beam radiotherapy (EBRT) with weekly concurrent chemotherapy. All patients tolerated the procedure well with minimal acute and late toxicities. Duodenal ulceration was observed in 1 patient. At a mean follow up of 10.25 months (5-24 months), stents were patent in 10/12 subjects and stent patency duration of 9.8 months (5-22) was reported. Intraluminal brachytherapy post PTCD is feasible and effective in preventing stent occlusion with minimal acute and late toxicities.


An insight into the sialotranscriptome of Triatoma rubida (Hemiptera: Heteroptera).
The kissing bug Triatoma rubida (Uhler, 1894) is found in southwestern United States and parts of Mexico where it is found infected with Trypanosoma cruzi, invades human dwellings and causes allergies from their bites. Although the protein salivary composition of several triatomine species is known, not a single salivary protein sequence is known from T. rubida. Furthermore, the salivary diversity of related hematophagous arthropods is very large probably because of the immune pressure from their hosts. Here we report the sialotranscriptome analysis of T. rubida based on the assembly of 1,820 high-quality expressed sequence tags, 51% of which code for putative secreted peptides, including lipocalins, members of the antigen five family, apyrase, hemolysin, and trialysin families. Interestingly, T. rubida lipocalins are at best 40% identical in primary sequence to those of T. protracta, a kissing bug that overlaps its range with T. rubida, indicating the diversity of the salivary lipocalins among species of the same hematophagous genus. We additionally found several expressed sequence tags coding for proteins of clear Trypanosoma spp. origin. This work contributes to the future development of markers of human and pet exposure to T. rubida and to the possible development of desensitization therapies. Supp. Data 1 and 2 (online only) of the transcriptome and deducted protein sequences can be obtained from http://exon.niaid.nih.gov/transcriptome/Trubida/Triru-S1-web.xlsx and http://exon.niaid.nih.gov/transcriptome/Trubida/Triru-S2-web.xlsx.


Clinical significance of serum soluble HLA class I antigens in systemic lupus erythematosus.
Serum soluble HLA (SHLA) class I antigens in patients with systemic lupus erythematosus (SLE) were examined by enzyme-linked immunosorbent assay. The primary antibody used was W6/32, anti-HLA class I mouse monoclonal antibody, and the secondary antibody was peroxidase-conjugated anti-mouse IgG antibody. The reactivity of SHLA class I antigens, the number of peripheral lymphocytes, and the clinical state of SLE were compared. The results were as follows; 1) Sixteen of 55 samples from SLE patients were SHLA class I antigen-positive (29%). 2) The reactivity of SHLA class I antigens was closely related to disease activity. 3) The number of lymphocytes and the reactivity of SHLA class I antigens were negatively correlated. 4) Nine of 23 samples from the active SLE patients were SHLA class I antigen-negative. Of these, 7 were from patients with a nephrotic syndrome due to lupus nephritis. It is concluded that SHLA class I antigens can be an useful marker for monitoring the clinical state of SLE.


Patterning-induced surface chirality and modulation of director twist in a nematic cell.
A substrate coated with a polyimide alignment layer is scribed bidirectionally with the stylus of an atomic force microscope to create an easy axis for liquid-crystal orientation. The resulting noncentrosymmetric topography breaks two-dimensional inversion symmetry and results in a spatial amplitude modulation of an imposed twisted nematic state. This is observed optically as spatially periodic light and dark stripes. When the alignment layer is scribed unidirectionally the centrosymmetric topography maintains inversion symmetry, and no stripes are observed. The appearance of the twist modulation is consistent with a chiral term in the free energy.


[The possible effect of basic periodontal treatment on the outcome of pregnancy].
More publications support the statement that maternal periodontitis may be a risk factor of preterm delivery and low birth-weight, than the number of those that suggest opposing evidence. The proving or rejecting of the questionable relationship may be carried out--among other methods--by analyzing the results of interventional studies. The aim of our study was to investigate whether or not it is possible to decrease the chance of preterm birth and low birth-weight in the case of healthy non-smoking women through the elimination of periodontal inflammation by providing professional oral hygiene treatment during pregnancy. After periodontal examination 80 pregnant women received professional oral hygiene treatment including plaque and calculus removal, root planing, motivation and instruction (treatment group); only the periodontal status was recorded in 79 cases (control group). The mean length of pregnancy and birth-weight in the treatment group were compared with similar data of the control group. Data were assessed using statistical methods. The mean weight of newborns was 3005.3 grams in the treatment group, while 2644.2 grams in the control group (p < 0.0001). The delivery occurred later in the treatment group (37.0 week), than among the control group (36.4 week), although the difference was not significant (p = 0.059). In the periodontal treated group, which had a statistically suitable number of members, the periodontal treatment might have contributed to a more optimal date of delivery and to achieving a larger birth-weight.


[Mortality following sustained ventricular tachycardia treated according to the results of programmed ventricular stimulation].
The purpose of this study was to evaluate longterm mortality in 44 patients undergoing electrophysiologically guided therapy for sustained monomorphic ventricular tachycardia. We applied the following modified response criteria: non-inducibility or slowing of induced ventricular tachycardia to < or = 150 bpm. On this basis, 25/44 (57%) patients were classified as responders, and 19/44 (43%) as non-responders. Responders had palpitations significantly more often as the leading clinical sign (68% vs. 21%, p < 0.05). Non-responders showed a lower mean ejection fraction (36 +/- 15% vs. 46 +/- 17%, p < 0.05) and a trend to higher incidence of syncopal ventricular tachycardia (58% vs. 12%, p = 0.15). All responders and 3/19 non-responders continued on drug therapy. 11 non-responders received an implantable cardioverter-defibrillator (ICD), 3 underwent surgery and 2 catheter ablation. 14/44 patients (32%) died during a mean follow-up of 3.2 years. Sudden arrhythmic death occurred in 3/28 patients on drug therapy (1/25 responders, 2/3 non-responders), and in 2/16 non-responders with invasive therapy (one with an ICD and one after catheter ablation). The cumulative all-cause mortality at 3 years was similar in both groups (26% vs 22%, n.s.), but the mortality from sudden arrhythmic death showed a trend to be lower in the responder-group (0% vs 17%, p = 0.09).(ABSTRACT TRUNCATED AT 250 WORDS)


Orofacial Mycoses in Coronavirus Disease-2019 (COVID-19): A Systematic Review.
Studies reviewing orofacial mycoses in coronavirus disease-2019 (COVID-19) caused by severe acute respiratory syndrome 2 (SARS-CoV-2) infection are sparse. Here we review the major oral and maxillofacial mycoses of COVID-19, the associated comorbidities, and the probable precipitating factors. English-language manuscripts published between March 2020 and October 2021 were searched using PubMed, OVID, SCOPUS, and Web of Science databases, using appropriate keywords. We identified 30 articles across 14 countries, which met the inclusion criteria of PRISMA guidelines. These yielded a total of 292 patients with laboratory-confirmed COVID-19, 51.4% (n = 150) of whom presented with oral and maxillofacial fungal infections, mainly comprising candidosis, mucormycosis, and aspergillosis. Candida infections were the most prevalent, present in 64% (n = 96), followed by mucormycosis, and only a single case of aspergillosis was noted. Oral and maxillofacial mycoses were predominantly seen in those with comorbidities, especially in those with diabetes (52.4%). Oral mucormycosis was noted in 8.6% (n = 13) and mainly manifested on the hard palate. An overall event rate of oral/maxillofacial mucormycosis manifestation in patients with COVID-19 with diabetes mellitus type 1/2 was about 94% (49/52; 95% confidence interval, 0.73%-0.89%), implying a very high association between diabetes mellitus and the latter condition. All fungal infections appeared either concurrently with COVID-19 symptoms or during the immediate recovery period. SARS-CoV-2 infection-related immunosuppression, steroid therapy, as well as comorbidities such as diabetic hyperglycemia appear to be the major predisposing factors for the onset of oral and maxillofacial mycoses in patients with COVID-19 across all age groups.


Prevalence and characteristics of ocular pain in non-infectious uveitis: a quality of life study.
To survey the frequency, character, severity and impact of ocular pain on quality of life in adult patients with non-infectious uveitis (NIU). This patient-requested cross-sectional survey study describes the results of three self-administered questionnaires (the National Eye Institute Visual Function Questionnaire, the 36-Item Short Form Health Survey (SF-36) and the McGill Pain Questionnaire Dutch Language Version) from 147 patients with NIUs from a university-based tertiary referral centre in Utrecht. The mean Visual Function Questionnaire (VFQ) Ocular Pain Score of all patients with NIU was 72 (±24), which is significantly lower than an ocular disease-free reference group (90±15, P<0.0001), indicating more ocular pain. This was true for all types of NIU, regardless of the localisation: although Ocular Pain Scores were lower in patients with anterior uveitis (AU) compared with patients with non-AU (mean 62 (±24) vs 74 (±24), P=0.04), patients with non-AU still scored substantially lower than the reference group that had no ocular history (P<0.0001). Patients with NIU also scored significantly lower on all other VFQ subscales as well as on the SF-36 subscales 'Role Limitations due to physical problems', 'Vitality', 'General health' and 'Bodily Pain' compared with controls. The VFQ Ocular Pain subscale correlated with other quality of life subscales (both VFQ-25 and SF-36), indicating a relationship between pain and quality of life. This study shows that ocular pain is highly prevalent in patients with NIU, regardless of the localisation. Furthermore, ocular pain has an impact on quality of life.


Epithelial cells as alternative human biomatrices for comet assay.
The comet assay is a valuable experimental tool aimed at mapping DNA damage in human cells in vivo for environmental and occupational monitoring, as well as for therapeutic purposes, such as storage prior to transplant, during tissue engineering, and in experimental ex vivo assays. Furthermore, due to its great versatility, the comet assay allows to explore the use of alternative cell types to assess DNA damage, such as epithelial cells. Epithelial cells, as specialized components of many organs, have the potential to serve as biomatrices that can be used to evaluate genotoxicity and may also serve as early effect biomarkers. Furthermore, 80% of solid cancers are of epithelial origin, which points to the importance of studying DNA damage in these tissues. Indeed, studies including comet assay in epithelial cells have either clear clinical applications (lens and corneal epithelial cells) or examine genotoxicity within human biomonitoring and in vitro studies. We here review improvements in determining DNA damage using the comet assay by employing lens, corneal, tear duct, buccal, and nasal epithelial cells. For some of these tissues invasive sampling procedures are needed. Desquamated epithelial cells must be obtained and dissociated prior to examination using the comet assay, and such procedures may induce varying amounts of DNA damage. Buccal epithelial cells require lysis enriched with proteinase K to obtain free nucleosomes. Over a 30 year period, the comet assay in epithelial cells has been little employed, however its use indicates that it could be an extraordinary tool not only for risk assessment, but also for diagnosis, prognosis of treatments and diseases.


Renal MR angiography.
Three-dimensional gadolinium MR angiography (3D-Gd-MRA) accurately visualizes the renal arteries with almost no degradation from inplane saturation or motion artifacts. The diagnostic accuracy for detecting and grading of renal artery stenosis, as well as the assessment of other vascular pathology, approaches that of conventional x-ray angiography. For the renovascular system, this technique requires precise contrast media bolus timing since multiple successively enhancing structures are present. Details on performing renal MR angiography, strategies for image analysis, and examples of common renal vascular pathology are reviewed. The 3D-Gd-MRA protocol can be easily combined with other MR imaging techniques to provide a comprehensive assessment of the hemodynamic and functional significance of renal artery stenosis.


Tobacco mosaic virus infection disproportionately impacts phloem associated translatomes in Arabidopsis thaliana and Nicotiana benthamiana.
In this study we use vascular specific promoters and a translating ribosome affinity purification strategy to identify phloem associated translatome responses to infection by tobacco mosaic virus (TMV) in systemic hosts Arabidopsis thaliana ecotype Shahdara and Nicotiana benthamiana. Results demonstrate that in both hosts the number of translatome gene alterations that occurred in response to infection is at least four fold higher in phloem specific translatomes than in non-phloem translatomes. This finding indicates that phloem functions as a key responsive tissue to TMV infection. In addition, host comparisons of translatome alterations reveal both similarities and differences in phloem responses to infection, representing both conserved virus induced phloem alterations involved in promoting infection and virus spread as well as host specific alterations that reflect differences in symptom responses. Combined these results suggest phloem tissues play a disproportion role in the mediation and control of host responses to virus infection.


Platelet function changes as monitored by cone and plate(let) analyzer during beating heart surgery.
Off-pump coronary artery bypass (OPCAB) is believed to reduce cardiopulmonary bypass (CPB)-related complications, including platelet damage. A hypercoagulable state instead of coagulopathy has been reported following OPCAB surgeries due to CPB. Whether platelet function is changed when the injurious effect of CPB is eliminated was investigated. Platelet function was determined with the cone and plate(let) analyzer (CPA) method. The 2 parameters, average size (AS) and surface coverage (SC) of platelet aggregates, were measured with the CPA method to assess platelet aggregation and adhesion. These parameters were evaluated, and their values were compared at several stages of OPCAB surgery. The correlations of postoperative bleeding with platelet function at different stages of the surgery and with other factors, such as platelet count, hematocrit, and transfusions, were studied. Both AS and SC increased during several stages of the operation, and postoperative values (mean +/- SD) were significantly higher than preoperative values (30.4 +/- 8.1 microm 2 versus 23.3 +/- 6.9 microm 2 for AS [ P =.02] and 7.6% +/- 3.6% versus 5.2% +/- 1.8% for SC [ P =.04]). The mean total bleeding volume was 875 micro 415 mL. Preoperative AS and SC were the only parameters significantly ( P =.01) and linearly ( r = 0.7) related to postoperative bleeding. An increased platelet function, as determined by the CPA method, is found following OPCAB surgery. This phenomenon is probably at least partially responsible for the thrombogenic state after OPCAB surgery. Lack of platelet injury attributed to CPB may divert the system toward a more thrombogenic state. Preoperative platelet function, as evaluated by the CPA method, is an independent risk factor determining postoperative bleeding.


A Novel Gait Phase Recognition Method Based on DPF-LSTM-CNN Using Wearable Inertial Sensors.
Gait phase recognition is of great importance in the development of rehabilitation devices. The advantages of Long Short-Term Memory (LSTM) and Convolutional Neural Network (CNN) are combined (LSTM-CNN) in this paper, then a gait phase recognition method based on LSTM-CNN neural network model is proposed. In the LSTM-CNN model, the LSTM layer is used to process temporal sequences and the CNN layer is used to extract features A wireless sensor system including six inertial measurement units (IMU) fixed on the six positions of the lower limbs was developed. The difference in the gait recognition performance of the LSTM-CNN model was estimated using different groups of input data collected by seven different IMU grouping methods. Four phases in a complete gait were considered in this paper including the supporting phase with the right hill strike (SU-RHS), left leg swimming phase (SW-L), the supporting phase with the left hill strike (SU-LHS), and right leg swimming phase (SW-R). The results show that the best performance of the model in gait recognition appeared based on the group of data from all the six IMUs, with the recognition precision and macro-F1 unto 95.03% and 95.29%, respectively. At the same time, the best phase recognition accuracy for SU-RHS and SW-R appeared and up to 96.49% and 95.64%, respectively. The results also showed the best phase recognition accuracy (97.22%) for SW-L was acquired based on the group of data from four IMUs located at the left and right thighs and shanks. Comparably, the best phase recognition accuracy (97.86%) for SU-LHS was acquired based on the group of data from four IMUs located at left and right shanks and feet. Ulteriorly, a novel gait recognition method based on Data Pre-Filtering Long Short-Term Memory and Convolutional Neural Network (DPF-LSTM-CNN) model was proposed and its performance for gait phase recognition was evaluated. The experiment results showed that the recognition accuracy reached 97.21%, which was the highest compared to Deep convolutional neural networks (DCNN) and CNN-LSTM.


Acyl coenzyme A binding protein. Conformational sensitivity to long chain fatty acyl-CoA.
Cellular unbound long chain fatty acyl-CoAs (>14 carbon) are potent regulators of gene transcription and intracellular signaling. Although the cytosolic acyl-CoA binding protein (ACBP) has high affinity for medium chain fatty acyl-CoAs, direct interaction of ACBP with >14-carbon fatty acyl-CoAs has not been established. Steady state, photon counting fluorescence spectroscopy directly established that rat liver ACBP bound 18-carbon cis- and trans-parinaroyl-CoA, Kd = 7.03 +/- 0.95 and 4.40 +/- 0.43 nM. Time-resolved fluorometry revealed that ACBP-bound parinaroyl-CoAs had high rotational freedom within the single, relatively hydrophobic (epsilon <32), binding site. Tyr and Trp fluorescence dynamics demonstrated that apo-ACBP was an ellipsoidal protein (axes of 15 and 9 A) whose conformation was altered by oleoyl-CoA in the holo-ACBP as shown by a 2-A decrease of ACBP hydrodynamic diameter and increased Trp segmental motions. Thus, native liver ACBP binds >14-carbon fatty acyl-CoAs with nanomolar affinity at a single binding site. Acyl-CoA-induced conformational alterations in ACBP may be significant to its putative functions in lipid metabolism and regulation of processes sensitive to unbound long chain fatty acyl-CoAs.


A novel DR beta 1*03 allele identified by polymerase chain reaction-restriction fragment length polymorphism.
This report describes a new MHC class II allele, HLA-DR beta 1*0306, discovered in a 31-year-old Norwegian male. The allele typed serologically as DRw52 (DR3) and amplified in PCR using DR52-associated group primers. This product could not be identified using established restriction digests, however. Use of Asp 700, Msp I, Hha I, Bse RI, Mnl I, Hph I, and Bsrb I gave banding patterns expected for a DR beta 1*03011 allele, but Rsa I had an additional site at codon 47. Sequencing showed a single base change at this position, with the substitution of tyrosine for phenylanine in this new allele. The biologic impact of this substitution remains to be determined.


Development of a Langmuir probe array for radial potential profile measurement in the collisional merging formation FRC.
The radial electric field in a field-reversed configuration (FRC) plasma plays an important role in the global stability and confinement properties. Herein, we developed a new Langmuir probe array named "Skewered probe" employed in measuring the radial potential profile in the collisional merging formation of an FRC in the FAT-CM (FRC Amplification via Translation - Collisional Merging) device. Because an FRC has a strong toroidal flow, the skewered probe consists of alternately skewered ring electrodes and ceramic beads on a thin stainless-steel tube to neutralize the effect of plasma flow. The developed array has nine electrodes, one every 2 cm from r = 9-25 cm, and it measures the FRC boundary in the case when the radius of the excluded flux ranges from 10 to 20 cm. The skewered probe also has one additional electrode that measures the potential near the chamber wall as a reference for the other electrodes. The radial potential profile of the FRC formed by the collisional merging method in the FAT-CM device was measured using the probe, and the results showed that the region of negative potential gradually changed to a positive potential after merging the FRCs. It was also shown that a strong outward electric field is formed near the separatrix at n = 2 rotational instability.


[Acute intestinal obstructions in adults: quantified semiology (signs and their value) and surgical treatment].
The prevalence of signs and symptoms of acute intestinal obstruction in the adult was studied in a prospective study of 600 cases extracted from a data base on acute abdomen in 7,000 patients. This study of prevalence allows a precise definition of intestinal obstruction syndrome and to differentiate two types of presentations according to the site of obstruction on the small bowel or the colon. However, it is more difficult to differentiate simple bowel obturation from vascular strangulation. Surgical treatment depends mainly on the cause of obstruction.


Characterization of Erysipelothrix rhusiopathiae isolates from laying hens and poultry red mites (Dermanyssus gallinae) from an outbreak of erysipelas.
Infection with the zoonotic bacterium Erysipelothrix rhusiopathiae causes severe disease outbreaks (erysipelas) in poultry flocks. As this bacterium has been isolated from the poultry red mite (Dermanyssus gallinae), this parasite has been suggested as a possible means of transmission of E. rhusiopathiae on and between poultry farms. To further elucidate the capacity of the mite as a reservoir, we analysed and compared 56 bacterial isolates from laying hens and nine isolates from mites by pulsed-field gel electrophoresis (PFGE), using the restriction enzyme SmaI. The isolates originated from one outbreak in a laying hen flock housed in an indoor litter-based aviary system. Except for two isolates, a homogeneous banding pattern was obtained from all isolates analysed, suggesting that a single strain was the cause of the outbreak. Another finding was that isolates from individual hens could exhibit slightly different PFGE patterns. Mites collected from the same house at the end of the production period of the following flock were negative for the presence of E. rhusiopathiae. An increasing number of erysipelas outbreaks as well as escalating problems with D. gallinae are expected in other European countries related to the forthcoming changes in housing systems for laying hens. Consequently, further studies are needed to investigate the importance of erysipelas in poultry and the importance of D. gallinae in the transmission of E. rhusiopathiae.


Visuospatial mnemonic load modulates event-related slow potentials.
Electroencephalographic slow wave potentials were recorded during the performance of visuospatial working memory tasks. The aim was to study the effects of varying mnemonic loads on slow potentials, and to dissociate the contribution of mnemonic and motor components. Subjects were tested with three spatial delayed matching-to-sample tasks in which the mnemonic load varied while the preparatory motor demands remained constant. The delay-related slow potential was more negative during the tasks in which the subjects had to memorize the locations of six or four stimuli than when only one location had to be memorized. Significant differences between the slow potentials in the tasks with different mnemonic loads were recorded at frontal and temporal recording sites. Since the preparatory motor requirements were similar in all tasks, the modulation of slow potentials reflects working memory processing rather than motor preparatory activity.


The Nucleus Accumbens and Ketamine Treatment in Major Depressive Disorder.
Animal models of depression repeatedly showed stress-induced nucleus accumbens (NAc) hypertrophy. Recently, ketamine was found to normalize this stress-induced NAc structural growth. Here, we investigated NAc structural abnormalities in major depressive disorder (MDD) in two cohorts. Cohort A included a cross-sectional sample of 34 MDD and 26 healthy control (HC) subjects, with high-resolution magnetic resonance imaging (MRI) to estimate NAc volumes. Proton MR spectroscopy (1H MRS) was used to divide MDD subjects into two subgroups: glutamate-based depression (GBD) and non-GBD. A separate longitudinal sample (cohort B) included 16 MDD patients who underwent MRI at baseline then 24 h following intravenous infusion of ketamine (0.5 mg/kg). In cohort A, we found larger left NAc volume in MDD compared to controls (Cohen's d=1.05), but no significant enlargement in the right NAc (d=0.44). Follow-up analyses revealed significant subgrouping effects on the left (d⩾1.48) and right NAc (d⩾0.95) with larger bilateral NAc in non-GBD compared to GBD and HC. NAc volumes were not different between GBD and HC. In cohort B, ketamine treatment reduced left NAc, but increased left hippocampal, volumes in patients achieving remission. The cross-sectional data provided the first evidence of enlarged NAc in patients with MDD. These NAc abnormalities were limited to patients with non-GBD. The pilot longitudinal data revealed a pattern of normalization of left NAc and hippocampal volumes particularly in patients who achieved remission following ketamine treatment, an intriguing preliminary finding that awaits replication.


Watermelon stomach in a patient with primary Sjögren's syndrome.
Watermelon stomach (WS) or gastric antral vascular ectasia (GAVE) is a rare cause of upper gastrointestinal bleeding described in a variety of autoimmune disorders. Association of watermelon stomach with Sjögren's syndrome is extremely rare. CASE REPORT; We presented a 67-year old female with primary Sjögren's syndrome (SS) who had developed a persistent severe iron-deficiency anemia. An upper gastric endoscopy revealed the presence of gastric antral vascular ectasia (GAVE) as a cause of occult gastrointestinal bleeding. The treatment with argon-plasma coagulation was postponed as the conservative therapy with iron substitution and proton pump inhibitor led to improvement of anemia and hemoglobin levels normalization. This is the first report of WS in a patient with primary SS without the presence of coexisting autoimmune disorder. Recognition of this rare, but clinically important, cause of gastrointestinal bleeding may decrease comorbidity in patients with autoimmune disorders including primary Sjögren's syndrome.


[Haemostatic changes related to extracorporeal circulation (ECC) (author's transl)].
After-bleeding not due to surgery following extracorporeal circulation (ECC) in open heart surgery has its origin in disorders of platelet number and function as well as in changes of the haemostatic balance. Not only are the plasmatic factors of the coagulation and the fibrinolytic system of importance but also their inhibitors which tend to avoid dysfunctions and overshooting reactions. To detect early the danger of an excessive bleeding tendency assays of fibrinogen (factor I), prothrombin (factor II), antithrombin III (AT III) and of alpha 2-macroglobulin (A 2 M) are recommended. A substitution therapy can be oriented by the results of these coagulation parameters.


The magnitude of undernutrition and associated factors among adult chronic kidney disease patients in selected hospitals of Addis Ababa, Ethiopia.
Undernutrition is a common comorbidity in chronic kidney disease patients which augments the progression of the disease to an end-stage renal disease, renal dysfunction and related morbidity and mortality. However, in Ethiopia, there is a dearth of research evidence in this regard. Therefore, this study aimed to assess the magnitude of undernutrition and its associated factors among adult chronic kidney disease patients. An institution-based cross-sectional study was conducted in selected hospitals of Addis Ababa from May to August 2018. Data were collected by structured and pretested questionnaires. Patients' charts were reviewed from their medical profiles. Body mass index was calculated from anthropometric measurements using calibrated instruments. Serum albumin level was determined by reference laboratory standard procedure. Data were entered into Epi- data version 3.1 and exported to SPSS version 21 for analysis. Descriptive statistics were calculated and presented by tables, graphs and texts. Binary and multivariable logistic regression analyses were computed and the level of statistical significance was declared at p-value <0.05. From the total sample size of 403 participants, 371 were involved in the study. The prevalence of undernutrition (BMI<18.5) among adult chronic kidney disease patients was 43.1% (95% CI: 38%-48%). Undernutrition (BMI<18.5) was significantly higher among patients with diabetic nephropathy [AOR = 2.00, 95% CI, 1.09-2.66], serum albumin value less than 3.8g/dl [AOR = 4.21: CI, 2.07-5.07], recently diagnosed with diabetes mellitus [AOR = 2.36, 95% CI, 1.03-3.14] and stage V chronic kidney disease [AOR = 3.25:95% CI, 1.00-3.87]. Undernutrition in chronic kidney disease patients was significantly higher among patients with diabetic nephropathy, patients on stage V chronic kidney disease, recently diagnosed with diabetes mellitus and serum albumin value less than 3.8g/dl.


Fe biomineralization mirrors individual metabolic activity in a nitrate-dependent Fe(II)-oxidizer.
Microbial biomineralization sometimes leads to periplasmic encrustation, which is predicted to enhance microorganism preservation in the fossil record. Mineral precipitation within the periplasm is, however, thought to induce death, as a result of permeability loss preventing nutrient and waste transit across the cell wall. This hypothesis had, however, never been investigated down to the single cell level. Here, we cultured the nitrate reducing Fe(II) oxidizing bacteria Acidovorax sp. strain BoFeN1 that have been previously shown to promote the precipitation of a diversity of Fe minerals (lepidocrocite, goethite, Fe phosphate) encrusting the periplasm. We investigated the connection of Fe biomineralization with carbon assimilation at the single cell level, using a combination of electron microscopy and Nano-Secondary Ion Mass Spectrometry. Our analyses revealed strong individual heterogeneities of Fe biomineralization. Noteworthy, a small proportion of cells remaining free of any precipitate persisted even at advanced stages of biomineralization. Using pulse chase experiments with (13)C-acetate, we provide evidence of individual phenotypic heterogeneities of carbon assimilation, correlated with the level of Fe biomineralization. Whereas non- and moderately encrusted cells were able to assimilate acetate, higher levels of periplasmic encrustation prevented any carbon incorporation. Carbon assimilation only depended on the level of Fe encrustation and not on the nature of Fe minerals precipitated in the cell wall. Carbon assimilation decreased exponentially with increasing cell-associated Fe content. Persistence of a small proportion of non-mineralized and metabolically active cells might constitute a survival strategy in highly ferruginous environments. Eventually, our results suggest that periplasmic Fe biomineralization may provide a signature of individual metabolic status, which could be looked for in the fossil record and in modern environmental samples.


Three new carbazole alkaloids and biological activities of Murraya koenigii.
Three new carbazole alkaloids, mukoenigatin (1), bikoeniquinonine (2) and murrayadinal (3), were isolated from the aerial parts of Murraya koenigii, along with mukeonine-B (4). Their molecular structures were determined on the basis of spectral analysis including UV, IR, MS, and 2D NMR spectroscopy. The antimicrobial activity of different fractions of plant extract was also determined.


Swapping of The N-Terminal Domain of Human Topoisomerase 1B with the Corresponding Plasmodium Falciparum Counterpart Strongly Impairs Enzyme Activity.
DNA topoisomerases 1B are a class of ubiquitous enzyme that solves the topological problems associated with biological processes such as replication, transcription and recombination. Numerous sequence alignment of topoisomerase 1B from different species shows that the lengths of different domains as well as their amino acids sequences are quite different. In the present study a hybrid enzyme, generated by swapping the N-terminal of Plasmodium falciparum into the corresponding domain of the human, has been characterized. The chimeric enzyme was generated using different sets of PCR. The in vitro characterization was carried out using different DNA substrate including radio-labelled oligonucleotides. The chimeric enzyme displayed slower relaxation activity, cleavage and re-ligation kinetics strongly perturbed when compared to the human enzyme. These results indicate that the N-terminal domain has a crucial role in modulating topoisomerase activity in different species.


The effect of anti-thyroid antibodies on endometrial volume, embryo grade and IVF outcome.
Thyroid auto-immunity (TAI) has been implicated as the most common cause of hypothyroidism in general population, especially in women. Many studies revealed that increased infertility incidences with TAI. The aim of the present article was to evaluate the effect of thyroid auto-antibody (TAA) positivity on embryological parameters, IVF-outcome and endometrial volume (EnV) in infertile patients who were applied for routine artificial reproductive technologies (ART) programme. This study included prospective, sequential, cross-sectional analyses of parameters obtained from 69 patients with unexplained infertility. It was the first ART application of patients. Patients were homogenous for age, body mass index, basal hormone measurements and underwent same ovulation induction protocol. They were evaluated for thyroid hormone profile and TAAs and divided into three groups; TAA negative group (n = 31), TAA positive group (n = 23) and TAA positive and euthyroid with medication group (n = 15). There were no differences among groups for the number of Grade-1 and Grade-2 embryos, distribution of embryo-grades, number of oocytes retrieved and fertilised, biochemical pregnancy ratios (PR), EnV and miscarriage ratio. However, the clinical PR was significantly lower in the TAA positive group (p = 0.024). In conclusion, the embryo grades and EnV did not differ among groups. But the clinical PR differs and the anti-thyroid peroxides positivity, above the cut-off point, affects the clinical PR.


A nurse-led pathway to treat self-harm injuries.
Self-harm occurs frequently in psychiatric hospitals and other mental health and outpatient settings. Many patients at the Ayr Clinic are at risk of self-harm and often access the local accident and emergency service for minor injury care. The medical and nursing response to people who repeatedly self-harm, given increasing pressures and dwindling A&E resources, can often be one of impatience, frustration and hostility. A nurse-led pathway was developed by the Ayr Clinic and NHS Ayrshire and Arran to fast-track the assessment and treatment of such injuries and improve health professionals' attitudes towards these patients. This project was winner of the Nursing Times emergency and critical care awrad 2014.


Dietary fibre and lipid metabolism.
The influence of dietary fibre on lipid metabolism can be assessed at several levels. In animal studies it is possible to investigate the effects of fibre on serum and tissue lipid levels, on lipid absorption and excretion and, in specific instances, on experimental atherosclerosis. In man we are limited to examination of blood lipid and lipoprotein levels and on patterns of lipid excretion. Ershoff and his co-workers (eg) were among the first to demonstrate beneficial effects of fibre on serum and liver cholesterol levels in cholesterol-fed rats. They also showed that not all that we call fibre is hypocholesterolaemic. Cellulose, for instance, caused increases in liver cholesterol levels in rats. Cellulose also leads to increased levels of carcass cholesterol. Experiments in chickens, rabbits and monkeys have shown cellulose to exert the least beneficial effect on atherosclerosis or aortic sudanophilia. In man the insoluble fibres such as bran or cellulose are not hypocholesterolaemic, whereas soluble fibres such as guar gum or pectin are. The effects are also reflected in levels of high and low density lipoproteins. Studies relating to mechanism(s) of fibre action have shown that fibre may increase cholesterol and bile acid excretion and, in some cases, may affect faecal bile acid composition. Fibres have been shown to bind bile acids in vitro, the extent of binding being a function of both the type of fibre used and the type of bile acid or bile salt being studied. Fibre has also been shown to bind other lipids. Fibre also influences the rate of cholesterol absorption. To date most results can be classified according to the type of fibre used, ie soluble or insoluble; ionic or non-ionic. Our added understanding of modes of action of fibre will depend on better ability to relate structure to function.


A new mechanism of action for the anticancer drug mitomycin C: mechanism-based inhibition of thioredoxin reductase.
Mitomycin C (MMC) is a chemotherapeutic drug that requires an enzymatic bioreduction to exert its biological effects. Upon reduction, MMC is converted into a highly reactive bis-electrophilic intermediate that alkylates cellular nucleophiles. Alkylation of DNA is the most favored mechanism of action for MMC, but other modes of action, such as redox cycling and inhibition of rRNA, may also contribute to the biological action of the drug. In this work, we show that thioredoxin reductase (TrxR) is also a cellular target for MMC. We show that MMC inhibits TrxR in vitro, using purified enzyme, and in vivo, using cancer cell cultures. The inactivation presents distinctive parameters of mechanism-based inhibitors: it is time- and concentration-dependent and irreversible. Additionally, spectroscopic experiments (UV, circular dichroism) show that the inactivated enzyme contains a mitosene chromophore. On the basis of kinetic and spectroscopic data, we propose a chemical mechanism for the inactivation of the enzyme that starts with a reduction of the quinone ring of MMC by the selenolthiol active site of TrxR and a subsequent alkylation of the active site by the activated drug. We also report that MMC inactivates TrxR in cancer cell cultures and that this inhibition correlates directly with the cytotoxicity of the drug, indicating that inhibition of TrxR may play a major role in the biological mode of action of the drug.


Fibrinogen, t-PA, and PAI-1 plasma levels in patients with pulmonary hypertension.
We measured fibrinogen levels as well as the fibrinolytic parameters tissue-type plasminogen activator (t-PA) and plasminogen activator inhibitor 1 (PAI-1) in plasma samples obtained at basal conditions and after stimulating the fibrinolytic system by venous occlusion (VO). Samples were taken from patients with primary pulmonary hypertension (PPH), with secondary thromboembolic pulmonary hypertension (SPHTH), with secondary pulmonary hypertension due to congenital heart disease with Eisenmenger's reaction (SPHCD), and from healthy control individuals (CON). Fibrinogen levels were not significantly different between the groups with PPH and SPHTH or between SPHCD and CON. The latter groups, however, exhibited significantly lower fibrinogen plasma levels compared with PPH or SPHTH (p < 0.01). Basal plasma levels of t-PA antigen, t-PA activity, and PAI-1 activity, respectively, did not differ significantly between the study groups. After VO, mean t-PA activity levels increased to a higher extent in control subjects compared with patients with PPH, or SPHTH, or SPHCD, with significant differences only between CON and SPHTH or CON and PPH (p < 0.03). Patients with PPH and SPHTH exhibit both increased fibrinogen plasma levels and a diminished fibrinolytic response compared with healthy subjects. Moreover, the fibrinogen plasma levels in patients with SPHCD are in normal range, and the fibrinolytic response is similar to CON compared with PPH and SPHTH, thus indicating the existence of a comparable prothrombotic situation in patients with PPH and SPHTH.


SOX2 is essential for in vivo reprogramming of seminoma-like TCam-2 cells to an embryonal carcinoma-like fate.
Type II germ cell cancers (GCC) are divided into seminomas, which are highly similar to primordial germ cells and embryonal carcinomas (EC), often described as malignant counterparts to embryonic stem cells.Previously, we demonstrated that the development of GCCs is a highly plastic process and strongly influenced by the microenvironment. While orthotopic transplantation into the testis promotes seminomatous growth of the seminoma-like cell line TCam-2, ectopic xenotransplantation into the flank initiates reprogramming into an EC-like fate.During this reprogramming, BMP signaling is inhibited, leading to induction of NODAL signaling, upregulation of pluripotency factors and downregulation of seminoma markers, like SOX17. The pluripotency factor and EC-marker SOX2 is strongly induced.Here, we adressed the molecular role of SOX2 in this reprogramming. Using CRISPR/Cas9-mediated genome-editing, we established SOX2-deficient TCam-2 cells. Xenografting of SOX2-deficient cells into the flank of nude mice resulted in maintenance of a seminoma-like fate, indicated by the histology and expression of OCT3/4, SOX17, TFAP2C, PRDM1 and PRAME. In SOX2-deficient cells, BMP signaling is inhibited, but NODAL signaling is not activated. Thus, SOX2 appears to be downstream of BMP signaling but upstream of NODAL activation. So, SOX2 is an essential factor in acquiring an EC-like cell fate from seminomas.A small population of differentiated cells was identified resembling a mixed non-seminoma. Analyses of these cells revealed downregulation of the pluripotency and seminoma markers OCT3/4, SOX17, PRDM1 and TFAP2C. In contrast, the pioneer factor FOXA2 and its target genes were upregulated, suggesting that FOXA2 might play an important role in induction of non-seminomatous differentiation.


Dangerous asthma.
Greater awareness of the morbid condition in severe asthma attacks would decrease the still considerable number of deaths which occur in otherwise fit, young asthmatics. Failure to recognise serious attacks in time may be due to the presumption that the disorder is one of bronchoconstriction. More dangerous is the hyperaemia and exudation of plasma into the bronchial lumen where it mixes with mucus to form plugs. Bronchial epithelium damaged during an attack may take weeks to regenerate, increasing the risk from another attack. Patients should be instructed to consult a doctor when expected bronchodilator relief decreases in either degree or duration.


Clinical characteristics, risk factors, and cardiac manifestations of cancer patients with COVID-19.
Coronavirus disease 2019 (COVID-19) due to severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has been associated with cardiovascular features, which may be deteriorated in patients with cancer. However, cardiac outcomes of cancer patients with COVID-19 have not been closely examined. We retrospectively assessed 1,244 patients with COVID-19 from February 1 to August 31, 2020 (140 cancer and 1,104 noncancer patients). Demographic and clinical data were obtained and compared between cancer and noncancer groups. Including the cardiac biomarkers, we also analyzed laboratory findings between these two groups. Risk factors for in-hospital mortality were identified by multivariable Cox regression models. For cancer group, 56% were in severe and critical status with more diabetes and immune deficiency, whereas the proportion was 10% for noncancer group. Patients with cancer had increased levels of leukocyte, neutrophil count, and blood urea nitrogen (BUN) (all P < 0.01), whereas lymphocyte count was significantly lower (P < 0.001). The most common solid tumor types were gastrointestinal cancer (26%), lung cancer (21%), and breast and reproductive cancer (both 19%). There is a rising for cardiac biomarkers, including pro-B-type natriuretic peptide (Pro-BNP), sensitive troponin I (cTnI), myoglobin (MYO), creatine kinase-MB (CK-MB), as well as D-Dimer in COVID-19 cancer population, especially in deceased subjects with cancer. The 30-day in-hospital mortality in cancer group was dramatically raised than that in noncancer group (12.9% vs. 4.0%, P < 0.01). In multivariable Cox regression models, fever, disease severity status, and underlying diseases were risk factors for mortality. COVID-19 patients with cancer relate to deteriorating conditions and poor cardiac outcomes accompanied by a high in-hospital mortality, which warrants more aggressive treatment.NEW & NOTEWORTHY Our study indicates that the 30-day mortality is higher in COVID-19 patients with cancer; more COVID-19 patients with cancer are in severe and critical status; age, respiratory rate, neutrophil count, AST, BUN, MYO, Pro-BNP, disease severity status, underlying diseases, and fever are risk factors for in-hospital mortality among COVID-19 cancer cases; COVID-19 patients with cancer display severely impaired myocardium, damaged heart function, and imbalanced homeostasis of coagulation; what is more, those with both cancer and CVD have more significantly increased Pro-BNP and D-Dimer level.


Analysis of pharmaceutical formulations using atmospheric pressure ion mobility spectrometry combined with liquid chromatography and nano-electrospray ionisation.
The hyphenation of liquid chromatography with atmospheric pressure ion mobility spectrometry is reported using a custom-made dynamic nano-electrospray ionisation (nano-ESI) interface. The analysis of pharmaceutical actives is described, including beta blocker (timolol), antidepressant (paroxetine), analgesic (paracetamol) and opiate (codeine) preparations. On-line ultraviolet diode array (UV) spectroscopic detection was used prior to sample ionisation, to evaluate chromatographic and nano-ESI interface performance. Active drug responses were characterised by chromatographic retention time and electrophoretic ion mobility drift time, and selected ion mobility responses were used to evaluate method performance. Limits of detection for active drugs were in the low-nmol to pmol range. Quantitative responses were investigated using a series of standard solutions of caffeine, showing good linearity (R(2) = 0.9982, n = 6) and reproducibility (RSD = 2.3 %, n = 6). The analysis of an over the counter pharmaceutical formulation demonstrates the potential of ion mobility spectrometry combined with liquid chromatography and nano-electrospray ionisation for the rapid determination of active drugs, as a result of the electrophoretic separation and selectivity afforded by IMS.


Spotlight on brimonidine topical gel 0.33% for facial erythema of rosacea: safety, efficacy, and patient acceptability.
Brimonidine tartrate is a highly selective alpha 2 agonist that induces direct vasoconstriction of small arteries and veins, thereby reducing vasodilation and edema. To review the current literature regarding the safety, efficacy, and patient acceptability of brimonidine 0.33% gel. A PubMed search was performed using the terms brimonidine 0.33% gel, rosacea, safety, efficacy, and acceptability. Peer-reviewed clinical trials and case reports from 2012 to 2016 were screened for inclusion of safety, efficacy, and/or patient acceptability data. Brimonidine topical gel 0.33% is associated with mild, transient skin-related adverse reactions. Efficacy may be achieved within 30 minutes of administration with maximal reductions in erythema 3-6 hours after administration. Patient satisfaction with use of brimonidine topical gel is superior to vehicle gel for facial appearance, treatment effect, facial redness, and daily control of facial redness. Studies were typically limited to 1-year follow-up. Only one study has examined the use of brimonidine topical gel in combination with other rosacea and acne medications. Brimonidine topical gel 0.33% is a safe, effective, and patient-accepted treatment for facial erythema of rosacea.


Toxicity of emerging energetic soil contaminant CL-20 to potworm Enchytraeus crypticus in freshly amended or weathered and aged treatments.
We investigated the toxicity of an emerging polynitramine energetic material hexanitrohexaazaisowurtzitane (CL-20) to the soil invertebrate species Enchytraeus crypticus by adapting then using the Enchytraeid Reproduction Test (ISO/16387:2003). Studies were designed to develop ecotoxicological benchmark values for ecological risk assessment of the potential impacts of accidental release of this compound into the environment. Tests were conducted in Sassafras Sandy Loam soil, which supports relatively high bioavailability of CL-20. Weathering and aging procedures for CL-20 amended into test soil were incorporated into the study design to produce toxicity data that better reflect soil exposure conditions in the field compared with the toxicity in freshly amended soils. Concentration-response relationships for measurement endpoints were determined using nonlinear regressions. Definitive tests showed that toxicities for E. crypticus adult survival and juvenile production were significantly increased in weathered and aged soil treatments compared with toxicity in freshly amended soil, based on 95% confidence intervals. The median effect concentration (EC50) and EC20 values for juvenile production were 0.3 and 0.1 mg kg-1, respectively, for CL-20 freshly amended into soil, and 0.1 and 0.035 mg kg-1, respectively, for weathered and aged CL-20 soil treatments. These findings of increased toxicity to E. crypticus in weathered and aged CL-20 soil treatments compared with exposures in freshly amended soils show that future investigations should include a weathering and aging component to generate toxicity data that provide more complete information on ecotoxicological effects of emerging energetic contaminants in soil.


The physical state of nafcillin sodium in frozen aqueous solutions and freeze-dried powders.
The purpose of this study was to develop a better understanding of the physical chemistry of freeze drying of lyotropic liquid crystals using nafcillin sodium as a model solute. Solutions and freeze-dried powders of nafcillin sodium were studied by polarized light microscopy, differential scanning calorimetry, x-ray powder diffraction, and water vapor adsorption. Differential scanning calorimetry thermograms of nafcillin sodium solutions contain a melting endotherm at approximately -5.5 degrees C and, depending on the concentration and heating rate, a crystallization exotherm immediately after this endotherm followed by the melting endotherm of ice. When the sample is annealed at -4 degrees C, both the endotherm and exotherm are eliminated, and a new endotherm appears at approximately -1 degree C on the shoulder of the ice-melting endotherm. The data are interpreted as melting of a liquid crystalline phase, followed by crystallization. X-ray powder diffractograms of unannealed freeze-dried nafcillin sodium are consistent with a lamellar liquid crystal. Diffractograms of annealed freeze-dried nafcillin sodium indicate crystalline material which is a different crystal form than the monohydrate starting material. Moisture adsorption isotherms of the freeze-dried annealed (crystalline) and unannealed (liquid crystalline) nafcillin sodium show different affinities for moisture compared to the crystalline starting material. Solid-state stability data demonstrate that the freeze-dried liquid crystalline form of nafcillin sodium is much less stable than the freeze-dried crystal-line material. The literature recognizes two types of solute behavior on freezing, where the solute either crystallizes from the freeze concentrate or remains amorphous. Lyotropic liquid crystal formation during freezing represents a separate category of freezing behavior, the physical chemistry of which is worthy of further investigation.


[Effects of HS1-associated protein X-1 on the lupus activities: experiment with MRL/lpr lupus-like mice].
To investigate the effects of HS1-associated protein X-1 (HAX-1) on the lupus activity of MRL/lpr lupus-like mice. Fifteen MRL/lpr mice were divided into 3 equal groups: Group A, injected with phosphate-buffered saline, Group B, injected intraperitoneally with control virus AdEGFP and Group C, injected intraperitoneally with recombinant AdHAX-1 twice a week for 4 weeks. Peripheral blood samples were collected before the injection, and 2 and 4 weeks after the injection to be detected the white blood cell count, antinuclear antibody (ANA), anti-double strand-DNA antibodies, circulating immune complex (CIC), anti-histone antibodies, and interferon (IFN)-gamma. The level of urine protein was measured, too. Then the mice were killed, a kidney underwent direct immunofluorescence (DIF) to observe the deposition of Immune complexes, and the other kidney underwent periodic acid-Schiff (PAS) staining and pathological examination. MTT method was used to detect the proliferation of the lymphocytes in the spleen. Splenocytes were isolated from the other 15 MRL/lpr mice and then divided into 4 groups: Group, transfected with DMRIE-C without plasmid; Group E, as negative control group; Group F, transfected with blank plasmid pGenesil-1; and Group G, transfected with pGenesil-HAX-1. Forty-eight hours later MTT method was used to detect the proliferation rateof the spleen lymphocytes. The urine protein level of Group C was significantly higher than those of Groups A and B (both P < 0.01). Four weeks later the levels of ANA, anti-double strand DNA antibodies, and IFN-gamma were all significantly higher than those of Groups A and B (all P < 0.01). Hypercellularity and increased deposition of IgG in glomeruli were also observed in Group C. The score of glomeruli lesion of Group C (1.50 +/- 0.34) was significantly higher than those of Groups A (0.67 +/- 0.14) and Groups B (0.81 +/- 0.26) (both P < 0.01). MTT method showed that the growth curve of the spleen lymphocytes of Group C was higher than those of Groups A and B. The spleen lymphocyte proliferation rate and the levels of IFN-gamma of Group G was significantly lower than that of Group F (both P < 0.05). One of the important factors in apoptosis regulation of SLE, HAX-1 may be involved in the pathogenesis of SLE, and the silence of HAX-1 may be beneficial for the improvement of SLE.


[Desmoplastic fibroblastoma].
We report a light microscopic and ultrastructural appearance, and immunohistochemical profile of two benign tumours of soft tissue recently designated as desmoplastic fibroblastoma (collagenous fibroma). The tumours were located in the foot and well demarcated. Microscopically, they were composed of stellate--or spindle--shaped cells embedded in a hypovascular fibrous of fibromyxoid stroma. Immunohistochemical examination showed that tumours were diffusely positive for vimentin and focaly positive for smooth muscle actin and muscle specific actin. Ultrastructurally, the neoplastic cells had features of fibroblasts and myofibroblasts.


Role of the laparoscopic approach to cytoreductive nephrectomy in metastatic renal-cell carcinoma: does size matter?
Retrospective studies have shown laparoscopic cytoreductive nephrectomy (LCN) to be a safe procedure in selected patients. The objective of this article is to identify characteristics that may predict when a laparoscopic procedure may offer improved postoperative outcome and whether it affects the timing of postoperative systemic therapy compared with open surgery. A cohort of 43 LCN cases were matched with 43 open cytoreductive nephrectomy (OCN) cases based on both pathologic size of tumor and stage. Eleven cases were laparoscopic converted to open nephrectomy. Cases excluded from the analysis were adjacent organ involvement, inferior vena cava involvement, and bulky lymphadenopathy. Data analysis of 11 variables was performed using the t test, log-rank, and Wilcoxon tests. Significance was at P = 0.05. Survival data were calculated using the Kaplan-Meier estimate. Significant differences between LCN vs OCN were estimated blood loss (mean 277 ml vs 816 ml) and length of hospitalization (3.2 days vs 5.1 days). The median size of tumor for LCN cases was 7.5 cm and for OCN, 9.5 cm. The mean size of tumor of LCN vs laparoscopic converted to open cases was 6.8 cm vs 11.2 cm, and this difference was significant. There was no significant difference in postoperative performance status, time to commencement of systemic treatment, or in survival time between both groups. This study provides further evidence that a laparoscopic approach with cytoreductive nephrectomy in metastatic renal-cell carcinoma is a safe option for tumors 10 cm and smaller. The approach (laparoscopic vs open) had no effect on postoperative complications or time to systemic therapy. Procedures with tumors larger than 10 cm were more likely to be converted to an open procedure. Tumors larger than 10 cm may be best approached via an open procedure, especially in the presence of involvement of adjacent organs or bulky lymphadenopathy.


[Suicidal behavior in the aged].
Suicide in the elderly has been underemphasized in gerontological research. This deficit is even more surprising as the aged, especially the very old male group, have the highest suicide rate in the population. This pattern has not markedly changed over time. The very old male is at highest risk of suicide, a fact still waiting for a conclusive explanation. Epidemiologists have predominantly investigated four groups of risk factors for suicide in old age; (1) chronic and painful illnesses, (2) psychiatric disorders (mainly depression), (3) conflicts and stress in interpersonal relationships, and (4) social isolation and loneliness. Suicide finally results from stressful life events, losses and conflicts, which, through an accumulating effect, end up in hopelessness and despair. The research literature does not offer a convincing explanation for the characteristics of suicide in old age. Sociologists (e.g. Durkheim) have tried to explain suicide in the aged by stressing the "anomic" situation in old age, characterized by disintegration, the "roleless role" decreasing influence and weakening ties to groups and institutions in society. Psychiatrists on the other hand (e.g. Ringel, Henseler) offer explanations derived from a psychodynamic point of view. In their opinion, suicide is the final solution to a severe neurotic crisis. The psychiatric explanation for suicide tends to neglect stressful life events, losses and environmental influences which result in an intolerable life situation, and finally in a self-destroying reaction. The life-event paradigm and the cognitive personality theory give the opportunity to develop a model in which environmental and psychological (personality) variables can be exemplified. Furthermore, their interdependence can be described. The function of this model is primarily heuristic. It can stimulate more specific research in the field of suicide and suicide attempts in old age.


Serum neuron-specific enolase and immunohistochemical markers of neuroendocrine differentiation in lung cancer.
An enzyme immunoassay for serum neuron-specific enolase (NSE) was evaluated with respect to analytical performance and clinical utility and compared with immunohistochemical evaluation of neuroendocrine differentiation. Values obtained agreed well with values obtained using a radioimmunoassay method giving a correlation coefficient of 0.934. Analytical performance of the enzyme immunoassay was good but the diagnostic sensitivity of 82% in extensive and 67% in limited disease was insufficient for serum NSE to be of value in the diagnosis of small cell lung cancer (SCLC). Serum NSE decreased significantly in 11 of 15 patients with SCLC following institution of chemotherapy. Classification of lung cancers into SCLC and non small cell lung cancer (NSCLC) types is largely based on tumour morphology. Neuroendocrine differentiation may not be morphologically evident. Immunohistochemical staining of tumour tissue with markers of neuroendocrine differentiation, i.e. NSE (both monoclonal and polyclonal antibodies) Leu 7, Chromogranin A and P G P 9.5 was performed in both patients with SCLC and NSCLC. 38 per cent of patients with NSCLC had both raised serum NSE and positive NSE (polyclonal) immunoperoxidase staining of lung tissue. A further 35 per cent of patients showed a raised serum NSE or positive immunohistochemistry but not both. The presence of two positive immunoperoxidase markers in lung tissue has been suggested as an indicator of responsiveness to chemotherapy in NSCLC patients. A number of factors may affect immunohistochemical positivity in tissue sections and the additional use of a serum marker may better define chemotherapy responsive groups.


Generation of Isthmic Organizer-Like Cells from Human Embryonic Stem Cells.
The objective of this study was to induce the production of isthmic organizer (IsO)-like cells capable of secreting fibroblast growth factor (FGF) 8 and WNT1 from human embryonic stem cells (ESCs). The precise modulation of canonical Wnt signaling was achieved in the presence of the small molecule CHIR99021 (0.6 μM) during the neural induction of human ESCs, resulting in the differentiation of these cells into IsO-like cells having a midbrain-hindbrain border (MHB) fate in a manner that recapitulated their developmental course in vivo. Resultant cells showed upregulated expression levels of FGF8 and WNT1. The addition of exogenous FGF8 further increased WNT1 expression by 2.6 fold. Gene ontology following microarray analysis confirmed that IsO-like cells enriched the expression of MHB-related genes by 40 fold compared to control cells. Lysates and conditioned media of IsO-like cells contained functional FGF8 and WNT1 proteins that could induce MHB-related genes in differentiating ESCs. The method for generating functional IsO-like cells described in this study could be used to study human central nervous system development and congenital malformations of the midbrain and hindbrain.


Patient monitoring: the big picture.
Increased provision of patient monitoring equipment can assist in the process of raising standards of care. The range of such equipment is extensive, and estates professionals including planners, engineers and facilities managers need to be aware of what items are appropriate to different clinical areas and the sort of back-up support the equipment will require. This article is formed from a report of an independent multi-disciplinary working group which reviewed patient monitoring technology and looked at factors involved in purchasing monitoring equipment.


Using Fish Population Metrics to Compare the Effects of Artificial Reef Density.
Artificial reefs continue to be added as habitat throughout the world, yet questions remain about how reef design affects fish diversity and abundance. In the present study, the effects of reef density were assessed for fish communities and sizes of economically valuable Lutjanus campechanus 13 km off Port Mansfield, Texas, at a reef composed of more than 4000 concrete culverts. The study spanned from May to June in 2013 and 2014, and sites sampled included natural reefs, bare areas, and varying culvert patch density categories, ranging from 1-190 culverts. Abundances of adults and species evenness of juvenile populations differed between the years. Fish communities did not significantly differ among density categories; however, highest species richness and total abundances were observed at intermediate culvert densities and at natural reefs. Whereas the abundance of L. campechanus did not differ among density categories, mean total lengths of L. campechanus were greatest at the lower density. Our findings suggest that reefs should be deployed with intermediate patch density of 71-120 culverts in a 30-m radius to yield the highest fish abundances.


Human cytidine deaminase: purification of enzyme, cloning, and expression of its complementary DNA.
Cytidine (CR) deaminase was purified 47,000-fold to homogeneity from human placenta. The molecular mass of CR deaminase was estimated to be 48.7 kDa by gel filtration and 16.1 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis, suggesting that it contains three or four identical subunits. We determined the amino acid sequence of several peptide fragments and designed 5'-primers to amplify, by the polymerase chain reaction, a specific 364-base pair DNA fragment using human liver complementary DNA (cDNA) as the template. This DNA fragment, which contains the codons of one peptide, was used as a probe to screen a cDNA library from human liver. We isolated and sequenced a cDNA clone of 910 base pairs that contained a 5' nontranslated region, a 438-base pair coding region, and a 3' nontranslated region with a polyadenylated tail. The translated region of the clone contained a deduced sequence of 146 amino acids, with a predicted molecular mass of 16.2 kDa and the sequences of our peptides. The cDNA was ligated in pGEX vector and expressed in Escherichia coli. The expressed protein had a high CR deaminase activity and molecular mass of 16.3 kDa. These data demonstrate clearly that the open reading frame of our cDNA clone codes for a functional human CR deaminase. Polymerase chain reaction amplifications of gene-specific DNA fragments from human/rodent hybrid cells indicate the localization of CR deaminase gene to human chromosome 1. The cDNA for CR deaminase will be a useful molecular probe to investigate the importance of this enzyme in chemotherapy.


NRG Oncology-Radiation Therapy Oncology Group Study 1014: 1-Year Toxicity Report From a Phase 2 Study of Repeat Breast-Preserving Surgery and 3-Dimensional Conformal Partial-Breast Reirradiation for In-Breast Recurrence.
To determine the associated toxicity, tolerance, and safety of partial-breast reirradiation. Eligibility criteria included in-breast recurrence occurring >1 year after whole-breast irradiation, <3 cm, unifocal, and resected with negative margins. Partial-breast reirradiation was targeted to the surgical cavity plus 1.5 cm; a prescription dose of 45 Gy in 1.5 Gy twice daily for 30 treatments was used. The primary objective was to evaluate the rate of grade ≥3 treatment-related skin, fibrosis, and/or breast pain adverse events (AEs), occurring ≤1 year from re-treatment completion. A rate of ≥13% for these AEs in a cohort of 55 patients was determined to be unacceptable (86% power, 1-sided α = 0.07). Between 2010 and 2013, 65 patients were accrued, and the first 55 eligible and with 1 year follow-up were analyzed. Median age was 68 years. Twenty-two patients had ductal carcinoma in situ, and 33 had invasive disease: 19 ≤1 cm, 13 >1 to ≤2 cm, and 1 >2 cm. All patients were clinically node negative. Systemic therapy was delivered in 51%. All treatment plans underwent quality review for contouring accuracy and dosimetric compliance. All treatment plans scored acceptable for tumor volume contouring and tumor volume dose-volume analysis. Only 4 (7%) scored unacceptable for organs at risk contouring and organs at risk dose-volume analysis. Treatment-related skin, fibrosis, and/or breast pain AEs were recorded as grade 1 in 64% and grade 2 in 7%, with only 1 (<2%) grade ≥3 and identified as grade 3 fibrosis of deep connective tissue. Partial-breast reirradiation with 3-dimensional conformal radiation therapy after second lumpectomy for patients experiencing in-breast failures after whole-breast irradiation is safe and feasible, with acceptable treatment quality achieved. Skin, fibrosis, and breast pain toxicity was acceptable, and grade 3 toxicity was rare.


Catechol-O-methyltransferase (COMT) polymorphisms modulate working memory in individuals with schizophrenia and healthy controls.
Cognitive impairment is a core feature of schizophrenia, related to dopaminergic dysfunction in the prefrontal cortex (PFC). It is hypothesized that functional single nucleotide polymorphism (SNP) rs4680 of the catechol-O-methyltransferase (COMT) gene could mediate the relationship between cognition and dopamine activity in the PFC. Other COMT SNPs could also play a role. We evaluated the role of three COMT SNPs (rs737865, rs165599, and rs4680) in schizophrenia and their impact on three working memory tasks. For genetic association analyses, 212 individuals with schizophrenia and 257 healthy controls (HCs) were selected. The Visual Working Memory (VWM) Task, Keep Track Task, and Letter Memory Task were administered to 133 schizophrenics and 93 HCs. We found a significant association of rs737865, with the GG genotype exerting a protective effect and the GA haplotype (rs4680/rs165599) exerting a risk effect for schizophrenia. COMT rs4680 AA carriers and rs737865 AA carriers scored lowest on the Keep Track Task. When the genotype*group interaction effect was evaluated, rs165599 exerted opposite effects for VWM and Keep Track task performance in patients and controls, with AA carriers scoring lowest on both tests among controls, but highest among patients. These data support the hypothesis that COMT polymorphisms may be associated with schizophrenia and modulate cognition in patients and controls.


High-risk age window for mortality in children with cystic fibrosis after lung transplantation.
LTx in children with CF remains controversial. The UNOS database was queried from 1987 to 2013 for CF patients <18 yr of age at time of transplant. PCHR model was used to quantify hazard of mortality. 489 recipients were included in the survival analysis. The hazard function of post-transplant mortality was plotted over attained age to identify age window of highest risk, which was 16-20 yr. Unadjusted PCHR model revealed ages immediately after the high-risk window were characterized by lower hazard of mortality (HR = 0.472; 95% CI = 0.302, 0.738; p = 0.001). After adjusting for potential confounders, the decline in mortality hazard immediately after the high-risk window remained statistically significant (HR = 0.394; 95% CI: 0.211, 0.737; p = 0.004). Hazard of mortality in children with CF after LTx was highest between 16 and 20 yr of attained age and declined thereafter.


What do Cochrane systematic reviews say about interventions for autism spectrum disorders?
Autism spectrum disorders (ASDs) include autistic disorder, Asperger's disorder and pervasive developmental disorder. The manifestations of ASDs can have an important impact on learning and social functioning that may persist during adulthood. The aim here was to summarize the evidence from Cochrane systematic reviews on interventions for ASDs. Review of systematic reviews, conducted within the Discipline of Evidence-Based Medicine, Escola Paulista de Medicina, Universidade Federal de São Paulo. We included and summarized the results from Cochrane systematic reviews on interventions for ASDs. Seventeen reviews were included. These found weak evidence of benefits from acupuncture, gluten and casein-free diets, early intensive behavioral interventions, music therapy, parent-mediated early interventions, social skill groups, Theory of Mind cognitive model, aripiprazole, risperidone, tricyclic antidepressants and selective serotonin reuptake inhibitors (SSRI); this last only for adults. No benefits were found for sound therapies, chelating agents, hyperbaric oxygen therapy, omega-3, secretin, vitamin B6/magnesium and SSRI for children. Acupuncture, gluten and casein-free diets, early intensive behavioral interventions, music therapy, parent-mediated early interventions, social skill groups and the Theory of Mind cognitive model seem to have benefits for patients with autism spectrum disorders (very low to low-quality evidence). Aripiprazole, risperidone, tricyclic antidepressants and SSRI (this last only for adults) also showed some benefits, although associated with higher risk of adverse events. Experimental studies to confirm a link between probable therapies and the disease, and then high-quality long-term clinical trials, are needed.


Comparison of the BD Veritor System for Flu A+B with the Alere BinaxNOW influenza A&B card for detection of influenza A and B viruses in respiratory specimens from pediatric patients.
The performance characteristics of two commercially available rapid tests for influenza, the BD Veritor System for Flu A+B (BD) and the Alere BinaxNOW influenza A&B card (BN), were evaluated using 200 frozen clinical specimens collected from January 2011 to June 2012 from pediatric patients. Real-time reverse transcriptase PCR (RT-PCR) was used as the gold standard to evaluate the results obtained by the two different assays. Of the 200 specimens tested, real-time RT-PCR assay detected influenza A or B virus in 116 samples, while BD detected 104 samples and BN detected 84 samples as positive. The overall sensitivity and specificity for detection of both influenza A and B virus in comparison to those of real-time RT-PCR were 89.6% (95% confidence interval [CI], 82.2 to 94.3) and 98.8% (95% CI, 92.6 to 99.9) for BD Veritor and 72.4% (95% CI, 63.2 to 80.0) and 100% (95% CI, 94.5 to 100.0) for BinaxNOW. Workflow analysis indicated that overall processing times for a batch size of 10 specimens were virtually identical between both systems. Overall, these results indicate that the BD Veritor assay was more sensitive than the BinaxNOW assay in detection of influenza A and B viruses in respiratory specimens from pediatric patients.