Cell stemness is maintained upon concurrent expression of RB and the mitochondrial ribosomal protein S18-2.
Stemness encompasses the capability of a cell for self-renewal and differentiation. The stem cell maintains a balance between proliferation, quiescence, and regeneration via interactions with the microenvironment. Previously, we showed that ectopic expression of the mitochondrial ribosomal protein S18-2 (MRPS18-2) led to immortalization of primary fibroblasts, accompanied by induction of an embryonic stem cell (ESC) phenotype. Moreover, we demonstrated interaction between S18-2 and the retinoblastoma-associated protein (RB) and hypothesized that the simultaneous expression of RB and S18-2 is essential for maintaining cell stemness. Here, we experimentally investigated the role of S18-2 in cell stemness and differentiation. Concurrent expression of RB and S18-2 resulted in immortalization of Rb1-/- primary mouse embryonic fibroblasts and in aggressive tumor growth in severe combined immunodeficiency mice. These cells, which express both RB and S18-2 at high levels, exhibited the potential to differentiate into various lineages in vitro, including osteogenic, chondrogenic, and adipogenic lineages. Mechanistically, S18-2 formed a multimeric protein complex with prohibitin and the ring finger protein 2 (RNF2). This molecular complex increased the monoubiquitination of histone H2ALys119, a characteristic trait of ESCs, by enhanced E3-ligase activity of RNF2. Furthermore, we found enrichment of KLF4 at the S18-2 promoter region and that the S18-2 expression is positively correlated with KLF4 levels. Importantly, knockdown of S18-2 in zebrafish larvae led to embryonic lethality. Collectively, our findings suggest an important role for S18-2 in cell stemness and differentiation and potentially also in cancerogenesis.


An Update on Refractory Hypertension.
To update on definition, diagnosis, prevalence, patient characteristics, pathophysiology, and treatment of refractory hypertension (RfHTN). Refractory hypertension (RfHTN) is defined as blood pressure (BP) that is uncontrolled despite using ≥ 5 antihypertensive medications of different classes, including a long-acting thiazide diuretic and a mineralocorticoid receptor antagonist (MRA) at maximal or maximally tolerated doses. This new phenotype is different from resistant hypertension (RHTN), defined as BP that is uncontrolled despite using ≥ 3 medications, commonly a long-acting calcium channel blocker (CCB), a blocker of the renin-angiotensin system (angiotensin-converting enzyme [ACE] inhibitor or angiotensin receptor blocker [ARB]), and a diuretic. The RHTN phenotype includes controlled RHTN, BP that is controlled on 4 or more medications. RfHTN is largely attributable to increased sympathetic activity, unlike RHTN, which is mainly due to increased intravascular fluid volume frequently caused by hyperaldosteronism and chronic excessive sodium ingestion. Compared to those with controlled RHTN, patients with RfHTN have a higher prevalence of target organ damage and do not have elevated aldosterone levels. Ongoing clinical trials are assessing the safety and efficacy of using devices to aid with BP control in patients with RfHTN. RfHTN is a separate entity from RHTN and is generally attributable to increased sympathetic activity.


Effects of Dipole Potential Modifiers on Heterogenic Lipid Bilayers.
In this work, we examine the ability of dipole modifiers, flavonoids, and RH dyes to affect the dipole potential (φ d) and phase separation in membranes composed of ternary mixtures of POPC with different sphingolipids and sterols. Changes in the steady-state conductance induced by cation-ionophore complexes have been measured to evaluate the changes in dipole potential of planar lipid bilayers. Confocal fluorescence microscopy has been employed to investigate lipid segregation in giant unilamellar vesicles. The effects of flavonoids on φ d depend on lipid composition and dipole modifier type. The effectiveness of RH dyes to increase φ d depends on sphingolipid type but is not influenced by sterol content. Tested modifiers lead to partial or complete disruption of gel domains in bilayers composed of POPC, sphingomyelin, and cholesterol. Substitution of cholesterol to ergosterol or 7-dehydrocholesterol leads to a loss of fluidizing effects of modifiers except phloretin. This may be due to various compositions of gel domains. The lack of influence of modifiers on phase scenario in vesicles composed of ternary mixtures of POPC, cholesterol, and phytosphingosine or sphinganine is related to an absence of gel-like phase. It was concluded that the membrane lateral heterogeneity affects the dipole-modifying abilities of the agents that influence the magnitude of φ d by intercalation into the bilayer and orientation of its own large dipole moments (phloretin and RH dyes). The efficacy of modifiers that do not penetrate deeply and affect φ d through water adsorption (phlorizin, quercetin, and myricetin) is not influenced by lateral heterogeneity of membrane.


Structural effects of an LQT-3 mutation on heart Na+ channel gating.
Computational methods that predict three-dimensional structures from amino acid sequences have become increasingly accurate and have provided insights into structure-function relationships for proteins in the absence of structural data. However, the accuracy of computational structural models requires experimental approaches for validation. Here we report direct testing of the predictions of a previously reported structural model of the C-terminus of the human heart Na(+) channel. We focused on understanding the structural basis for the unique effects of an inherited C-terminal mutation (Y1795C), associated with long QT syndrome variant 3 (LQT-3), that has pronounced effects on Na(+) channel inactivation. Here we provide evidence that this mutation, in which a cysteine replaces a tyrosine at position 1795 (Y1795C), enables the formation of disulfide bonds with a partner cysteine in the channel. Using the predictions of the model, we identify the cysteine and show that three-dimensional information contained in the sequence for the channel protein is necessary to understand the structural basis for some of the effects of the mutation. The experimental evidence supports the accuracy of the predicted structural model of the human heart Na(+) channel C-terminal domain and provides insight into a structural basis for some of the mutation-induced altered channel function underlying the disease phenotype.


Aspects of inattention in low birth weight children.
The objective of this study was to explore the nature of impaired attention in low birth weight children. A population-based sample (n = 129) of 11-year-old low birth weight children was compared with a randomized group of normal birth weight children (n = 128). Continuous Performance Test, a Cue-Target Task, and a Stroop Color-Word test were used to assess the following dimensions of attentional function: inattention, impulsivity, vigilance, selected attention, and alternating attention. The attention subscale from the Child Behavior Check List questionnaire was included to assess maternal report of inattention. Low birth weight children had generally slower reaction time than normal birth weight children and manifested impaired vigilance, but this could be attributed to group differences in confounding parental factors. Impairment was not a function of birth weight within the low birth weight group, but low birth weight boys manifested impaired vigilance compared with normal birth weight boys. Every fourth low birth weight child was reported with inattention, but small differences were found between the inattentive and attentive low birth weight children. No specific attentional dysfunction was observed in the low birth weight study group. The nature of reported impaired attention among low birth weight children is still uncertain.


A miniaturized analyzer for the catalytic determination of iodide in seawater and pharmaceutical samples.
A monolithic flow conduit was especially designed and coupled to a multisyringe unit (MSFIA) in order to develop a kinetic analytical method. The new device, denoted CHIP, integrates different functions in a reduced size including confluent mixing, reaction coil, and thermostating, which allowed minimizing the dimensions of the entire analyzer system. The CHIP-MSFIA was satisfactorily applied to the determination of iodide using the Sandell-Kolthoff reaction. The resulting system allows fast, simple and automatic analysis in seawater samples and a pharmaceutical preparation. By the use of an additional syringe, reagent blank, sample blank, unspecific interferences, and sample analyte concentration was evaluated by simply changing the mode of operation. The instrumentation and analytical procedures were optimized in respect of sensitivity. A limit of detection of 4.7 μg L(-1) and linear working range of 4.3-70 μg L(-1) were achieved. Two new modes of air bubble elimination were developed and employed in this work, leading to improved reproducibility (RSD %=1.5%) and method robustness. The accuracy of analysis for seawater samples was evaluated by an Add-Recovery test obtaining recoveries between 97 and 106%. For the quantification of iodide in the pharmaceutical sample, standard addition calibration was required. No significant differences between the found and the certified value were found. The entire analytical procedure lasted about 150 s enabling a measuring frequency of 23 h(-1).


Models for the prediction of successful induction of labor based on pre-induction sonographic measurement of cervical length.
To examine the effect of pre-induction cervical length, parity, gestational age at induction, maternal age and body mass index (BMI) on the possibility of successful delivery in women undergoing induction of labor. In 822 singleton pregnancies, induction of labor was carried out at 35 to 42(+6) weeks of gestation. The cervical length was measured by transvaginal sonography before induction. The effect of cervical length, parity, gestational age, maternal age and BMI on the interval between induction and vaginal delivery within 24 hours was investigated using Cox's proportional hazard model. The likelihood of vaginal delivery within 24 hours and risk for cesarean section overall and for failure to progress was investigated using logistic regression analysis. Successful vaginal delivery within 24 hours of induction occurred in 530 (64.5%) of the 822 women. Cesarean sections were performed in 161 (19.6%) cases, 70 for fetal distress and 91 for failure to progress. Cox's proportional hazard model indicated that significant prediction of the induction-to-delivery interval was provided by the pre-induction cervical length (HR=0.89, 95% CI 0.88-0.90, p<0.0001), parity (HR=2.39, 95% CI 1.98-2.88, p<0.0001), gestational age (HR=1.13, 95% CI 1.07-1.2, p=or<0.0001) and birth weight percentile (HR=0.995, 95% CI 0.99-0.995, p=0.001), but not by maternal age or BMI. Logistic regression analysis indicated that significant prediction of the likelihood of vaginal delivery within 24 hours was provided by pre-induction cervical length (OR=0.86, 95% CI 0.84-0.88, p<0.0001), parity (OR=3.59, 95% CI 2.47-5.22, p<0.0001) and gestational age (OR=1.19, 95% CI 1.07-1.32, p=or<0.0001) but not by BMI or maternal age. The risk of cesarean section overall was significantly associated with all the variables under consideration, i.e., pre-induction cervical length (OR=1.09, 95% CI 1.06-1.11, p<0.0001), parity (OR=0.25, 95% CI 0.17-0.38, p<0.0001), BMI (OR=1.85, 95% CI 1.24-2.74, p=0.0024), gestational age (OR=0.88, 95% CI 0.78-0.98, p=0.0215) and maternal age (OR=1.04, 95% CI 1.01-1.07, p=0.0192). The risk of cesarean section for failure to progress was also significantly associated with pre-induction cervical length (OR=1.11, 95% CI 1.07-1.14, p<0.0001), parity (OR=0.26, 95% CI 0.15-0.43, p<0.0001), gestational age (OR=0.83, 95% CI 0.73-0.96, p=0.0097) and BMI (OR=2.07, 95% CI 1.27-3.37, p=0.0036). In women undergoing induction of labor, pre-induction cervical length, parity, gestational age at induction, maternal age and BMI have a significant effect on the interval between induction and delivery within 24 hours, likelihood of vaginal delivery within 24 hours and the risk of cesarean section.


Sanguinarine, a phototoxic H2O2-producing alkaloid.
Sanguinarine chloride, a quaternary salt of a benzophenanthrene alkaloid, was phototoxic to catalase-deficient strains of Escherichia coli but not to Trichoplusia ni (cabbage looper moth larvae), an insect with high levels of catalase activity. Chemical analyses confirm that sanguinarine is an efficient producer of H2O2. This differential toxicity suggests that the mode of phototoxic action involves production of H2O2 which could be detoxified in many organisms by catalase.


Carboxymethyl-selenopyruvic acid as the product of the oxidative deamination of carboxymethyl-selenocysteine.
CMSeC labeled with 14C in the carboxymethyl moiety was incubated with snake venom L-aminoacid oxidase. As the product of the reaction only one ketoacid was detected, which retained all the radioactivity of the oxidized substrate. This clearly shows that no breakdown of the C-Se bond of CMSeC occurs during its oxidation, and confirms previously reported data indicating that the ketoacid arising from CMSeC is CMSeP.


Implications of second-look laparotomy in the context of optimally resected stage III ovarian cancer: a non-randomized comparison using an explanatory analysis: a Gynecologic Oncology Group study.
A non-randomized comparison of outcome in women undergoing second-look laparotomy (SLL) or clinical follow-up, after receiving six cycles of combination chemotherapy with paclitaxel plus either cisplatin or carboplatin, for optimally resected stage III ovarian cancer. Prior to chemotherapy randomization, patients chose whether or not to undergo SLL; this was a stratification factor to insure balance of treatment assignment. Any subsequent therapy was physician-directed. Explanatory analysis replaced intent-to-treat because of a higher likelihood of detecting SLL effect in the presence of noncompliance. There were 393 patients (median age: 54) who Elected SLL and 399 (median age: 59) who Elected No SLL. The former group was more likely to have gross residual disease at initial surgery than the latter group (69% versus 60%, respectively). In the Elected SLL group, 59 (15%) patients subsequently refused surgery, in nine (2%) surgery was contraindicated, and 31 (8%) relapsed or died prior to the procedure. Cancer was found in 46% of 294 (75%) patients undergoing SLL. Since early failures (prior to SLL) do not address benefit, such patients (SLL: 32; No SLL: 33), defined as progression-free survival (PFS) < 6 months, were excluded from analysis. The adjusted relative risk of progression is 0.89 (95% confidence interval: 0.75, 1.07); the difference in median PFS is 1.0 month (SLL: 23.9 months; No SLL: 22.9 months). The survival rate curves are superimposable. In the context of a non-randomized comparison, the performance of a SLL was not associated with longer survival.


Severe Obesity in Children May Not Pose Independent Risk for Influenza Complications.
Subsets of children are targeted for influenza vaccination due to known conditions that increase the risk of influenza complications. The purpose of this study was to determine if severe obesity in children suggests targeted vaccination. A retrospective chart review of a large Midwestern pediatric hospital identified 188 cases of influenza complications (defined as requiring hospitalization or death) aged 2 to <20 years old from August 1, 2010 through June 30, 2013. Severe obesity was defined as body mass index (BMI) ≥99% for age and gender, with patients grouped by severe obesity status (yes vs. no). Cases were reviewed for previously identified risk conditions for influenza complications (e.g., asthma, pneumonia, diabetes), and were classified as having or not having a known high risk condition. Of 188 cases, 174 (93%) had a high-risk condition, while only 14 (7%) had no known condition. All 14 (100%) with no known high-risk condition had a BMI <99%. All 15 (100%) with BMI ≥99% had a known high-risk condition. The association between severe obesity status and influenza complications was not statistically significant (p = 0.61). This suggests that severe obesity in children is not an independent high-risk condition for influenza complications defined as requiring hospitalization or resulting in death, once other known influenza risk factors are considered. Based on this data, clinicians should not target children for influenza vaccination based on weight status. We cannot comment about whether severe obesity represents increased risk for less severe cases of influenza.


Development and application of a spray tip that enables electrocoagulation of a variety of tissues.
Spray hemostasis is possible using a high-frequency power source from the tip of an electric scalpel; however, the difficulties regarding the uniformity and rapidity of the hemostasis surface remain. This study reports the development of a novel electrocoagulation device tip that can be used in endoscopic and robotic surgeries and can quickly coagulate and hemostat and easily adjust the extent of cauterization and hemostasis while minimizing the depth of thermal injury. The safety and efficacy of the hemostatic device were verified in a porcine model. A liver surface transection was conducted in vivo and the rapidity of the hemostatic effect of the device was observed. An extracted stomach, kidney, and liver were cauterized ex vivo by three operators with different surgical skills and the effects were analyzed pathologically. In addition, a sacrificed pig cadaver was used to achieve hemostasis at a renal transection site using the multi-spray endoscope tip. An increase in the number of tip terminals expanded the cauterization surface and shortened the cauterization time. In parenchymatous organs, uniform cauterization was possible without increasing the depth of thermal injury. The cauterization depth did not depend on the operator's skill, and the spray coagulation was safe. The variable spray tip allowed for simple hemostasis during open and laparoscopic surgeries. This novel electrocoagulation device tip can be developed as a forceps that can change the spray range and can be used during laparoscopic and robotic surgeries.


Host monitoring by aphid migrants: do gynoparae maximise offspring fitness?
Gynoparae (autumn migrants) of the bird cherry-oat aphid,Rhopalosiphum padi (L.), were not found to land randomly on their primary host,Prunus padus L. Some trees, although within a few metres of heavily infested trees, were not colonized at all. This phenomenon occurs regularly from year to year.The reasons for this non-random landing pattern are examined in terms of the success of subsequent generations, special attention being paid to the reproductive rate of the oviparae and fundatrices and the success of the developing populations in the spring of the following year.Trees in Scotland and Finland were examined in this manner and the hypothesis developed that the gynoparae ofR. padi show maternal care in selecting hosts that favour their offspring's survival and reproduction.Aphids were reared under controlled conditions in the laboratory on twigs cut from trees of known previous infestation levels. Aphids on twigs from preferred trees had better survival rates and produced more offspring than those aphids on twigs from non-preferred trees.


"From food waste to food supervision"-Cuttlefish Ink Natural Nanoparticles-Driven Dual-mode Lateral Flow Immunoassay for Advancing Point-of-Care Tests.
Apart from the obvious benefit of "trash-to-treasure", the acquisition of natural nanomaterials from cheap and renewable waste has been intensively researched because of various bioactivities and physical-chemical features. Herein, for the first time, we employed natural cuttlefish ink nanoparticles (CINPs) as a multifunctional label and designed colorimetric-photothermal dual-mode lateral flow immunoassays (CINPs-mediated CPLFIA) for sensitive detection of clenbuterol (CL). The accessibility and renewability of CINPs overcome barriers that artificial nanomaterials face, such as complex manufacturing and relatively high costs. Additionally, inspired by the mussel adhesion, the bio-affinity of CINPs, such as antibody coupling and preservation, was investigated and showed to be considerably superior to Au NPs, leading to significantly increased immunosensor sensitivity. Meanwhile, CINPs exhibit excellent photothermal conversion efficiency for dual-signal production, avoiding the effect of environmental elements (particularly light) for colorimetric mode. Besides, the biosensor was integrated with a smartphone and a thermal imager for portable sensing. After optimization, the detection limit of CINPs-mediated CPLFIA was 0.179 ng mL-1 (colorimetric mode) and 0.076 ng mL-1 (photothermal mode), which were significantly lower than traditional gold nanoparticles-based LFIA (0.786 ng mL-1). This research attempted to explain the rise in sensitivity. From food waste to food supervision, this research explores the hidden value of natural resources.


FKBP1A rs6041749 polymorphism is associated with allograft function in renal transplant patients.
To investigate the potential impact of single-nucleotide polymorphisms (SNPs) in the FK506-binding protein (FKBP)-calcineurin (CaN)-nuclear factor of activated T cells (NFAT) signaling pathway on the efficacy and safety of tacrolimus (TAC) in Chinese renal transplant patients. Seventy-seven tag SNPs were detected in 146 patients who were on TAC-based maintenance immunosuppression and who followed up for at least 2 years. The relationships of these polymorphisms with clinical outcomes such as acute rejection, acute nephrotoxicity, pneumonia, and estimated glomerular filtration rate (eGFR) were explored. For the FKBP1A rs6041749 polymorphism, which has a significant association with renal function over time, a preliminary functional analysis was performed using a dual-luciferase reporter gene system. The patients with FKBP1A rs6041749 TT genotype had a more stable eGFR level than CC and CT carriers (P = 2.08 × 10-8) during the 2 years following transplantation. Dual-luciferase reporter assay results showed that the rs6041749 C variant could enhance the relative luciferase activity compared with the T variant, which indicated that the rs6041749 C allele may increase the FKBP1A gene transcription. In addition, we did not find any association between these genetic variants and the risk of acute rejection, acute nephrotoxicity, and pneumonia in renal transplant patients receiving TAC-based immunosuppression. FKBP1A rs6041749 C allele carriers are at higher risk for eGFR deterioration. The variant might serve as a biomarker to predict allograft function in renal transplant patients.


Lactobacillus salivarius and Lactobacillus agilis feeding regulates intestinal stem cells activity by modulating crypt niche in hens.
Previously, we found that Lactobacillus salivarius, Lactobacillus agilis, and Lactobacillus aviarius were associated with excellent egg nutrition in native chicken. Next, the optimal Lactobacillus combination is worth studying. Here, a total of 120 HyLine hens (30 hens per group contained 3 replicate cells, 10 hens/cell) in the laying peak period were randomly divided into (1) control, (2) L. salivarius + L. agilis, (3) L. salivarius + L. aviarius, and (4) L. agilis + L. aviarius groups, fed with diet only or with corresponding Lactobacilli (108 colony-forming units/hen/day) for 30 days. As a result, L. salivarius + L. agilis feeding could (1) improve egg-laying rate, egg weight, and albumen's amino acid levels; (2) increase Lactobacillus abundance, decrease Escherichia coli abundance, upregulate the tryptophan metabolism pathway-related molecules, and downregulate the primary bile acid biosynthesis pathway-related molecules in intestinal contents; and (3) upregulate oxidative-phosphorylation pathway-related genes, reactive oxygen species levels, and mRNA abundance of Wnt3a, Dll1, Lgr5, CCDN1, and CDK2 in the crypt. Collectively, L. salivarius + L. agilis feeding in hens could improve intestinal microflora and metabolism profile, promote crypt's local energy metabolism and reactive oxygen species levels, and thus enhance Paneth cells and intestinal stem cells activity.Key points• Lactobacilli co-feeding could improve laying performance and egg nutrition.• Lactobacilli co-feeding could improve intestinal microflora and metabolism profile.• Lactobacilli co-feeding could enhance Paneth cells and intestinal stem cells activity.


Real-time burn depth assessment using artificial networks: a large-scale, multicentre study.
Early judgment of the depth of burns is very important for the accurate formulation of treatment plans. In medical imaging the application of Artificial Intelligence has the potential for serving as a very experienced assistant to improve early clinical diagnosis. Due to lack of large volume of a particular feature, there has been almost no progress in burn field. 484 early wound images are collected on patients who discharged home after a burn injury in 48 h, from five different levels of hospitals in Hunan Province China. According to actual healing time, all images are manually annotated by five professional burn surgeons and divided into three sets which are shallow(0-10 days), moderate(11-20 days) and deep(more than 21 days or skin graft healing). These ROIs were further divided into 5637 patches sizes 224 × 224 pixels, of which 1733 shallow, 1804 moderate, and 2100 deep. We used transfer learning suing a Pre-trained ResNet50 model and the ratio of all images is 7:1.5:1.5 for training:validation:test. A novel artificial burn depth recognition model based on convolutional neural network was established and the diagnostic accuracy of the three types of burns is about 80%. The actual healing time can be used to deduce the depth of burn involvement. The artificial burn depth recognition model can accurately infer healing time and burn depth of the patient, which is expected to be used for auxiliary diagnosis improvement.


Perceived Mistreatment in Health Care Settings and its Relationship with HIV Clinical Outcomes in HIV-positive People who Use Drugs in Vancouver, Canada.
People living with HIV (PLWH) often experience stigma and discrimination by health care professionals. We investigated the prevalence of perceived mistreatment in health care settings and its associations with HIV clinical outcomes and health care-seeking behaviour using data from a long-running prospective cohort of PLWH who use drugs. Of the 857 participants included, 19% reported at least one instance of perceived mistreatment during the study period. In adjusted longitudinal analyses, perceived mistreatment was positively associated with not being on ART in the same follow-up period, and participants who reported perceived mistreatment were less likely to report seeing a physician in the subsequent follow-up period. Daily use of injection drugs was positively associated with reporting perceived mistreatment. These findings demonstrate the implications of negative health care interactions in a population that must consistently engage with the health care system, and the need for stigma-reducing educational interventions for health care professionals.


pH-, sugar-, and temperature-sensitive electrochemical switch amplified by enzymatic reaction and controlled by logic gates based on semi-interpenetrating polymer networks.
Phenylboronic acid (PBA) moieties are grafted onto the backbone of poly(acrylic acid) (PAA), forming the PAA-PBA polyelectrolyte. The semi-interpenetrating polymer network (semi-IPN) films composed of PAA-PBA and poly(N,N-diethylacrylamide) (PDEA) were then synthesized on electrode surface with entrapped horseradish peroxidase (HRP), designated as PDEA-(PAA-PBA)-HRP. The films demonstrated reversible pH-, fructose-, and thermo-responsive on-off behavior toward electroactive probe K(3)Fe(CN)(6) in its cyclic voltammetric (CV) response. This multiswitchable CV behavior of the system could be further employed to control and modulate the electrochemical reduction of H(2)O(2) catalyzed by HRP immobilized in the films with K(3)Fe(CN)(6) as the mediator in solution. The responsive mechanism of the system was also explored and discussed. The pH-sensitive property was attributed to the electrostatic interaction between the PAA component of the films and the probe at different pH; the thermo-responsive behavior originated from the structure change of PDEA hydrogel component of the films with temperature; the fructose-sensitive property was ascribed to the structure change of the films induced by the complexation between the PBA constituent and the sugar. This smart system could be used as a 3-input logic network composed of enabled OR (EnOR) gates in chemical or biomolecular computing by combining the multiresponsive property of the films and the amplification effect of bioelectrocatalysis and demonstrated the potential perspective for fabricating novel multiswitchable electrochemical biosensors and bioelectronic devices.


Synthesis, prototropic tautomerism studies of indenocarbothioamides and their conversion to biologically active indenothiazole derivatives.
Indenocarbothioamides 2, obtained from reaction of indane-1,3-dione with aryl isothiocyanates, on condensation with 3-chloropentane-2,4-dione and chloroacetyl chloride furnished 1,3-indanedione coupled thiazole and thiazolidin-5-one derivatives, respectively. The structure of the obtained products was assigned on the basis of spectral data (IR, NMR and Mass). Prototropic tautomerism studies of carbothioamides 2 were carried out in solution (1H NMR, 13C NMR and UV-vis), gas phase (Mass) and solid state (IR and X-ray). X-ray diffraction studies of carbothioamide 2a have revealed the existence of enolic form in the solid state. DFT studies of various possible tautomeric forms in gas phase as well as in solution state corroborated by the experimental results. Antibacterial studies of indenothiazole and indenothiazolidin-5-one derivatives have been reported.


Association between asthma self-management knowledge and asthma control in the elderly.
Considerable evidence points to the importance of patient education in achieving better asthma control. However, little is known about the effect of older adults' asthma knowledge on asthma control. To identify the relation between asthma self-management knowledge and short-term asthma control in older adults. The study included 82 patients with asthma undergoing usual asthma care who were interviewed by the same trained allergist. At the same time, the authors conducted an asthma knowledge questionnaire, an Asthma Control Test, skin prick testing, spirometry, and evaluation of inhaler device technique. The Turkish version of the asthma knowledge questionnaire was administered to all participants in face-to-face interviews. The relation between asthma knowledge and asthma control was tested by regression analysis. The education level was below the secondary level in 79% of patients; 59% of patients were obese; and 44% of patients had mild, 41% had moderate, and 5% had severe persistent asthma. The atopy rate was 21%. The most common sensitization was to Dermatophagoides pteronyssinus. Fifty-two percent of patients had uncontrolled asthma; 22 patients showed proper use of the inhaler device. Forty patients (48%) had limited asthma knowledge. The main source of asthma knowledge was from physicians for 81% of patients. There was no significant association between patients' asthma knowledge and asthma control level (P = .991). Knowledge of asthma was very low in elderly patients and usual asthma care was largely insufficient. These findings suggest that asthma education programs should be developed for older adults based on their education level and clinical asthma characteristics.


The role of JAK inhibitors in hematopoietic cell transplantation.
The Janus Kinase (JAK)/Signal Transducers and Activators of Transcription (STAT) pathway is essential for both the regulation of hematopoiesis and the control of inflammation. Disruption of this pathway can lead to inflammatory and malignant disease processes. JAK inhibitors, designed to control the downstream effects of pro-inflammatory and pro-angiogenic cytokines, have been successfully used in pre-clinical models and clinical studies of patients with autoimmune diseases, hematologic malignancies, and the hematopoietic cell transplantation (HCT) complication graft versus host disease (GVHD). In the last decade, JAK inhibitors Ruxolitinib, Fedratinib, and most recently Pacritinib have been United States Federal Drug Administration (FDA) approved for the treatment of myelofibrosis (MF). Ruxolitinib was also recently approved for the treatment of steroid refractory acute as well as chronic GVHD; JAK inhibitors are currently under evaluation in the pre-HCT setting in MF and for the prevention of GVHD. This review will focus on the role of JAK inhibitors in the treatment of hematologic malignancies, the potential function of pre-HCT JAK inhibitors in patients with MF, and the role of JAK inhibitors in the prevention and treatment of acute and chronic GVHD.


An improved method to detect ethyl glucuronide in urine using reversed-phase liquid chromatography and pulsed electrochemical detection.
Pulsed electrochemical detection (PED) following reversed-phase liquid chromatography (LC) has been applied recently to the detection of ethyl glucuronide (EtG) in the urine of live and deceased individuals. In this paper, several key improvements to the method are made to enhance sensitivity, reproducibility, and accuracy. These improvements include (i) further optimization of the sample preparation procedure that has increased the recovery from ca. 50% to 84+/-3% in synthetic urine matrix; (ii) changing the internal standard from methyl glucuronide (MetG) to propyl glucuronide (ProG), which does not elute within the interference of the matrix; and (iii) altering the mobile phase of the separation from acetonitrile to t-butanol to virtually eliminate signal suppression in PED. As a consequence, detection limits have been reduced to 0.01 microg mL(-1), reproducibility has been improved by a factor of two, and sample size has been reduced five-fold. Blind studies in synthetic urine showed no significant difference between the amount recovered and the true value determined at the 95% confidence level for all samples. Importantly, PED requires no derivatization, and it can detect virtually all glucuronides.


Modulation of peptide binding by HLA-B27 polymorphism in pockets A and B, and peptide specificity of B*2703.
The results in this study address three aspects of peptide binding to the disease-associated antigen HLA-B27 and its modulation by polymorphism: the contribution of major anchor residues 2 and 9, the role of pocket B polymorphism in modulating peptide specificity, and the binding properties of B*2703, a subtype not found to be associated with spondyloarthropathy. Synthetic analogs of peptides naturally presented by B*2705 were used to demonstrate that residue 2 is essential, since Ala2 analogs bound marginally to B*2705, but the specificity of B*2705 for Arg2 is not absolute, and show that the contribution of basic residue 9 to binding was significant, but less than Arg2. The effect of single mutations in the B pocket was to decrease or--with the Glu > Met-45 mutation--totally shift pocket B specificity for Arg2 towards other residues at this position. This was shown by quantitating the relative binding of Gln2 and Ala2 analogs, and by pool-sequencing of the peptides bound in vivo to these mutants. Peptides naturally presented by B*2705 apparently bound with a lower affinity to pocket A variants with altered hydrogen bonding to the peptide N terminus, including B*2703. Binding of peptide analogs with changes at positions 2 or 9 suggested that in B*2703 pocket A, interactions are weaker and pocket B interactions are stronger than in B*2705. This can be explained by the effect of the unique His59 change in B*2703 in both pockets. Thus, B*2703 is probably the HLA-B27 sub-type with the most stringent specificity for the Arg2 peptide motif.


Characterisation and evaluation of metal-loaded iminodiacetic acid-silica of different porosity for the selective enrichment of phosphopeptides.
Silica particles of different porosity were functionalised with iminodiacetic acid (IDA) and loaded with Fe(III) to yield immobilised metal affinity chromatography stationary phases (Fe(III)-IDA-silica) for phosphopeptide enrichment. The elution step of bound phosphopeptides was optimised with a 32P radioactive labelled peptide by a comprehensive study. Several elution systems, including phosphate buffers of different pH and concentration and ethylenediaminetetraacetic acid solutions were employed. Furthermore the effect of support porosity on elution behaviour was investigated. Under best conditions recoveries higher than 90% were achieved. A solid-phase extraction (SPE) protocol was developed for fractionation of phosphorylated and non-phosphorylated peptides and desalting of the fractions which is essential for subsequent mass spectrometric analysis by the combination of Fe(III)-IDA-silica and C18-silica particles. The pH of the loading buffer was found to be a critical parameter for the efficiency of the SPE protocol. As tryptic digests of alpha-lactalbumin, lysozyme and ribonuclease A mixed with three synthetic phosphopeptides were fractionated, pH 2.5 provided minimal proportion of unspecific bound peptides when comparing the fractions after mu-LC-electrospray ionization MS separation. The effect of a sample derivatisation reaction (methylation) on the efficiency of phosphopeptide enrichment was further investigated. Blocking carboxylate groups by methyl ester formation totally prevented unspecific interaction with the immobilised Fe(III) ions, but generated partially methylated phosphopeptides that increased the complexity of the phosphorylated fraction.


Frequency of caseous lymphadenitis (CLA) in sheep slaughtered in an abattoir in Tabriz: comparison of bacterial culture and pathological study.
From January to February 2008, 468 sheep carcasses (335 male and 133 female) in a Khosroshahr (suburb of Tabriz, East Azerbaijan province, Iran) abattoir were randomly selected for inspection. The aim of the study was to estimate the frequency of caseous lymphadenitis (CLA) in sheep and to compare the results of bacterial cultures and histopathology of suspected cases. The mean age of the population was 2.5 years. One hundred ninety-seven cases containing 153 (77.7%) males and 44 (22.3%) females had prominent enlargement of one of the lymph nodes (i.e., prescapular, prefemoral, inguinal, supramammary, or midiastinal); these were removed with the surrounding tissue for further evaluation. For confirmed diagnosis of CLA, samples were sent for microbiology and pathology analysis. Standard bacteriological culture methods for isolation of Corynebacterium pseudotuberculosis and tissue preparations for histopathological sections were performed. To evaluate the effect of age on the frequency of CLA, animals were categorized in four groups: under 1, 1-2, 2-3, and over 3 years of age. Based on the results, in 59 (12.60%) carcasses C. pseudotuberculosis was isolated, and in 94 (20.08%) of the cases histopathological studies revealed pathognomonic signs (lamellated exudates or onion ring) of CLA. The frequency of CLA based on bacteriological culture was 12.60% and on histopathological study 20.08%. In 37 (18.8%) of the carcasses, both bacteriological and histopathological studies confirmed CLA. The frequency of CLA following microscopic examination (20.08%) presented a more precise diagnosis compared to bacteriological culture (12.60%) and macroscopic evaluation of the lymph nodes (P < 0.05). Furthermore, there was a positive correlation rate between the bacteriological culture and histopathological study (r = 0.196, P = 0.006). The prescapular lymph node had the highest infection rate with 54 (1.70 ± 0.97) and supramammary lymph node had the lowest with two (0.07 ± 0.41) (P < 0.05). There was an increase in CLA detection with increasing age (P < 0.05), the mean age of animals with a positive CLA test were 2.92 years and in the oldest age group 31 (47%) cases had the highest infection.


Tumor-targeted gene therapy using Adv-AFP-HRPC/IAA prodrug system suppresses growth of hepatoma xenografted in mice.
Clinical efficacy of current therapies for hepatocellular carcinoma (HCC) treatment is limited. Indole-3-acetic acid (IAA) is non-toxic for mammalian cells. Oxidative decarboxylation of IAA by horseradish peroxidase (HRP) leads to toxic effects of IAA. The purpose of this study was to investigate the effects of a novel gene-targeted enzyme prodrug therapy with IAA on hepatoma growth in vitro and in vivo mouse hepatoma models. We generated a plasmid using adenovirus to express HRP isoenzyme C (HRPC) with the HCC marker, alpha-fetoprotein (AFP), as the promoter (pAdv-AFP-HRPC). Hepatocellular cells were infected with pAdv-AFP-HRPC and treated with IAA. Cell death was detected using MTT assay. Hepatoma xenografts were developed in mice by injection of mouse hepatoma cells. The size and weight of tumors and organs were evaluated. Cell death in tumors was assessed using hematoxylin and eosin-stained tissue sections. HRPC expression in tissues was detected using Reverse Transcriptase-Polymerase Chain Reaction. IAA stimulated death of hepatocellular cells infected with pAdv-AFP-HRPC, in a dose- and time-dependent manner, but not in control cells. Growth of hepatoma xenografts, including the size and weight, was inhibited in mice treated with pAdv-AFP-HRPC and IAA, compared with that in control group. pAdv-AFP-HRPC/IAA treatment induced cell death in hepatoma xenografts in mice. HRPC gene expressed only in hepatoma, but not in other normal organs of mice. pAdv-AFP-HRPC/IAA treatment did not cause any side effects on normal organs. These findings suggest that pAdv-AFP-HRPC/IAA enzyme/prodrug system may serve as a strategy for HCC therapy.


Risk Factors for Actinic Keratoses: A Systematic Review and Meta-Analysis.
To integrate evidence and assess the risk factors associated with actinic keratosis (AK). Unrestricted searches were conducted on five electronic databases, with an end-date parameter of September 2021. We summarized the study characteristics and pooled the results from individual studies by using a random-effects model. The risk of bias was estimated using the Cochrane Risk of Bias Tool, and the quality of evidence was estimated according to the Newcastle-Ottawa Scale. Sixteen studies were included in final analysis, and we assessed the AK risk among a variety of risk factors. Overall, the male sex (odds ratio (OR): 2.51; 95% confidence interval (CI): 1.94-3.25; P < 0.01), age >45 years (OR = 7.65, 95% CI: 2.95-19.86; P < 0.01), light Fitzpatrick skin phototype (OR = 2.32, 95% CI: 1.74-3.10; P < 0.01), light hair color (OR = 2.17, 95% CI: 1.40-3.36; P < 0.01), light eye color (OR = 1.67, 95% CI: 1.03-2.70; P = 0.04), freckles on face/arms (OR = 1.88, 95% CI: 1.37-2.58; P < 0.01), suffered positive history of other types of non-melanoma skin cancer (OR = 4.46, 95% CI: 2.71-7.33; P < 0.01), sunburns in childhood (OR = 2.33, 95% CI: 1.47-3.70; P < 0.01) and adulthood (OR = 1.50, 95% CI: 1.12-2.00; P < 0.01), severe sunburn (OR = 1.94, 95% CI: 1.62-2.31; P < 0.01), and chronic occupational and/or recreational sun exposure (OR = 3.22, 95% CI: 2.16-4.81; P < 0.01) increased the risk of AK. Moreover, sunscreen use (OR = 0.51, 95% CI: 0.34-0.77; P < 0.01) and history of atopy reduced the risk of AK. Sensitivity analysis yielded consistent results. The included studies showed a high risk of bias. We confirm several well-known AK risk factors and their quantitative data, and summarized the uncommon risk factors and protective factors. Our results may inform on the design and implementation of AK screening and educational programs.


Papillary carcinoma of the breast: diagnostic agreement and management implications.
Papillary carcinoma (PC), which is a rare type of breast cancer, comprises a heterogeneous group of tumours. The diagnostic categorization of PC as in-situ and invasive disease remains a matter of debate with respect to interpretation of its overlapping histological features, and with respect to the uncertainty in clinical behaviour that this dilemma raises. The aim of this study was to assess the diagnostic agreement regarding PC among reporting breast pathologists. Six cases of PC included in the UK National Health Service Breast Screening Programme breast pathology interpretive external quality assurance scheme in the last 10 years were reviewed. In this scheme, one representative haematoxylin and eosin-stained slide from each case is circulated to an average of 600 participants. Data on diagnostic categories were collected and slides were reviewed according to the World Health Organization (WHO) diagnostic criteria. The number of final diagnoses of malignancy (in situ or invasive) was highest for invasive PC (99% of the participants diagnosed it as malignant), followed by solid PC (94% and 95%, respectively), encapsulated PC (92% and 92%, respectively), and papillary ductal carcinoma in situ (DCIS) (88%). Most cases of papillary DCIS were correctly classified as in-situ disease (77%), but 28% of the participants classified invasive PC cases as in-situ disease. Of the participants, 24% reported encapsulated PC as invasive disease. Of the two solid PC cases, one showed some features consistent with the WHO description of invasive solid PC, whereas the other showed features of classic (non-invasive) solid PC. Both cases were reported as invasive by 75% and 77% of participants, respectively. Breast specialists more frequently classified PC as an in-situ carcinoma than did non-specialist participants, and the difference was significant (P = 0.013). Recognition of PC as a malignant entity (in situ or invasive) is high, but concordance of its classification into in-situ and invasive disease is low. Histological features that can define invasion in PC should be better defined. These rare lesions require additional diagnostic work-up, and difficult cases should trigger consensus opinion or expert referral.


ICSI outcomes for infertile men with severe or complete asthenozoospermia.
Severe or complete asthenozoospermia is a rare entity that can lead to male infertility. In this study, we explored whether different extents of severe or complete asthenozoospermia could affect intracytoplasmic sperm injection (ICSI) outcomes and compared the ICSI outcomes using testicular spermatozoa with those using ejaculated spermatozoa in couples with complete asthenozoospermia. Ninety-seven couples with severe or complete asthenozoospermia who underwent ICSI between January 2014 and December 2018 were included. According to the sperm category used in ICSI, patients were categorized into four groups: ejaculated progressive motile sperm group (Ep-group), ejaculated non-progressive motile sperm group (En-group), ejaculated immotile sperm group (Ei-group), and testicular sperm group (TESE-group). We compared the baseline characteristics, hormone profile, semen parameters, normal fertilization, good-quality embryos on day 3, transferred embryos, and ICSI outcomes in the four groups. The clinical pregnancy rate was significantly increased in the Ep-group (65.4%, P = 0.019) and TESE-group (63.6%, P = 0.035) compared with that in the Ei-group (23.1%). The ongoing pregnancy rate in the Ei-group was significantly lower than that in the Ep-group (23.1% vs. 61.5%, P = 0.041). Moreover, the biochemical pregnancy rate, ongoing pregnancy rate, and live birth rate were much lower in the Ei-group than in the TESE-group (30.8% vs. 63.6%, 23.1% vs. 40.4% and 23.1% vs. 40.4%, respectively). In couples with complete asthenozoospermia, testicular spermatozoa should be preferred to ejaculated spermatozoa for obtaining a better ICSI outcome. With the appropriate selection of testicular spermatozoa, the extent of severe or complete asthenozoospermia may not affect the ICSI outcomes. Future studies with a larger sample size are warranted to validate these findings.


Impact of molecular biology studies on the understanding of brain tumors in childhood.
Pediatric brain tumors are the second most common form of childhood malignancy. Brain tumors are a very heterogenous group of tumors and the pathogenesis of many of these tumors is yet to be clearly elucidated. Current diagnostic tools include histopathology and immunohistochemistry, but classification based on these means has significant limitations. As our understanding of the molecular biology of individual tumors continues to increase it has led to the identification of reliable and increasingly available molecular biomarkers. Molecular techniques are likely to complement current standard means of investigation and help not only overcome diagnostic challenges but may also result in better disease classification and risk stratification, leading to more personalized therapeutic approaches.


Author Correction: Optimization of Molecules via Deep Reinforcement Learning.
An amendment to this paper has been published and can be accessed via a link at the top of the paper.


Functional analysis of the 1p34.3 risk locus implicates GNL2 in high-grade serous ovarian cancer.
The high-grade serous ovarian cancer (HGSOC) risk locus at chromosome 1p34.3 resides within a frequently amplified genomic region signifying the presence of an oncogene. Here, we integrate in silico variant-to-function analysis with functional studies to characterize the oncogenic potential of candidate genes in the 1p34.3 locus. Fine mapping of genome-wide association statistics identified candidate causal SNPs local to H3K27ac-demarcated enhancer regions that exhibit allele-specific binding for CTCF in HGSOC and normal fallopian tube secretory epithelium cells (FTSECs). SNP risk associations colocalized with eQTL for six genes (DNALI1, GNL2, RSPO1, SNIP1, MEAF6, and LINC01137) that are more highly expressed in carriers of the risk allele, and three (DNALI1, GNL2, and RSPO1) were upregulated in HGSOC compared to normal ovarian surface epithelium cells and/or FTSECs. Increased expression of GNL2 and MEAF6 was associated with shorter survival in HGSOC with 1p34.3 amplifications. Despite its activation of β-catenin signaling, RSPO1 overexpression exerted no effects on proliferation or colony formation in our study of ovarian cancer and FTSECs. Instead, GNL2, MEAF6, and SNIP1 silencing impaired in vitro ovarian cancer cell growth. Additionally, GNL2 silencing diminished xenograft tumor formation, whereas overexpression stimulated proliferation and colony formation in FTSECs. GNL2 influences 60S ribosomal subunit maturation and global protein synthesis in ovarian cancer and FTSECs, providing a potential mechanism of how GNL2 upregulation might promote ovarian cancer development and mediate genetic susceptibility of HGSOC.


Interaction between sodium ascorbate and dopamine.
The interaction between sodium ascorbate and dopamine was investigated by three different parameters: radical intensity, prooxidant action, and cytotoxicity induction. Sodium ascorbate and dopamine produced the doublet and quartet ESR signals under alkaline conditions (pH 8.0-9.5), respectively. Addition of increasing concentrations of sodium ascorbate completely scavenged the dopamine radical and replaced the latter with its own radical. Similarly, dopamine slightly, but significantly reduced the radical intensity of sodium ascorbate. These two compounds stimulated the methionine oxidation and hydrogen peroxide generation in culture medium, but in combination, their stimulation activities were weakened. Both of these two compounds dose-dependently reduced the viable cell number of human oral squamous carcinoma HSC-4 cells, and their cytotoxic activity was significantly reduced by catalase. When these two compounds were mixed together before adding to HSC-4 cells, both of their cytotoxic activities were diminished. The present study demonstrates the interaction between sodium ascorbate and dopamine, which might modify their biological activities and generation of nerve disorders such as Parkinson's disease.


The impact on coagulation of an intravenous loading dose in addition to a subcutaneous regimen of low-molecular-weight heparin in the initial treatment of acute coronary syndromes.
We sought to quantify the impact of adding an intravenous loading dose to a subcutaneous regimen of enoxaparin in patients with an acute coronary syndrome (ACS). It is unclear whether an intravenous (i.v.) loading dose of enoxaparin should be added to a subcutaneous (s.c.) regimen in patients with ACS. Patients admitted with ACS were randomized to i.v.+s.c.(n = 14) or s.c. alone (n = 11) enoxaparin treatment. Coagulation markers were measured at nine time points during the first 24 h of treatment. The i.v.+s.c. therapy immediately resulted in therapeutic anti-Xa levels, which remained significantly higher for 6 h compared with s.c. alone, without reaching excessively high levels. A rapid decrease of plasma prothrombin fragments 1+2 (F(1+2)) levels was observed as soon as 5 min after the i.v. injection (33% lower; p = 0.007), and these levels remained lower up to 2 h after the start of treatment compared with SQ alone. The ex vivo thrombin generation time was maximally prolonged at 5 min post-injection in the i.v.+s.c. group and remained significantly prolonged up to 6 h post-injection compared with s.c. alone. The tissue factor pathway inhibitor plasma activity was immediately increased by 194% with i.v.+s.c., whereas the maximum increase with s.c. alone was 47% at 3 h. Therapeutic plasma levels of enoxaparin are achieved significantly earlier by an i.v.+s.c. regimen compared with s.c. alone, without leading to unacceptably high levels. As the risk of thrombotic complications is greatest early after admission, the observed differences in antithrombotic effects may translate into a clinical benefit. However, this remains to be established.


Effect of serotonin on cyclic AMP level in rat hypothalamus slices during development.
The effect of serotonin (5-HT) on cyclic AMP levels in rat hypothalamic slices was age-dependent. The sensitivity of the cyclic AMP system to 5 X 10(-5) M 5-HT already existed in the foetus at 21 days of pregnancy. It reached a maximum on the 7th postnatal day and then decreased with age. In adult tissue the response was still present and was antagonized by two serotonergic antagonists (methiothepin and metergoline). When compared to the progressive increase of 5-HT level in the hypothalamus the data suggest a different evolution of serotonergic innervation and of 5-HT receptors.


External validation of Pittsburgh Cardiac Arrest Category illness severity score.
Early prognostication post-cardiac arrest can help determine appropriate medical management and help evaluate effectiveness of post-arrest interventions. The Pittsburgh Cardiac Arrest Category (PCAC) severity score is a 4-level illness severity score found to strongly predict patient outcomes in both in- (IHCA) and out-of-hospital cardiac arrests (OHCA). We aimed to validate the PCAC severity score in an external cohort of cardiac arrest patients. We retrospectively assigned PCAC scores to both IHCA and OHCA patients treated by our hypothermia team from July 1, 2009 to July 1 2016. Our primary outcome was survival to hospital discharge. Secondary outcomes were favorable functional status defined as favorable discharge disposition (home or acute rehabilitation), discharge Cerebral Performance Category (CPC); and discharge modified Rankin Scale (mRS). We tested the association of PCAC and outcomes using a multivariable adjusted logistic regression model. We included 317 subjects in our model. PCAC was strongly associated with survival I Reference; II adjusted odds ratio (OR) 0.20 95% confidence interval (CI) 0.35-0.66, III (OR 0.14 CI 0.3-0.73, p < 0.05); IV (OR 0.05 CI 0.01-0.24, p < 0.01). PCAC was similarly associated with favorable functional outcomes: favorable discharge disposition II (OR 0.12 CI 0.02-0.68), III (OR 0.19 CI 0.05-0.74, p < 0.05) IV (OR 0.05 CI 0.01-0.22, p < 0.01); favorable CPC score II (OR 0.25 CI 0.06-1.03), III (OR 0.14 CI 0.03-0.57, p < 0.01), IV (OR 0.05 CI 0.01-0.20, p < 0.01) and favorable mRS (OR 0.47 CI (0.33-0.68)). Early (<6 h post-arrest) PCAC severity scoring strongly predicts patient outcomes from cardiac arrest in both OHCA and IHCA.


Altered whole-brain resting-state functional connectivity and brain network topology in typhoon-related post-traumatic stress disorder.
Altered resting-state functional connectivity has been found in patients with post-traumatic stress disorder (PTSD). However, the alteration of resting-state functional connectivity at whole-brain level in typhoon-traumatized individuals with PTSD remains largely unknown. To investigate changes in whole-brain resting-state functional connectivity and brain network topology in typhoon-traumatized subjects with and without PTSD. Cross-sectional study. Twenty-seven patients with typhoon-related PTSD, 33 trauma-exposed controls (TEC), and 30 healthy controls (HC) underwent resting-state functional MRI scanning. The whole brain resting-state functional connectivity network was constructed based on the automated anatomical labeling atlas. The graph theory method was used to analyze the topological properties of the large-scale resting-state functional connectivity network. Whole-brain resting-state functional connectivity and the topological network property were compared by analyzing the variance. There was no significant difference in the area under the curve of γ, λ, σ, global efficiency, and local efficiency among the three groups. The PTSD group showed increased dorsal cingulate cortex (dACC) resting-state functional connectivity with the postcentral gyrus (PoCG) and paracentral lobe and increased nodal betweenness centrality in the precuneus relative to both control groups. Compared with the PTSD and HC groups, the TEC group showed increased resting-state functional connectivity between the hippocampus and PoCG and increased connectivity strength in the putamen. In addition, compared with the HC group, both the PTSD and TEC groups showed increased connectivity strength and nodal efficiency in the insula. Aberrant resting-state functional connectivity and topology were found in all trauma-exposed individuals. These findings broaden our knowledge of the neuropathological mechanisms of PTSD.


Assays and activities of glycosidic enzymes in bovine peripheral blood leukocytes.
Methods are described for the quantitative measurement of N-acetyl-beta-D-glucosaminidase, alpha-mannosidase and beta-glucuronidase in peripheral blood leukocytes of the bovine. Enzyme kinetics and stability were determined. Activities of the glycosidases in polymorphonuclear leukocytes and mononuclear leukocytes were determined using the optimized assays. Polymorphonuclear leukocytes had greater activities of N-acetyl-beta-D-glucosaminidase and alpha-mannosidase, and similar levels of beta-glucuronidase, when compared to mononuclear leukocytes.


A new amperometric glucose microsensor: in vitro and short-term in vivo evaluation.
For biosensor fabrication, it is important to optimize materials and methods in order to create predictable function in vitro and in vivo. For this reason, we designed a new glucose sensor ('revised protocol') that utilized an outer permselective membrane made of amphiphobic polyurethane which allows glucose passage through hydrophilic segments. An inner polyethersulfone membrane, stabilized with a trimethoxysilane, provided specificity. Before application of the inner membrane, it was necessary to etch the platinum electrode with a radio frequency oxygen plasma. The revised protocol sensors (n=185) were compared with sensors fabricated with an earlier ('original') protocol (n=204) which used an outer polyurethane without hydrophilic segments and a complex inner membrane of cellulose acetate and Nafion. The function of revised protocol sensors was more predictable in vitro as evidenced by a much lower variation of glucose sensitivity than the original protocol sensors. Revised and original protocol sensors were nearly linear up to a glucose concentration of 20 mM. In vitro interference from 0.1 mM acetaminophen was minimal in both groups of sensors and would be expected to represent about 2% of the total sensor response at normal glucose levels for revised protocol sensors. Prolonged testing of the revised protocol sensors for 11 days during immersion in buffer revealed stable sensitivities (day 1: 6.12+/-1.34 nA/mM; day 3: 6.33+/-1.40; day 8: 7.13+/-1.39; and day 11: 7.56+/-1.47; sensitivity for day 1 vs. each other day: not significant) and no critical loss of glucose oxidase activity. The response of the revised protocol sensors (n=7) to intraperitoneal glucose was tested in rats approximately one day after subcutaneous implantation and the sensors tracked glucose closely with a slight lag of 3-6 min.


Melatonin use and the risk of 30-day mortality among US veterans with sepsis: A retrospective study.
Prior research suggests melatonin has beneficial effects that could improve survival among sepsis patients. This exploratory analysis sought to compare 30-day survival among melatonin treated and untreated patients with sepsis. A retrospective cohort study was conducted among patients with a primary inpatient admission diagnosis for sepsis utilizing the International Classification of Diseases, versions 9 and 10, Clinical Modification (ICD-9-CM and ICD-10-CM) diagnosis codes between 2000 and 2021. Propensity score weighting was utilized, accounting for demographic, clinical, and laboratory factors. Weighted Cox models were estimated for 30-day in-hospital and 30-day overall survival. A total of 9386 patients were included in the study with 593 exposed to melatonin within the first day of hospitalization. Propensity score weighted Cox models reveal melatonin was associated with a 37.9% decreased risk of 30-day in-hospital mortality (HR = 0.621; 95% CI = [0.415-0.931]) and a 33.5% decreased risk of 30-day overall mortality (HR = 0.665; 95% CI = [0.493-0.897]). Factors associated with higher risk of both in-hospital and overall mortality include male sex, white race, age, higher Charlson comorbidity burden, sodium and potassium levels, intensive care unit stay, invasive ventilation, and vasopressor use. Higher serum albumin levels are associated with lower mortality risks. Among patients diagnosed with sepsis, exposure to melatonin was associated with a lower in-hospital and 30-day mortality. Additional research is warranted to fully understand the role of melatonin in sepsis.


[Current trends and first clinical studies of therapy for chronic hepatitis C in children using pegylated interferon alpha and ribavirin].
Our aim was to evaluate the recent clinical data concerning the course of chronic hepatitis C in children and the rationale indications for therapy. There is no doubt that in adult group the pegylated interferon is better than standard interferon and results in higher response rate in combination with ribavirin. Actually complicating the issues surrounding hepatitis C in children is the lack of information on the efficacy and safety of this therapy in pediatric age group. The first preliminary data of our clinical study concerning the therapy with pegylated interferon alpha 2b and ribavirin in 10 children with chronic hepatitis C (genotype 1) suggest a good tolerance of this drug and the end of treatment virologic response in 8 children (80%).


Elemental analysis of urinary calculi by laser induced plasma spectroscopy.
Laser induced plasma spectroscopy (LIPS) has been applied to analyse and identify elemental constituents of urinary calculi. Measurements on seven different urinary stone samples were conducted and the concentrations of some key elemental species were estimated. The elements detected with the present system were: Calcium, Magnesium, Sodium, Samarium, Potassium and Lead. Absolute concentrations of the species were derived from pre-calibration of the system for each element. Their concentrations were found to be widely different in different samples. It was observed that the samples containing a significant amount of lead have large proportion of calcium. It has been established that LIPS would allow real time clinic measurements of elemental contents and the concentrations in the biomaterials without sample preparation. The technique has the potential for routine clinic applications in urological disorder diagnosis.


Tonofilament differentiation in human epidermis, isolation and polypeptide chain composition of keratinocyte subpopulations.
Epidermal tonofilaments are intermediate filaments with an unusually complex polypeptide chain composition which undergoes profound changes during differentiation. Assessment of the possible functional significance of these changes requires their correlation with defined stages of keratinocyte differentiation. Methods have, therefore, been developed for the isolation of the four keratinocyte subpopulations from human epidermis in high yield and purity without detectable damage to tonofilament chains. Trypsinization at 4 degrees C yielded sheets of granular and horny layers and a suspension of basal and spinous cells. Granular cells were removed from the horny layer sheets by mechanical action and basal cells separated from spinous cells by selective attachment to collagen. The living cell fractions were representative, by morphological and ultrastructural criteria, of their counterparts in situ. Analysis of the keratinocyte subpopulations by SDS-PAGE revealed directly their tonofilament chain composition. Basal cells contain two quantitatively major tonofilament chains (Mr 58000 and 50000). A third major chain (Mr 70000) is only synthesised suprabasally when the cells lose their ability to divide. The Mr 50000 chain is removed before the granular cell stage and several minor chains are present in the spinous and granular cells. Two chains appearing for the first time in horny cells probably result from processing during the terminal stages of differentiation. It is suggested that synthesis of the Mr 70000 chain in suprabasal keratinocytes is normally linked with their loss of mitotic ability. This may be significant in the skin disease psoriasis in which synthesis of the Mr 70000 chain is defective and the epidermis hyperproliferates.


Using Return on Investment Operational and Monte Carlo Modeling Techniques to Predict Financial Performance in a Tertiary Care Outpatient Clinic.
The vast majority of health care quality improvement studies provide inadequate financial analysis to accurately predict a return on investment. We hypothesized that using return on invested capital operational mapping combined with a Monte Carlo simulation financial model could accurately predict institutional costs and operational metrics within an outpatient urology clinic. A process map of a typical outpatient clinic visit was developed, and time studies were performed by following a sample of patients while considering all operational and financial variables that contributed to patient care. this process map was adapted into a return on invested capital-tree for financial modeling. Stochastic modeling using Monte Carlo simulation was performed to estimate financial metrics based on these operational and financial inputs for both the 2017-2018 and 2018-2019 fiscal years. These were then compared to the actual performance measures of those fiscal years. Combined return on invested capital-Monte Carlo simulation modeling generated financial and operational estimates that characterized the clinic's performance based on multivariable inputs. Most financial estimates for 2017-2018 differed by <4.31% from the actual financial values from that year. In predicting financial performance for 2018-2019, most of the estimated values were <7.67% different from their actual financial statement line items. As a proof of concept, this study demonstrated that a combined return on invested capital-operational mapping and Monte Carlo simulation modeling can predict key financial metrics in a tertiary care clinic. As such, common business tools can be useful in a health care setting when clinicians are evaluating how investments in quality improvement will influence their financial and operational performance.


[Effect of xylitol and sorbitol on metabolic indices in diabetic children].
The influence of readily available carbohydrates (sugar, xylitol and sorbitol) on the metabolic processes was studied in 80 children with diabetes mellitus. It was found that even low doses (20 g) of sorbitol and xylitol produced an unfavourable effect on the lipid metabolism and liver function. It is recommended that low doses (20 g) of sugar be included into the diet for children with diabetes mellitus.


Minimization of the adverse environmental effects of discarded onions by avoiding disposal through dehydration and food-use.
Onion is a commonly used vegetable in the Cuyo Region, Argentina, and important in the world global vegetable production ranking. Production levels often exceed immediate markets of fresh sale, and excesses, although still edible, are currently discarded (mostly incinerated and disposed in landfill, creating numerous environmental hazards). To minimize these adverse environmental problems, this research investigates upgrading the discarded products by dehydration, hence guaranteeing their ultimate food-use. The dehydration process of discarded onion cv. Crioula Roxa was hence studied at 60 and 70 °C, temperatures selected to maintain the main physicochemical characteristics of the onions, while also creating optimum heat and mass transfer coefficients while significantly reducing the energy consumption and CO2 emissions. When using an electrically-heated convective dryer, the Specific Energy Consumption (SEC) values and the CO2 emissions during dehydration at 70 °C are 738.89 kWh.kg-1 and 264.74 kg of CO2 kg-1, respectively. These values are only 41.61 kWh.kg-1 and 2-4 kg of CO2 kg-1 if a solar dryer is applied. The thermal diffusivities were 1.86✕10-10 m2 s-1 (dehydration) and 1.08✕10-10 m2 s-1 (rehydration), showing a weak effect of the dehydration process on the solid structure and properties.


A role for ADP-ribosylation factor 6 in the processing of G-protein-coupled receptors.
After agonist-induced internalization, the vasopressin V2 receptor (V2R) does not recycle to the plasma membrane. The ADP-ribosylation factor (ARF) proteins initiate vesicular intracellular traffic by promoting the recruitment of adaptor proteins; thus, we sought to determine whether ARF6 could promote V2R recycling. Neither the agonist-induced internalization nor the recycling of the V2R was regulated by ARF6, but a constitutively active mutant of ARF6 reduced cell-surface V2Rs 10-fold in the absence of agonist treatment. Visualization of the ARF6 mutant-expressing cells revealed a vacuolar-staining pattern of the V2R instead of the normal plasma membrane expression. Analysis of V2R maturation revealed that reduced cell-surface expression was due to the diminished ability of the newly synthesized receptor to migrate from the endoplasmic reticulum to the Golgi network. The same mechanism affected processing of the V1aR and acetylcholine M2 receptors. Therefore, ARF6 controls the exit of the V2 and other receptors from the endoplasmic reticulum in addition to its established role in the trafficking of plasma-membrane-derived vesicles.


The yolk sac in early pregnancy failure.
An attempt was made to visualize the yolk sac in 845 patients scheduled for chorionic villi sampling. The distribution of yolk sac diameters and the interpolating growth curve up to 11 weeks of development were analyzed in 239 pregnant women who were delivered of normal infants. The highest visualizing rate of the yolk sac in normal pregnancies was 97 at 7 weeks of gestation. A total of 130 miscarriages occurred before chorionic villi sampling. In these cases, the diameter of the yolk sac versus crown-rump length tended to be larger than found in normal pregnancies. The visualizing rate of the yolk sac in miscarriages after the embryo had been formed was significantly higher in those women who demonstrated fetal heart activity (82.1%) than in those who did not (54.5%). On the other hand, the yolk sac was observed in 44% of miscarriages without a visible embryo. These findings suggest different types of missed abortion. An abnormal karyotype was observed in 23 of 29 chromosomal analyses performed on aborted specimens. An abnormal karyotype was observed in all eight cases with only a yolk sac-like structure within the gestational sac.


Defining the clinical phenotype of COVID-19 in children.
On January 7, 2020, a novel coronavirus (SARS-CoV-2) was identified as the causative agent of a cluster of pneumonia of unknown origin detected in Wuhan City by Chinese authorities. Since SARS-CoV-2 discovery, the corresponding disease (COVID-19) has rapidly expanded throughout the globe, making as a consequence the World Health Organization (WHO) declaring a pandemic. As of May 19, 2020, over 4.806.299 cases of COVID-19 had been confirmed worldwide, with more than 318.599 deaths.


Emerging Implications of the Concept of Hydricity in Energy-Relevant Catalytic Processes.
The "hydricity" of a species refers to its hydride-donor ability. Similar to how the pKa is useful for determining the extent of dissociation of an acid, the hydricity plays a vital role in understanding hydride-transfer reactions. A large number of transition-metal-catalyzed processes involve the hydride-transfer reaction as a key step. Among these, two key reactions-proton reduction to evolve H2 and hydride transfer to CO2 to generate formate/formic acid-represent a promising solution to build a sustainable and fossil-fuel-free energy economy. Therefore, it is imperative to develop an in-depth relationship between the hydricity of transition-metal hydrides and its influencing factors, so that efficient and suitable hydride-transfer catalysts can be designed. Moreover, such profound knowledge can also help in improving existing catalysts, in terms of their efficiency and working mechanism. With this broad aim in mind, some important research has been explored in this area in recent times. This Minireview emphasizes the conceptual approaches developed thus far, to tune and apply the hydricity parameter of transition-metal hydrides for efficient H2 evolution and CO2 reduction/hydrogenation catalysis focusing on the guiding principles for future research in this direction.


A time-gated fluorescence detector using a tuning fork chopper.
We describe an affordable gated fluorescence detection system to measure fluorescent compounds with long-lifetimes that uses a tuning fork chopper to block the intense excitation pulse from a flash lamp or short-lived fluorescence. A conventional non-gated inexpensive photosensor module is used to collect the luminescence signal. Using the long-lived luminescence from the terbium(III)-dipicolinic acid (DPA) chelate, we demonstrate a limit of detection (LOD) of 120pM for DPA. This system is not only an order of magnitude less expensive than an electronically gateable phototomultiplier tube (GPMT), it exhibits no evidence of gradual loss of sensitivity, due likely to photocathode fatigue and deterioration, observed with a GPMT.


Brain Functional Basis of Subjective Well-being During Negative Facial Emotion Processing Task-Based fMRI.
Subjective well-being (SWB) is the eternal pursuit of all mankind. Individual differences in SWB may reflect the way of emotional processing. Neuroimaging studies have partly examined the neural mechanism of the individual differences in SWB using resting-state functional magnetic resonance imaging (rs-fMRI). However, few studies have investigated the relationship between the individual differences in SWB and brain functional connectivity during facial emotion processing task. Here, we applied negative facial emotion processing task-based fMRI to explore the brain functional basis of the individual differences in SWB. Results showed that SWB is positively correlated with the activation of right posterior cingulate cortex, left interior temporal gyrus and left angular gyrus for the comparison of negative stimulus and neutral stimulus, revealing that happy individuals may be more proactive to use attention transfer and behavioral inhibition strategies to decrease negative experiences during negative emotional processing. In addition, high SWB is associated with strong functional connectivity between high-level cognitive networks (e.g., frontal-parietal network) and low-level perceptual networks (e.g., sensorimotor network), and weak functional connectivity within default mode network and within low-level perceptual networks, indicating that the self-reflection, emotional regulation and cognitive control during negative facial emotion processing underlies the individual differences in SWB. These findings provide a novel insight to characterize the brain functional basis of the individual differences in SWB.


Serum copper as a marker of inflammation in prediction of short term outcome in high risk patients with chronic heart failure.
Oxidative process and inflammation are regarded as important factors in the pathogenesis of chronic heart failure. Our study was aimed at investigating the prognostic value of serum copper levels in high risk subjects with chronic heart failure. Serum copper levels and other prognostic indicators were determined in the group of 60 patients with chronic heart failure due to ischemic heart disease: 30 consecutive subjects with acute decompensation of chronic heart failure (acute group A) and 30 patients with chronic stable heart failure (group B). Patients were followed prospectively 12 months. Primary end-point was the mean time to death and/or heart failure hospital admission. The mean time to death was in the group A 279.4+/-18.9 days and 351.7+/-13.6 days in the group B (p<0.0001). Cox proportional hazard model revealed that the time to death for all subjects (n=60) was affected by cardiothoracic ratio (p<0.001). The time to combined end-point death or hospital admission was affected by serum copper concentration (p<0.0001). Serum copper levels predicted short term outcome in high risk patients with chronic heart failure.


Tracing metal footprints via global renewable power value chains.
The globally booming renewable power industry has stimulated an unprecedented interest in metals as key infrastructure components. Many economies with different endowments and levels of technology participate in various production stages and cultivate value in global renewable power industry production networks, known as global renewable power value chains (RPVCs), complicating the identification of metal supply for the subsequent low-carbon power generation and demand. Here, we use a multi-regional input-output model (MRIO) combined with a value chain decomposition model to trace the metal footprints (MFs) and value-added of major global economies' renewable power sectors. We find that the MFs of the global renewable power demand increased by 97% during 2005-2015. Developed economies occupy the high-end segments of RPVCs while allocating metal-intensive (but low value-added) production activities to developing economies. The fast-growing demand for renewable power in developed economies or developing economies with upper middle income, particularly China, is a major contributor to the embodied metal transfer increment within RPVCs, which is partly offset by the declining metal intensities in developing economies. Therefore, it is urgent to establish a metal-efficient and green supply chain for upstream suppliers as well as downstream renewable power installers for just transition in the power sector across the globe.


Role of condensates in modulating DNA repair pathways and its implication for chemoresistance.
For cells, it is important to repair DNA damage, such as double-strand and single-strand DNA breaks, because unrepaired DNA can compromise genetic integrity, potentially leading to cell death or cancer. Cells have multiple DNA damage repair pathways that have been the subject of detailed genetic, biochemical, and structural studies. Recently, the scientific community has started to gain evidence that the repair of DNA double-strand breaks may occur within biomolecular condensates and that condensates may also contribute to DNA damage through concentrating genotoxic agents used to treat various cancers. Here, we summarize key features of biomolecular condensates and note where they have been implicated in the repair of DNA double-strand breaks. We also describe evidence suggesting that condensates may be involved in the repair of other types of DNA damage, including single-strand DNA breaks, nucleotide modifications (e.g., mismatch and oxidized bases), and bulky lesions, among others. Finally, we discuss old and new mysteries that could now be addressed considering the properties of condensates, including chemoresistance mechanisms.


Metoprolol for prevention of bucking at orotracheal extubation: a double-blind, placebo-controlled randomised trial.
Respiratory responses to extubation can cause serious postoperative complications. Beta-blockers, such as metoprolol, can interfere with the cough pathway. However, whether metoprolol can effectively control respiratory reflexes during extubation remains unclear. The objective of this study is to evaluate the efficacy of intravenous metoprolol in attenuating respiratory responses to tracheal extubation. Randomized, double-blinded, placebo-controlled trial. Tertiary referral center located in Brasília, Brazil. Recruitment: June 2021 to December 2021. 222 patients of both sexes with an American Society of Anesthesiologists (ASA) physical status I-III aged 18-80 years. Patients were randomly assigned to receive intravenous metoprolol 5 mg IV or placebo at the end of surgery. The primary outcome was the proportion of patients who developed bucking secondary to endotracheal tube stimulation of the tracheal mucosa during extubation. Secondary outcomes included coughing, bronchospasm, laryngospasm, Mean Blood Pressure (MAP), and Heart Rate (HR) levels. Two hundred and seven participants were included in the final analysis: 102 in the metoprolol group and 105 in the placebo group. Patients who received metoprolol had a significantly lower risk of bucking (43.1% vs. 64.8%, Relative Risk [RR = 0.66], 95% Confidence Interval [95% CI 0.51-0.87], p = 0.003). In the metoprolol group, 6 (5.9%) patients had moderate/severe coughing compared with 33 (31.4%) in the placebo group (RR = 0.19; 95% CI 0.08-0.43, p < 0.001). Metoprolol reduced the risk of bucking at extubation in patients undergoing general anesthesia compared to placebo.


Epi-direction detected multimodal imaging of an unstained mouse retina with a Yb-fiber laser.
In this work, we present all epi-direction detected images of an unstained mouse retina using multiphoton microscopy with a sub-50 fs Yb-fiber laser centered at 1.07 μm. This wavelength is particularly interesting as the fundamental wavelength is transparent to the anterior segment of the eye and the higher harmonics are above DNA-damaging UV wavelengths. We present a characterization of the multimodal signals emitted from the different retinal layers, as well as from the choroid and the sclera. By characterizing native multiphoton signals from the retina, we move closer to having Yb-fiber considered for in vivo diagnosis of retinal disease through multiphoton microscopy as well as for corrective therapies.


A heterotrinuclear bioinspired coordination complex capable of binding to DNA and emulation of nuclease activity.
The investigation of compounds capable of strongly and selectively interacting with DNA comprises a field of research in constant development. In this work, we demonstrate that a trinuclear coordination complex based on a dinuclear Fe(III)Zn(II) core designed for biomimicry of the hydrolytic enzyme kidney bean purple acid phosphatase, containing an additional pendant arm coordinating a Pd(II) ion, has the ability to interact with DNA and to promote its hydrolytic cleavage. These results were found through analysis of plasmid DNA interaction and cleavage by the trinuclear complex 1 and its derivatives 2 and 3, in addition to the analysis of alteration in the DNA structure in the presence of the complexes through circular dichroism and DNA footprinting techniques. The suggested covalent interaction of the palladium-containing complex with DNA was analysed using an electrophoretic mobility assay, circular dichroism, high resolution gel separation techniques and kinetic analysis. This is a new and promising metal complex targeted to nucleic acids and acting in two separate ways: strong DNA interaction and hydrolytic cleavage.


Metal-catalyzed oxidation of 2-alkenals generates genotoxic 4-oxo-2-alkenals during lipid peroxidation.
Lipid peroxidation products react with cellular molecules, such as DNA bases, to form covalent adducts, which are associated with aging and disease processes. Since lipid peroxidation is a complex process and occurs in multiple stages, there might be yet unknown reaction pathways. Here, we analyzed comprehensively 2'-deoxyguanosine (dG) adducts with oxidized arachidonic acid using liquid chromatography-tandem mass spectrometry and found the formation of 7-(2-oxo-hexyl)-etheno-dG as one of the major unidentified adducts. The formation of this adduct was reproduced in the reaction of dG with 2-octenal and predominantly with 4-oxo-2-octenal (OOE). We also found that other 2-alkenals (with five or more carbons) generate corresponding 4-oxo-2-alkenal-type adducts. Importantly, it was found that transition metals enhanced the oxidation of C4-position of 2-octenal, leading to the formation of OOE-dG adduct. These findings demonstrated a new pathway for the formation of 4-oxo-2-alkenals during lipid peroxidation and might provide a mechanism for metal-catalyzed genotoxicity.


Fracture resistance and failure patterns of endodontically treated mandibular molars with and without glass fiber post in combination with a zirconia-ceramic crown.
The aim of this study was to investigate the influence of a fiber post on the fracture mechanics of zirconia crowns inserted over endodontically treated teeth with different extent of coronal damage. Endodontically treated human molars with three types of coronal damage received fiber posts before cementation of zirconia-veneered crowns. Controls received composite resin cores without fiber posts. The specimens were loaded to failure and fractographically examined using a scanning electron microscope (SEM). Statistical analysis revealed that specimens with fiber posts demonstrated significantly higher failure loads and favorable fracture pattern compared to the controls. At fractographic analysis, specimens with fiber posts demonstrated delamination of the veneer ceramic from intact zirconia under structure. Meanwhile, the specimens that were restored without a fiber post demonstrated micro-cracking of the composite core build-up resulting in loss of the support under the zirconia crowns which was responsible for the initiation of radial crack and catastrophic damage. Within the limitation of this study, the insertion of fiber post improved the support under zirconia crowns which resulted in higher fracture loads and favorable failure type compared to composite core build-up.


Inhibition of hemorrhagic and edematogenic activities of snake venoms by a broad-spectrum protease inhibitor, murinoglobulin; the effect on venoms from five different genera in Viperidae family.
In order to obtain basic data on the effect of broad-spectrum protease inhibitor against local symptoms of Viperidae snake envenomation, inhibitory capacity of rat murinoglobulin on local hemorrhagic and edematogenic activities of venoms from Crotalus atrox, Bothrops jararaca, Lachesis muta muta, Trimeresurus flavoviridis and Echis carinatus sochureki were examined. Murinoglobulin, pre-incubated with the crude venoms at 37 degrees C for 15 min, inhibited hemorrhagic activity of all five venoms to various extents. The activity of C. atrox was almost completely inhibited at the murinoglobulin/venom ratio (w/w) of 20. The activity of B. jararaca, Lachesis muta muta and T. flavoviridis venoms was considerably inhibited at the ratio of 20 (77.2, 80.0 and 86.2% inhibition, respectively), however some of the activity still remained even at the ratio of 40 (84.2, 79.8 and 86.2% inhibition, respectively). Among the five venoms, E. c. sochureki venom is quite resistant to murinoglobulin treatment and statistically significant inhibition was only found at the ratio of 40 (64.1% inhibition). Fibrinolytic and gelatinase activities were more susceptible to murinoglobulin inhibition. The treatment at the ratios of 10 and 20 almost completely inhibited respectively the fibrinolytic and the gelatinase activities of all the venoms. Murinoglobulin treatment also significantly inhibited the edematogenic activity of L. muta muta, T. flavoviridis and Echis carinatus sochureki. The treatment of murinoglobulin at the ratio of 40 considerably suppressed the swelling up to 60 min after subcutaneous injection of L. muta muta and E. c. sochureki venoms, and up to 30 min after T. flavoviridis venom injection. Murinoglobulin is a potent inhibitor against local effects of multiple snake venoms in Viperidae family.


Activity of medullary respiratory neurons regenerating axons into peripheral nerve grafts in the adult rat.
Autologous segments of peroneal nerve were implanted into the medulla oblongata of young adult rats. To investigate activity of medullary respiratory neurons regenerating axons into these grafts, unitary recording from single fibers was performed on small strands teased from the grafts. Spontaneous activity was observed in teased fibers in 7 of 9 grafts recorded 2-5 months after graft implantation. Respiratory-related activity was found in 5 of these grafts and could in most cases be characterized as emanating from medullary respiratory neurons other than cranial motoneurons. The integrity of the input connections to the neurons that had regenerated axons was manifested by normal patterns of unitary respiratory-related activity and by the responsiveness of firing patterns of these neurons to lung hyperinflation and to the inspiratory off-switch effect induced by vagal stimulation. No spontaneous respiratory activity was found in fibers teased from any of the 10 grafts studied 9-11 months after implantation. Five of these grafts were blind-ended as were the 2-5-month grafts; the other 5 grafts formed bridges between the medulla and C4 ventral horn. No physiologic evidence of functional connections with phrenic motoneurons was found in these bridge grafts. These experiments indicate that physiologic function is maintained or regained in some respiratory neurons regenerating axons into peripheral nerve grafts but that this function is not indefinitely preserved in the absence of functional reconnection with an appropriate target.


Genetics of rheumatoid arthritis: a new boost is needed in Latin American populations.
Rheumatoid arthritis (RA) is an autoimmune inflammatory rheumatic disease which affects several organs and tissue, predominantly the synovial joints. Like many other autoimmune diseases, RA is a complex disease, where genetic variants, environmental factors and random events interact to trigger pathological pathways. Genetic implication in RA is evident, and recent advances have expanded our knowledge about the genetic factors that contribute to RA. An exponential increment in the number of genes associated with the disease has been described, mainly through gene wide screen studies (GWAS) involving international consortia with large patient cohorts. However, there are a few studies on Latin American populations. This article describes what is known about the RA genetics, the future that is emerging, and how this will develop a more personalized approach for the treatment of the disease. Latin American RA patients cannot be excluded from this final aim, and a higher collaboration with the international consortia may be needed for a better knowledge of the genetic profile of patients from this origin.


Antiplasmodial activity of bacilosarcin A isolated from the octocoral-associated bacterium Bacillus sp. collected in Panama.
This study was designed for isolating and characterizing antiplasmodial compounds from marine octocoral-associated bacteria. The organic extract of the Bacillus sp. was subjected to purification using several chromatography techniques guided by bioassays to yield three isocoumarin derivatives (1-3). Chemical structures of the compounds were elucidated on the basis of HRMS spectra and NMR spectroscopy. The antiplasmodial activity of the isolated compounds was evaluated in vitro against the chloroquine-resistant Plasmodium falciparum strain W2. Isolated compounds were identified as bacilosarcin A (1), AI77-F (2), and AI77-H (3). Bacilosarcin A (1) displayed a low micromolar activity (IC(50) = 2.2 μM) against P. falciparum while compounds 2 and 3 showed no activity. Bacilosarcin A was found to be responsible for the antiplasmodial activity observed in the crude extract obtained from the Bacillus sp.


The scaffold MyD88 acts to couple protein kinase Cepsilon to Toll-like receptors.
Mice lacking protein kinase Cepsilon (PKCepsilon) are hypersensitive to both Gram-positive and Gram-negative bacterial infections; however, the mechanism of PKCepsilon coupling to the Toll-like receptors (TLRs), responsible for pathogen detection, is poorly understood. Here we sought to investigate the mechanism of PKCepsilon involvement in TLR signaling and found that PKCepsilon is recruited to TLR4 and phosphorylated on two recently identified sites in response to lipopolysaccharide (LPS) stimulation. Phosphorylation at both of these sites (Ser-346 and Ser-368) resulted in PKCepsilon binding to 14-3-3beta. LPS-induced PKCepsilon phosphorylation, 14-3-3beta binding, and recruitment to TLR4 were all dependent on expression of the scaffold protein MyD88. In mouse embryo fibroblasts and activated macrophages from MyD88 knock-out mice, LPS-stimulated PKCepsilon phosphorylation was reduced compared with wild type cells. Acute knockdown of MyD88 in LPS-responsive 293 cells also resulted in complete loss of Ser-346 phosphorylation and TLR4/PKCepsilon association. By contrast, MyD88 overexpression in 293 cells resulted in constitutive phosphorylation of PKCepsilon. A general role for MyD88 was evidenced by the finding that phosphorylation of PKCepsilon was induced by the activation of all TLRs tested that signal through MyD88 (i.e. all except TLR3) both in RAW cells and in primary human macrophages. Functionally, it is established that phosphorylation of PKCepsilon at these two sites is required for TLR4- and TLR2-induced NFkappaB reporter activation and IkappaB degradation in reconstituted PKCepsilon(-/-) cells. This study therefore identifies the scaffold protein MyD88 as the link coupling TLRs to PKCepsilon recruitment, phosphorylation, and downstream signaling.


Central retinal vein thrombosis associated with resistance to activated protein C.
Activated protein C resistance was recently described as a major cause of venous thrombosis. We observed a central retinal vein thrombosis in a woman with activated protein C resistance. DNA analysis showed the patient to be heterozygous for the factor V gene mutation, which is related to activated protein C resistance. Patients with retinal vein thrombosis should be examined for activated protein C resistance.


A fiber optic point quadrat system for improved accuracy in vegetation sampling.
An automated, fiber optic point quadrat system for vegetation sampling is described. Because the effective point diameter of this system never exceeds 25 μm it minimizes the substantial errors which can arise with conventional point quadrats. Automatic contact detection eliminates operator subjectivity, and permits work in dense canopies. Additionally, sampling speed is increased over that of conventional systems.


Sovereign wealth fund investments and financial performance of target firms: The disciplinary role of debt in political agenda theory.
Using a sample of Indian firms acquired by the Norwegian Sovereign Wealth Funds, we investigate whether Sovereign Wealth Fund investments affect the capital structure of firms. We also examine whether leverage serves as a disciplinary mechanism in attenuating the political agenda effects of Sovereign Wealth Fund investments. Our findings reveal that Sovereign Wealth Fund ownership and size reduce leverage. We also find that sovereign wealth fund ownership of 2% and below increases financial performance, supporting the monitoring hypothesis. At above 2% sovereign wealth fund ownership stake, profitability drops significantly, supporting the political agenda hypothesis. We also find that leverage reduces the negative impacts of sovereign wealth fund ownership on the firm's financial performance when the firm's sovereign wealth fund ownership exceeds 2%, suggesting that at certain sovereign wealth fund ownership thresholds, the firm must decide on taking further debt to attenuate government opportunism behaviour and political agendas. Our findings are robust to an alternative measure of sovereign wealth funds, financial constraints, and endogeneity concerns.


An immunohistochemical method for the study of aminoglycoside ototoxicity in the guinea pig cochlea using decalcified frozen sections.
An immunohistochemical technique with decalcified frozen sections was used to study aminoglycoside ototoxicity. Decalcified guinea pig cochleas were cut with a fine blade parallel to the plane of the modiolus to facilitate the penetration of inclusion material and the manipulation of frozen sections. Light microscopy was carried out and additional frozen sections were employed for an immuno-electron microscopic study. Twenty-four hours after a single transtympanic injection of 10 mg gentamicin, there was a definite distribution of the drug in only type I hair cells of the ampullae as well as in both inner and outer hair cells along the length of the cochlea. In those animals treated intraperitoneally with 200 mg/kg amikacin for 8 days, the drug was located in the outer hair cells of the cochlea, with a tendency to decrease from base to apex and in the inner hair cells towards the apex.