Penetration of substances into tumour tissue. Model studies using saccharides, thymidine and thymidine-5'-triphosphate in cellular spheroids.
In order to achieve a better understanding of factors involved in drug penetration into poorly vascularized tumour tissue, the penetration of some model substances was studied in vitro. Multicellular human tumour spheroids were used as model system. The test substances were [3H]thymidine and [14C]glucose, both of which are capable of passing easily through cell membranes, and [3H]thymidine-5'-triphosphate, [3H]sucrose and [3H]inulin, all of which are unable to pass directly through cell membranes. The penetration of these substances was studied using a dry histological and autoradiographical method preserving the distribution of water-soluble substances. The two thymidine compounds penetrated very efficiently into the spheroids, and their penetration patterns were rather similar. The saccharides differed somewhat in their penetration properties. Glucose had the fastest penetration and inulin the slowest. After 15 min, however, inulin was also found isotropically distributed within the spheroids. Thus, extracellular penetration seemed to be a possible way for a substance to reach the central parts of a spheroid. The differences between the saccharides could be due to some extent to differences in molecular weight and solubility.


Clinical significance of NDRG3 in patients with breast cancer.
The expression level of NDRG3 gene is investigated among breast cancer (BC) patients. Real-time quantitative PCR was performed. NDRG3 was downregulated in BC patients particularly in advanced stage of the disease. HER2 status was significantly correlated with the expression of NDRG3. Also, triple-negative BC patients showed low levels of NDRG3 expression in comparison to other subtypes. Lastly, the expression of NDRG3 had significant impact on survival, with NDRG3 downregulated patients having the worst event-free survival rate among others. We have presented that NDRG3 might be a tumor suppressor candidate. NDRG3 downregulation might be involved in the tumorigenesis and development of invasive BC in an advanced phase of the disease.


Antistaphylococcal activity of Inula helenium L. root essential oil: eudesmane sesquiterpene lactones induce cell membrane damage.
The purpose of this study was to investigate the inhibitory/bactericidal activity and cell membrane effects of the hydrodistilled essential oil of Inula helenium L. roots against Staphylococcus aureus. Additionally, detailed chemical investigation was done in order to pinpoint the most active oil constituents and also the parts of these molecules responsible for their antimicrobial effect. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined using the broth microdilution method. The membrane-active nature of this oil was investigated by measuring the culture turbidity, leakage of phosphates, and 260-nm-absorbing material, together with lysis of the exposed cells. Finally, the effect of the oil on the cells was visualized using scanning electron microscopy (SEM). The chemical composition of the essential oil was analyzed using gas chromatography-mass spectrometry (GC-MS) and preparative medium-pressure liquid chromatography (MPLC). Chemical modification of the oil was performed using catalytic hydrogenation (H(2), Pd/C) and reduction with NaBH(4). The MIC and MBC values were 0.01 μl mL(-1) and 0.02 μl mL(-1), respectively. Membrane damage was demonstrated through increased permeability (phosphates and nucleic acid leakage), followed by lysis of the exposed cells, captured on SEM images. The most active constituents were alantolactone, isoalantolactone, and diplophyllin. The essential oil showed very potent antistaphylococcal activity, with obvious membrane-damaging effects. Sesquiterpene lactones were found to be the most active principles of the oil, whose eudesmane core olefinic bonds, along with the α,β-methylene-lactone ring, are essential structural parts responsible for the exhibited antimicrobial activity.


Incidence of superficial sylvian vein compromise and postoperative effects on CT imaging after surgical clipping of middle cerebral artery aneurysms.
Dissection and retraction of the sylvian fissure can cause venous insufficiency and may be an important contributor to postoperative edema or hemorrhage after clipping of a middle cerebral artery (MCA) aneurysm. The incidence of changes in the superficial middle cerebral vein (SMCV) and adjacent veins and whether such changes increase the amount of edema or hemorrhage on postoperative CT is the focus of this study. Pre- and postoperative angiograms of 100 consecutive patients with MCA aneurysms treated by craniotomy and clipping were compared to determine the postoperative incidence of changes involving the SMCV. CTs from the normal and abnormal postoperative venous groups were compared to determine the amount of edema or presence of parenchymal hemorrhage. Postoperatively, 31 (31%) SMCVs were altered, 20 to a minor or moderate degree. Eleven cases were pronounced. In 9 (9%) cases, the SMCV was completely obscured or failed to fill on postoperative angiography. More edema (observer 1, P < .0002; observer 2, P < .0006) and small brain parenchymal hemorrhages (observer 1, P < .00003; observer 2, P < .00001) were found on the postoperative CT images of the group whose SMCVs were altered than those that were unchanged. Neurosurgeons and neuroradiologists should be attentive to changes in the SMCV and adjacent venous structures to optimize outcomes of procedures involving the sylvian fissure.


Curcumin protects rat liver from streptozotocin-induced diabetic pathophysiology by counteracting reactive oxygen species and inhibiting the activation of p53 and MAPKs mediated stress response pathways.
Curcumin (CUR) is a highly pleiotropic molecule and possesses anti-inflammatory, hypoglycemic, antioxidative, wound-healing and antimicrobial activities. The present study was carried out to investigate whether CUR plays any beneficial role in streptozotocin (STZ) induced hepatic pathophysiology in diabetic rats. STZ exposure increased hepatic damage associated serum markers (ALT, ALP and LDH) as well as NO production in the liver tissue. Moreover, the same exposure enhanced ROS generation and lipid peroxidation; reduced GSH levels and antioxidant enzyme activities. Hyperglycemia induced hepatic pathophysiology also activated stress response pathways (involving phosphorylation of p38, ERK1/2 MAPKs and p53) and reduced mitochondrial membrane potential which in turn led to cellular apoptosis as evidenced from increased hepatic DNA fragmentation as well as FACS analysis. However, treatment with CUR effectively counteracts diabetes-induced, oxidative stress mediated hepatic damage and could act as a therapeutic in lessening liver dysfunction in diabetic subjects.


Proteomic analysis of brain mitochondrial proteome and mitochondrial complexes.
We describe various complementary techniques to achieve multidimensional mitochondrial proteome fractionation and analysis. Previously described methods for 2D-DIGE/mass spectrometry and 1D-SDS-PAGE/Western techniques and protein complex analysis by BN-PAGE/Western and sucrose gradient ultracentrifugation/SDS-PAGE/mass spectrometry are optimized to characterize the brain mitochondrial proteome. This approach allows for a comprehensive identification of mitochondrial proteomic differences between health and disease conditions.


Microbiological transformation of cannabinoids.
Microorganisms were screened for their ability to modify 2 synthetic cannabinoid substrates (I and II). Structure analyses revealed that microorganisms transformed the substrates by (a) primary oxidation of the side chain, beta-oxidation of the side chain, ketone formation on the side chain or cyclohexene ring, (b) secondary hydroxylation on the side chain, (c) aromatization of the cyclohexene ring, and (d) tertiary hydroxylation at the b/c ring juncture.


Effect of new biologically active polypeptides on dihexadecyl phosphate vesicles.
Effect of new biologically active polypeptides on hexadecyl phosphate vesicles. The effect that monomeric and trimeric chemotactic peptides may have on the permeation of DHP vesicles was studied by means of electronic absorption spectroscopy. The results show that attaching covalently two or more of the same chemotactic peptides to a suitable carrier molecule produced changes in the permeation of DHP vesicles that deviate from simple additivity rule. Thus a parallelism between the biological potency of the peptides and DHP permeation exists.


Structure, catalysis, chitin transport, and selective inhibition of chitin synthase.
Chitin is one of the most abundant natural biopolymers and serves as a critical structural component of extracellular matrices, including fungal cell walls and insect exoskeletons. As a linear polymer of β-(1,4)-linked N-acetylglucosamine, chitin is synthesized by chitin synthases, which are recognized as targets for antifungal and anti-insect drugs. In this study, we determine seven different cryo-electron microscopy structures of a Saccharomyces cerevisiae chitin synthase in the absence and presence of glycosyl donor, acceptor, product, or peptidyl nucleoside inhibitors. Combined with functional analyses, these structures show how the donor and acceptor substrates bind in the active site, how substrate hydrolysis drives self-priming, how a chitin-conducting transmembrane channel opens, and how peptidyl nucleoside inhibitors inhibit chitin synthase. Our work provides a structural basis for understanding the function and inhibition of chitin synthase.


Reviewing the neurobiology of electroconvulsive therapy on a micro- meso- and macro-level.
Electroconvulsive therapy (ECT) remains the one of the most effective of biological antidepressant interventions. However, the exact neurobiological mechanisms underlying the efficacy of ECT remain unclear. A gap in the literature is the lack of multimodal research that attempts to integrate findings at different biological levels of analysis METHODS: We searched the PubMed database for relevant studies. We review biological studies of ECT in depression on a micro- (molecular), meso- (structural) and macro- (network) level. ECT impacts both peripheral and central inflammatory processes, triggers neuroplastic mechanisms and modulates large scale neural network connectivity. Integrating this vast body of existing evidence, we are tempted to speculate that ECT may have neuroplastic effects resulting in the modulation of connectivity between and among specific large-scale networks that are altered in depression. These effects could be mediated by the immunomodulatory properties of the treatment. A better understanding of the complex interactions between the micro-, meso- and macro- level might further specify the mechanisms of action of ECT.


Tobramycin absorption from the peritoneal cavity.
Previous studies have found serum tobramycin concentrations greater than 8 mg/L among patients undergoing treatment for peritonitis using hourly dialysate exchanges containing 10 mg/L of tobramycin. In this study, tobramycin absorption from the peritoneal cavity was evaluated among patients treated with exchanges of 180 min. intraperitoneal duration. Six patients in each group were studied. Mass transfer coefficients [MTC (mL/min.)] were calculated for noninfected (C) and peritonitis (P) periods. The mean MTC for C was 6.5 +/- 2.1 (SD) mL/min., and for P, 18.5 +/- 8.2 (SD) mL/min. (p less than 0.01). Peritonitis increased the rate of uptake from the peritoneal cavity. In these acute studies, the total dose of tobramycin delivered was small, so that serum concentrations remained less than 0.2 mg/L among controls and 0.5 mg/L among patients with peritonitis. Prolonged exposure to tobramycin, in association with more rapid absorption from the peritoneal cavity, may lead to serum concentrations greater than 8 mg/L.


Follicular helper T cell signature of replicative exhaustion, apoptosis, and senescence in common variable immunodeficiency.
Common variable immunodeficiency (CVID) is the most frequent primary antibody deficiency whereby follicular helper T (Tfh) cells fail to establish productive responses with B cells in germinal centers. Here, we analyzed the frequency, phenotype, transcriptome, and function of circulating Tfh (cTfh) cells in CVID patients displaying autoimmunity as an additional phenotype. A group of patients showed a high frequency of cTfh1 cells and a prominent expression of PD-1 and ICOS as well as a cTfh mRNA signature consistent with highly activated, but exhausted, senescent, and apoptotic cells. Plasmatic CXCL13 levels were elevated in this group and positively correlated with cTfh1 cell frequency and PD-1 levels. Monoallelic variants in RTEL1, a telomere length- and DNA repair-related gene, were identified in four patients belonging to this group. Their blood lymphocytes showed shortened telomeres, while their cTfh were more prone to apoptosis. These data point toward a novel pathogenetic mechanism in CVID, whereby alterations in DNA repair and telomere elongation might predispose to antibody deficiency. A Th1, highly activated but exhausted and apoptotic cTfh phenotype was associated with this form of CVID.


Membrane-altering effects of velnacrine and N-methylacridinium: relevance to tacrine and Alzheimer's disease.
The interaction of pharmacological agents potentially useful in Alzheimer's disease, 9-amino-1,2,3,4-tetrahydroacridine (THA or tacrine) and its major metabolite velnacrine (or HP-029), along with related compounds with cytoskeletal proteins in human erythrocyte membrane was investigated using electron paramagnetic resonance spin labeling techniques. The results suggest that: (1) the position of the positive charge of tacrine may be important in the mechanism of its interaction with the membrane cytoskeleton; (2) like tacrine, velnacrine also strengthens cytoskeletal protein-protein interactions in erythrocyte membranes, but appears to be only about half as potent as tacrine. These results are discussed with relevance to therapeutic use of these agents in Alzheimer's disease.


The truncation-by-death problem: what to do in an experimental evaluation when the outcome is not always defined.
Although experiments are viewed as the gold standard for evaluation, some of their benefits may be lost when, as is common, outcomes are not defined for some sample members. In evaluations of marriage interventions, for example, a key outcome--relationship quality--is undefined when a couple splits up. This article shows how treatment-control differences in mean outcomes can be misleading when outcomes are not defined for everyone and discusses ways to identify the seriousness of the problem. Potential solutions to the problem are described, including approaches that rely on simple treatment-control differences-in-means as well as more complex modeling approaches.


Monitoring and descriptive analysis of radon in relation to seismic activity of Northern Pakistan.
Earthquakes are one of the major causes of natural disasters and its forecasting is challenging task. Some precursory phenomenon exists in theory in relation to earthquakes occurrence. The emission of radioactive gas named 'radon' before the earthquakes is a potential earthquake precursory candidate. The study aims to monitor and to analyze the radon in relation to seismic activity in Northern Pakistan. For this purpose RTM-2200 has been used to monitor the changes in radon concentration from August 01, 2014 to January 31, 2015 in Northern Pakistan. Significant temporal variations has been observed in radon concentration. The bivariate analysis of radon with other variables manifests its positive relationship with air pressure and relative humidity and negative relationship with temperature. 2σ upper control limit on monthly basis are computed for detection of anomalous trends in the data. Overall increasing trend is detected in radon concentration. Five earthquakes from August 01, 2014 to January 31, 2015 have been selected from earthquake catalogue, depending upon their magnitude and distance from monitoring station and out of which radon concentration can be associated with only two earthquakes correlated with tectonic effect of radon concentration. Both of events have same magnitude 5.5 and occurred on September 13 and October 14, 2014 respectively. Very large variations have been observed in radon for the last two months of the study period, which may be occurred due to some other geological and environmental changes, but are not related to the earthquake activity.


Serotonin transporter occupancy with TCAs and SSRIs: a PET study in patients with major depressive disorder.
The aim of the present clinical positron emission tomography study was to examine if the 5-HTT is a common target, both for tricyclic antidepressants (TCAs) and selective serotonin reuptake inhibitors (SSRIs). Serotonin transporter (5-HTT) occupancy was estimated during treatment with TCA, SSRI and mirtazapine in 20 patients in remission from depression. The patients were recruited from out-patient units and deemed as responders to antidepressive treatment. The radioligand [¹¹C]MADAM was used to determine the 5-HTT binding potential. The mean 5-HTT occupancy was 67% (range 28-86%). There was no significant difference in 5-HTT occupancy between TCA (n=5) and SSRI (n=14). 5-HTT affinity correlated with the recommended clinical dose. Mirtazapine did not occupy the serotonin transporter. The results support that TCAs and SSRIs have a shared mechanism of action by inhibition of 5-HTT.


A clinical-genetic approach to assessing cardiovascular risk in patients with CKD.
Coronary heart disease (CHD) is the primary cause of death in individuals with chronic kidney disease (CKD), but current equations for assessing coronary risk have low accuracy in this group. We have reported that the addition of a genetic risk score (GRS) to the Framingham risk function improved its predictive capacity in the general population. The aims of this study were to evaluate the association between this GRS and coronary events in the CKD population and to determine whether the addition of the GRS to coronary risk prediction functions improves the estimation of coronary risk at the earliest possible stages of kidney disease. A total of 632 CKD patients, aged 35-74 years, who had Stage 4-5 CKD, were on dialysis, had a functioning renal transplant or had returned to dialysis after transplant failure were included and followed up for a mean of 9.3 years. The transitions between disease states and the development of coronary events were registered. The increase in predictive ability that was obtained by including the GRS was measured as the improvement in the C-statistic and as the net reclassification index. The GRS was independently associated with the risk of CHD (hazards ratio 1.34; 95% confidence interval 1.04-1.71; P = 0.022), especially in Stages 4 and 5 CKD, and kidney transplant patients. A coronary risk prediction function that incorporated chronic kidney disease (CKD) disease state, age, sex and the GRS had significantly greater predictive capacity (AUC 70.1, P = 0.01) and showed good reclassification (net reclassification improvement 28.6). This new function, combining genetic and clinical data, identifies CKD patients with a high risk of coronary events more accurately, allowing us to prevent such events more effectively.


A comparison of concentric needle electromyography, quantitative EMG and single fibre EMG in the diagnosis of neuromuscular diseases.
Conventional concentric needle electromyography, quantitative electromyography using automatic analysis (Quant-EMG) and single fibre electromyography (SFEMG) were compared in the right biceps muscle of 10 patients with anterior horn cell disease and 20 patients with primary muscle disease. Abnormalities were demonstrable by all 3 techniques in the majority of cases irrespective of whether the biceps was weak. SFEMG was found to be more sensitive than Quant-EMG in neurogenic cases and it also provided an indication of disease activity. In cases of myopathy Quant-EMG was of more diagnostic help and was easier to perform than SFEMG. Large amplitude potentials found in some cases of myopathy were associated with increase in the mean amplitude on Quant-EMG and these probably emanate from hypertrophied muscle fibres.


Optimization of transmission near infrared spectrometry procedures for quality control of pesticide formulations.
The use of different response functions to be optimized in the frame of the use of near infrared spectrometry for quality control of active principles in agrochemical formulations has been evaluated. Both, simple functions, based on parameters like sensitivity, repeatability, accuracy, signal to noise ratio, limit of detection or sample throughput, and a complex function, considering all the aforementioned aspects, were employed in the development of a new method for Iprodione determination in agrochemicals. Optimization strategies were based on the previous screening of the most important instrumental factors like number of cumulated scans, nominal resolution, mirror velocity and zero filling factor, based on a two-level full factorial design and on the search for the optimum conditions using central composite designs. Data found evidenced the influence of the response function on the optimum values of experimental conditions and could be employed as a general guide to evaluate the experimental factors in routine use of near infrared spectrometry. Finally the optimized method for Iprodione has been applied to the determination of Diuron and results found compared with those obtained by a conventional approach.


Oxalosis: cause of degenerative spinal stenosis. A case report and review of the literature.
The case presented illustrates an unusual entity. The differential diagnosis of degenerative spinal stenosis is voluminous, although incomplete without including oxalosis. Oxalosis as a cause of spinal stenosis has not yet been reported in the orthopedic literature.


Apoptosis-associated protein expression in human salivary gland morphogenesis.
Salivary gland (SG) development is based on branching morphogenesis, in which programmed cell death has been proposed to play a role in cell signalling and organ shaping. In the mouse salivary gland apoptosis has been suggested to play a key role in lumen formation, removing the central cells of the epithelial stalks. Here we analyse the expression of several anti- and pro-regulators of apoptosis during human SG development in a range of developmental stages. Foetal SGs obtained from the University of São Paulo were analysed by immunohistochemistry to assess the expression of apoptosis-associated proteins: caspases (caspase-6, -7, -9 and cleaved caspase-3), Bcl-2 family members (Bax, Bak, Bad, Bid, Bcl-2, Bcl-x and Bcl-xL), Survivin (BIRC5), Cytochrome C and Apaf-1. Nuclear expression of Bax and Bak was identified in presumptive luminal areas at initial stages, while Bcl-xL showed the most relevant anti-apoptotic activity. Caspase-6, -7 and -9 were expressed during all stages, while interestingly cleaved caspase-3 showed no prominent expression, indicating that caspase-7 is the main effector. Apoptosome complex components Apaf-1 and Cytochrome C, as well as survivin were all positive in developing glands. The particular expression pattern of several apoptotic regulators in human SG development suggests the existence of a fundamental role for apoptosis during duct formation. The absence of Bad and Bid expressions indicates that the instrinsic pathway is more active then the extrinsic during human gland formation. The subcellular localisation of intrinsic-apoptosis proteins correlated with apoptotic activity, but also suggested additional non-apoptotic functions.


The delivery of thrombi-specific nanoparticles incorporating oligonucleotides into injured cerebrovascular endothelium.
In acute vascular events, the endothelium derived tissue factor (TF) is the trigger of the coagulation cascade. In this study, EGFP-EGF1 protein-conjugated PEG-PLGA nanoparticle was employed as a TF targeting vehicle, the NF-κB decoy oligonucleotides (ODNs) was incorporated into it and the resulting EGF1-EGFP-NP-ODNs were evaluated as a vector for therapy of cortex infarction. At 2 h after transfection of TF expressed rat brain capillary endothelial cell, EGF1-EGFP-NP-ODNs was more efficiently internalized and located in the cytoplasm than NP-ODNs. At 4 h and 6 h after administration, ODNs were present in the nuclei and obviously inhibited the TF expression. At 6 h after i.v. administration in vivo, most EGF1-EGFP-NP were accumulated in the embolism vessels, distributed in the damaged endothelial cells and lowered the TF expression. At 24 h after i.v. administration, MR imaging of cortex infarcts were predominantly dwindled.


Assessment of temperature, proximity, and course of the esophagus during radiofrequency ablation within the left atrium.
Left atrioesophageal fistula is a devastating complication of atrial fibrillation ablation. There is no standard approach for avoiding this complication, which is caused by thermal injury during ablation. The objectives of this study were to evaluate the course of the esophagus and the temperature within the esophagus during pulmonary vein antrum isolation (PVAI) and correlate these data with esophagus tissue damage. Eight-one patients presenting for PVAI underwent esophagus evaluation that included temperature probe placement. Esophagus course was obtained with computed tomography, 3D imaging (NAVX), or intracardiac echocardiography. For each lesion, the power, catheter and esophagus temperature, location, and presence of microbubbles were recorded. Lesion location and esophagus course were defined with 6 predetermined left atrial anatomic segments. Endoscopy evaluated tissue changes during and after PVAI. Of 81 patients, the esophagus coursed near the right pulmonary veins in 23 (28.4%), left pulmonary veins in 31 (38.3%), and mid-posterior wall in 27 (33%). Esophagus temperature was significantly higher during left atrial lesions along its course than with lesions elsewhere (38.9+/-1.4 degrees C, 36.8+/-0.5 degrees C, P<0.01). Lesions that generated microbubbles had higher esophagus temperatures than those without (39.3+/-1.5 degrees C, 38.5+/-0.9 degrees C, P<0.01). Power was not predictive of esophagus temperatures. Distance between the esophagus and left atrium was 4.4+/-1.2 mm. Lesions near the course of the esophagus that generated microbubbles significantly increased esophagus temperature compared with lesions that did not. Power did not correlate with esophagus temperatures. Esophagus variability makes the avoidance of lesions along its course difficult. Rather than avoiding posterior lesions, emphasis could be placed on better esophagus monitoring for creation of safer lesions.


Does diabetes mellitus have an impact on the prognosis for patients with cervical cancer?
To evaluate the impact of comorbid diabetes mellitus (DM) on prognoses among patients with cervical cancer. We analyzed cervical cancer outcomes in patients who treated in two hospitals retrospectively. Patients were divided into those with DM and those without. Clinicopathologic parameters, disease-free survival (DFS), and overall survival (OS) rates were evaluated. Of the 494 patients, 50 had DM. These were more likely to be older than those in the non-DM group and their body mass index (BMI) was higher. They showed higher levels of tumor markers and had more combined diseases. They were less likely to have had surgical treatment. Among these patients, 12 (24%) experienced a recurrence (hazard ratio, HR, 1.484; 95% confidence interval, CI, 0.746-2.951). Differences in DFS did not show statistical significance. In the OS analysis, 11 (22%) in the DM group and 62 (14%) in the non-DM group died (HR, 1.239; 95% CI, 0.606-2.533). No statistically significant differences were also observed for cancer-specific death (HR, 1.246; 95% CI, 0.567-2.737). Those with DM and an adenocarcinoma tended to have an increased risk of dying compared with the non-DM patients with an adenocarcinoma (HR, 3.673; 95% CI, 0.990-13.625), but this difference was not statistically significant (p=0.0518). Diabetes mellitus did not have an impact on the prognosis for patients with cervical cancers. In those with an adenocarcinoma, patients with diabetes tended to have an increased risk of dying compared with the non-DM group, but this difference was not statistically significant.


Symmetry-breaking of animal cytokinesis.
Cytokinesis is a mechanism that separates dividing cells via constriction of a supramolecular structure, the contractile ring. In animal cells, three modes of symmetry-breaking of cytokinesis result in unilateral cytokinesis, asymmetric cell division, and oriented cell division. Each mode of cytokinesis plays a significant role in tissue patterning and morphogenesis by the mechanisms that control the orientation and position of the contractile ring relative to the body axis. Despite its significance, the mechanisms involved in the symmetry-breaking of cytokinesis remain unclear in many cell types. Classical embryologists have identified that the geometric relationship between the mitotic spindle and cell cortex induces cytokinesis asymmetry; however, emerging evidence suggests that a concerted flow of compressional cell-cortex materials (cortical flow) is a spindle-independent driving force in spatial cytokinesis control. This review provides an overview of both classical and emerging mechanisms of cytokinesis asymmetry and their roles in animal development.


Fractures of the distal radius treated with a nonbridging external fixation technique using multiplanar k-wires.
Joint-bridging external fixation is a minimally invasive treatment option for distal radius fractures. Although radial length can be restored easily the anatomic reduction of articular fragments and restoration of the normal volar tilt proves to be more difficult. A method of nonbridging hybrid fixation of distal radius fractures facilitates fracture reduction and allows for free wrist movement. Twenty-five consecutive patients with fractures of the distal radius were treated with nonbridging external fixation for 6 weeks. The stepwise surgical technique comprised a preliminary joint-bridging construction for reduction purposes, the subsequent insertion of 3 to 4 K-wires in the distal fragment, the assembling of wires to a bar nearly parallel to the fracture line, and lastly the removal of the joint-bridging part. Clinical and radiologic evaluation was performed on the first and seventh days and at 6 weeks and 2 years after surgery. All fractures united. Palmar tilt (> or =0 degrees ) and articular surface (articular step-off < 2 mm) were restored in all patients whereas loss of radial length occurred in 4 patients having the distal fracture fragment secured with 3 K-wires. No radial shortening was seen in fractures with 4 K-wires inserted in the distal fragment. Functional results at 2 years after surgery showed an average extension of 55 degrees and flexion of 64 degrees without significant differences between extra-articular and intra-articular fractures. There was no extensor tendinitis or pin loosening in the distal fragment; however, 3 pin track infections of proximal pins occurred. This surgical technique of nonbridging external fixation is a good treatment option for distal radius fractures: it permits wrist movement. We recommend the insertion of 4 K-wires in the distal fracture fragment.


Incorporation of dipalmitoyl phosphatidycholine into human plasma low density lipoproteins: effects on composition and structure.
Bovine liver phosphatidylcholine exchange protein was used to transfer di[14C]palmitoyl phosphatidylcholine from sonicated vesicles to human plasma low density lipoproteins (LDL). The incorporation of di[14C]-palmitoyl phosphatidylcholine into LDL was associated with an increased percentage of total phospholipid and increased lipid rigidity as shown by an increase in the fluorescence polarization of 12-(9-anthroyloxy)-stearic acid and 1,6-diphenyl-1,3,5-hexatriene.


Regarding time-sharing with convergent operations.
This study contrasts the structural bottleneck and the resource view of attentional limits in time-sharing performance. The research incorporated features of the psychological refractory period (PRP) and the relative priority paradigm designed to maximize joint performance. A main distinction between the two attention views was their prediction on the extent that graded performance tradeoff was possible with graded priority changes. Detailed analysis of the performance and time-sharing strategies called into question the conclusions based exclusively on the PRP paradigm.


Recombinant chimeric enzymes for lignocellulosic biomass hydrolysis.
Chimeras with improved properties can be designed by synergistically incorporating multiple proteins into one peptide chain. Such design enhances hydrolysis efficiency and lowers enzyme production costs for numerous applications, particularly in the fields of lignocellulosic biomass hydrolysis, plant-based industries, food additives, waste treatment, and biofuels. In this minireview, we summarize a brief, common approach to construct recombinant enzymes, especially in the Pichia pastoris host. We present the design strategies and the desired properties of chimeras and provide examples of recent studies in the area of lignocellulosic biomass hydrolysis. Subsequently, we describe the progress in structure prediction tools that facilitate the design of chimeric proteins toward identifying the best combinations of fusion partners and linkers.


Factors associated with self-perceived burden to the primary caregiver in older patients with hematologic malignancies: an exploratory study.
Although cancer patients frequently experience self-perceived burden to others, this perception has not been enough studied. The aim of this study was to investigate the prevalence of self-perceived burden to the primary caregiver (SPB-PC) and associated factors in an older patient population with hematologic malignancies at the time of chemotherapy initiation. In total, 166 consecutive patients with hematologic malignancies aged ≥65 years were recruited at the time of chemotherapy initiation. Patients' SPB-PC was assessed using a 100-mm visual analogue scale (VAS). Characteristics potentially associated with SPB-PC, including sociodemographic and medical characteristics, physical functioning status (Karnofsky performance score, activities of daily living (ADL)/instrumental ADL), symptoms (fatigue, pain, nausea, quality of life), psychological distress (Hospital Anxiety and Depression Scale (HADS)), perceived cognitive function (Functional Assessment of Cancer Therapy Cognitive (FACT-Cog) Scale), and patients'/primary caregivers' personal relationship characteristics (family tie, support), were assessed. Thirty-five percent of patients reported moderate to severe SPB-PC (VAS ≥ 50 mm). Patients' SPB-PC was associated with lower Karnofsky performance (β = -0.135, p = 0.058) and ADL (β = -0.148, p = 0.037) scores, and higher HADS (β = 0.283, p < 0.001) and FACT-Cog perceived cognitive impairments subscale (β = 0.211, p = 0.004) scores. The proportion of explained variance was 23.5%. Health care professionals should be aware that about one third of older cancer patients experience moderate to severe SPB-PC at the time of chemotherapy initiation. They should adapt their support of patients who report such a feeling. Copyright © 2016 John Wiley & Sons, Ltd.


Mitotic kinase cascades orchestrating timely disjunction and movement of centrosomes maintain chromosomal stability and prevent cancer.
Centrosomes are microtubule-organizing centers that duplicate in S phase to form bipolar spindles that separate duplicated chromosomes faithfully into two daughter cells during cell division. Recent studies show that proper timing of centrosome dynamics, the disjunction and movement of centrosomes, is tightly linked to spindle symmetry, correct microtubule-kinetochore attachment, and chromosome segregation. Here, we review mechanisms that regulate centrosome dynamics, with emphasis on the roles of key mitotic kinases in the proper timing of centrosome dynamics and how aberrancies in these processes may cause chromosomal instability and cancer.


Vitrectomy with internal limiting membrane removal for clinically significant macular oedema.
Little has been published on internal limiting membrane (ILM) removal for clinically significant macular oedema (CSME) in eyes not improving following maximal laser treatment. Retrospective review of the charts and intraoperative video films of 30 consecutive eyes of 27 patients undergoing vitrectomy, ILM peeling, and gas tamponade by a single surgeon. The average patient was 52 years old; 56% were males. The oedema was caused by diabetes in 87% of the eyes, 42% of which had proliferative disease. The oedema was diffuse in 16 eyes (53%) and cystoid (CME) in 14 (47%). In 21% of eyes, spontaneous vitreous detachment was present or the eye had already undergone vitrectomy. All but one eye had at least 6 months of follow-up (mean 12 months). The oedema resolved completely in 26 eyes (90%) and partially in 3 eyes (10%). The visual acuity improved at least two Snellen lines in 19 eyes (66%) and one line in 4 eyes (14%); the average improvement was 4 lines. The acuity was unchanged in three eyes (10%) and worsened in three eyes (10%), all due to cataract. Among the 14 eyes with CME, 11 (79%) became completely dry and the visual acuity improved in 11 eyes (79%). No ILM-related complication was encountered in any eye. Vitrectomy with ILM peeling is a promising approach to eyes with CSME. It appears to offer visual improvement at a much higher rate than laser therapy and with a longer-lasting effect than that of intravitreal triamcinolone. A larger study is necessary to confirm these preliminary findings.


BalI and MspI polymorphisms of the dopamine D3 receptor gene in African Blacks and Caucasians.
An exonic BalI polymorphism and an intronic MspI polymorphism of the dopamine D3 gene were genotyped in 101 Caucasians from the Alsace and in 56 people from the Congo. This is the first study of the BalI polymorphism in sub-Saharan Africa and the first population study of the MspI site. BalI allele 1 was rare in the Congo (0.12) whereas it is the most frequent allele in all studies in Europe and Asia. MspI allele 1 was also significantly less frequent in the Congolese (0.24) than in Caucasians (0.52). D3 gene alleles show different frequencies in sub-Saharan Africa and may be useful for population studies.


Oxidative stress effect on progesterone-induced blocking factor (PIBF) binding to PIBF-receptor in lymphocytes.
Receptor-ligand binding is an essential interaction for biological function. Oxidative stress can modify receptors and/or membrane lipid dynamics, thus altering cell physiological functions. The aim of this study is to analyze how oxidative stress may alter receptor-ligand binding and lipid domain distribution in the case of progesterone-induced blocking factor/progesterone-induced blocking factor-receptor. For membrane fluidity regionalization analysis of MEC-1 lymphocytes, two-photon microscopy was used in individual living cells. Lymphocytes were also double stained with AlexaFluor647/progesterone-induced blocking factor and Laurdan to evaluate -induced blocking factor/progesterone-induced blocking factor-receptor distribution in the different membrane domains, under oxidative stress. A new procedure has been developed which quantitatively analyzes the regionalization of a membrane receptor among the lipid domains of different fluidity in the plasma membrane. We have been able to establish a new tool which detects and evaluates lipid raft clustering from two-photon microscopy images of individual living cells. We show that binding of progesterone-induced blocking factor to progesterone-induced blocking factor-receptor causes a rigidification of plasma membrane which is related to an increase of lipid raft clustering. However, this clustering is inhibited under oxidative stress conditions. In conclusion, oxidative stress decreases membrane fluidity, impairs receptor-ligand binding and reduces lipid raft clustering.


A novel splice site mutation in the SDCCAG8 gene in an Iranian family with Bardet-Biedl syndrome.
Bardet-Biedl syndrome (BBS: OMIM 209,900) is a rare ciliopathic human genetic disorder that affects many parts of the body systems. BBS is a genetically heterogeneous disorder with a wide spectrum of clinical manifestations which makes its diagnosis and management more challenging. RetNet reports 18 genes that cause BBS and each of genes has had several known mutations. Genetic studies suggesting that serologically defined colon cancer antigen 8 (SDCCAG8) gene mutations are a major cause of BBS. In this section, we investigated the consanguineous Iranian family members with BBS. Whole-exome sequencing and Sanger sequencing, were performed to screen and confirm the suspicious pathogenic mutations. The identified mutation was investigated using bioinformatics tools to predict the effect of the mutation on protein structure. Sequential analysis identified a novel splice site mutation c.1221 + 2 T > A in the SDCCAG8 gene in BBS patients. Structure-based approaches have predicted significant structural alterations in SDCCAG8 protein. This study was conducted to show the aberrant alternative splicing as one of the single splicing mutations identified can cause BBS by affecting the function of SDCCAG8 protein.


Parental genome separation and asynchronous centromere division in interspecific F1 hybrids in Lathyrus.
Chromosomes were studied in root-tip metaphase cells of several F1 interspecific Lathyrus hybrids including: L. hirsutus x L. cassius (H x C), L. cassius x L. hirsutus (C x H), L. cassius x L. odoratus (C x O), and their parents, all with 2n = 2x = 14. Two types of morphologically distinct centromeres were identified in the hybrids on the basis of the degree of contraction of the primary constriction. At least 12 well-defined centromeres were seen in all cells of L. hirsutus, L. cassius and L. odoratus, and about 80% of cells had 14. The hybrids were more variable than the species. H x O contained between six and 14 well-defined centromeres, while cells of H x C, C x H and C x O all had seven well-defined and seven weakly defined centromeres. These were used as markers to plot their spatial disposition in two dimensions on metaphase spreads. In H x C, C x H and C x O the two types of centromeres showed a significant tendency to occupy two spatially distinct and concentrically arranged domains on the metaphase plate (P < 0.005). Owing to shortage of material subsequent work was restricted to H x C and C x H. Six or seven chromosomes of one parental genome were selectively labelled by in situ hybridization using biotinylated total genomic DNA from either parent as a probe. Moreover, there was a very strong correlation between centromere type and genomic origin (P < 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)


On the development of midpiece mitochondria of mouse spermatozoa.
As seen by scanning electron microscopy, the mitochondrial helix in the developing midpiece of mouse testicular spermatozoa is dextral in direction and consists of spherical mitochondrial units arranged in an orderly array of four units per gyre: three appearing in face view and a fourth hidden from view at the back of the gyre. As the spermatozoa mature, the dextral helix is transformed into a sinistral helix. Its constituent spherical mitochondria either change direction abruptly without changing shape; or having first become semilunar or diamond-shaped, they change direction gradually. Mitochondrial division follows the change in helical pitch producing a double sinistral helix. The spherical (or semilunar/diamond-shaped) mitochondria presumably elongate to form the units of the mature midpiece.


Autonomic control of ventricular function in health and disease: current state of the art.
Cardiac autonomic dysfunction is one of the main pillars of cardiovascular pathophysiology. The purpose of this review is to provide an overview of the current state of the art on the pathological remodeling that occurs within the autonomic nervous system with cardiac injury and available neuromodulatory therapies for autonomic dysfunction in heart failure. Data from peer-reviewed publications on autonomic function in health and after cardiac injury are reviewed. The role of and evidence behind various neuromodulatory therapies both in preclinical investigation and in-use in clinical practice are summarized. A harmonic interplay between the heart and the autonomic nervous system exists at multiple levels of the neuraxis. This interplay becomes disrupted in the setting of cardiovascular disease, resulting in pathological changes at multiple levels, from subcellular cardiac signaling of neurotransmitters to extra-cardiac, extra-thoracic remodeling. The subsequent detrimental cycle of sympathovagal imbalance, characterized by sympathoexcitation and parasympathetic withdrawal, predisposes to ventricular arrhythmias, progression of heart failure, and cardiac mortality. Knowledge on the etiology and pathophysiology of this condition has increased exponentially over the past few decades, resulting in a number of different neuromodulatory approaches. However, significant knowledge gaps in both sympathetic and parasympathetic interactions and causal factors that mediate progressive sympathoexcitation and parasympathetic dysfunction remain. Although our understanding of autonomic imbalance in cardiovascular diseases has significantly increased, specific, pivotal mediators of this imbalance and the recognition and implementation of available autonomic parameters and neuromodulatory therapies are still lagging.


Circadian rhythms in blood lactate concentration during incremental ergometer rowing.
This study examined whether circadian rhythms affect lactate threshold (Th(lac)) during rowing exercise. Eleven male, endurance-trained athletes [mean (SD) age 29.5 (6.1) years] rowed at 0200, 0600, 1000, 1400, 1800 and 2200 hours under the same experimental conditions. Capillary blood (25 microl) was obtained from the tip of the toe during the last 30 s of a continuous, multi-stage, 3-min, incremental protocol on the Concept II ergometer. To determine Th(lac), a curve-fitting procedure (the D(max) method), a visual method (Th(lac-vis)) and the fixed blood lactate concentration of 4.0 mmol l(-1) (Th(lac-4 mM)) were used. Circadian rhythms were apparent for oxygen consumption and heart rate at Th(lac) using the D(max) method (P=0.02 and P=0.04 respectively), with the acrophases at 2139 hours and 2032 hours respectively coinciding in phase with that of core body temperature. The conclusion is that tests should be completed at the same time of day at which the athlete usually trains, to ensure precision of Th(lac) determination, especially when the D(max )method is used to determine Th(lac).


Cancer in xeroderma pigmentosum and related disorders of DNA repair.
Nucleotide-excision repair diseases exhibit cancer, complex developmental disorders and neurodegeneration. Cancer is the hallmark of xeroderma pigmentosum (XP), and neurodegeneration and developmental disorders are the hallmarks of Cockayne syndrome and trichothiodystrophy. A distinguishing feature is that the DNA-repair or DNA-replication deficiencies of XP involve most of the genome, whereas the defects in CS are confined to actively transcribed genes. Many of the proteins involved in repair are also components of dynamic multiprotein complexes, transcription factors, ubiquitylation cofactors and signal-transduction networks. Complex clinical phenotypes might therefore result from unanticipated effects on other genes and proteins.


An aptamer based competition assay for protein detection using CNT activated gold-interdigitated capacitor arrays.
An aptamer can specifically bind to its target molecule, or hybridize with its complementary strand. A target bound aptamer complex has difficulty to hybridize with its complementary strand. It is possible to determine the concentration of target based on affinity separation system for the protein detection. Here, we exploited this property using C-reactive protein (CRP) specific RNA aptamers as probes that were immobilized by physical adsorption on carbon nanotubes (CNTs) activated gold interdigitated electrodes of capacitors. The selective binding ability of RNA aptamer with its target molecule was determined by change in capacitance after allowing competitive binding with CRP and complementary RNA (cRNA) strands in pure form and co-mixtures (CRP:cRNA=0:1, 1:0, 1:1, 1:2 and 2:1). The sensor showed significant capacitance change with pure forms of CRP/cRNA while responses reduced considerably in presence of CRP:cRNA in co-mixtures (1:1 and 1:2) because of the binding competition. At a critical CRP:cRNA ratio of 2:1, the capacitance response was dramatically lost because of the dissociation of adsorbed aptamers from the sensor surface to bind when excess CRP. Binding assays showed that the immobilized aptamers had strong affinity for cRNA (K(d)=1.98 μM) and CRP molecules (K(d)=2.4 μM) in pure forms, but low affinity for CRP:cRNA ratio of 2:1 (K(d)=8.58 μM). The dynamic detection range for CRP was determined to be 1-8 μM (0.58-4.6 μg/capacitor). The approach described in this study is a sensitive label-free method to detect proteins based on affinity separation of target molecules that can potentially be used for probing molecular interactions.


Effect of immunoglobulin G on membrane-bound enzyme activity of sarcoma 180 cells.
Changes in the activity of membrane bound ATPase of Sarcoma 180 cells caused by immunoglobulin G (IgG) of anti-Sarcoma 180 was investigated in relation to the incorporation of amino acid by the cells. Enzymatic activity of ATPase was increased up to 160% of the original activity upon incubation of the cell with IgG. Kinetic studies showed that IgG did not change the affinity of this enzyme for the substrate, but exerted influence upon catalytic efficiency of the enzyme. The rate of incorporation of leucine into Sarcoma 180 cells was also affected by IgG, as observed in the effect of IgG on the enzymatic reaction of the cells.


Binding of eucaryotic elongation factor Tu to nucleic acids.
The binding of eucaryotic elongation factor Tu (eEF-Tu) to nucleic acids was investigated. eEF-Tu binds to a variety of different nucleic acids with high affinity, showing a strong preference for 18 S and 28 S rRNA over transfer RNA and for ribose-containing polymers over polydeoxyribonucleotides. The factor binds at multiple sites on 28 S rRNA without strong cooperativity. eEF-Tu binds strongly to poly(G) and poly(U) but weakly, if at all, to poly(A) and poly(C). Experiments employing an airfuge demonstrate that eEF-Tu can form a quaternary complex containing the factor, 28 S rRNA, aminoacyl-tRNA, and GTP. The existence of two distinct RNA binding sites on eEF-Tu suggests that rRNA may play a role in the recognition of eEF-Tu.aminoacyl-tRNA.GTP complexes by polysomes. Support for this suggestion comes from experiments which show that poly(G) inhibits the factor-dependent binding of aminoacyl-tRNA to mRNA-programmed 80 S ribosomes. In addition, it is shown that eEF-Tu possesses an intrinsic GTPase activity which is stimulated significantly by 28 S rRNA, poly(G), and poly(U). The binding of eEF-Tu to poly(G) lowers the activation energy for eEF-Tu GTPase from 74.3 to 65.9 kJ . mol-1 and approximately doubles the Vmax of the enzymatic reaction. The results are discussed in relation to the binding of eEF-Tu to ribosomes during protein synthesis.


Complexes of photosynthetic redox proteins studied by NMR.
In the photosynthetic redox chain, small electron transfer proteins shuttle electrons between the large membrane-associated redox complexes. Short-lived but specific protein:protein complexes are formed to enable fast electron transfer. Recent nuclear magnetic resonance (NMR) studies have elucidated the binding sites on plastocyanin, cytochrome c (6) and ferredoxin. Also the orientation of plastocyanin in complex with cytochrome f has been determined. Based on these results, general features that enable the formation of such transient complexes are discussed.


Implementation of a Statewide Policy Mandating School-Based Fitness Assessment Screening, Georgia: 2018.
Objectives. To evaluate the statewide implementation of childhood fitness assessment and reporting in Georgia.Methods. We collected survey data from 1683 (919 valid responses from a random-digit-dialed survey and 764 valid responses from a Qualtrics panel) parents of public school students in Georgia in 2018.Results. Most parents reported that their child participated in fitness assessments at school, yet only 31% reported receiving results. If a child was identified as needing improvement, parents were significantly more likely to change the diet and exercise of both the child and the family.Conclusions. A state-level mandatory fitness assessment for children may be successful in state-level surveillance of fitness levels; parental awareness of the policy, receipt of the fitness assessment information, and action on receiving the screening information require more efforts in implementation.


The risk of getting worse: surgically acquired deficits, perioperative complications, and functional outcomes after primary resection of glioblastoma.
Gross total resection (GTR) prolongs survival but is unfortunately not achievable in the majority of patients with glioblastoma multiforme (GBM). Cytoreductive debulkings may relieve symptoms of mass effect, but it is unknown how long such effects sustain and to what degree the potential benefits exceed risks. We explore the impact of surgical morbidity on functional outcome and survival in unselected GBM patients. We retrospectively included 144 consecutive adult patients operated on for primary GBM at a single institution between 2004 and 2009. Reporting of adverse events was done in compliance with Good Clinical Practice Guidelines. A total of 141 (98%) operations were resections and 3 (2%) were biopsies. A decrease in Karnofsky performance status (KPS) scores was observed in 39% of patients after 6 weeks. There was a significant decrease between pre- and postoperative KPS scores (P < 0.001). Twenty-two (15.3%) patients had surgically acquired neurological deficits. Among patients who underwent surgical resection, those with surgically acquired neurological deficits were less likely to receive radiotherapy (P < 0.001), normofractioned radiotherapy (P = 0.010), and chemotherapy (P = 0.003). Twenty-eight (19.4%) patients had perioperative complications. Among patients who underwent surgical resection, those with perioperative complications were less likely to receive normofractioned radiotherapy (P = 0.010) and chemotherapy (P = 0.009). Age (P = 0.019), surgically acquired neurological deficits (P < 0.001), and surgical complications (P = 0.006) were significant predictors for worsened functional outcome after 6 weeks. GTR (P = 0.035), perioperative complications (P = 0.008), radiotherapy (P < 0.001), and chemotherapy (P = 0.045) were independent factors associated with 12-month postoperative survival. Patients with perioperative complications and surgically acquired deficits were less likely to receive adjuvant therapy. While cytoreductive debulking may not improve survival in GBM, it may decrease the likelihood of patients receiving adjuvant therapy that does.


[Multiple sclerosis disclosed by spinal anesthesia].
A case is reported of an acute onset of previously undiagnosed multiple sclerosis, revealed by an oculomotor paralysis following spinal anaesthesia performed for minor orthopaedic surgery. The need for a complete preoperative physical examination is underlined by this case, looking for latent neuromuscular disorders before undertaking such techniques, and for a thorough neurological work-up should such a complication arise. The harmlessness of regional anaesthesia in multiple sclerosis patients is controversial; without entering into such a debate, the direct relationship between spinal anaesthesia and acute exacerbation of the disease in our patient seemed more than likely.


Theoretical investigation on the diatomic ligand migration process and ligand binding properties in non-O2-binding H-NOX domain.
The Nostoc sp (Ns) H-NOX (heme-nitric oxide or OXygen-binding) domain shares 35% sequence identity with soluble guanylate cyclase (sGC) and exhibits similar ligand binding property with the sGC. Previously, our molecular dynamic (MD) simulation work identified that there exists a Y-shaped tunnel system hosted in the Ns H-NOX interior, which servers for ligand migration. The tunnels were then confirmed by Winter et al. [PNAS 2011;108(43):E 881-889] recently using x-ray crystallography with xenon pressured conditions. In this work, to further investigate how the protein matrix of Ns H-NOX modulates the ligand migration process and how the distal residue composition affects the ligand binding prosperities, the free energy profiles for nitric oxide (NO), carbon monooxide (CO), and O2 migration are explored using the steered MDs simulation and the ligand binding energies are calculated using QM/MM schemes. The potential of mean force profiles suggest that the longer branch of the tunnel would be the most favorable route for NO migration and a second NO trapping site other than the distal heme pocket along this route in the Ns H-NOX was identified. On the contrary, CO and O2 would prefer to diffuse via the shorter branch of the tunnel. The QM/MM (quantum mechanics/molecular mechanics) calculations suggest that the hydrophobic distal pocket of Ns H-NOX would provide an approximately vacuum environment and the ligand discrimination would be determined by the intrinsic binding properties of the diatomic gas ligand to the heme group.


Overexpression of miR-32 in Chinese hamster ovary cells increases production of Fc-fusion protein.
The demand for industrial genetically modified host cells were increased with the growth of the biopharmaceutical market. Numerous studies on improving host cell productivity have shown that altering host cell growth and viability through genetic engineering can increase recombinant protein production. During the last decades, it was demonstrated that overexpression or downregulation of some microRNAs in Chinese Hamster Ovary (CHO) cells as the host cell in biopharmaceutical manufacturing, can improve their productivity. The selection of microRNA targets has been based on their previously identified role in human cancers. MicroRNA-32 (miR-32), which is conserved between humans and hamsters (Crisetulus griseus), was shown to play a role in the regulation of cell proliferation and apoptosis in some human cancers. In this study, we investigated the effect of miR-32 overexpression on the productivity of CHO-VEGF-trap cells. Our results indicated that stable overexpression of miR-32 could dramatically increase the productivity of CHO cells by 1.8-fold. It also significantly increases cell viability, batch culture longevity, and cell growth. To achieve these results, following the construction of a single clone producing an Fc-fusion protein, we transfected cells with a pLexJRed-miR-32 plasmid to stably produce the microRNA and evaluate the impact of mir-32 overexpression on cell productivity, growth and viability in compare with scrambled control. Our findings highlight the application of miRNAs as engineering tools and indicated that miR-32 could be a target for engineering CHO cells to increase cell productivity.


Engineered matrices for skeletal muscle satellite cell engraftment and function.
Regeneration of traumatically injured skeletal muscles is severely limited. Moreover, the regenerative capacity of skeletal muscle declines with aging, further exacerbating the problem. Recent evidence supports that delivery of muscle satellite cells to the injured muscles enhances muscle regeneration and reverses features of aging, including reduction in muscle mass and regenerative capacity. However, direct delivery of satellite cells presents a challenge at a translational level due to inflammation and donor cell death, motivating the need to develop engineered matrices for muscle satellite cell delivery. This review will highlight important aspects of satellite cell and their niche biology in the context of muscle regeneration, and examine recent progresses in the development of engineered cell delivery matrices designed for skeletal muscle regeneration. Understanding the interactions of muscle satellite cells and their niche in both native and engineered systems is crucial to developing muscle pathology-specific cell- and biomaterial-based therapies.


Spectral properties of phosphopyridoxyl-Lys-7(-41)-ribonuclease A.
We have studied the spectral properties of RNAase A containing a phosphopyridoxyl residue at the epsilon-NH2 group of Lys-7 or Lys-14. The overall conformations of the native and modified enzymes were shown to be rather similar. All three proteins have similar circular dichroism spectra within the 220-300-nm region, and similar thermal transition temperatures. All the changes in the RNAase A molecule modified are located in close proximity to the alkylated lysine residue. The phosphopyridoxyl group of (P-Pxy)-epsilon-Lys-41-RNAase A is situated directly at the enzyme active site and is 25% butied in the protein globule. The P-pyridoxyl group of (P-Pxy)-epsilon-Lys-7-RNAase A was shown to be located in the vicinity of the active site and to be more exposed to the solvent. In the pyridoxyl phosphate absorption band, optical activity is induced in both proteins. Study of the pH dependence of the changes occurring in the circular dichroism and absorption spectra has shown that in the modified proteins, the pyridoxyl phosphate chromophore is rather sensitive to the ionic state of the surrounding medium and serves as a "reporter" group when the relationship between structure and function of the RNAase A active site is being investigated.


Sympathetic denervation caused by long-term noradrenaline infusions; prevention by desipramine and superoxide dismutase.
1. The effects of continuous intravenous infusion of noradrenaline (0.01 and 0.1 microgram kg-1 h-1) were studied in both the infused lateral saphenous vein and the contralateral saphenous vein of normal dogs. Noradrenaline, saline, noradrenaline + desipramine or noradrenaline + superoxide dismutase were infused using Alzet osmotic minipumps. 2. After a 5 day infusion period, the noradrenaline content in plasma and in both saphenous veins was determined, and the venous tissues submitted to light microscope morphometry and ultrastructural study and used for the determination of their O-methylation capacity (with [3H]-isoprenaline as a substrate). 3. Noradrenaline caused dose-dependent damage to the sympathetic nerve endings of the lateral saphenous veins. Concomitant changes in extraneuronal structure and function were observed (hypertrophy of smooth muscle cells, nuclear dysmorphy, thickening of the vessel wall, impairment in O-methylation capacity). 4. Desipramine and superoxide dismutase prevented or reduced the effects of noradrenaline on both the morphological and the biochemical parameters; the protection afforded by superoxide dismutase was more marked than that by desipramine. 5. It is concluded that moderately high doses of noradrenaline exert a 6-hydroxydopamine-like effect and that this chemical sympathectomy is partially or totally prevented by desipramine or superoxide dismutase. The data suggest that a substance derived from noradrenaline, in the formation of which free oxygen radicals are involved and which is subject to neuronal uptake, is the chemical entity responsible for the neurotoxic effect observed.


[Application of fasciocutaneous flap in repairing III degree burn wound in facio-cervical region].
To evaluate the clinical efficacy of fasciocutaneous flap in the repair of the III degree burn wound in the facio-cervical region. From January 1997 to October 2001, 11 cases of III degree burn wounds in the facio-cervical region were repaired with the cervical-thoracic fasciocutaneous flap ranging from 18 cm x 8 cm to 13 cm x 5 cm. Donor sites were covered with "Z" plasty or skin grafting. Except for partial necrosis of distal end of the flap in 1 case, the flaps in the other 10 cases all survived and presented satisfactory appearance and function during the following up. The fasciocutaneous flap in the cervical-thoracic region may provide a large area of flap, and can be easily transferred. It's an ideal flap for the repair of skin defect in the facio-cervical region, especially for the repair of the wounds with infection or exposure of vessel, nerve and tendon.


Specific in vitro IgG subclass synthesis and lymphocyte proliferation responses in herpes virus encephalitis.
Cerebrospinal fluid, peripheral blood lymphocytes (PBL) and sera from 5 patients with herpes simplex encephalitis (HSVE), 3 with varicellae zoster (VZV) meningoencephalitis and 5 with encephalitis of unknown origin (NUD) were analyzed. Lymphocytes from both blood and CSF were shown to synthesize anti-VZV IgG subclasses in VZV meningoencephalitis and anti-HSV IgG subclasses in HSVE. The subclass patterns of CSF and in vitro synthesized anti-viral IgG were similar, suggesting that a considerable portion of the antiviral IgG subclasses detected are synthesized in the CNS compartment. Antigen presentation in vitro seemed to produce a heterologous IgG4 and/or 3 response in 3 patients. Lymphocyte proliferation was detectable in response to HSV and VZV, respectively.


Traumatic peripheral nerve injuries: a classification proposal.
Peripheral nerve injuries (PNIs) include several conditions in which one or more peripheral nerves are damaged. Trauma is one of the most common causes of PNIs and young people are particularly affected. They have a significant impact on patients' quality of life and on the healthcare system, while timing and type of surgical treatment are of the utmost importance to guarantee the most favorable functional recovery. To date, several different classifications of PNIs have been proposed, most of them focusing on just one or few aspects of these complex conditions, such as type of injury, anatomic situation, or prognostic factors. Current classifications do not enable us to have a complete view of this pathology, which includes diagnosis, treatment choice, and possible outcomes. This fragmentation sometimes leads to an ambiguous definition of PNIs and the impossibility of exchanging crucial information between different physicians and healthcare structures, which can create confusion in the choice of therapeutic strategies and timing of surgery. The authors retrospectively analyzed a group of 24 patients treated in their center and applied a new classification for PNI injuries. They chose (a) five injury-related factors, namely nerve involved, lesion site, nerve type (whether motor, sensory or mixed), surrounding tissues (whether soft tissues were involved or not), and lesion type-whether partial/in continuity or complete. An alphanumeric code was applied to each of these classes, and (b) four prognostic codes, related to age, timing, techniques, and comorbidities. An alphanumeric code was produced, similar to that used in the AO classification of fractures. The authors propose this novel classification for PNIs, with the main advantage to allow physicians to easily understand the characteristics of nerve lesions, severity, possibility of spontaneous recovery, onset of early complications, need for surgical treatment, and the best surgical approach. according to the Oxford 2011 level of evidence, level 2.


Detection of superoxide radical in all biological systems by Thin Layer Chromatography.
The study presents a new method that detects O2•-, via quantification of 2-hydroxyethidium (2-ΟΗ-Ε+) as low as ∼30 fmoles by High-Performance Thin Layer Chromatography (HPTLC). The method isolates 2-ΟΗ-Ε+ after its extraction by the anionic detergent SDS (at 18-fold higher than its CMC) together with certain organic/inorganic reagents, and its HPTLC-separation from di-ethidium (di-Ε+) and ethidium (Ε+). Quantification of 2-OH-E+ is based on its ex/em maxima at 290/540 nm, and of di-E+ and E+ at 295/545 nm. The major innovations of the present method are the development of protocols for (i) efficient extraction (by SDS) and (ii) sensitive quantification (by HPTLC) for 2-OH-E+ (as well as di-E+ and E+) from most biological systems (animals, plants, cells, subcellular compartments, fluids). The method extracts 2-ΟΗ-Ε+ (by neutralizing the strong binding between its quaternary N+ and negatively charged sites on phospholipids, DNA etc) together with free HE, while protects both from biological oxidases, and also extracts/quantifies total proteins (hydrophilic and hydrophobic) for expressing O2•- levels per protein quantity. The method also uses SDS (at 80-fold lower than its CMC) to extract/remove/wash 2-ΟΗ-Ε+ from cell/organelle exterior membrane sites, for more accurate internal content quantification. The new method is applied on indicative biological systems: (1) artificially stressed (mouse organs and liver mitochondria and nuclei, ±exposed to paraquat, a known O2•- generator), and (2) physiologically stressed (cauliflower plant, exposed to light/dark).


Antihypertensive drugs in blood donations in Serbia - Observational study.
Most of the blood banks worldwide do not defer donors for their medication, with the exception of the teratogenic and platelet aggregation-inhibiting drugs use. In Serbia, where around 245.000 units of blood are collected each year, it is not common practice to consider the medication of potential blood donors. Therefore, the aim of this study was to quantify the presence of antihypertensive drugs in blood samples collected from blood donors treated for hypertension and to raise the issue of the recipient risks posed by drug residues in blood products. Serum samples were obtained from 450 volunteer blood donors collected during the year 2017 who reported the use of antihypertensive drugs. All blood donors were required to interrupt regular antihypertensive therapy for 24 h before blood donation and LCMS determination of antihypertensive drugs was performed. Beta blockers were detected in 81 out of 203 samples which tested positive for the presence of antihypertensive drugs. Concentrations above the limit of quantification were determined in 58% of samples positive on beta blockers, containing metoprolol and bisoprolol in amounts sufficient to produce a therapeutic effect in the recipient. Therefore, the obtained results suggested that the safety of blood donation from individuals with treated hypertension should not be neglected. A solution for this problem might be the establishment of a standard LCMS screening procedure as a tool for testing the blood of donors taking drugs.


Insulin-degrading enzyme is capable of degrading receptor-bound insulin.
In the investigation of the intracellular sites of insulin degradation, it might be important whether receptor-bound insulin could be a substrate for insulin-degrading enzyme (IDE). Insulin receptor and IDE were purified from rat liver using a wheat germ agglutinin column and monoclonal anti-IDE antibody affinity column, respectively. [125I]insulin-receptor complex was incubated with various amounts of IDE at 0 degree C in the presence of disuccinimidyl suberate and analyzed by reduced 7.5% SDS-PAGE and autoradiography. With increasing amounts of IDE, the radioactivity of 135 kd band (insulin receptor alpha-subunit) decreased, whereas that of 110 kd band (IDE) appeared then gradually increased, suggesting that IDE could bind to receptor-bound insulin. During incubation of insulin-receptor complex with IDE at 37 degrees C, about half of the [125I]insulin was dissociated from the complex. However, the time course of [125I]insulin degradation in this incubation was essentially identical to that of free [125I]insulin degradation. Cross-linked, non-dissociable receptor-bound [125I]insulin was also degraded by IDE. Rebinding studies to IM-9 cells showed that the receptor binding activity of dissociated [125I]insulin from insulin-receptor complex incubated with IDE was significantly (p less than 0.001) decreased as compared with that without the enzyme. These results, therefore, show that IDE could recognize and degrade receptor-bound insulin, and suggest that IDE may be involved in insulin metabolism during receptor-mediated endocytosis through the degradation of receptor-bound insulin in early neutral vesicles before their internal pH is acidified.


Possible urate nephropathy of the newborn infant as a cause of transient renal insufficiency.
Oliguric renal failure, bilateral renal enlargement, and hyperuricemia were noted in three term infants. The findings on intravenous pyelography and radionuclide renography were consistent with intratubular obstruction to urinary outflow in two of the infants. The onset of diuresis appeared to follow intravenous pyelography in these infants. Urinary urate crystalluria was prominent in each infant in the first few days after the onset of diuresis, during which normal serum urate concentrations and normal renal function were established.


Prevalence and anatomical characteristics of developmental venous anomalies: an MRI study.
Developmental venous anomalies (DVAs) are extreme anatomical venous variations formed by multiple radiating medullary veins, which converge centripetally into a single collecting vein. Their coexistence with symptomatic cavernous malformations (CMs) has been reported in the literature. The aim of this study was to assess the characteristics of DVAs using MRI. A total of 6948 head MRIs of adult Caucasian patients were retrospectively analyzed to determine the number and locations of DVAs. We collected the data on the termination of the collecting vein, the prevalence of DVA-related CMs, and MRI FLAIR signal-hyperintensity corresponding to the location of the DVA. At least one DVA was identified in 7.46% of the patients. The prevalence decreased with age, with a Pearson correlation coefficient of - 0.7328. A total of 599 DVAs were identified. Multiple DVAs were found in 10.92% of the patients with DVAs. The DVAs were identified more often in the supratentorial region (73.12%, p < 0.0001), and the most common location was the frontal lobe (35.23%). The collecting vein usually drained into the superficial cerebral veins (68.78%). CMs were observed in 4.14% of the patients with DVAs, and the prevalence showed a positive correlation with age. Signal-intensity abnormalities were identified in the vicinity of 5.18% DVAs. Knowledge about characteristics of DVAs and associated anomalies is essential for neuroradiologists and neurosurgeons. The large number of currently available diagnostic studies enables us to assess anatomical variants on a great number of subjects.


Radiative effects of aerosols over Indo-Gangetic plain: environmental (urban vs. rural) and seasonal variations.
Aerosol radiative effects over two environmentally distinct locations, Kanpur (urban site) and Gandhi College (rural location) in the Indo-Gangetic plain (IGP), a regional aerosol hot spot, utilizing the measured optical and physical characteristics of aerosols, an aerosol optical properties model and a radiative transfer model, are examined. Shortwave aerosol radiative forcing (ARF) at the top of the atmosphere (TOA) is < -12 W m( - 2) over Kanpur and Gandhi College. ARF at the surface is ≥ -30 W m( - 2). Atmospheric warming is maximum during premonsoon (>30 W m( - 2)). Shortwave atmospheric heating due to aerosols is >0.4 K/day over IGP and peaks during premonsoon at >0.6 K/day due to lower single scattering albedo (SSA) and higher surface albedo. TOA forcing is always less negative over Kanpur when compared to Gandhi College due to lower surface albedo except in postmonsoon owing to higher SSA. This happens as TOA forcing depends on SSA and surface albedo in addition to aerosol optical depth. The magnitude of longwave forcing and atmospheric cooling in an absolute sense is significantly small and contributes only about 20% or less to the net (shortwave + longwave) forcing. Aerosol radiative effects over these two locations, despite differences in aerosol characteristics, are similar, thus confirming that aerosols and their radiative influence get transported due to circulation. ARF over Kanpur and Gandhi College is an order of magnitude higher when compared to greenhouse gas forcing. A large reduction in surface reaching solar irradiance accompanied by large atmospheric warming can have implications on precipitation and hydrological cycle, and these aerosol radiative effects should be included while performing regional-scale aerosol climate assessments.


Analysis of mutagenic DNA repair in a thermoconditional mutant of Saccharomyces cerevisiae. IV. Influence of DNA replication and excision repair on REV2 dependent UV-mutagenesis and repair.
A double mutant being thermoconditionally defective in mutation induction as well as in repair of pre-lethal UV-induced DNA damage (rev2ts) and deficient in excision repair (rad3-2) was studied in temperature-shift experiments. The influence of inhibitors of DNA replication (hydroxyurea, aphidicolin) was determined. Additionally, an analysis of the dose-response pattern of mutation induction ("mutation kinetics") at several ochre alleles was carried out. It was concluded that the UV-inducible REV2 dependent mutagenic repair process is not induced in excision-deficient cells. In excision-deficient cells, REV2 dependent mutation fixation is slow and mostly post-replicative though not dependent on DNA replication. The REV2 mediated mutagenic process could be separated from the repair function.


Ascites.
Ascites is the pathologic accumulation of fluid in the peritoneum. It is the most common complication of cirrhosis, with a prevalence of approximately 10%. Over a 10-year period, 50% of patients with previously compensated cirrhosis are expected to develop ascites. As a marker of hepatic decompensation, ascites is associated with a poor prognosis, with only a 56% survival 3 years after onset. In addition, morbidity is increased because of the risk of additional complications, such as spontaneous bacterial peritonitis and hepatorenal syndrome. Understanding the pathophysiology of ascites is essential for its proper management.


Discovering cytokines as targets for chemotherapy-induced painful peripheral neuropathy.
Chemotherapy-induced peripheral neuropathy (CIPN), a dose-limiting neurotoxic effect of chemotherapy, is the most common reason for early cessation of cancer treatment. This can result in an increased risk of recurrence and decreased survival rate. Inflammatory cascade activation, proinflammatory cytokine upregulation, and neuro-immune communication pathways play essential roles in the initiation and progression of CIPN. Most notably, TNF-α, IL-1β, IL-6, and CCL2 are involved in neuropathic pain. Further elucidation of the role of these cytokines could lead to their development and use as biomarkers for predicting the onset of painful peripheral neuropathy and early axonal damage. In this review, we provide evidence for the involvement of cytokines in CIPN, the possible underlying mechanisms, and their use as potential therapeutic targets and biomarkers to prevent and improve the painful peripheral neuropathy related to chemotherapeutic agents.


Ultrastructural study of in vitro larval development of Echinococcus granulosus protoscoleces.
Through ultrastructural study of the morphological forms developed in vitro during protoscolex culture, we describe larval E. granulosus histogenesis. The transformation of the spined microtriches in the protoscolex into truncated microtriches that develop within the hydatid cyst is discussed. The paper also describes the mitochondria location change that occurs during the evolution; the mitochondria pass from the most internal area of the distal cytoplasm along the cytoplasmic extensions into the cytoplasm of tegumental cells. The ultrastructures of both the vesiculated protoscolex and the posterior bladder demonstrate that each state corresponds to the initial step on one of the two paths of in vitro vesicular development.


Biomaterials and nanomedicine for bone regeneration: Progress and future prospects.
Bone defects pose a heavy burden on patients, orthopedic surgeons, and public health resources. Various pathological conditions cause bone defects including trauma, tumors, inflammation, osteoporosis, and so forth. Auto- and allograft transplantation have been developed as the most commonly used clinic treatment methods, among which autologous bone grafts are the golden standard. Yet the repair of bone defects, especially large-volume defects in the geriatric population or those complicated with systemic disease, is still a challenge for regenerative medicine from the clinical perspective. The fast development of biomaterials and nanomedicine favors the emergence and promotion of efficient bone regeneration therapies. In this review, we briefly summarize the progress of novel biomaterial and nanomedical approaches to bone regeneration and then discuss the current challenges that still hinder their clinical applications in treating bone defects.


The gut metabolite, trimethylamine N-oxide inhibits protein folding by affecting cis-trans isomerization and induces cell cycle arrest.
Trimethylamine N-Oxide (TMAO) is an important metabolite, which is derived from choline, betaine, and carnitine in various organisms. In humans, it is synthesized through gut microbiota and is abundantly found in serum and cerebrospinal fluid (CSF). Although TMAO is a stress protectant especially in urea-rich organisms, it is an atherogenic agent in humans and is associated with various diseases. Studies have also unveiled its exceptional role in protein folding and restoration of mutant protein functions. However, most of these data were obtained from studies carried on fast-folding proteins. In the present study, we have investigated the effect of TMAO on the folding behavior of a well-characterized protein with slow folding kinetics, carbonic anhydrase (CA). We discovered that TMAO inhibits the folding of this protein via its effect on proline cis-trans isomerization. Furthermore, TMAO is capable of inducing cell cycle arrest. This study highlights the potential role of TMAO in developing proteopathies and associated diseases.


Drilling Deeper into tooth brushing skills: Is proactive interference an under-recognized factor in oral hygiene behavior change?
Proper tooth brushing is a seemingly simple motor activity that can promote oral health. Applying health theories, such as the Information-Motivation-Behavioral Skills (IMB) model, Motivational Interviewing (MI) and Integrated Health Coaching (IHC), may help optimize tooth brushing technique in those with suboptimal skills. Some motor activities, including tooth brushing, may over time become rote and unconscious actions, such that an existing habit can inhibit new learning, i.e., exert proactive interference on learning the new skill. Proactive interference may impede the acquisition of new tooth brushing skills; thus, in this report, we: (1) Review how the habit of tooth brushing is formed; (2) Postulate how proactive interference could impede the establishment of proper tooth brushing retraining; (3) Discuss the merits of this hypothesis; and (4) Provide guidance for future work in this topic within the context of an approach to behavior change that integrates IMB, MI and IHC methodology.


Sequence determination and analysis of three plasmids of Pseudomonas sp. GLE121, a psychrophile isolated from surface ice of Ecology Glacier (Antarctica).
Pseudomonas sp. GLE121 (a psychrophilic Antarctic strain) carries three plasmids: pGLE121P1 (6899 bp), pGLE121P2 (8330 bp) and pGLE121P3 (39,583 bp). Plasmids pGLE121P1 and pGLE121P2 show significant sequence similarity to members of the IncP-9 and IncP-7 incompatibility groups, respectively, while the largest replicon, pGLE121P3, is highly related to plasmid pNCPPB880-40 of Pseudomonas syringae pathovar tomato NCPPB880. All three plasmids have a narrow host range, limited to members of the genus Pseudomonas. Plasmid pGLE121P3 encodes a conjugal transfer system, while pGLE121P1 carries only a putative MOB module, conserved in many mobilizable plasmids. Plasmid pGLE121P3 contains an additional load of genetic information, including a pair of genes with homology to the rulAB operon, responsible for ultraviolet radiation (UVR) tolerance. Given the increasing UV exposure in Antarctic regions, the expression of these genes is likely to be an important adaptive response.


Biologic considerations for drug targeting in cancer patients.
There is a great need for more specific targeting of chemotherapeutic agents, but development of specific therapy will be difficult in light of the barriers which separate a tumor from the vasculature, tumor cell heterogeneity and instability, technological advances necessary for drug delivery design and introduction into the clinic and mechanisms for assessment of efficacy (Table 5). While imposing, these problems are not insurmountable. A clearer understanding of the specific goal of a drug delivery/drug targeting approach will make the expectations more realistic and the chances for success greater. Primary to improving drug targeting is a better understanding of the biology of tumors. There are a number of limitations to drug targeting technology mentioned above, but, at present, the more difficult limitations are imposed by tumors themselves and the host's response to a tumor. Currently available technology does not offer Erlich's magic bullet and it does not appear that a single entity will suffice for cancer. Perhaps a mixture of drug delivery systems, each reducing the toxicity of a particular component of therapy will provide the first realistic goal in drug targeting. Each form of drug targeting is limited by biochemical and biophysical properties of the host, tumor cells, drug delivery system and interactions between them. Successes in vitro are moot without corresponding data in the (unfortunately) more complex organismic level. Development of drug targeting approaches will also require a critical, thorough evaluation of the ability to specifically deliver drug in vivo.


Retrospective evaluation of epidermal skin prick tests in patients living in Aegean region.
Sensitization to aeroallergens of Aegean region is not well decumented. In this study we evaluated the epidermal skin prick test results of the patients who applied to allergy outpatient department retrospectively. Epidermal skin prick test of the 5055 patients were evaluated. Of these patients 2638 (52 %) were female, 2417 (48 %) male, 1213 (24 %) adult and 3842 (76 %) pediatric patient, 1163 (23 %) patients were allergic rhinitis, 2477 (49 %) were bronchial asthma, 505 (10 %) were allergic rhinitis with bronchial asthma, 556 (11 %) were chronic urticaria, 253 (5 %) were wheezy infant and 101 (2 %) patients were atopic dermatitis, 2932 (58 %) had atopy history in their first and second degree relatives. Patients were aged between 3.5 months and 79 years (mean 14.1 3.2 years and median 11 years). In epidermal skin prick tests sensitization to house dust mites (D. farinae, D. pteroniyssinus), pollens (grass, cereals and trees), moulds, animal danders, foods (especially in early childhood) and cockroach were evaluated. Our data indicate that allergens that may be the cause of the high prevalence of allergic diseases in Izmir are probably produced by pollens and mites.


Reduced haloperidol plasma concentration and clinical response in acute exacerbations of schizophrenia.
Twenty-nine hospitalized patients suffering acute exacerbations of schizophrenia were treated for 2 weeks with fixed daily oral doses of haloperidol prospectively calculated to achieve a haloperidol plasma concentration of either 8-18 ng/ml or 25-35 ng/ml. Reduced haloperidol as well as haloperidol concentrations were assayed to determine if the former enhanced the predictability of response. Wee 2 haloperidol plasma concentrations were negatively correlated to clinical response as measured by the percentage change in the BPRS score from baseline (r = -0.43, P less than 0.05). In contrast, week 2 plasma concentrations of reduced haloperidol, total haloperidol (haloperidol + reduced haloperidol), and reduced haloperidol/haloperidol ratio did not correlate with the change in the BPRS score. Chi-square analysis concluded that patients with ratios greater than one were no less likely to be treatment responders (less than 25% improvement in BPRS from baseline and week 2 BPRS less than 55) than those with ratios less than one. Although these data lend additional support to reports of a curvilinear relationship between haloperidol plasma concentration and clinical response, they also suggest that reduced haloperidol plasma concentrations are of no value in predicting treatment response.


Iliac arteriovenous fistulas after lumbar spinal surgery.
Iatrogenic arteriovenous fistula (AVF) is an unusual and potentially fatal complication of lumbar spinal surgery. The presentation of these injuries is usually late, with symptoms such as leg swelling or cardiac failure. It is crucial to suspect AVF in the patient which presents to emergency with lumbar spinal surgery history. The diagnosis is often based on imaging studies such as computed tomography (CT) or magnetic resonance (MR) angiography. Surgery was the first choice of treatment, but with recent advances in stent technology endovascular approach has become widely popular. We present two cases of AVF secondary to lumbar spinal surgery, one of them presenting with overt heart failure and the other one with leg swelling.


Robust bipartite tracking consensus of multi-agent systems via neural network combined with extended high-gain observer.
In this paper, the robust bipartite tracking consensus problem for second-order multi-agent systems has been addressed in the presence of lumped disturbances and unknown velocity information. The leader's dynamics are modeled by uncertain fractional-order equality based on the neural network (NN), and the followers can only obtain the position information of the leader under the signed networks. A novel robust bipartite tracking consensus control scheme is developed by combining the NN approximators, the continuous sliding mode control (SMC) strategy, and the improved extended high-gain observer (EHGO). The improved EHGO is used to compensate for and estimate each agent's lumped disturbances and unknown velocity information in the controller design process. Moreover, NN is constructed to approximate the velocity and acceleration of the uncertain leader's dynamics for generating the feedforward signal of controllers, and the adaptive update laws of estimation parameters are generated online based on the Lyapunov function. Finally, the effectiveness of the proposed control strategy is verified by some numerical simulations.


Quantitative comparison of various methods for detecting eggs of Toxocara canis in samples of sand.
Six techniques for recovering unembryonated Toxocara canis eggs from sand samples were tested for efficiency and suitability for routine use. The tests were done under standardised conditions on 50g of sand samples contaminated experimentally with 10, 100 and 500 eggs of T. canis. Best result was achieved by the method of Dunsmore et al. [Dunsmore, J.D., Thompson, R.C.A., Bates, I.A., 1984. Vet. Parasitol. 16, 303-311]. The results were expressed as the number of T. canis eggs recorded and percentage rates of recovery in sand samples.


Dermatitis to captopril.
reports on delated cutaneous reactions to captopril have been seldom reported. Captopril is an angiotensin-converting enzyme (ACE) inhibitor and their cutaneous side-effects are documented, but little has been published concerning the usefulness of patch test when they occur. We presented the case of a patient who developed a cutaneous reaction induced by captopril with positive patch test. patch testing was performed with captopril, other ACE (enalapril, lisinopril ramipril), and European standard series. Following, we performed a double-blind oral challenge test with drugs who results was negative. Positive reaction were obtained to captopril at 4 days and the others test being negative. The same test were negative in five control patients. The patient tolerated enalapril, and lisinopril without problems. the allergological studies confirmed sensitisation to captopril and tolerance to lisinopril, and enalapril. When patch test are performed with several drugs of the same family, results seem to indicate an absence of cross-sensitivity, but in several patients, oral provocation test were needed because patch test gave no conclusive information.


Necrotizing Fasciitis? A Mimic Produced by an Unusual Presentation of Necrotizing Neutrophilic Dermatosis of the Hand: A Case Report.
A 50-year-old woman presented to our emergency department with hand pain and a laceration. Despite administration of antibiotics, the patient progressively worsened leading to a concern for a necrotizing soft-tissue infection. She underwent multiple debridements yet continued to worsen clinically. Tissue biopsy was eventually consistent with a necrotizing neutrophilic dermatosis. The patient was then started on oral steroids with rapid improvement of her clinical status and hand examination. While neutrophilic dermatoses are rare, the orthopaedic surgeon treating soft-tissue infections should maintain a broad differential when treatment of a potential necrotizing infection has not led to diagnostic closure and clinical improvement.


Recent advances in catalytic decomposition of ozone.
Ozone (O3), as a harmful air pollutant, has been of wide concern. Safe, efficient, and economical O3 removal methods urgently need to be developed. Catalytic decomposition is the most promising method for O3 removal, especially at room temperature or even subzero temperatures. Great efforts have been made to develop high-efficiency catalysts for O3 decomposition that can operate at low temperatures, high space velocity and high humidity. First, this review describes the general reaction mechanism of O3 decomposition on noble metal and transition metal oxide catalysts. Then, progress on the O3 decomposition performance of various catalysts in the past 30 years is summarized in detail. The main focus is the O3 decomposition performance of manganese oxides, which are divided into supported manganese oxides and non-supported manganese oxides. Methods to improve the activity, stability, and humidity resistance of manganese oxide catalysts for O3 decomposition are also summarized. The deactivation mechanisms of manganese oxides under dry and humid conditions are discussed. The O3 decomposition performance of monolithic catalysts is also summarized from the perspective of industrial applications. Finally, the future development directions and prospects of O3 catalytic decomposition technology are put forward.


Activation of Gab1 in pancreatic acinar cells: effects of gastrointestinal growth factors/hormones on stimulation, phosphospecific phosphorylation, translocation and interaction with downstream signaling molecules.
The scaffolding/adapter protein, Gab1, is a key signaling molecule for numerous stimuli including growth factors and G protein-coupled-receptors (GPCRs). A number of questions about Gab1 signaling remain and little is known about the ability of gastrointestinal (GI) hormones/neurotransmitters/growth factors to activate Gab1. Therefore, we examined their ability to activate Gab1 and explored the mechanisms involved using rat pancreatic acini. HGF and EGF stimulated total Gab1 tyrosine phosphorylation (TyrP) and TyrP of Gab1 phospho-specific sites (Y307, Y627), but not other pancreatic growth factors, GI GPCRs (CCK, bombesin, carbachol, VIP, secretin), or agents directly activating PKC or increasing Ca2+. HGF-stimulated Y307 Gab1 TyrP differed in kinetics from total and Y627. Neither GF109203X, nor inhibition of Ca2+ increases altered HGF's effect. In unstimulated cells>95% of Gab1 was cytosolic and HGF stimulated a 3-fold increase in membrane Gab1. HGF stimulated equal increases in pY307 and pY627 Gab1 in cytosol/membrane. HGF stimulated Gab1 association with c-Met, Grb2, SHP2, PI3K, Shc, Crk isoforms and CrkL, but not with PLCgamma1. These results demonstrate that only a subset of pancreatic growth factors (HGF/EGF) stimulates Gab1 signaling and no pancreatic hormones/neurotransmitters. Our results with Gab1 activation with different growth factors, the role of PKC, and its interaction with distant signaling molecules suggest the cellular mechanisms of Gab1 signaling show important differences in different cells. These results show that Gab1 activation plays a central role in HGF's ability to stimulate intracellular transduction cascades in pancreatic acinar cells and this action likely plays a key role in HGF's ability to alter pancreatic cell function (i.e., growth/regeneration).


Expression, purification, crystallization and preliminary X-ray crystallographic analysis of human β-galactosidase.
β-D-Galactosidase (β-Gal) is an exoglycosidase that cleaves β-galactosides from glycoproteins, sphingolipids and keratan sulfate. This study reports the expression, purification, crystallization and preliminary X-ray crystallographic analysis of human lysosomal β-Gal. The sitting-drop vapour-diffusion method was used to crystallize β-Gal in complexes with its product galactose and with the inhibitor 1-deoxygalactonojirimycin. The resulting crystals were isomorphous and belonged to space group P2(1). The crystals of the β-Gal-galactose and the β-Gal-inhibitor complexes had unit-cell parameters a = 94.8, b = 116.1, c = 140.3 Å, β = 92.2° and a = 94.8, b = 116.0, c = 140.3 Å, β = 92.2°, respectively. Diffraction data were collected to 1.8 Å resolution for both crystals.