Cloning, expression and growth promoting action of Red tilapia (Oreochromis sp.) neuropeptide Y.
Neuropeptide Y, a 36 amino acid peptide abundantly expressed in the brain, is the most potent orexigenic factor known to date in mammals. It has been shown to be one of the most conserved neuropeptides in vertebrate evolution. It seems that neuropeptide Y functions, in addition to sequence conservation, are also well conserved in fish. In the present study, we cloned and reported the cDNA sequence coding for tilapia 36 aminoacid neuropeptide Y. We express the tilapia neuropeptide Y gene in Escherichia coli driven by T7 promoter. The recombinant neuropeptide Y was purified up to 80% by affinity chromatography. We developed both, a food intake and a growth performance experiment to evaluate the effects of neuropeptide Y administration. Juvenile tilapia receiving recombinant neuropeptide Y (1 microg/g of body weight) by intraperitoneal injection increased food intake compared to controls (p < 0.05). Similarly, in the growth performance experiment, we observed an increase in body weight (p < 0.05) of tilapia fry receiving the same dose of the peptide. Neuropeptide Y treatment had no significant effect on hepatosomatic index and muscle moisture content. On the other hand, muscle protein content was increased in treated animals. These results demonstrate that administration of biologically active E. coli-derived neuropeptide Y resulted in a growth promoting action in fish.


Somataglycan-S: a neuronal surface proteoglycan defines the spinocerebellar system.
The formation and maintenance of functionally specific neuronal networks may depend on specific proteoglycans localized to the surface membranes of a subset of neurons. Monoclonal antibody (MAb) 6A2 labeled a distinct subset of CNS neurons: the somas and proximal dendrites of cells making up the spinocerebellar and reticular systems. These pathways contribute to proprioceptive and exteroceptive functions. Ultrastructurally, MAb 6A2 immunoreactivity was distributed focally along the cell surface membranes and the adjacent extracellular space. On western blots of immunoaffinity-purified preparations from cerebellar homogenates, a major, broad band of approximately 400 kDa is labeled by MAb 6A2. Increased electrophoretic mobility of the purified antigen after digestion with chondroitinase ABC and keratanase suggests that the antigen is a proteoglycan bearing chondroitin sulfate and keratan sulfate glycosaminoglycans. Unsulfated N-acetyl-galactosamine residues linked to unsaturated uronic acid constituted the initial disaccharide in the chondroitin sulfate glycosaminoglycan chains. N- and O-linked oligosaccharides on the core protein were detected by the biotinylated lectins wheat germ agglutinin and Jacalin, respectively, and by MAb anti-HNK-1. Lyase and glycosidase digests result in a 280-kDa band. This proteoglycan, somataglycan-S, may provide a key to the role of glycoconjugates in determining neuronal diversity and system specificity.


Transcatheter Versus Surgical Pulmonary Valve Replacement in Repaired Tetralogy of Fallot.
Transcatheter pulmonary valve replacement (TC-PVR) is an alternative to surgical PVR (S-PVR) in repaired Tetralogy of Fallot (TOF). The purpose of this study is to compare in-hospital outcomes, hospital costs, and projected 5-year total costs of S-PVR to TC-PVR in patients with repaired TOF. We performed a multicenter, retrospective cohort study of children and adults with TOF ≥ 8 years of age who underwent PVR from January 1, 2010 to December 31, 2016 at 46 centers contributing to the Pediatric Health Information Systems database. Baseline characteristics, in-hospital outcomes, and costs were compared between the two groups. Projected 5-year costs were calculated by combining cost data with published reintervention rates. A total of 194 TC-PVR and 1,072 S-PVR were performed. The baseline characteristics of the TC-PVR and S-PVR groups were not significantly different with the exception of greater age in the TC-PVR group (median age of 17 years vs 15 years, p value <0.001). Discharge mortality, hospital charges and estimated cost, surgical complication rates, and acute kidney failure were not significantly different between the groups. Intensive care unit use, intensive care unit length of stay (LOS), mechanical ventilation use, extracorporeal membrane oxygenation use, and total LOS were lower with TC-PVR than S-PVR. Projected 5-year costs were greater with TC-PVR compared with S-PVR ($64,762 vs $56,536) due to the cost of the transcatheter pulmonary valve and higher reintervention rates. In conclusion, despite longer LOS and greater in-hospital resource utilization for patients with TOF who underwent S-PVR compared with TC-PVR, mortality, and in-hospital costs are the same, and projected 5-year costs are less.


Spinal arteriovenous malformations.
Spinal cord arteriovenous malformations encompass a variety of distinct lesions with aberrant connections between the arterial and venous circulation. Symptomatology and clinical presentation of such lesions are directly related to their location and particular pathophysiology. In the authors' surgical experience, 90% of patients have had the same or improved neurologic function and long-term follow-up.


High-frequency ventilation and conventional mechanical ventilation in newborn babies with respiratory distress syndrome: a prospective, randomized trial.
Morbidity and mortality remain high amongst babies ventilated for a respiratory distress syndrome (RDS). Whether newly developed ventilators allowing high frequency ventilation such as high frequency flow interrupted ventilation (HFFIV) could decrease the morbidity and the mortality was investigated in a randomized study. Preterm babies weighing < or = 1800 g suffering from RDS and ventilated by conventional mechanical ventilation (CMV) were randomized to be further ventilated either by CMV (group CMV) or by HFFIV (group HFFIV) when peak inspiratory pressure (PIP) on CMV was > or = 20 cmH2O. The study was undertaken in the neonatal intensive care unit of the Erasmus Hospital. 24 patients entered into the investigation and were randomized but 2 patients were removed from the study because the switch over to HFFIV failed. Eight of the 12 CMV patients and 5 of the 10 HFFIV patients completed the study. Clinical variables, blood gas analysis and ventilatory variables were looked at. There were no differences in mortality, in incidence of air leaks and pulmonary complications or in blood gas analysis. Bronchopulmonary dysplasia was not decreased by the use of HFFIV. It is concluded that HFFIV is safe although it offers no concrete advantages over CMV when applied as we did in a low pressure approach.


A Systematic Review of Measures of Dysarthria Severity in Stroke Patients.
To define methods to measure dysarthria due to stroke and guide physicians in delineating a diagnostic protocol using the best current strategies. Systematic review. A search was conducted on PubMed, EMBASE, the Cochrane Library, and Web of Science to identify measurement methods for dysarthria severity in adults after stroke. Two reviewers independently reviewed articles and came to a consensus about which ones to include. The authors excluded all duplicates, articles involving individuals with aphasia or other speech problems other than dysarthria, and articles unrelated to stroke. Articles were included if diagnostic measures were used to examine the effectiveness of speech rehabilitation in stroke patients. The search identified 1154 articles with the keywords "stroke" OR "ictus" OR "cerebral vascular accident" AND "dysarthria" OR "Speech and Language Disorders" AND "diagnosis" OR "assessment." The reviewers analyzed 86 full texts. There were 37 publications that met the criteria and were included in the systematic review. These articles were used to describe the main methods used for measuring the severity of stroke-related dysarthria before and after speech rehabilitation. Despite the range of diagnostic tools available, robust trials are lacking, and the diagnostic approaches are always different. More research is needed to find the best diagnostic methodologies and delineate a definitive diagnostic protocol.


Bone mineral content, cortical thickness and fracture rate in osteoporotic women after withdrawal of treatment with nandrolone decanoate, 1-alpha hydroxyvitamin D3, or intermittent calcium infusions.
Twenty-six post-menopausal osteoporosis patients were studied prospectively over a 2-yr period after the cessation of 2 yr of treatment with either nandrolone decanoate (anabolic steroid), 1 alpha hydroxyvitamin D3 or intermittent calcium infusions, the total observation period being 4 yr. Serial evaluations of bone mineral content in the radius, cortical thickness of the metacarpals and spinal roentgenograms were obtained. Nine patients who had been treated with 50 mg nandrolone decanoate every three weeks did not lose bone from the radius or the metacarpals during the 2 yrs following withdrawal of therapy. Eight patients who had received 1 microgram/day of oral 1 alpha hydroxyvitamin D3 retained a positive bone mineral content in the radius but lost cortical thickness from the metacarpals after withdrawal of therapy. Nine patients who had been treated with intermittent calcium infusions over 12 consecutive days annually did not lose further bone mineral content from the radius, but did lose cortical thickness at the expected rate during the 2-yr post-treatment period. The fracture rate at the end of the 4-yr observation period was 40% lower in the nandrolone decanoate group than in the 1-alpha-hydroxyvitamin D3 and calcium infusion groups. The results of this study indicate that, in contrast to oestrogen and calcium therapy, nandrolone decanoate and 1-alpha-hydroxyvitamin D3 have lasting beneficial effects, and that data obtained by photon absorptiometry do not always coincide with those obtained by radiogrammetry. These findings are highly relevant to the design of future therapeutic trials.


pH-Based Detection of Target Analytes in Diluted Serum Samples Using Surface Plasmon Resonance Immunosensor.
Detection of minute quantities of target antigens in serum samples (consisting of a mixture of proteins/biomolecules) can be achieved by enhancement of the capture efficiencies of heterogeneous immunosensors. An important process parameter which affects the capture of target analytes in such immunosensors is the pH of the solution as the target proteins present in the serum samples are charged molecules. Here, we investigated the capture of prostate-specific antigens (PSAs), first in a mixed-analyte system wherein the solution contained two other non-specific proteins along with the target analyte, using the surface plasmon resonance spectroscopy. There are no reports on the detection of antigens in a mixed system based on the optimization of the pH values of the carrier fluid, and this is the motivation of the present work. Further, we studied interference effects caused by the presence of these non-specific proteins in the mixed-analyte systems by artificially increasing the ratio of the interfering proteins to that of the target protein. Eventually PSA spiked into the rabbit serum samples was captured through the optimization of the pH of the solution. We could detect PSA in the serum samples when diluted to 100 times or more, where the amounts of other interfering proteins were ~ 66 times that of the amount of PSA. This study proposes a heterogeneous immunosensor to detect the target analytes in the diluted serum samples by tuning pH the of solution mixture, which can be utilized to detect disease biomarkers in serum samples.


The changing face of HR professionals' expectations amidst COVID-19: a comparison in between Sri Lanka and foreign context.
The COVID-19 has brought significant transformation to organizations throughout the world in expediting the sudden transition to digital business operations for business continuity. Thus, businesses need to examine the expectations of Human Resource (HR) professionals to adapt to the unexpected changes aroused by the novel COVID-19 pandemic. The study aimed to compare HR professionals' expectations from the workplace in Sri Lanka and foreign countries during the COVID-19 pandemic. The researchers utilized the qualitative research method and applied the thematic analysis in order to analyze the gathered data from 28 semi-structured interviews. According to the findings, HR professionals in Sri Lanka anticipate returning to work at office premises, and if this work transition continues in the future, they expect more incentives and strategies for work-life balance. In contrast, HR professionals in foreign countries anticipate a hybrid work culture with employee wellbeing sessions. Accordingly, the study implies that organizations should facilitate the expected requirements to continue work in the current and future crises. The HR policymakers will be able to carry out more precise planning activities in the future when developing policies related to managing HR functions in crises.


Precise determination of U isotopic compositions in low concentration carbonate samples by MC-ICP-MS.
We developed a fast and simple analytical procedure for precise determination of U isotopic compositions in low concentration natural samples. The main advantage of the new method is that it requires only 12ng U and can obtain all U isotopic ratios without using spike. Five carbonate reference materials (JCp-1, RKM-4, RKM-5, GBW04412 and GBW04413) and 3 international standards with different matrices (IAPSO, IRMM-3184 and CRM-U010) were analyzed for ((234)U/(238)U) and (238)U/(235)U ratios by MC-ICPMS. Using our method, the results for these standards are in close agreement with the certified values, 1.144 ± 0.004, 0.966 ± 0.004 and 0.990 ± 0.003 for ((234)U/(238)U) and 137.72 ± 0.13, 137.64 ± 0.15 and 98.63 ± 0.04 for (238)U/(235)U, in IAPSO, IRMM-3184 and CRM-U010, respectively. The long-term reproducibility of ((234)U/(238)U) and (238)U/(235)U is 0.970 ± 0.002 and 137.56 ± 0.09; 1.144 ± 0.004 and 137.72 ± 0.13, respectively, for in-house U solution and IAPSO. The new ((234)U/(238)U) results for carbonates show much better precision than previous studies and also reflect their age variability. The obtained (238)U/(235)U ratios, representing the first measurements in these carbonate specimens, are rather constant. The method described here requires only 12 ng of U for analysis and can be completed in 5.2 min. The approach provides a fast method to measure ((234)U/(238)U) and (238)U/(235)U ratios in sample matrices commonly encountered in studies of chemical weathering, oceanography and paleoclimatology.


Single sweep analysis of visual evoked potentials through a model of parametric identification.
An original method is presented for the single sweep analysis of visual evoked potentials (VEP's). The introduced algorithm bases upon an AutoRegressive with eXogenous input (ARX) modeling. A Least Squares procedure estimates the coefficients of the model and allows to obtain a complete black-box description of the signal generation mechanism, besides providing a filtered version of the single sweep potential. The performance of the algorithm is verified on proper simulation tests and the experimental results put into evidence the noticeable improvement of signal-to-noise ratio with a consequent better recognition of the classical parameters of the peaks (latencies and amplitudes). The possibility of measuring these parameters on a single sweep basis enables to evaluate the dynamics of the Central Nervous System response during the entire course of the examination. A classification of the estimated evoked potentials in a small number of subsets, on the basis of their morphology, is also possible.


Further evidence for the regulation of bacterial populations in soil by protozoa.
After the addition to soil of large numbers of a cowpea Rhizobium strain, the population declined steadily until the numbers reached about 10(7)/g, and the protozoa rose to about 10(4)/g. When indigenous protozoa were suppressed by the addition of actidione to the soil, the density of the test rhizobium did not fall initially, but its abundance declined to about 10(7)/g when actidione-resistant protozoa arose in significant numbers. The addition to actidione-treated soil of an antibiotic-resistant strain of Paramecium led to a rapid decrease in the population of the rhizobium, the density reaching essentially the same value as in soil receiving neither the drug nor the paramecia. The same changes occurred with Xanthomonas campestris as test prey except that its numbers fell to about 10(5)/g of soil. These data provide further evidence for the key role of protozoa in controlling the abundance of populations of certain bacteria introduced into soil.


Detection of G119S ace-1 (R) mutation in field-collected Anopheles gambiae mosquitoes using allele-specific loop-mediated isothermal amplification (AS-LAMP) method.
Malaria vectors have developed resistance to the four families of insecticides available for public health purposes. For example, the kdr mutation is associated with organochlorines and pyrethroids resistance. It is of particular concern that organophosphate and carbamate resistance associated with the G119S ace-1 (R) mutation has recently increased in West Africa in extent and frequency, and is now spreading through the Anopheles gambiae malaria vector population. There is an urgent need to improve resistance management using existing insecticides and new tools to quickly assess resistance level for rapid decision-making. DNA extracted from field-collected mosquitoes was used to develop the method. Specific primers were designed manually to match the mutation region and an additional mismatched nucleotide in the penultimate position to increase specificity. Other primers used are common to both wild and mutant types. The allele specific (AS)-LAMP method was compared to the PCR restriction fragment length polymorphism (PCR-RFLP) and real-time PCR (RT-PCR) methods using the genomic DNA of 104 field-collected mosquitoes. The primers designed for LAMP were able to distinguish between the wild type (ace-1 (S) ) and mutated type allele (ace-1 (R) ). Detection time was 50 min for the wild type homozygous and 64 min for the heterozygous. No amplification of the resistant allele took place within the 75-min test period when using the wild type primers. For the ace-1 (R) resistant type, detection time was 51 min for the resistant homozygous and 55 min for the heterozygous. No amplification of the wild type allele took place within the 75-min test period when using the resistant type primers. Gel electrophoresis of LAMP products confirmed that amplification was primer-DNA specific, i.e., primers could only amplify their target specific DNA. AS-LAMP, PCR-RFLP, and RT-PCR showed no significant difference in the sensitivity and specificity of their ace-1 (R) detection ability. The AS-LAMP method could detect the ace-1 (R) mutation within 60 min, which is faster than conventional PCR-RFLP. This method may be used to quickly detect the ace-1 (R) mutation for rapid decision-making, even in less well-equipped laboratories.


Involvement of vimentin in neurite outgrowth damage induced by fipronil in SH-SY5Y cells.
Fipronil, a phenylpyrazole insecticide, is more selective in its potency towards insects than humans and is thus commonly used. In this study, we demonstrated that exposure to fipronil may pose a human health risk. We observed in vitro the shortening of neurite outgrowths of SH-SY5Y neuroblastoma cells upon treatment with fipronil, even at a non-cytotoxic concentration. Fipronil induced apoptosis involving caspase-6, which is an apoptotic effector highly implicated in neurodegenerative diseases. Moreover, at a concentration that did not induce apoptosis, mitochondrial dysfunction and autophagic vacuole formation were detected. Interestingly using proteomics, we identified vimentin to be dramatically expressed by SH-SY5Y cells as a response to fipronil treatment. Not only did the expression of total vimentin increase, different isoforms were observed, indicating alterations in post-translational modifications. Vimentin was localized at the neurite outgrowth, possibly to repair the damage in cellular structure. However at high concentrations of fipronil, vimentin was found in less defined fibrils, in bridge-like formation, and dense surrounding vacuoles. In all, our results indicate that vimentin plays an important role in fipronil-induced neurotoxicity in SH-SY5Y cells.


Antigen-antibody interface properties: composition, residue interactions, and features of 53 non-redundant structures.
The structures of protein antigen-antibody (Ag-Ab) interfaces contain information about how Ab recognize Ag as well as how Ag are folded to present surfaces for Ag recognition. As such, the Ab surface holds information about Ag folding that resides with the Ab-Ag interface residues and how they interact. In order to gain insight into the nature of such interactions, a data set comprised of 53 non-redundant 3D structures of Ag-Ab complexes was analyzed. We assessed the physical and biochemical features of the Ag-Ab interfaces and the degree to which favored interactions exist between amino acid residues on the corresponding interface surfaces. Amino acid compositional analysis of the interfaces confirmed the dominance of TYR in the Ab paratope-containing surface (PCS), with almost two fold greater abundance than any other residue. Additionally TYR had a much higher than expected presence in the PCS compared to the surface of the whole antibody (defined as the occurrence propensity), along with aromatics PHE, TRP, and to a lesser degree HIS and ILE. In the Ag epitope-containing surface (ECS), there were slightly increased occurrence propensities of TRP and TYR relative to the whole Ag surface, implying an increased significance over the compositionally most abundant LYS>ASN>GLU>ASP>ARG. This examination encompasses a large, diverse set of unique Ag-Ab crystal structures that help explain the biological range and specificity of Ag-Ab interactions. This analysis may also provide a measure of the significance of individual amino acid residues in phage display analysis of Ag binding.


10 practical tips for successful outsourcing.
Preparation and professional help are keys to success in negotiating outsourcing contracts: Resist internal and external pressures to rush. Know what you're signing. Insist on accountability. Leave yourself an out. Always sweat the details.


Transvenous revision of leads with cardiac perforation following device implantation-Safety, outcome, and complications.
Cardiac perforation is a rare complication of cardiac implantable electronic device (CIED) implantation. Transvenous revision of perforated leads is associated with the risk of cardiac tamponade and death. Little is known about periprocedural complications and outcome of these patients. All patients referred to our department with evidence or suspicion of cardiac perforation following CIED implantation underwent chest X-ray, transthoracic echocardiography, device interrogation, and, if necessary, a cardiac computed tomography (CT)-scan to diagnose lead perforation and associated complications. Transvenous lead revision (TLR) was performed in all patients with evidence of lead perforation. Patient characteristics, procedural complications, and outcome were recorded and analyzed. Fifty-six patients (75 ± 10 years, 43% male) were diagnosed with cardiac perforation, 34 patients (61%) early within 30 days post-implantation, and 22 patients (39%) thereafter. The most frequent perforation site was the right ventricular (RV) apex (75%), followed by the RV free wall (16%) and the right atrial appendage (9%). A total of 16 patients (29%) presented with severe complications; 12 patients (21%) with pericardial effusion treated by pericardiocentesis before lead revision and four patients (7%) with hematothorax requiring drainage. Late perforations showed significantly more frequent cardiac tamponades (P = .041). TLR was performed without further complications in 54 patients (96%). None of the patients required surgical treatment or experienced in-hospital death. Cardiac perforation following CIED implantation is associated with severe complications in nearly one-third of the cases. Transvenous revision of the perforated lead can safely be performed with a very low complication rate.


The effect of Paf antagonists on bronchial hyperresponsiveness induced by Paf, propranolol or indomethacin.
1. Intravenous administration of platelet activating factor, Paf (600 ng kg-1 h-1) to ventilated anaesthetised guinea-pigs induced bronchial hyperresponsiveness to i.v. acetylcholine. 2. Pretreatment of ventilated, anaesthetised guinea-pigs with the beta-adrenoceptor antagonist, propranolol, or the non-steroidal anti-inflammatory drug, indomethacin, induced bronchial hyperresponsiveness to i.v. histamine. 3. Paf-induced bronchial hyperresponsiveness was significantly attenuated by pretreatment with three different Paf antagonists, CV-3988, BN 52021 and WEB 2086. 4. Pretreatment of guinea-pigs with CV-3988, BN 52021 or WEB 2086 at doses inhibiting Paf-induced bronchial hyperresponsiveness, had no significant effect on propranolol or indomethacin-induced bronchial hyperresponsiveness. 5. It is suggested that bronchial hyperresponsiveness induced by propranolol or indomethacin is not secondary to Paf release in the guinea-pig.


External load can alter the energy cost of prolonged exercise.
The present study was undertaken to examine the energy cost of prolonged walking while carrying a backpack load. Six trained subjects were tested while walking for 120 min on a treadmill at a speed of 1.25 m.s-1 and 5% elevation with a well fitted backpack load of 25 and 40 kg alternately. Carrying 40 kg elicited a significantly higher (p less than 0.01) energy cost than 25 kg. Furthermore, whereas carrying 25 kg resulted in a constant energy cost, 40 kg yielded a highly significant (p less than 0.05) increase in energy cost over time. The study implies that increase in load causes physical fatigue, once work intensity is higher than 50% maximal work capacity. This is probably due to altered locomotion biomechanics which in turn lead to the increase in energy cost. Finally, the prediction model which estimates energy cost while carrying loads should be used with some caution when applied to heavy loads and long duration of exercise, since it might underestimate the actual energy cost.


Ligament tissue engineering: an evolutionary materials science approach.
The anterior cruciate ligament (ACL) is important for knee stabilization. Unfortunately, it is also the most commonly injured intra-articular ligament. Due to poor vascularization, the ACL has inferior healing capability and is usually replaced after significant damage has occurred. Currently available replacements have a host of limitations, this has prompted the search for tissue-engineered solutions for ACL repair. Presently investigated scaffolds range from twisted fiber architectures composed of silk fibers to complex three-dimensional braided structures composed of poly (L-lactic acid) fibers. The purpose of these tissue-engineered constructs is to apply approaches such as the use of porous scaffolds, use of cells, and the application of growth factors to promote ligament tissue regeneration while providing mechanical properties similar to natural ligament.


Molecular basis for interaction of the protein tyrosine kinase ZAP-70 with the T-cell receptor.
The crystal structure of the tandem SH2 domains of human ZAP-70 in complex with a peptide derived from the zeta-subunit of the T-cell receptor reveals an unanticipated interaction between the two domains. A coiled coil of alpha-helices connects the two SH2 domains, producing an interface that constitutes one of the two critical phosphotyrosine binding sites. These and other unique features provide the molecular basis for highly selective association of ZAP-70 with the T-cell receptor.


High-dose fluconazole therapy for cancer patients with solid tumors and candidemia: an observational, noncomparative retrospective study.
Response rates for candidemia treated with standard-dose fluconazole (400 mg/day) are approximately 70%. Higher doses of fluconazole have been recommended for susceptible dose-dependent Candida isolates. Herein, we describe the outcome of 20 patients with solid tumors and candidemia treated with high-dose fluconazole (HDF) at The University of Texas M.D. Anderson Cancer Center (1998-2002). Patients were identified either by searching the microbiology laboratory database or through direct referral from primary oncology services to the Infectious Diseases Consultative Services. A retrospective review of cases was performed. HDF was defined as > or =600 mg/day. Five patients were treated with 600 mg/day, whereas 15 patients received 800 mg/day. Only one patient was neutropenic. The median APACHE II score at the onset of candidemia was 12 (range 6-24). The most common species identified were Candida albicans (eight patients, 40%) and Candida parapsilosis (seven patients, 35%). Of 19 patients whose quantitative data were available, eight (42%) had high-grade candidemia [> or =200 colony forming units (CFU)/ml]. Fifteen (83%) of 18 isolates were fluconazole susceptible, and two (both Candida glabrata) were fluconazole resistant (MIC 64 each) in vitro. Nineteen patients (95%) responded to HDF therapy. The only HDF failure occurred in a patient with C. glabrata (MIC 64.0) infection. The other patient with C. glabrata (MIC 64.0) infection responded to HDF. Central venous catheters were removed from all patients with > or =10 CFU/ml candidemias. All patients with high-grade candidemias responded to HDF. The median duration of HDF therapy was 16 (range 6-42) days. No significant toxicity occurred. Although our data are limited, HDF appears to be well tolerated and may be associated with higher response rates than standard-dose fluconazole in a selected group of patients with solid tumors and candidemia caused by species that are susceptible to this triazole.


Folding and activity of cAMP-dependent protein kinase mutants.
The catalytic subunit of cAMP-dependent protein kinase (PKA) can easily be expressed in Escherichia coli and is catalytically active. Four phosphorylation sites are known in PKA (S10, S139, T197 and S338), and the isolated recombinant protein is a mixture of different phosphorylated forms. Obtaining uniformly phosphorylated protein requires separation of the protein preparation leading to significant loss in protein yield. It is found that the mutant S10A/S139D/S338D has similar properties as the wild-type protein, whereas additional replacement of T197 with either E or D reduces protein expression yield as well as folding propensity of the protein. Due to its high sequence homology to Akt/PKB, which cannot easily be expressed in E. coli, PKA has been used as a surrogate kinase for drug design. Several mutations within the ATP binding site have been described to make PKA even more similar to Akt/PKB. Two proteins with Akt/PKB-like mutations in the ATP binding site were made (PKAB6 and PKAB8), and in addition S10, S139 and S338 phosphorylation sites have been removed. These proteins can be expressed in high yields but have reduced activity compared to the wild-type. Proper folding of all proteins was analyzed by 2D 1H, 15N-TROSY NMR experiments.


Two-level bilateral ultrasound-guided erector spinae plane block for kyphosis corrective surgery - A case report.
Ultrasound-guided erector spinae plane (ESP) block has been used to provide lumbar analgesia. The craniocaudal spread of local anesthetic in this block can be unpredictable in patients with kyphosis due to backward curvature of the spine, which might prevent longitudinal spread. This is a case of a 33-year-old male (60 kg) diagnosed with type 3b kyphosis of the thoracolumbar region at the level of L1 who underwent extended pedicle subtraction osteotomy. ESP block was administered at two different levels, one at T12 (above the angle of the kyphosis L1) and another at L3 (below the angle of kyphosis L1), as the curvature of kyphosis can hinder the longitudinal spread of local anesthetic. Two-level ESP block results in better craniocaudal spread of local anesthetic in a patient undergoing kyphosis correction surgery.


Regulation of mast cell-mediated innate immunity during early response to bacterial infection.
Although the area of research on the role of MCs in innate immunity is relatively new, a number of studies that are reviewed here provide substantial evidence that MCs play a critical role in host immune defense against gram-negative bacteria. The studies show that mast cells have the ability to recognize and engulf bacteria and they release a number of inflammatory mediators including interleukin (IL)-4, IL-6, IL-10, TNF alpha, and leukotrienes in response to bacterial challenge. MC-derived TNF alpha and leukotrienes are shown to be important for bacterial clearance and early recruitment of phagocytic help at the site of infection. Studies directed at elucidating the molecular mechanisms associated with mast cell recognition of bacteria and subsequent events leading to mast cell mediator release revealed that GPI anchored CD48 molecule present on the cell surface of mast cells serves as a receptor for the bacterial adhesion molecule, FimH. The ligation of CD48 receptor by FimH-expressing bacteria results in bacterial uptake into caveolar chambers. This distinct mechanism of bacterial uptake promotes bacterial survival inside the cytosol of the mast cells. Although the exact mechanism(s) of how MC-dependent inflammatory responses are regulated is currently not known, recent studies have shown that complement, CD11 beta/CD18 (Mac-1) and protein tyrosine kinase JAK3, and TLR4 are important for the full expression of MC-dependent innate immunity in mice.


Disappearing abdominal mass: a case report.
The remarkable parallel of the clinical course and scintiscans of an abdominal lymphoma in a 52-year-old man are reported.


3D LiDAR-Based Precision Vehicle Localization with Movable Region Constraints.
This paper discusses a high-performance similarity measurement method based on known map information named the cross mean absolute difference (CMAD) method. Applying the conventional normalized cross-correlation (NCC) feature registration method requires sufficient numbers of feature points, which must also exhibit near-normal distribution. However, Light Detection and Ranging (LiDAR) ranging point cloud data scanned and collected on-site are scarce and do not fulfill near-normal distribution. Consequently, considerable localization errors occur when NCC features are registered with map features. Thus, the CMAD method was proposed to effectively improve the NCC algorithm and localization accuracy. Because uncertainties in localization sensors cause deviations in the localization processes, drivable moving regions (DMRs) were established to restrict the range of location searches, filter out unreasonable trajectories, and improve localization speed and performance. An error comparison was conducted between the localization results of the window-based, DMR⁻CMAD, and DMR⁻NCC methods, as well as those of the simultaneous localization and mapping methods. The DMR⁻CMAD method did not differ considerably from the window-based method in its accuracy: the root mean square error in the indoor experiment was no higher than 10 cm, and that of the outdoor experiment was 10⁻30 cm. Additionally, the DMR⁻CMAD method was the least time-consuming of the three methods, and the DMR⁻NCC generated more localization errors and required more localization time than the other two methods. Finally, the DMR⁻CMAD algorithm was employed for the successful on-site instant localization of a car.


Characterization of cortisol binding sites in chicken liver plasma membrane.
1. The presence of sites specifically binding [3H]cortisol in plasma membrane isolated from chicken liver has been determined. The kinetic parameters of this binding are: Kd = 4.5 nM and Bmax = 2225 fmol/mg protein in presence of 10(-6) M progesterone. 2. The affinities of several natural and synthetic steroids for the membrane binding site respect to the binding of 4 nM [3H]cortisol without competitor increased in the following order: Testosterone less than pregnenone less than dexamethasone less than progesterone less than prednisolone less than corticosterone less than deoxycorticosterone. 3. Other steroids such as estradiol, ouabain and triamcinolone acetonide does not bind to the plasma membrane. 4. Metal ions such as Ca2+ and Mg2+ did not modify the binding of [3H]cortisol. 5. Neither propranolol nor phentolamine, beta- and alpha-adrenergic antagonists affected [3H]cortisol binding to the plasma membranes. 6. The result suggest that the binding site detected is more specific for glucocorticoids and it is different of nuclear glucocorticoid receptor and progesterone receptor.


Reduction of thyrotropin-releasing hormone concentrations in central nervous system of African lungfish during estivation.
Thyrotropin-releasing hormone (TRH) has been implicated as an important modulator of arousal state in mammals. Changes in the content of TRH in several brain regions accompany hibernation in the ground squirrel. In the present study, the involvement of TRH in the regulation of arousal was further investigated in the African lungfish, Protopterus annectens, which contain high concentrations of TRH throughout its central nervous system and enter a hibernation-like state, estivation. Lungfish were divided into three groups. Group 1 was fed normally, group 2 was starved while aquatic, and group 3 was allowed to enter into a state of estivation. After 3 months, the lungfish were sacrificed and the concentrations of TRH, norepinephrine, dopamine, and serotonin were determined in the telencephalon, diencephalon, medulla, and spinal cord. In estivation, there was a significant decline in the concentration of TRH in the diencephalon, with no alteration in other regions. Starvation had no effect on regional TRH concentrations. The concentration of norepinephrine, dopamine, and serotonin did not change in estivation; however, a significant elevation of norepinephrine in the diencephalon and dopamine in the telencephalon was observed in starvation. Starvation and estivation were associated with significant declines in the protein content of the diencephalon and medulla. The estivation-linked decline in TRH in the diencephalon of the lungfish is similar to the decrease in TRH content in the hypothalamus in hibernating ground squirrels. These findings lend further support to the importance of TRH in the regulation of arousal state.


Effect of aging on polyethylene microfiber surface properties and its consequence on adsorption characteristics of 17alpha-ethynylestradiol.
This study assessed the interactive changes to the endocrine disruptor 17 alpha-ethynylestradiol (EE2) triggered by photoaging onto fibrous microplastics frequently found in the environment. The physicochemical property change of the polyethylene (PE) microfiber according to irradiation (i.e. 14 d UV-C (254 nm)) was studied through Fourier transform infrared spectroscopy, scanning electron microscope, and contact angle analysis. Additionally, the EE2 adsorption kinetics experiment was performed for the PE microfiber before and after UV irradiation to assess the change in adsorption characteristics. After UV irradiation, the PE microfiber surface roughness increased, the oxygen-containing functional group (e.g. carbonyl group) increased, and the contact angle (virgin PE: 80.02°, aged PE: 65.13°) decreased. A decrease in the surface hydrophobicity led to a decrease in the adsorption rate of EE2 (virgin PE: k = 0.0105 h-1, aged PE: not calculated). The hydrophobic interaction significantly affects the adsorption behavior of hydrophobic organic pollutants such as EE2 onto MPs, and continuous photo-aging of MPs may cause a new pattern of ecological risk. Therefore, there is a greater necessity for additional research relevant to this issue.


Determination of ketamine and norketamine in hair by micropulverized extraction and liquid chromatography-high resolution mass spectrometry.
Ketamine (KT), primarily used as a general anaesthetic agent in clinical practice, has become very popular in recent years as a recreational drug, due to its dissociative and hallucinogenic effects. A liquid chromatography-high resolution mass spectrometry (LC-HRMS) method has been developed and validated for the quantification of KT and its main metabolite norketamine (NK) in 2.0mg of hair. Sample preparation consisted of a rapid, simultaneous pulverization and extraction step in acidic solution, followed by centrifugation and filtration. Gradient elution was performed by an Atlantis T3 analytical column, and deuterated KT was used as the internal standard. Positive ion electrospray ionization and HRMS determination in full-scan mode were achieved with an Orbitrap mass spectrometer. The method has a linear range of 0.05-50 ng/mg, a limit of quantisation of 0.05 ng/mg and a limit of detection of 0.02 ng/mg for both KT and NK. The validated method was applied for the determination of KT and NK in two authentic hair samples from subjects suspected of taking psychoactive substances. The detection of the metabolite at low concentration gave proof for systemic drug origin and an investigation into the possible presence of further metabolites was performed by means of retrospective screening.


Normo-or slightly overcorrection show better results after medial closing wedge high tibial osteotomy.
The objectives of this study were to evaluate functional results, revision-free survival, and the influence of postoperative alignment on outcomes after MCWHTO. This retrospective study included 27 MCWHTO operated on from 2009 to 2021. Radiographic measurements were performed pre- and postoperatively. The HKA (Hip-Knee-Ankle angle), MPTA (Medial Proximal Tibial angle), LDFA (Lateral Distal Femoral Angle), JLO (Joint Line Obliquity), and JLCA (Joint Line Convergence Angle) were evaluated. The Knee injury and Osteoarthritis Outcome Score (KOOS), the International Knee Society (IKS) Function and Knee Score, and the Subjective Knee Value (SKV) as well as revision-free survival were evaluated. Postoperative alignment and its influence on clinical outcomes were also analysed. The mean follow-up was 61.9 months ± 31.4 (13-124). The HKA, MPTA, and JLCA angles were decreased post-operatively (respectively, Δ = 5.9° ± 2.6, p < 0.001; Δ = 6.1° ± 3.2, p < 0.001 and Δ = 2.5° ± 1.9, p < 0.001). LDFA and JLO were unchanged, post-operatively (respectively, Δ = 0.1° ± 2.2, p = 0.93 and Δ = 1.2° ± 3.3, p = 0.23). Postoperative HKA correlated with knee IKS (R = - 0.15, p = 0.04) and function IKS (R = - 0.44, p = 0.03). Postoperative LDFA correlated with knee IKS(R = 0.8, p < 0.01). Patients with postoperative HKA ≤ 180° had better KOOS (Δ = 12.3, p = 0.04) and IKS function (Δ = 28.1, p < 0.01) than those with HKA > 180°. Functional results and revision-free survival after MCWHTO are satisfactory when the deformity is located in the proximal tibia. The joint line obliquity is not significantly altered with small tibial correction and, obtaining an overall neutral or slightly varus alignment under the conditions of this study allowed an improvement in the postoperative clinical scores. The literature is still inconclusive on the ideal alignment for valgus deformities and larger series are needed to draw definitive conclusions. IV, case series.


Automated classification of RNA 3D motifs and the RNA 3D Motif Atlas.
The analysis of atomic-resolution RNA three-dimensional (3D) structures reveals that many internal and hairpin loops are modular, recurrent, and structured by conserved non-Watson-Crick base pairs. Structurally similar loops define RNA 3D motifs that are conserved in homologous RNA molecules, but can also occur at nonhomologous sites in diverse RNAs, and which often vary in sequence. To further our understanding of RNA motif structure and sequence variability and to provide a useful resource for structure modeling and prediction, we present a new method for automated classification of internal and hairpin loop RNA 3D motifs and a new online database called the RNA 3D Motif Atlas. To classify the motif instances, a representative set of internal and hairpin loops is automatically extracted from a nonredundant list of RNA-containing PDB files. Their structures are compared geometrically, all-against-all, using the FR3D program suite. The loops are clustered into motif groups, taking into account geometric similarity and structural annotations and making allowance for a variable number of bulged bases. The automated procedure that we have implemented identifies all hairpin and internal loop motifs previously described in the literature. All motif instances and motif groups are assigned unique and stable identifiers and are made available in the RNA 3D Motif Atlas (http://rna.bgsu.edu/motifs), which is automatically updated every four weeks. The RNA 3D Motif Atlas provides an interactive user interface for exploring motif diversity and tools for programmatic data access.


Incontinentia pigmenti: clinical and neuroradiologic features.
Developmental brain malformations and destructive processes of unknown etiology were described in incontinentia pigmenti (IP). Two patients, a male and a female, with characteristic skin lesions and central nervous system (CNS) involvement are reported. Neuroradiological examinations revealed hypoplasia of corpus callosum, neuronal heterotopias, and periventricular white matter damage. No specific infectious, inflammatory, or metabolic abnormalities were identified. These neuroradiographic findings may suggest that an ischemic pathogenetic mechanism occurred prenatally. We speculate that the brain damage in IP may occur during CNS development and in successive stages. Magnetic resonance imaging appears more useful to detect white matter lesions and brain malformations in patients with IP.


Ocular toxoplasmosis in the immunocompromised host.
Disseminated toxoplasmosis is a well-known complication of immunodeficiency states, including those induced by malignancies, steroid and cytotoxic drug therapy, and AIDS. In immunodeficient patients, toxoplasmic infections of the eye are less common than toxoplasmic infections of other organs for unknown reasons. When ocular toxoplasmosis does occur in the immunodeficient host, or if immunosuppressive therapy is administered to patients with active disease, widespread tissue destruction by proliferating organisms may result. Immunodeficiency alone may not be sufficient, however, to cause reactivation of encysted organisms in retinochoroidal scars. Ocular toxoplasmosis in the immunocompromised host presents difficult problems in diagnosis and management. There may be a variety of clinical lesions, including single foci of retinochoroiditis in one or both eyes, multifocal lesions, or diffuse areas of retinal necrosis. The majority of lesions do not arise from the borders of preexisting scars, which suggests that they result from acquired infection or dissemination of organisms from nonocular sites of disease. Toxoplasma gondii may infect iris, choroid, and vitreous-tissues that are not usually infected in the immunocompetent host. Ocular lesions appear to respond to standard antiparasitic drug therapies, but continued treatment is probably necessary to prevent reactivation of disease in the most immunocompromised patients. The best treatment regimens have yet to be determined. Histopathologic studies show little retinal inflammation; therefore anti-inflammatory drugs, such as oral steroids, probably have no role in the management of infection.


Examining appearance pressures, thinness and muscularity internalizations, and social comparisons as correlates of drive for muscularity and thinness-oriented disordered eating in Chinese heterosexual men and women: Testing an integrated model.
Given existing empirical support for the tripartite influence and social comparison models to explain variance in body dissatisfaction and disordered eating across diverse populations in the Western context, research is needed to describe these models in non-Western populations. The present study tested an integrated model inclusive of appearance pressures, thinness and muscularity internalizations, and social comparisons in relation to drive for muscularity and thinness-oriented disordered eating in heterosexual Chinese adult men (n = 510) and women (n = 473). Separated by gender, two integrated models had good fit statistics per structural equation modeling. In men and women, higher appearance pressures were uniquely related to higher drive for muscularity and thinness-oriented disordered eating. In men and women, higher thinness and muscularity internalizations were uniquely related to higher thinness-oriented disordered eating and drive for muscularity, respectively. In men and women, higher upward body image comparisons were uniquely related to higher drive for muscularity, and in men only, higher downward body image comparisons were uniquely related to higher thinness-oriented disordered eating. The present findings, which extend theories of eating pathology, may aid in improving treatment efforts for body dissatisfaction and disordered eating in heterosexual Chinese adult men and women.


[Plastic surgery of postoperative defects in patients with tumors of the maxillofacial area].
Defects resulting from surgical treatment for maxillo-facial tumors of various origin were studied. Procedures for reconstruction versus site are discussed and immediate results are presented. Reconstruction should be performed in early postoperative period to assure normal food passage.


Persistent Atrial Fibrillation Ablation With Contact Force-Sensing Catheter: The Prospective Multicenter PRECEPT Trial.
This study sought to evaluate the safety and effectiveness of catheter ablation of persistent atrial fibrillation (PsAF) using a porous tip contact force-sensing catheter. Although the safety and effectiveness of catheter ablation of paroxysmal atrial fibrillation are established, there are limited data on outcomes in patients with PsAF. As such, no ablation catheter is currently approved by the Food and Drug Administration for PsAF ablation. The prospective, multicenter, nonrandomized PRECEPT (Prospective Review of the Safety and Effectiveness of the THERMOCOOL SMARTTOUCH SF Catheter Evaluated for Treating Symptomatic PersistenT AF) study was conducted at 27 sites in the United States and Canada. Enrollment criteria included documented symptomatic PsAF and nonresponse or intolerance to ≥1 antiarrhythmic drug (Class I or III). An individualized treatment approach was used including pulmonary vein isolation with ablation of additional targets permitted at the investigators' discretion. To optimize treatment outcomes, a 3-month post-ablation medication adjustment period followed by a 3-month therapy consolidation period were included. Arrhythmia recurrences were stringently monitored by monthly and symptomatic transtelephonic monitoring, electrocardiography, and Holter monitoring for up to 15 months after ablation. Of 381 enrolled participants, 348 had the investigational catheter inserted and underwent ablation. The primary adverse event rate was 4.1% (15 events in 14 participants). Kaplan-Meier analyses estimated a primary effectiveness success rate of 61.7% and a clinical success rate of 80.4% at 15 months. The results demonstrate the clinical safety and effectiveness of PsAF ablation using contact force-sensing technologies. The primary adverse event was within the expected range and similar to those reported in historical studies of paroxysmal AF ablation. (Prospective Review of the Safety and Effectiveness of the THERMOCOOL SMARTTOUCH SF Catheter Evaluated for Treating Symptomatic PersistenT AF; NCT02817776).


The COVID-19 pandemic and its repercussions in our interventional radiology unit.
This paper aims to describe our experience in an interventional radiology unit in a hospital in Spain that was severely affected by the COVID-19 pandemic. To this end, we did a prospective observational study of 20 consecutive patients with COVID-19 who underwent 21 interventional radiology procedures between March 13, 2020 and May 11, 2020. We describe the measures taken to reorganize the work and protective measures, as well as the repercussions of the situation on our unit's overall activity and activity in different phases. The COVID-19 pandemic has represented a challenge in our daily work, but learning from our own experience and the recommendations of the Spanish radiological societies (SERVEI and SERAM) has enabled us to adapt successfully. Our activity dropped only 22% compared to the same period in 2019.


Diagnoses received by narcolepsy patients in the year prior to diagnosis by a sleep specialist.
Narcolepsy is a neurological disorder whose clinical features include excessive daytime sleepiness, hypnagogic hallucinations, cataplexy, sleep paralysis, and disrupted nocturnal sleep. It has been shown that there may be quite a long interval between the onset of symptoms, and the correct diagnosis. We tested the hypothesis that given their severe symptomatology, these patients would have been diagnosed more often with a variety of psychiatric and neurologic conditions than controls in the year prior to confirmation of their narcolepsy diagnosis. Using the Province of Manitoba Health database, we compared the diagnoses made in the year prior to initial sleep disorder center evaluation of 77 patients with narcolepsy (33 males, 44 females) and 1,155 matched control subjects from the general population. Sleep disorders center in University-based teaching hospital N/A. N/A. Patients were much more likely than controls to be diagnosed with mental disorders (Odds ratio (OR) = 4.0645; 95% confidence limit (CL) = 2.4671-6.6962; p<0.0001) and nervous system disorders (OR= 5.0495; CL = 3.0606 -8.3309; p<0.0001) and there was a trend towards more injuries in these patients (OR =1.6316; CL = 0.9857-2.7007; p=0.0514). We found that cases were statistically much more likely than controls to have received a diagnosis for neurotic disorders (17% of cases), depression (16%), personality disorders (3%) and adjustment reaction (4%). Although the cases had twice as many doctor visits as the controls (9.3 +/- 0.97 (sem) vs. 4.8 +/- 0.17 p<0.0001), only 38% of them had received a diagnosis of narcolepsy in the year prior to sleep specialist evaluation. Neurologists had the highest "success rate" for correct diagnosis: neurologists diagnosed narcolepsy in 55% of the cases they had seen. The other medical practitioners diagnosed narcolepsy in a much smaller percentage of the cases they had seen: 23.5% for internists (excluding neurologists), 21.9% for general practitioners, 11.1% for psychiatrists, and 0% for pediatricians. In the year prior to documentation of narcolepsy in a sleep disorders center, patients with narcolepsy were diagnosed with a wide variety of mental and neurologic disorders. Our findings are supportive of either the coexistence of these disorders in narcolepsy patients or a high frequency of missed diagnosis by their clinicians. The latter may help explain the very long interval between onset of symptoms and correct diagnosis.


Genetic Diversity and Population Structure of Juniperus seravschanica Kom. Collected in Central Asia.
Juniperus seravschanica Kom. is a species that grows widely in the mountain ranges from Central Asia to Oman. It is an important tree for the formation of shrub-forest massifs in mountainous areas and for draining and fixing soils from middle to high altitudes. A comprehensive study of the species' genetic diversity and population structure is a basic approach to understanding the current status of J. seravschanica resources for the development of future conservation strategies. Samples from 15 populations of J. seravschanica were collected from the mountain ranges of Uzbekistan, Kyrgyzstan, and Kazakhstan. The genetic diversity and population structure of 15 Central Asian populations of J. seravschanica were assessed using 11 polymorphic simple sequence repeat (SSR) markers. Genetic diversity parameters, including the number of alleles (na), the effective number of alleles (ne), Shannon's information index (I), the percentage of polymorphic loci (PPL), Nei's genetic diversity index (Nei), principal coordinate analysis (PCoA), etc., were evaluated. The analysis of 15 J. seravschanica populations based on 11 polymorphic SSRs detected 35 alleles. The average PIC value was 0.432, and the highest value (0.662) was found in the JT_40 marker. Nei's genetic diversity index for the J. seravschanica populations was 0.450, ranging from 0.407 (population 14) to 0.566 (population 4). The analysis of molecular variance (AMOVA) showed that 90.3% of total genetic variation is distributed within the population. Using the alleles of all the populations, the gene flow (Nm) was found to be 4.654. Population structure analysis revealed poor clustering in the studied populations and confirmed our AMOVA results. The output of this work can be efficiently used for the maintenance of the species across the Central Asian region.


Pineal-induced alterations in reproductive function and pituitary prolactin in the female rat: the effects of bilateral superior cervical ganglionectomy and nervi conarii transection.
The reproductive organs of female rats subjected to blinding and and anosmia were hypotrophic while pituitary prolactin stores were markedly depressed in these animals. All of the effects of dual sensory deprivation were reversed by either superior cervical ganglionectomy or nervi conarii transection. It was concluded that both of these surgical procedures were as effective as pinealectomy in reversing the pineal-induced alterations in the reproductive physiology of the blind-anosmic female rat.


Chronic Lyme disease: a review.
Studies have shown that most patients diagnosed with chronic Lyme disease either have no objective evidence of previous or current infection with Borrelia burgdorferi or are patients who should be classified as having post-Lyme disease syndrome, which is defined as continuing or relapsing nonspecific symptoms (such as fatigue, musculoskeletal pain, and cognitive complaints) in a patient previously treated for Lyme disease. Despite extensive study, there is currently no clear evidence that post-Lyme disease syndrome is caused by persistent infection with B burgdorferi. Four randomized placebo-controlled studies have shown that antibiotic therapy offers no sustained benefit to patients who have post-Lyme disease syndrome. These studies also showed a substantial placebo effect and a significant risk of treatment-related adverse events. Further research to elucidate the mechanisms underlying persistent symptoms after Lyme disease and controlled trials of new approaches to the treatment and management of these patients are needed.


Study of the physiological mechanism of delaying cucumber senescence by wheat intercropping pattern.
This paper investigates the physiological mechanism of the effect of delaying cucumber senescence on protein levels under the experimental model of monoculture and wheat intercropping. We analyzed cucumber roots for differential protein expression, and protein types were identified by core technology of proteomics. 45 differential proteins were identified as being differentially expressed between monoculture and intercropping of wheat, which were involved in carbohydrate metabolism, antioxidant and stress response, growth and development regulation, biological information transfer, and nucleic acid biosynthesis. The results showed the rate of photosynthesis of cucumber was increased under wheat intercropping pattern due to three enzymes being up-regulated. The respiration of cucumber was reduced when five enzymes were down-regulated. The antioxidant and resistant capacity of cucumber was enhanced significantly under wheat intercropping pattern because two enzymes were down-regulated while the activity of four other antioxidant enzymes was up-regulated. Intercropping wheat could delay the senescence of cucumber leaves by increasing the expression of IPT gene, reducing the expression of PAO and ETR2 gene, and inhibiting the expression of Cu/Zn-SOD and GS1 gene in later stages. Two proteins related to growth and development in cucumber were up-regulated, and one was down-regulated, while two proteins related to nucleic acid biosynthesis and chaperonin showed obvious down-regulation under wheat intercropping. Therefore, the growth and development was improved and senescence of cucumber could be delayed. Under intercropping pattern, the fresh weight, chlorophyll content, photosynthetic rate, and peroxidase activity of cucumber plants were higher than those of cucumber monoculture groups. Thus, the senescence of cucumber could be delayed under wheat intercropping by regulating its physiological mechanisms, such as by improving photosynthesis, reducing respiratory consumption, slowing the cell apoptosis rate, and enhancing the antioxidant and resistant capacity significantly, etc.


Aspergillus westerdijkiae polyketide synthase gene "aoks1" is involved in the biosynthesis of ochratoxin A.
Ochratoxin A (OTA) is a potential nephrotoxic, teratogenic, immunogenic, hepatotoxic and carcinogenic mycotoxin, produced by Aspergillus westerdijkiae NRRL 3174. Herein we describe the characterization of a putative OTA-polyketide synthase gene "aoks1", cloned by using gene walking approach. The predicted amino acid sequence of the 2kb clone display 34-60% similarities to different polyketide synthase genes including lovastatine biosynthesis gene "lovb" in A. terreus, compactin biosynthesis gene "mlcA" in Penicillium citrinum and OTA biosynthesis gene "otapksPN" in P. nordicum. Based on the reverse transcription PCR and kinetic secondary metabolites production studies, aoks1 expression was found to be associated with OTA biosynthesis. Further a mutant, in which the aoks1 gene was inactivated by Escherichia coli hygromycin B phosphotransferase gene, lost the capacity to produce OTA, but still producing mellein. To our knowledge this report describes for the first time characterization of a gene involved in OTA biosynthesis, with the information about mellein which was proposed in the literature to be an intermediate OTA. This study also suggests that aoks1 may be the second polyketide synthase gene required for OTA biosynthesis in A. westerdijkiae NRRL 3174.


Prevalence of feline immunodeficiency virus and other retroviral infections in sick cats in Italy.
Two hundred and seventy-seven sick pet cats living in Italy were tested for antibodies to feline immunodeficiency virus (FIV) and for feline leukemia virus (FeLV) antigen. Overall, 24% of the cats resulted positive for anti-FIV antibody and 18% for FeLV antigen. FIV was isolated from the peripheral mononuclear blood cells of ten out of 15 seropositive cats examined and from one out of eight saliva samples. No FIV isolations were obtained from six serum samples cultured. Feline syncytium forming virus (FeSFV) could be isolated from blood and/or saliva in ten out of 11 FIV seropositive cats examined, in six out of nine FeLV antigen positive cats, in two cats found positive for both infection markers, and in three out of 11 cats negative for both markers. Thus, the probability of isolating FeSFV was enhanced by infection with other exogenous retroviruses.


Food variety--a good indicator of nutritional adequacy of the diet? A case study from an urban area in Mali, West Africa.
This study assesses whether a simple count of food items and food groups can predict the nutritional adequacy of the diet in an economically poor country. A three-day weighed record of children. Koutiala town, in Southeastern Mali. Seventy-seven children, 13-58 months of age. One child was excluded owing to an extraordinarily low food variety. The study was conducted in April August 1995. Data from this study were used to create two different indices: Food Variety Score (FVS), a simple count of food items, and Dietary Diversity Score (DDS), a count of food groups. Mean Adequacy Ratio (MAR) was calculated as an indicator for nutrient adequacy, and used to validate FVS and DDS. Mean (s.d.) FVS was 20.5 (3.8) and mean (s.d.) DDS was 5.8 (1.1). A positive correlation was found both between FVS and MAR (Pearson 0.33, P < 0.001) and DDS and MAR (Pearson 0.39, P < 0.001). With cut-off points for FVS at 23 and for DDS at 6, the indices have high ability to identify those with a nutritionally inadequate diet. MAR increased with increasing FVS and DDS. FVS needs to be at least 15 or DDS at least 5 to give a satisfactory MAR. Although a simple count of food items or food groups cannot give a full picture of the adequacy of the nutrient intake, the results from this study show that the food scores can give a fairly good assessment of the nutritional adequacy of the diet, particularly if combined. Such indicators are important for identification of vulnerable groups in areas where people normally eat from a shared bowl, which makes detailed dietary intake studies difficult, time consuming and expensive.


Development of an integrated aerobic granular sludge MBR and reverse osmosis process for municipal wastewater reclamation.
The reclamation of municipal wastewater to obtain high-grade product water is a growing need due to the pressing global water shortage. However, the existing municipal wastewater treatment plants (WWTPs) with the conventional activated sludge process as a core is not a sustainable engineering solution towards future water sustainability. To tackle such an emerging water-wastewater nexus, a ferrous-assisted aerobic granular sludge membrane bioreactor and reverse osmosis (AGSMBR-RO) process was developed for municipal wastewater reclamation. Results show that about 99.9%, 99.7% and nearly 100% of dissolved organic carbon (DOC), ammonium-N and total phosphorus (TP), respectively, could be removed in the ferrous-assisted AGSMBR-RO process, while the product water could meet the typical NEWater quality of Singapore with respect to the parameters analysed in this study. Moreover, it was found that an addition of 6 mg/L of ferrous could improve the stability of aerobic granular sludge (AGS) through the coagulation and flocculation of suspended flocs as well as phosphorus removal. These in turn led to reduced membrane fouling in both AGSMBR and RO units. Consequently, the proposed process is a promising alternative for municipal wastewater reclamation.


Imaging and Diagnostic Challenges in a Patient With Refractory Hypoglycemia Caused by Insulinomas Related to Multiple Endocrine Neoplasia Type 1.
Insulinoma is a rare neuroendocrine tumor. It may occur sporadically or as part of the genetic tumor syndrome multiple endocrine neoplasia type 1 (MEN1). Diagnosis is challenging because of the small size of insulin producing tumors that lead to hyperinsulinemia. Advances in imaging modalities may provide more accurate diagnosis of primary tumors, metastasis, and tumor functional status. Advances allow for improved medical and surgical management with new tools for research of neuroendocrine tumors. Surgical excision of the primary tumor is often curative; however, insulinomas in MEN1 syndrome are often multifocal with a high rate of recurrence presenting unique challenges in management. Here, we present the case of a 34-year-old male with recurrent hypoglycemic episodes and hyperparathyroidism diagnosed with multiple pancreatic insulinomas secondary to MEN1. Furthermore, we provide a brief review of the literature and discuss the approach to diagnosis and management in patients with MEN1 syndrome and future areas of investigation.


Use of CPAP in patients with obstructive sleep apnea admitted to the general ward: effect on length of stay and readmission rate.
Obstructive sleep apnea (OSA) has been associated with multiple cardiovascular comorbidities. Despite increased awareness of OSA and its treatments, the management of OSA in the hospital setting remains below expectations. We retrospectively reviewed the demographics, clinical characteristics, and hospital course on 413 consecutive patients with a history of OSA on domiciliary CPAP therapy admitted to the general medical ward and analyzed the prevalence of CPAP use and its effect on length of stay (LOS), 30-day readmission rate, and time-to-readmission in our tertiary care teaching hospital. Of the 413 study participants, 264 (64.0 %) patients were receiving CPAP during their hospital admission. Patients who were receiving CPAP therapy during their hospitalization had a significantly higher body mass index (BMI) (41.4 vs. 36.8 kg/m(2), p < 0.001) and were more likely to be African-American (p = 0.01) and have congestive heart failure (CHF) (42.0 vs. 31.0 %, p = 0.038) peripheral vascular disease (PVD) (26.0 vs. 15.0 %, p = 0.013), and uncomplicated diabetes mellitus (p = 0.001) than those who were not. CPAP therapy in the hospital setting did not affect LOS (4.7 vs. 4.0 days, p = 0.291), readmission rate (11.0 % for both groups), or time-to-readmission (20.8 vs. 22.3 days, p = 0.762). The majority of patients who are on domiciliary CPAP therapy were receiving CPAP therapy while admitted to the general medical ward of a tertiary care academic hospital. Presence of comorbid conditions such as obesity and certain cardiovascular diseases may have increased the likelihood of prescribing CPAP therapy while in the hospital. In-hospital CPAP therapy did not appear to significantly influence short-term outcomes such as hospital LOS, readmission rate, or time-to-readmission.


Whole grain consumption and body mass index in adult women: an analysis of NHANES 1999-2000 and the USDA pyramid servings database.
To examine the relationship between whole grain consumption and body mass index (BMI) in a sample of American adult women. Dietary intake data from the National Health and Nutrition Examination Survey 1999-2000 were linked to the USDA Pyramid Servings Database. Women 19 years of age and older (n = 2,092) were classified into groups based on their average whole grain (WG) intake: 0 servings, more than 0 but less than 1 serving, and > or =1 servings per day. Within these classifications, mean BMI, mean waist circumference and percent overweight/obese (BMI > or = 25) were identified as primary dependent variables. Regression and logistic regression analyses were used to assess associations between BMI, waist circumference and percent of the population overweight/obese (BMI > or =25) and WG consumption. Women consuming at least one serving of WG had a significantly lower mean BMI and waist circumference than women with no WG consumption (p < 0.05). Multiple regression analysis showed a significant inverse relationship between BMI and whole grain intake after adjustment for age, energy intake, dietary fiber and alcohol intake (p = 0.004). This effect was mildly attenuated but remained significant after further adjustment for level of physical activity, smoking status, ethnicity and education (p = 0.018). The odds ratio for having a BMI > or = 25 was 1.47 (95% CI 1.12-1.94; p for trend 0.013) for women consuming no WG compared to those consuming at least one serving, after adjustment for all covariates. These data support other research suggesting increased WG intake may contribute to a healthy body weight in adult women.


Changes in qualitative composition of white muscle with nutritional status of Atlantic cod, Gadus morhua.
Proteins from white muscle are mobilized to cover energy requirements during long-term starvation in fish. Using SDS-polyacrylamide gel electrophoresis, we compared the soluble and insoluble fractions of white muscle proteins from fed and starved Atlantic cod, Gadus morhua, to establish whether preferential preservation or degradation of specific proteins occurred during starvation. While starvation induced no qualitative changes in the electrophoretic pattern of the myofibrillar fraction, our results document differential decreases in the levels of soluble proteins during fasting. Moreover, immunoblot analysis using a monoclonal antibody directed against actin, showed a marked accumulation of this protein in the sarcoplasmic fraction of starved individuals, most likely due to myofibrillar degradation.


Contested science: Individuals with higher metacognitive insight into interpretation of evidence are less likely to polarize.
Societal polarization over contested science has increased in recent years. To explain this development, political, sociological, and psychological research has identified societal macro-phenomena as well as cognitive micro-level factors that explain how citizens reason about the science. Here we take a radically different perspective, and highlight the effects of metacognition: How citizens reason about their own reasoning. Leveraging methods from Signal Detection Theory, we investigated the importance of metacognitive insight for polarization for the heavily contested topic of climate change, and the less heavily contested topic of nanotechnology. We found that, for climate change (but not for nanotechnology), higher insight into the accuracy of own interpretations of the available scientific evidence related to a lower likelihood of polarization over the science. This finding held irrespective of the direction of the scientific evidence (endorsing or rejecting anthropogenicity of climate change). Furthermore, the polarizing effect of scientific evidence could be traced back to higher metacognitive insight fostering belief-updating in the direction of the evidence at the expense of own, prior beliefs. By demonstrating how metacognition links to polarization, the present research adds to our understanding of the drivers of societal polarization over science.


Technologies for detecting metals in single cells.
In order to fully understand the metallomics of an organism, it is essential to know how much metal is present in each cell and, ideally, to know both the spatial and chemical distributions of each metal (i.e., where within the cell is a metal found, and in what chemical form). No single technique provides all of this information. This chapter reviews the various methods that can be used and the strengths and weaknesses of each.


Pleural effusion in a patient with COVID-19 pneumonia and lung cancer: A case report.
Coronavirus disease is a global infection presenting with a variety of clinical features, both pulmonary and extrapulmonary symptoms. Its diagnosis depend on clinical symptoms, laboratory tests, and the typical CT chest scan findings. In this report, a 72-year-old patient (known to have lung cancer) present with pleural effusion as possible sequel of COVID-19 pneumonia. Pleural effusion may present as a complication in a patient with coronavirus disease, but there is poor evidence to this claim in the medical literature.


Tapwater iontophoresis in the treatment of hyperhidrosis. Use of the Drionic device.
Tapwater iontophoresis is a recognized method of reducing sweat in various parts of the body. The Drionic Device is a battery-operated method of inducing tapwater iontophoresis. This simple device may be used at home and is effective in reducing hyperhidrosis for as long as 6 weeks.


Correction to: Optimal oil stockpiling, peak oil, and general equilibrium: case study of South Asia (oil importers) and Middle East (oil supplier).
The correct affiliation of the 1st Author is shown in this paper.


Sleepy driving: accidents and injury.
The study goals were to evaluate the associated risks of driving and to assess predictors of accidents and injury due to sleepiness. A cross-sectional Internet-linked survey was designed to elicit data on driving habits, sleepiness, accidents, and injuries during the preceding 3 years. Statistical analysis included logistic models with covariate-adjusted P values of <0.01 (odds ratios and 95% confidence intervals or limits). Independent accident predictors were sought. Responses from 10,870 drivers were evaluated. The mean +/- SD age was 36.9 +/- 13 years; 61% were women and 85% were white. The Epworth Sleepiness Scale overall baseline score was 7.4 +/- 4.2 (for drivers with no accidents) and ranged to 12.7 +/- 7.2 (for drivers with > or = 4 accidents) (P = < 0.0001). Twenty-three percent of all respondents experienced > or = 1 accident. Among respondents who reported > or = 4 accidents, a strong association existed for the most recent accident to include injury (P < 0.0001). Sleep disorders were reported by 22.5% of all respondents, with a significantly higher prevalence (35%, P = 0.002) for drivers who had been involved in > or = 3 accidents. Factors of sleepiness were strongly associated with a greater risk of automobile accidents. Predictors were identified that may contribute to accidents and injury when associated with sleepiness and driving.


Using PCDD/F congener patterns to determine the source of elevated TEQ concentrations in cows milk: a case study.
PCDD/F congener profiles have been used to determine the source of elevated TEQ concentrations in cows' milk collected in the vicinity of several industrial sites. Principal components analysis and modelling of the air-to-milk transfer of individual PCDD/F congeners have shown that the milk fingerprint was related to that of sediment taken from the River Rother adjacent to the farm where the cows had grazed. It is suggested that sediment from the river had been washed onto the grazing land during periods of flooding, and this had been ingested by the grazing cows. This pathway could represent an important route of contaminant intake for livestock grazing on the regularly inundated floodplains of rivers containing contaminated sediments.


Isotachophoresis evaluation of synthesis of intrathecal IgG subfractions in multiple sclerosis.
Cerebrospinal fluid (CSF) and serum samples from patients with multiple sclerosis and other neurological diseases were examined by capillary isotachophoresis (ITP). The percentage and the rate of synthesis of CSF IgG which migrated slowly with ITP were calculated. CSF specimens of most patients with multiple sclerosis contained increased percentages of slowly migrating IgG (slow IgG), corresponding to IgG oligoclonal bands in the "high-alkaline" region on isoelectric focusing. The patients with multiple sclerosis were found to have increased intrathecal synthesis of slow IgG, which correlated closely with the rate of intrathecal CNS IgG synthesis calculated by Tourtellotte's formula.


Vulvar pilonidal sinuses: treatment of a rare cause of vulvar pain.
It is hoped that if cases of vulvar pilonidal sinus (PS) are known to readers of this article, they will be called to the writer's attention and that further studies of the origin and cause of these anterior pathologic developments will result. A delay in diagnosis and treatment of this relatively harmless condition can lead to complications like abscess formation. Early diagnosis, complete excision and elimination of the anatomical factor that cause accumulation of hair fragments are followed by an excellent prognosis.


Herniamed: an internet-based registry for outcome research in hernia surgery.
Despite the high frequency of hernia surgery procedures and continuous improvements, thanks to new hernia meshes and fixation techniques, in Germany, for example, the recurrence rate and rate of chronic inguinal pain after inguinal surgery are more than 10% far too high. Introduction of a hernia register in Denmark led to a significant reduction in the recurrence rate. The aim of a hernia registry as an application-oriented outcome research tool is to monitor and evaluate (concomitant research) how the knowledge gleaned from evidence-based science is implemented in the everyday clinical setting and, ultimately, investigate its effectiveness (outcome research). The new Internet-based English- and German-language registry for the entire spectrum of inpatient and outpatient hernia surgery is designed to improve the quality of patient care and provide valid data on outcome research. Via the Internet, all relevant patient data (comorbidities, previous operations, staging, specific surgical technique, medical devices used, perioperative complications and follow-up data) can be entered into the registry database. The participating hospitals and surgeons can at any time view their own data by means of an evaluation statistics tool. The outcome research project Herniamed focuses on inguinal hernias, umbilical hernias, incisional hernias, epigastric hernias, parastomal hernias and hiatus hernias. The online-based outcome research registry meets the most stringent data protection criteria. With the Internet-based English- and German-language hernia register, a new instrument is now available for outcome research in hernia surgery.


Reduced antigenicity of formaldehyde-fixed arterial allografts and their potential clinical application.
This study was performed to examine the antigenicity of formaldehyde-fixed arterial allografts and their potential clinical application for hemodialysis access. Human femoral arteries were fixed with formaldehyde. New Zealand white rabbits were immunized with antigens prepared from fixed or unfixed grafts followed by anti-serum collection. The antigenicity of fixed grafts was evaluated using double-diffusion in two dimensions. Seventy-six fixed allografts were transplanted into 50 patients with end-stage renal diseases as hemodialysis access and monitored for acute rejection, blood flow, and patency rates. Anti-HLA antibody was evaluated using a panel-reactive antibody (PRA) assay in 6 patients. Formaldehyde-fixed grafts demonstrated reduced antigenicity as determined using double-diffusion in two dimensions when compared with unfixed grafts. Histological evaluation of allografts demonstrated absence of cell-mediated acute rejection after transplantation. Allografts demonstrated satisfactory blood flow with primary and secondary patency rates at 1, 2, and 3 years after allotransplantation. Serum collected from 6 patients after transplantation (mean, 3.3 +/- 3.2 years) did not demonstrate increased anti-HLA antibody levels. The allograft intimal hyperplasia was characterized by proliferation of actin-expressing smooth muscle cells. This study demonstrated that formaldehyde fixation reduced the antigenicity of arterial allografts. Acute rejection was absent after allotransplantation. The lack of increased PRA levels after allotransplantation suggested that allo-sensitization is not induced by fixed allografts. The use of formaldehyde-fixed allografts as hemodialysis access demonstrated blood flow and patency rates. Intimal hyperplasia of allografts is characterized by proliferation of actin-expressing cells. Our results supported the clinical use of formaldehyde-fixed arterial allografts for hemodialysis access.


Urea-induced modification of cytochrome c flexibility as probed by cyanide binding.
Cyanide binding to cytochrome c was monitored by absorption spectroscopy from neutral to acidic pH in the presence of urea. These results were compared with acid-induced unfolding at corresponding urea concentration monitored by absorption spectroscopy and circular dichroism. The association rate constant ka increased 20-fold when the concentration of urea was raised from 0M to 6M at neutral pH. However, the secondary structure of the protein was not affected, i.e. there was no striking conformational change in these urea concentrations at neutral pH. At the pH that was very close to the pK of acid-induced unfolding, the ka value reached its maximum (ka,max) in all urea concentrations. Interestingly, the ka,max value increased exponentially with increasing urea concentrations. These results are interpreted in terms of a change in the flexibility of the least stable part of the cyt c structure that is responsible for the Fe-S(Met80) bond disruption and for ligand binding to heme iron.


Analysis of the stimulatory factor of RNA polymerase II in the initiation and elongation complex.
The interaction between RNA polymerase II and its specific stimulatory factor S-II was studied using each component purified to near homogeneity. It was found that RNA polymerase II and S-II form a complex in the absence of DNA and nucleoside triphosphates, although their interaction was weak. This interaction became stable in the presence of a template and substrates, and a stoichiometric amount of S-II was found to remain in the transcription machinery during RNA chain elongation.


Influence of hydrodynamic interactions on lane formation in oppositely charged driven colloids.
The influence of hydrodynamic interactions on lane formation of oppositely charged driven colloidal suspensions is investigated using Brownian dynamics computer simulations performed on the Rotne-Prager level of the mobility tensor. Two cases are considered, namely sedimentation and electrophoresis. In the latter case the Oseen contribution to the mobility tensor is screened due to the opposite motion of counterions. The simulation results are compared to that resulting from simple Brownian dynamics where hydrodynamic interactions are neglected. For sedimentation, we find that hydrodynamic interactions strongly disfavor laning. In the steady state of lanes, a macroscopic phase separation of lanes is observed. This is in marked contrast to the simple Brownian case where a finite size of lanes was obtained in the steady state. For strong Coulomb interactions between the colloidal particles a lateral square lattice of oppositely driven lanes is stable similar to the simple Brownian dynamics. In an electric field, on the other hand, the behavior is found in qualitative and quantitative accordance with the case of neglected hydrodynamics.


A cell surface marker for neural crest and placodal cells: further evolution in peripheral and central nervous system.
The recent production of a monoclonal antibody (NC-1) recognizing migrating avian neural crest (NC) cells (M. Vincent, J. L. Duband , and J. P. Thiery , Dev. Brain Res. 9, 235-238, 1983) allowed us to detail their migration pathways at the trunk level of the chick embryo. Three routes can be recognized: NC cells facing the bulk of the somite accumulate to form a spinal ganglion, those facing the intersomitic space can readily reach periaortic areas to contribute to the primary sympathetic chain, and cells at intermediate levels between these two accumulate between the neural tube and the somite but some of them can escape between the sclerotome and the myotome and settle near the aorta. Histological and in vitro immunofluorescence patterns have demonstrated that the NC-1 antigen is a neuroectodermal feature. In addition to its presence on the great majority of NC cells, it persists at the surface of both neuronal and satellite cells of the peripheral ganglia. Moreover, it can be detected on neurogenic placodes and their derivatives. The appearance of the NC-1 antigen in the central nervous system coincides with the first noticeable morphological changes of the neutral tube and develops according to a rostro-caudal gradient which parallels its development: it seems, however, to be transiently expressed by the neuron cell bodies and to concentrate later on their processes. It is also present on non-neuronal cells derived from the neuroectoderm. The neuroectodermal character of NC-1 reactivity is further emphasized by its disappearance from the melanocytes and the mesectodermal derivatives of the NC. The loss by the latter, in ventral areas of the head, of the NC-1 epitope is discussed in relation to previous findings on the degree of commitment of the cephalic NC. The NC-1 epitope is associated with several high-molecular-weight polypeptides and may involve a carbohydrate moiety.


The impact of hydrocephalus and other CNS conditions on case management.
Hydrocephalus affects most people with spina bifida and can impact upon continence management adversely in two main areas. Firstly, increased intracranial pressure can cause a deterioration of continence and mental status, which sometimes can be insidious and at other times dramatic. Secondly, the effect of cognitive deficits secondary to central nervous system abnormalities can be subtle, but can be a profound barrier to achieving successful case management, including continence control and independent living.


Acute stress decreases serotonin transporter mRNA in the raphe pontis but not in other raphe nuclei of the rat.
In addition to elevated corticosterone levels, stress produces structural changes and neuronal damage especially in the hippocampus. In this line it has been shown, that in rats single or repeated immobilisation markedly reduces brain-derived neurotrophic factor (BDNF) mRNA levels in the hippocampal formation. Since this neurotrophin also controls the efficacy of serotonergic neurotransmission, the aim of the current study was to investigate the effect of acute immobilization stress on the expression of serotonin transporter (SERT) mRNA in the raphe nuclei as a parameter of serotonergic innervation. We have examined the expression of SERT mRNA and of BDNF mRNA in rats upon acute immobilisation by quantitative in situ hybridisation with a (35)S-labelled oligonucleotide probe. Elevated corticosterone levels in stressed animals confirmed as internal controls the effect of stress under our conditions. Acute stress led to a significant decrease of BDNF mRNA in the hippocampus and of SERT mRNA in the raphe pontis, but not in other raphe nuclei investigated. These data provide evidence for fast interactions between neurotrophins, corticosterone and serotonergic neurotransmission under stress conditions.


Fabrication of cell-laden microbeads and microcapsules composed of bacterial polyglucuronic acid.
In the past decades, the microencapsulation of mammalian cells into microparticles has been extensively studied for various in vitro and in vivo applications. The aim of this study was to demonstrate the viability of bacterial polyglucuronic acid (PGU), an exopolysaccharide derived from bacteria and composed of glucuronic acid units, as an effective material for cell microencapsulation. Using the method of dropping an aqueous solution of PGU-containing cells into a Ca2+-loaded solution, we produced spherical PGU microbeads with >93 % viability of the encapsulated human hepatoma HepG2 cells. Hollow-core microcapsules were formed via polyelectrolyte complex layer formation of PGU and poly-l-lysine, after which Ca2+, a cross-linker of PGU, was chelated, and this was accomplished by sequential immersion of microbeads in aqueous solutions of poly-l-lysine and sodium citrate. The encapsulated HepG2 cells proliferated and formed cell aggregates within the microparticles over a 14-day culture, with significantly larger aggregates forming within the microcapsules. Our results provide evidence for the viability of PGU for cell microencapsulation for the first time, thereby contributing to advancements in tissue engineering.


Platelet activation: a new biological activity of guinea-pig C3a anaphylatoxin.
3H-serotonin-release from labelled gp-platelets is established as a sensitive method for testing a new biological activity of gp-C3a anaphylatoxin in an autologous situation. Time-, dose- and temperature-dependent release reactions as well as specific inhibition by carboxypeptidase B and anti-C3a antibodies show that C3a is a potent and specific inducer of platelet activation. Inactive C3a does not induce 3H-serotonin-release but specifically inhibits the action of C3a on platelets.


Preconception health care and congenital disorders: mathematical modelling of the impact of a preconception care programme on congenital disorders.
This study aims to model the impact of preconception care on births with congenital disorders at a national level. Mathematical cross-sectional model based on life-table methodology. Research conducted in Cambridge, United Kingdom. Women aged 15-45 years in England, 2001. A mathematical model was constructed based on cross-sectional data from women aged 15-45 years in England undertaking one of three interventions, so as to reflect different strategies of preconception care: folic acid supplementation and fortification (representing national, universal interventions); alcohol intervention (reflecting primary care strategies); and diabetes management (targeting a population of high-risk women with a known chronic disease). Reduction in the prevalence of congenital disorders at birth. Between 585 (lower estimate) and 1085 (upper estimate) congenital disorders could be prevented with a national preconception programme, based on a single-year national cohort in England. This represents an 8-15% reduction in annual notifications of congenital disorders in live births annually. According to modelled estimates, folic acid fortification or supplementation, alcohol intervention, and diabetic management may result in a 46, 32-62, 53, and 54% reduction in the live birth prevalence of specific congenital disorders, respectively. In an ideal scenario, the application of this model decreases the total annual number of congenital disorder notifications by approximately one-sixth. A preconception care programme comprising three different strategies potentially can reduce the number of infants born with congenital disorders at a national level. This model provides strong support for preconception care to become a healthcare priority, and has major implications for healthcare planning.


Occurrence, structure, and evolution of nitric oxide synthase-like proteins in the plant kingdom.
Nitric oxide (NO) signaling regulates various physiological processes in both animals and plants. In animals, NO synthesis is mainly catalyzed by NO synthase (NOS) enzymes. Although NOS-like activities that are sensitive to mammalian NOS inhibitors have been detected in plant extracts, few bona fide plant NOS enzymes have been identified. We searched the data set produced by the 1000 Plants (1KP) international consortium for the presence of transcripts encoding NOS-like proteins in over 1000 species of land plants and algae. We also searched for genes encoding NOS-like enzymes in 24 publicly available algal genomes. We identified no typical NOS sequences in 1087 sequenced transcriptomes of land plants. In contrast, we identified NOS-like sequences in 15 of the 265 algal species analyzed. Even if the presence of NOS enzymes assembled from multipolypeptides in plants cannot be conclusively discarded, the emerging data suggest that, instead of generating NO with evolutionarily conserved NOS enzymes, land plants have evolved finely regulated nitrate assimilation and reduction processes to synthesize NO through a mechanism different than that in animals.


Profiling of cellular proteins in porcine reproductive and respiratory syndrome virus virions by proteomics analysis.
Porcine reproductive and respiratory syndrome virus (PRRSV) is an enveloped virus, bearing severe economic consequences to the swine industry worldwide. Previous studies on enveloped viruses have shown that many incorporated cellular proteins associated with the virion's membranes that might play important roles in viral infectivity. In this study, we sought to proteomically profile the cellular proteins incorporated into or associated with the virions of a highly virulent PRRSV strain GDBY1, and to provide foundation for further investigations on the roles of incorporated/associated cellular proteins on PRRSV's infectivity. In our experiment, sixty one cellular proteins were identified in highly purified PRRSV virions by two-dimensional gel electrophoresis coupled with mass spectrometric approaches. The identified cellular proteins could be grouped into eight functional categories including cytoskeletal proteins, chaperones, macromolecular biosynthesis proteins, metabolism-associated proteins, calcium-dependent membrane-binding proteins and other functional proteins. Among the identified proteins, four have not yet been reported in other studied envelope viruses, namely, guanine nucleotide-binding proteins, tyrosine 3-monooxygenase/tryptophan 5-monooxygenase, peroxiredoxin 1 and galectin-1 protein. The presence of five selected cellular proteins (i.e., β-actin, Tubulin, Annexin A2, heat shock protein Hsp27, and calcium binding proteins S100) in the highly purified PRRSV virions was validated by Western blot and immunogold labeling assays. Taken together, the present study has demonstrated the incorporation of cellular proteins in PRRSV virions, which provides valuable information for the further investigations for the effects of individual cellular proteins on the viral replication, assembly, and pathogenesis.


Seasonal prevalence of thermophilic Campylobacter infections in dairy cattle and a study of infection of sheep.
A total of 273 rectal swabs from dairy cows were cultured for Campylobacter jejuni/coli. The isolation rate was 17/72 (24%), 33/106 (31%) and 11/95 (12%) during summer, autumn and winter respectively. Approximately half of the isolates were C. jejuni and the other half C. coli. The isolates recovered from dairy cows were typed by bacterial restriction endonuclease DNA analysis (BRENDA) and compared with those of sheep. Seventeen different BRENDA patterns were produced by the isolates from dairy cows and six from 27 isolates of sheep. Of these 21 different BRENDA patterns only two were common to sheep and cattle.


Prevention of experimental anaphylactic ulcer by antigen prefeeding.
Studies were performed in animals to establish whether antigen prefeeding could present homocytotropic antibody synthesis and the induction of gastric ulcer resulting from mucosal anaphylaxis. A single digestive exposure to 1 or 10 mg ovalbumin induced a state of specific immunologic tolerance. Inhibition of the formation of specific reagins was shown by the study of mast cell degranulation. Tolerant animals presented a reduced incidence of gastric ulcer after subsequent mucosal challenge. These results are important for the development of methods of prevention and treatment of allergic diseases.