Effect of stimulus intensity on the sizes of chromatic perceptive fields.
The effects of intensity on chromatic perceptive field size were investigated along the horizontal meridian at 10 degrees temporal eccentricity by manipulating stimulus intensity from 0.3 to 3.3 log trolands. Following light adaptation, observers described the hue and saturation of monochromatic stimuli (440-660 nm, in 10 nm steps) for a series of test sizes (0.098-3 degrees) presented along the time period associated with the cone plateau of the dark-adaptation function. Perceptive field sizes of the four elemental hues (red, green, yellow, and blue) and the saturation component were estimated by three observers at each intensity level for each wavelength. In general, perceptive field sizes of blue and red are the smallest, and yellow and green are the largest. Furthermore, perceptive field sizes of all four hues decrease with increasing stimulus intensity, though the absolute change is largest for green and yellow. The decrease in size with increase in intensity cannot be completely explained in terms of saturation or rod signals and is likely, then, attributable to a cone-based mechanism.


Real-time detection of BRAF V600E mutation from archival hairy cell leukemia FFPE tissue by nanopore sequencing.
The MinION is a miniaturized high-throughput next generation sequencing platform of novel conception. The use of nucleic acids derived from formalin-fixed paraffin-embedded samples is highly desirable, but their adoption for molecular assays is hurdled by the high degree of fragmentation and by the chemical-induced mutations stemming from the fixation protocols. In order to investigate the suitability of MinION sequencing on formalin-fixed paraffin-embedded samples, the presence and frequency of BRAF c.1799T > A mutation was investigated in two archival tissue specimens of Hairy cell leukemia and Hairy cell leukemia Variant. Despite the poor quality of the starting DNA, BRAF mutation was successfully detected in the Hairy cell leukemia sample with around 50% of the reads obtained within 2 h of the sequencing start. Notably, the mutational burden of the Hairy cell leukemia sample as derived from nanopore sequencing proved to be comparable to a sensitive method for the detection of point mutations, namely the Digital PCR, using a validated assay. Nanopore sequencing can be adopted for targeted sequencing of genetic lesions on critical DNA samples such as those extracted from archival routine formalin-fixed paraffin-embedded samples. This result let speculating about the possibility that the nanopore sequencing could be trustably adopted for the real-time targeted sequencing of genetic lesions. Our report opens the window for the adoption of nanopore sequencing in molecular pathology for research and diagnostics.


A novel homozygous SLC25A1 mutation with impaired mitochondrial complex V: Possible phenotypic expansion.
SLC25A1 mutations are associated with combined D,L-2-hydroxyglutaric aciduria (DL- 2HGA; OMIM #615182), characterized by muscular hypotonia, severe neurodevelopmental dysfunction and intractable seizures. SLC25A1 encodes the mitochondrial citrate carrier (CIC), which mediates efflux of the mitochondrial tricarboxylic acid (TCA) cycle intermediates citrate and isocitrate in exchange for cytosolic malate. Only a single family with an SLC25A1 mutation has been described in which mitochondrial respiratory chain dysfunction was documented, specifically in complex IV. Five infants of two consanguineous Bedouin families of the same tribe presented with small head circumference and neonatal-onset encephalopathy with severe muscular weakness, intractable seizures, respiratory distress, and lack of psychomotor development culminating in early death. Ventricular septal defects (VSD) were demonstrated in three patients. Blood and CSF lactate were elevated with normal levels of plasma amino acids and free carnitine and increased 2-OH-glutaric acid urinary exertion. EEG was compatible with white matter disorder. Brain MRI revealed ventriculomegaly, thin corpus callosum with increased lactate peak on spectroscopy. Mitochondrial complex V deficiency was demonstrated in skeletal muscle biopsy of one infant. Homozygosity mapping and sequencing ruled out homozygosity of affected individuals in all known complex V-associated genes. Whole exome sequencing identified a novel homozygous SLC25A1 c.713A>G (p.Asn238Ser) mutation, segregating as expected in the affected kindred and not found in 220 control alleles. Thus, SLC25A1 mutations might be associated with mitochondrial complex V deficiency and should be considered in the differential diagnosis of mitochondrial respiratory chain defects.


The Yersinia enterocolitica infection in acute abdominal surgery. A clinical study with a 5-year follow-up period.
By estimation of antibody titres or possibly isolation of the micro-organism, the role of the Yersinia enterocolitica as an etiological agent in mesenterial lymphadenitis, ileal inflammation, or abdominal colics was evaluated in 94 consecutive patients (25 operated upon and 69 treated conservatively) during a 1-year period from 1975-76. Eighty four patients (24 operated upon and 60 treated conservatively) were re-examined 5 years later. Among the 25 patients operated upon, three presented acute terminal ileitis, two of them in association with acute Y. enterocolitica infection. A substantial part of the 22 patients with other peroperative findings also presented trace or positive Y. enterocolitica antibody titres. Some of these patients experienced complaints attributable to the Y. enterocolitica infection over the following years. At re-examination, previously elevated Y. enterocolitica antibody titres had generally normalized, and in the two patients who in 1975 presented acute terminal ileitis associated with acute Y. enterocolitica infection, radiological examination now demonstrated normal terminal ileum. Among 69 patients treated conservatively for abdominal colics, 36 in 1975 presented trace or positive titres. During the 5-year follow-up period, abdominal pain and joint complaints were significantly more commonly experienced by patients with trace or positive titres than by patients with negative titres (p = 0.007 and p = 0.004, respectively). The Y. enterocolitica is an important cause of abdominal disease in our region, and the infection commonly gives symptoms of long duration.


Selective non-enzymatic uric acid sensing in the presence of dopamine: electropolymerized poly-pyrrole modified with a reduced graphene oxide/PEDOT:PSS composite.
A highly selective electrochemical sensor based on a molecularly imprinted polymer (MIP) to be developed for uric acid detection in the presence of dopamine as an interference molecule was demonstrated in this study. This non-enzymatic uric acid sensor was developed by electropolymerizing poly-pyrrole onto a composite of electrochemically reduced graphene oxide (ErGO) and poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate) (PEDOT:PSS) on a glassy carbon electrode (GCE) to give MIP/ErGO/PEDOT:PSS electrodes. The structural properties, surface morphology, and electrochemical interface of this fabricated uric acid sensor was then characterized using infrared spectroscopy, scanning electron microscopy, atomic force microscopy, and electrochemical impedance spectroscopy. This fabricated non-enzymatic electrochemical sensor (MIP/ErGO/PEDOT:PSS modified GCE) also showed excellent analytical performance at the optimum ratio of monomer/template concentration and optimized electropolymerization conditions, for example in the low concentration range of 0.1-100 μM with a detection limit of 0.05 μM towards uric acid detection in the presence of dopamine. Furthermore, this fabricated uric acid sensor also exhibited good reproducibility and stability for uric acid measurements in the presence of dopamine for 5 consecutive days. In addition, this sensor demonstrates highly selective detection of uric acid in the presence of several interfering species such as dopamine, urea, glucose, magnesium ions, and ascorbic acid. This fabricated uric acid sensor ultimately showed satisfactory uric acid measurement in samples of human urine and is expected to be used in early-stage disease diagnosis.


Factors to consider when selecting animal models for postnatal teratology studies.
This presentation identifies some of the factors which should be considered when choosing a particular animal as a model to predict postnatal teratogenic hazards to man. The developmental pharmacologist-toxicologist must consider in concert the potential interactions of a xenobiotic on the mother, her placenta, her fetus, and her nursing young. Since the physiology of each of these components is in a state of dynamic flux during gestation and/or postnatal development, the critical time at which to expose the mother or her progeny to a chemical will vary widely from species to species and organ to organ. The identification of critical periods during the perinatal period may be quite difficult because the physiological or behavioral changes in the adult organism which signal that perinatal teratogenic effects have occurred are frequently subtle. Much research is still needed on the comparative aspects of maternal, placental, fetal, and neonatal pharmacokinetics and pharmacodynamics. We are not ready to intelligently evaluate large numbers of chemicals as potential postnatal teratogens for man.


Postsurgical decompression and immediate elemental feeding.
The paralytic ileus, poor nutritional state, and related complications that often develop following serious serious abdominal surgery have traditionally been accepted as unavoidable. Now, a nasogastric decompression and feeding tube is availabe that usually prevents paralytic ileus and also provides a complete liquid diet starting immedicately after the operation, thus speeding wound healing and recivery.


Multi-channel EEG recordings during a sustained-attention driving task.
We describe driver behaviour and brain dynamics acquired from a 90-minute sustained-attention task in an immersive driving simulator. The data included 62 sessions of 32-channel electroencephalography (EEG) data for 27 subjects driving on a four-lane highway who were instructed to keep the car cruising in the centre of the lane. Lane-departure events were randomly induced to cause the car to drift from the original cruising lane towards the left or right lane. A complete trial included events with deviation onset, response onset, and response offset. The next trial, in which the subject was instructed to drive back to the original cruising lane, began 5-10 seconds after finishing the previous trial. We believe that this dataset will lead to the development of novel neural processing methodology that can be used to index brain cortical dynamics and detect driving fatigue and drowsiness. This publicly available dataset will be beneficial to the neuroscience and brain-computer interface communities.


Pig H3K4me3, H3K27ac, and gene expression profiles reveal reproductive tissue-specific activity of transposable elements.
Regulatory sequences and transposable elements (TEs) account for a large proportion of the genomic sequences of species; however, their roles in gene transcription, especially tissue-specific expression, remain largely unknown. Pigs serve as an excellent animal model for studying genomic sequence biology due to the extensive diversity among their wild and domesticated populations. Here, we conducted an integrated analysis using H3K27ac ChIP-seq, H3K4me3 ChIP-seq, and RNA-seq data from 10 different tissues of seven fetuses and eight closely related adult pigs. We aimed to annotate the regulatory elements and TEs to elucidate their associations with histone modifications and mRNA expression across different tissues and developmental stages. Based on correlation analysis between mRNA expression and H3K27ac and H3K4me3 peak activity, results indicated that H3K27ac exhibited stronger associations with gene expression than H3K4me3. Furthermore, 1.45% of TEs overlapped with either the H3K27ac or H3K4me3 peaks, with the majority displaying tissue-specific activity. Notably, a TE subfamily (LTR4C_SS), containing binding motifs for SIX1 and SIX4, showed specific enrichment in the H3K27ac peaks of the adult and fetal ovaries. RNA-seq analysis also revealed widespread expression of TEs in the exons or promoters of genes, including 4 688 TE-containing transcripts with distinct development stage-specific and tissue-specific expression. Of note, 1 967 TE-containing transcripts were enriched in the testes. We identified a long terminal repeat (LTR), MLT1F1, acting as a testis-specific alternative promoter in SRPK2 (a cell cycle-related protein kinase) in our pig dataset. This element was also conserved in humans and mice, suggesting either an ancient integration of TEs in genes specifically expressed in the testes or parallel evolutionary patterns. Collectively, our findings demonstrate that TEs are deeply embedded in the genome and exhibit important tissue-specific biological functions, particularly in the reproductive organs.


Red cell distribution width and mortality in older adults: a meta-analysis.
Red cell distribution width (RDW) is a quantitative measure of variability in the size of circulating erythrocytes with higher values reflecting greater heterogeneity in cell sizes. Recent studies have shown that higher RDW is associated with increased mortality risk in patients with clinically significant cardiovascular disease (CVD). Whether RDW is prognostic in more representative community-based populations is unclear. Seven relevant community-based studies of older adults with RDW measurement and mortality ascertainment were identified. Cox proportional hazards regression and meta-analysis on individual participant data were performed. Median RDW values varied across studies from 13.2% to 14.6%. During 68,822 person-years of follow-up of 11,827 older adults with RDW measured, there was a graded increased risk of death associated with higher RDW values (p < .001). For every 1% increment in RDW, total mortality risk increased by 14% (adjusted hazard ratio [HR]: 1.14; 95% confidence interval [CI]: 1.11-1.17). In addition, RDW was strongly associated with deaths from CVD (adjusted HR: 1.15; 95% CI: 1.12-1.25), cancer (adjusted HR: 1.13; 95% CI: 1.07-1.20), and other causes (adjusted HR: 1.13; 95% CI: 1.07-1.18). Furthermore, the RDW-mortality association occurred in all major demographic, disease, and nutritional risk factor subgroups examined. Among the subset of 1,603 older adults without major age-associated diseases, RDW remained strongly associated with total mortality (adjusted HR: 1.32; 95% CI: 1.21-1.44). RDW is a routinely reported test that is a powerful predictor of mortality in community-dwelling older adults with and without age-associated diseases. The biologic mechanisms underlying this association merit investigation.


Quality of care in inflammatory bowel diseases: What is the best way to better outcomes?
Inflammatory bowel disease (IBD) is a lifelong, progressive disease that has disabling impacts on patient's lives. Given the complex nature of the diagnosis of IBD and its management there is consequently a large economic burden seen across all health care systems. Quality indicators (QI) have been created to assess the different façades of disease management including structure, process and outcome components. Their development serves to provide a means to target and measure quality of care (QoC). Multiple different QI sets have been published in IBD, but all serve the same purpose of trying to achieve a standard of care that can be attained on a national and international level, since there is still a major variation in clinical practice. There have been many recent innovative developments that aim to improve QoC in IBD including telemedicine, home biomarker assessment and rapid access clinics. These are some of the novel advancements that have been shown to have great potential at improving QoC, while offloading some of the burden that IBD can have vis-a-vis emergency room visits and hospital admissions. The aim of the current review is to summarize and discuss available QI sets and recent developments in IBD care including telemedicine, and to give insight into how the utilization of these tools could benefit the QoC of IBD patients. Additionally, a treating-to-target structure as well as evidence surrounding aggressive management directed at tighter disease control will be presented.


Inhibition of prostatic epithelial cell proliferation by a factor secreted specifically by prostatic stromal cells.
Stromal cells from the prostate were recently shown to inhibit clonal growth of the prostatic carcinoma cell lines PC-3 (hormone-independent) and LNCaP (hormone-sensitive) in coculture. Our study revealed that stromal cell-conditioned medium strongly inhibited proliferation of PC-3 and LNCaP cells when grown in monolayer culture. Antiproliferative activity was found to be reversible, and was produced specifically by prostatic stromal cells and not by stromal cells derived from skin, foreskin, uterus, kidney, and Wilms' tumor. Inhibition was not species-specific, since the cell lines AT-2.1 and MATLyLu, derived from the Dunning rat prostate tumor, were also sensitive. No inhibition, however, occurred on breast and renal carcinoma cell lines, suggesting a prostate-specific action. The putative inhibiting factor(s) could be concentrated and partially purified by ammonium sulfate precipitation. The possible role in stromal control of epithelial cell proliferation is discussed.


Stochastic assembly process dominates bacterial succession during a long-term microbial enhanced oil recovery.
Microbial enhanced oil recovery (MEOR) has been successfully used in oil exploitation to increase oil production. However, the mechanisms of microbial interactions and community assembly related to oil production performance along MEOR process are poorly understood. Here, we investigated the microbiome of an oil reservoir for a period of 5 years under three phases of different treatments with the injection of a mixture of microbes, nutrients, and air at different intensity. During the MEOR process, amplification of functional genes revealed an increase of genes related to hydrocarbon degradation linked to methanogenesis, supported by stable isotope analysis for confirmation of the methanogenesis activity. Meanwhile, a lower contribution of the ubiquitous/common taxa, closer and more positive associations, and lower modularity were observed in bacterial co-occurrence networks, with the rare taxa being the keystone taxa. The null model analysis and structural equation modeling revealed that the contribution of stochastic processes affected by functional groups and co-occurrence patterns to bacterial community increased significantly with the increase of oil production. This provides new insight that stochastic assembly in bacterial community increased along with MEOR process, and it is worthwhile paying attention to the uncertain consequences caused by random evolution since the treatment effect of MEOR is closely related to the in-situ community in oil reservoir.


Elective coniotomy. A prospective study.
The view current over the last 60 years or so that coniotomy often leads to subglottic stenosis has recently been called into question. In the present study 103 electively performed coniotomies have been analysed. The operation was found to be much easier than tracheotomy. There was no case of severe peroperative or postoperative complications. Six months after decannulation 28 patients were alive and accessible to follow-up examination. No evidence of subglottic stenosis was found. The main disadvantage of this operation would seem to be a tendency for the development of voice changes.


Microsurgical Reconstruction of the Jaws Using Vascularised Free Flap Technique in Patients with Medication-Related Osteonecrosis: A Systematic Review.
Osteonecrosis of the jaw (ONJ) has been reported to be associated with patients receiving primarily bisphosphonate (BP) therapies. However, lately it has been documented that other medications, such as RANK ligand inhibitor (denosumab) and antiangiogenic drug, can cause ONJ. Micro-osseous-vascular reconstruction of the jaws in patients affected by medication-related osteonecrosis of the jaw represents a viable option of treatment for patients affected by stage III of the disease. However, there are still considerable doubts about the success of this procedure in the short, medium, and long term. A multidatabase (PubMed/MEDLINE, EMBASE, and CENTRAL) systematic search was performed. Any type of studies considering human patients treated with antiresorptive and antiangiogenic drugs was considered. The aim of the research is to primarily understand the success rate of micro-osseous-vascular reconstruction in the short, medium, and long period of time. This review has also the goal of better understanding any perioperative and postoperative complications resulting from the use of the reconstruction techniques. Eighteen studies resulted eligible for the study. Fibula free flap is the most commonly utilised vascularised free flap reconstruction technique (80.76%). Ten out of eighteen studies reported no complications. Recurrence of osteonecrosis was registered in five cases (6.41%) after free flap reconstruction. The overall free flap success rate was 96.16%. Based on the limited data available in literature (Level 4 of the Oxford Evidence-based medicine scale), micro-osseous-vascular reconstruction of the jaws represents a valid treatment in patients with bisphosphonate-related osteonecrosis at stage III of the disease. However, additional data based on a larger cohort of patients are necessary to justify this type of intervention in patient affected by MRONJ.


Materials recovery from waste liquid crystal displays: A focus on indium.
In the present work the recovery of indium and of the polarizing film from waste liquid crystal displays was experimentally investigated in the laboratory. First of all, the polarizing film was removed by employing a number of different techniques, including thermal and chemical treatments. Leaching of indium was then performed with HCl 6N, which allowed solubilisation of approximately 90% In (i.e. 260 mg In per kg of glass) at room temperature, without shredding. Indium recovery from the aqueous phase was then investigated through solvent extraction with polyethylene glycol (PEG)-based aqueous biphasic systems. Indium extraction tests through the PEG-ammonium sulphate-water system were conducted as a function of PEG concentration, salt concentration and molecular weight of PEG, using 1,10 phenanthroline as a ligand. The experimental results demonstrated that indium partitioning between the bottom (salt-rich) and the top (PEG-rich) phase is quite independent on the composition of the system, since 80-95% indium is extracted in the bottom phase and 5-20% in the top phase; it was also found that when PEG concentration is increased, the ratio between the bottom and the upper phase volumes decreases, resulting in an increase of indium concentration in the bottom phase (at [PEG]=25% w/w, indium concentration in the bottom phase is ∼30% higher than the initial concentration before the extraction).


Role of efaproxiral in metastatic brain tumors.
Brain metastases from breast cancer are common and cause significant morbidity and mortality. Treatment with whole-brain radiotherapy provides a median survival of only 4-6 months. One mechanism affecting sensitivity to radiation is tumor oxygenation. Hypoxic tumor cells are more likely to be resistant to radiation and adversely affect prognosis. For nearly 70 years, all attempted modalities have failed to circumvent tumor hypoxia and increase tumor death. Abraham and colleagues developed a fibric acid derivative that would cause the displacement of oxygen from hemoglobin and improve tissue oxygenation. After modifications to enhance absorption into the red blood cell, RSR13 (efaproxiral) was developed. Efaproxiral was found to be safe and improved tumor oxygenation in preclinical studies. Early Phase I and II trials supported the earlier preclinical evidence while demonstrating that efaproxiral was safe and effective. A randomized Phase III study, Radiation Enhancing Allosteric Compounds of Hypoxic brain metastases (REACH, RT-009), failed to show a significant improvement in overall survival in all eligible patients treated with efaproxiral and whole-brain radiation. However, there was a statistically significant improvement in median survival and quality of life within a subset of metastatic breast cancer patients. Efaproxiral is currently being studied with radiotherapy in a confirmatory trial in the treatment of brain metastases from breast cancer. Furthermore, the combination of efaproxiral with radiotherapy is being investigated in other solid tumors.


Long-range order of organized oligonucleotide monolayers on Au(111) electrodes.
Oligonucleotides modified by a hexamethylene linker group adsorb on gold electrodes via Au-S bond formation. We have obtained novel data for adsorption of thiol-modified (HS) single-strand HS-10A and double-stranded HS-10AT oligonucleotides and for analogous thiol-free 10A (A = adenine) and 10T (T = thymine) nonspecifically adsorbed as reference molecules. Mercaptohexanol has served as a second reference molecule. The data are based on cyclic and differential pulse voltammetry, interfacial capacitance data, and in situ scanning tunneling microscopy (STM) directly in an aqueous buffer solution, with electrochemical potential control of both the sample electrode and the tip. All the data are based on single-crystal, atomically planar Au(111)-electrode surfaces. The high sensitivity of such surfaces provides accurate HS-10A and HS-10AT electrode coverages on the basis of the reductive desorption of the Au-S bond. The coverage is high and in keeping with dense monolayers of adsorbed HS-10A and HS-10AT in an upright or tilted orientation, with the oligonucleotide backbone repelled from the strongly negatively charged electrode surface. Adsorbed thiol-free 10A only gives a Au(111)-reconstruction peak, while 10T shows a subtle pattern involving pronounced voltammetric adsorption peaks indicative of both nonspecific adsorption via single thymine units and potential-dependent structural reorganization in the surface layer. In situ STM supports these findings at the molecular level. In situ STM of HS-10A discloses large, highly ordered domains at strongly negative sample potentials. Reversible domain formation and disordering could, moreover, be controlled by an electrochemical potential variation in the negative and positive directions, respectively. 10A and 10T did not form ordered adsorbate domains, substantiating that domain formation rests on adsorption of thiol-modified oligonucleotide adsorption in an upright or tilted orientation. The comprehensive, high-resolution information reported may hold prospects for single-molecule electronic conduction and molecular-scale mapping of oligonucleotide hybridization.


[Forgotten parasitic diseases].
The ecological flexibility of parasitic systems is observed in parasites, such as pentastomas of Linguatula serrata and Armillifer sp., cestodes of Spirometra erinacei europaei, and dirofilariae of D. repens, D. immitis, D. ursi in relation to ecological shifts in the areas and anthropological pressure.


Development of Rapid Extraction Method of Mycobacterium avium Subspecies paratuberculosis DNA from Bovine Stool Samples.
The rapid identification of Mycobacterium avium subspecies paratuberculosis (MAP) infected animals within the herd is essential for preventing the spread of the disease as well as avoiding human exposure. Although culture is seen as the gold standard, there are various molecular assays available i.e., polymerase chain reaction (PCR) or isothermal amplification technique (recombinase polymerase amplification (RPA)) for the detection of MAP. The accuracy of the molecular assays is highly dependent on the DNA extraction method. In order to establish a rapid point of need system for the detection of MAP DNA from stool samples, we developed a rapid DNA extraction protocol (MAP DNA SpeedXtract) specified for use in combination with the RPA. The whole procedure from "sample in" to "result out" was conducted in a mobile suitcase laboratory. The DNA extraction is based on reverse purification by magnetic beads, which reduces the required technical demand. The MAP DNA SpeedXtract was performed within 25 min and only three pipetting steps were needed. The amplification and detection time were 20 min in RPA. The sensitivity and specificity of the developed protocol in comparison with the lab-based silica membrane column extraction and real-time PCR were 90.9% (n = 22) and 100% (n = 23), respectively. In conclusion, we established a rapid and reliable protocol for the extraction and detection of MAP DNA. All reagents are cold chain independent. The entire setup is ideal for point of need identification of MAP infected cases.


Twins with progressive thoracic aortic aneurysm, recurrent dissection and ACTA2 mutation.
Thoracic aortic aneurysm (TAA) is a genetically mediated disease with variable age of onset. In the pediatric age range, nonsyndromic TAA frequently has a milder course than syndromic forms of TAA, such as Marfan syndrome or Loeys-Dietz syndrome. Herein, we describe 17-year-old identical twin brothers with severe progressive TAA due to a novel de novo ACTA2 mutation. Interestingly, both boys were diagnosed at age 11 with congenital mydriasis, a recently recognized manifestation of some ACTA2 mutations due to smooth muscle dysfunction. One of the brothers presented with acute-onset lower back pain that was identified as dissection of an abdominal aortic aneurysm. Imaging of the chest at this time showed severe fusiform TAA. Cardiac imaging in his twin showed similar TAA, but no abdominal aortic aneurysm. Both brothers underwent valve-sparing aortic root replacement, but have had progressive aortic disease with recurrent dissection requiring multiple surgeries. This case emphasizes the importance of identifying physical stigmata of smooth muscle dysfunction, such as mydriasis, as potential markers for associated aortopathy and vascular diseases.


[Analysis of DNA ploidy in 3,050 cases of malignant tumors].
DNA ploidy in paraffin-embedded sections collected from 3,050 cases of malignant tumors was analysed by flow cytometry. The result showed that DNA index distribution was ranged from 0.6 to 2.6, mostly at 1.10 to 1.85. DNA ploidy distribution showed that 536 cases (17.6%) were DNA diploid tumors; 2,227 cases (73.0%) were tumors with DNA aneuploid; 126 cases (4.1%) accompanied with tetraploid; and 161 cases (5.3%) with multiploid. Additionally, relationship between the histologic type as well as the grading and DNA ploid were also analysed in 1,117 cases of malignant tumors. The results indicated that DNA index was not correlated with the histologic typing, but correlated with the histologic grading (grade I DI = 1.24, grade II DI = 1.38, grade III DI = 1.51). It's considered that flow cytometry DNA grade index may be an objective and precise quantitative marker in judging the degree, grade and differentiation of tumor malignancy.


Pneumothorax in cystic fibrosis.
Pneumothorax is recognized as a common and life-threatening complication in cystic fibrosis (CF) patients, especially in those who are infected with P. aeruginosa, B. cepacia or Aspergillus, need enteral feeding, are diagnosed as suffering from allergic bronchopulmonary aspergillosis (ABPA), developed massive hemoptysis, and their respiratory function is seriously compromised. Structural impairment and altered airflow dynamics in the lungs of CF patients are considered as the main predisposing factors, but also inhaled medications and non-invasive positive pressure ventilation (NIPPV) could increase the risk of pneumothorax. Clinical presentation could range from dramatic to very mild. Management of spontaneous pneumothorax occurring to patients with CF is essentially similar to that for non-CF patients. Therapeutic options include intercostal tube drainage, video-assisted thoracoscopic surgery (VATS), and medical or surgical pleurodesis. Pneumothorax increases both short- and long-term morbidity and mortality in CF patients and causes significant deterioration of their quality of life.


[Uretero-vaginal fistulas: diagnosis and operative tactics. Apropos of 19 personal cases].
Personal experience of the treatment of 19 uretero-vaginal fistulae, observed from september 1987 to june 1991 on 17 patients in Africa is described and analysed. Occurring after gynecological or obstetrical surgery, the main cause is hysterectomy (82.3%). Elements for diagnosis were: the appearance of a urine leak, which in most cases came rapidly, echography, and most important, intravenous urography (IVU). Surgical management in all of the cases was mainly aimed at conservation. One end-to-end anastomosis and 18 cuff reimplantations were performed, combined in 2 cases with a sub-mucus tunnel (Politano), in 14 cases combined with a tubular Boari bladder flap (wherein 1 case was bilateral), 3 combined with a bladder psoas hitch and 2 with an isolated ileal graft (ileo-uretero-cystoplasty). One case of vesicouterine fistula and 4 associated vesicovaginal fistulae benefited from a simultaneous surgical operation. In this study of 17 patients treated, except for one patient deceased 14 months following a neoplastic recurrence, the results were excellent. 16 patients were cured. Follow-up extended to more than a year and have shown the perfect conservation of the excretory ducts in all of the cases studied. The cuffed ureterovesical reimplantation combined with a tubular bladder plasty definitively seem to be a most reliable management technique for all ureter injury encountered. While allowing the preservation of the kidney located just below, it has, in particular, prevented ureter stenosis. If the use of simpler methods (psoas bladder hitch) may be sufficient for some cases, the use of more complex methods (ileo-uretero-cystoplasty) for other cases, may be indispensable.


Familial bone marrow monosomy 7. Evidence that the predisposing locus is not on the long arm of chromosome 7.
Loss of expression of a tumor-suppressing gene is an attractive model to explain the cytogenetic and epidemiologic features of cases of myelodysplasia and acute myelogenous leukemia (AML) associated with bone marrow monosomy 7 or partial deletion of the long arm (7q-). We used probes from within the breakpoint region on 7q-chromosomes (7q22-34) that detect restriction fragment length polymorphisms (RFLPs) to investigate three families in which two siblings developed myelodysplasia with monosomy 7. In the first family, probes from the proximal part of this region identified DNA derived from the same maternal chromosome in both leukemias. The RFLPs in these siblings diverged at the more distal J3.11 marker due to a mitotic recombination in one patient, a result that suggested a critical region on 7q proximal to probe J3.11. Detailed RFLP mapping of the implicated region was then performed in two additional unrelated pairs of affected siblings. In these families, DNA derived from different parental chromosome 7s was retained in the leukemic bone marrows of the siblings. We conclude that the familial predisposition to myelodysplasia is not located within a consistently deleted segment on the long arm of chromosome 7. These data provide evidence implicating multiple genetic events in the pathogenesis of myelodysplasia seen in association with bone marrow monosomy 7 or 7q-.


ZBP-99 defines a conserved family of transcription factors and regulates ornithine decarboxylase gene expression.
Among transcription factors that regulate ornithine decarboxylase (ODC) gene expression are those that interact with GC-rich promoters, including Sp1 and ZBP-89. Sp1 functions as a transactivator and ZBP-89 as a transrepressor of both the ODC and gastrin promoters. This study reports the cloning and characterization of a second member of the ZBP family that also binds GC boxes. ZBP-99 contains four Krüppel-type zinc fingers that collectively share 91% amino acid sequence similarity and 79% sequence identity with those found in ZBP-89. In addition, there are highly conserved amino acid sequences in the carboxy-terminal segments of the two genes. In spite of their structural similarities, the two proteins are encoded at distinct loci, ZBP-89 on chromosome 3q21 and ZBP-99 on 1q32.1. The predicted open reading frame of ZBP-99 cDNA encodes a 99-kDa protein. Electrophoretic mobility shift assays showed that ZBP-99 protein specifically binds to the GC-rich promoter elements of gastrin and ODC genes. Northern blot analysis showed that a major ZBP-99 transcript of 5.6 kb is expressed ubiquitously at low levels, with elevated expression levels in placenta and in adult kidney, liver, and lymphocytes. Cotransfection of AGS gastric adenocarcinoma and HT-29 colon adenocarcinoma cells with a ZBP-99 expression construct and with an ODC reporter construct show that ZBP-99 repressed basal expression in the two cell lines by 80 and 60%, respectively. Collectively, the data suggest that ZBP-99 binds GC-rich promoters and may complement the activities mediated by ZBP-89.


[Effect of inhibitors of RNA and protein synthesis on audiogenic epileptiform reactions in rats of the KM line].
Injection of actinomycin D, puromycin and cycloheximide change the genetically determined reaction of rats to acoustic stimuli up to the appearance of resistance to sound. The change in the nature of audiogenic epileptiform seizures correlates with the disturbance of the RNA and protein synthesis in the experimental animals brain. The appearance of sound resistance does not correlate with the degree of inhibition of protein synthesis in the brain. It is assumed that the antiseizure effect of the drugs used is possibly due to their action on the neuromediator systems of the brain.


Analysis of authentication protocols in agent-based systems using labeled tableaux.
The study of multiagent systems (MASs) focuses on systems in which many intelligent agents interact with each other using communication protocols. For example, an authentication protocol is used to verify and authorize agents acting on behalf of users to protect restricted data and information. After authentication, two agents should be entitled to believe that they are communicating with each other and not with intruders. For specifying and reasoning about the security properties of authentication protocols, many researchers have proposed the use of belief logics. Since authentication protocols are designed to operate in dynamic environments, it is important to model the evolution of authentication systems through time in a systematic way. We advocate the systematic combinations of logics of beliefs and time for modeling and reasoning about evolving agent beliefs in MASs. In particular, we use a temporal belief logic called TML (+) for establishing trust theories for authentication systems and also propose a labeled tableau system for this logic. To illustrate the capabilities of TML (+), we present trust theories for several well-known authentication protocols, namely, the Lowe modified wide-mouthed frog protocol, the amended Needham-Schroeder symmetric key protocol, and Kerberos. We also show how to verify certain security properties of those protocols. With the logic TML (+) and its associated modal tableaux, we are able to reason about and verify authentication systems operating in dynamic environments.


The Small Molecule Alpha-Synuclein Aggregator, FN075, Enhances Alpha-Synuclein Pathology in Subclinical AAV Rat Models.
Animal models of Parkinson's disease, in which the human α-synuclein transgene is overexpressed in the nigrostriatal pathway using viral vectors, are widely considered to be the most relevant models of the human condition. However, although highly valid, these models have major limitations related to reliability and variability, with many animals exhibiting pronounced α-synuclein expression failing to demonstrate nigrostriatal neurodegeneration or motor dysfunction. Therefore, the aim of this study was to determine if sequential intra-nigral administration of AAV-α-synuclein followed by the small α-synuclein aggregating molecule, FN075, would enhance or precipitate the associated α-synucleinopathy, nigrostriatal pathology and motor dysfunction in subclinical models. Rats were given unilateral intra-nigral injections of AAV-α-synuclein (either wild-type or A53T mutant) followed four weeks later by a unilateral intra-nigral injection of FN075, after which they underwent behavioral testing for lateralized motor functionality until they were sacrificed for immunohistological assessment at 20 weeks after AAV administration. In line with expectations, both of the AAV vectors induced widespread overexpression of human α-synuclein in the substantia nigra and striatum. Sequential administration of FN075 significantly enhanced the α-synuclein pathology with increased density and accumulation of the pathological form of the protein phosphorylated at serine 129 (pS129-α-synuclein). However, despite this enhanced α-synuclein pathology, FN075 did not precipitate nigrostriatal degeneration or motor dysfunction in these subclinical AAV models. In conclusion, FN075 holds significant promise as an approach to enhancing the α-synuclein pathology in viral overexpression models, but further studies are required to determine if alternative administration regimes for this molecule could improve the reliability and variability in these models.


Advancing quality-of-life research by deepening our understanding of response shift: a unifying theory of appraisal.
Studies of response-shift phenomena in quality-of-life (QOL) research have grown steadily in the more than two decades of research. As a field, we have been calling a lot of different approaches "response shift" over the years, but the only approach that fully embodies the foundational (Sprangers and Schwartz, Soc Sci Med 48(11):1507-1515, 1999) theoretical model is appraisal. According to the definition proposed in this model, response shift is about individual cognitive change. This paper presents the argument that all response-shift research models need to be grounded in an understanding of cognitive appraisal. We present a reasoned argument for why appraisal is fundamental to QOL response-shift research. We describe current measurement and analytic methods for working with appraisal, and how these methods can be integrated into the current response-shift statistical 'tool box.' We propose future research directions on theory, methods, and cross-calibration of group- and individual-level methods. There are currently three tools available in multiple languages for assessing QOL appraisal processes. They have been tested and used to assess response shift in empirical studies of ~ 7000 people with chronic illness. The study findings illustrate how appraisal theory and methods can facilitate methodological investigations of and to enhance other response-shift detection methods. Future research directions are proposed to enrich QOL theory, response-shift methods, and interpretation of QOL change over time. Appraisal theory and methods are the closest approximation to a response-shift 'gold standard.' They provide the foundation for understanding response shift and point to a unified theory of QOL.


Four of the seven zinc fingers of the Evi-1 myeloid-transforming gene are required for sequence-specific binding to GA(C/T)AAGA(T/C)AAGATAA.
Expression of the Evi-1 gene is activated in murine myeloid leukemias by retroviral insertions and in human acute myelogenous leukemia by translocations and inversions involving chromosome band 3q26 where the gene resides. Aberrant expression of the Evi-1 gene has been shown to interfere with myeloid differentiation, which is proposed to be the basis for its role in leukemias. The Evi-1 gene encodes a 145-kDa DNA-binding protein containing two domains of seven and three Cys2-His2 zinc fingers. Previous studies identified a portion of the consensus DNA-binding sequence for the first domain of zinc fingers. The experiments presented here extend these studies and demonstrate that the first domain recognizes a consensus of 15 nucleotides consisting of GA(C/T)AAGA(T/C)AAGATAA. The first three fingers of the first domain do not detectably bind DNA but contribute to the binding by conferring a relative specificity for GACAA verses GATAA in the first position. The first three fingers also contribute to optimal binding of the 15-nucleotide consensus sequence.


In vitro eye irritation testing using the open source reconstructed hemicornea - a ring trial.
The aim of the present ring trial was to test whether two new methodological approaches for the in vitro classification of eye irritating chemicals can be reliably transferred from the developers' laboratories to other sites. Both test methods are based on the well-established open source reconstructed 3D hemicornea models. In the first approach, the initial depth of injury after chemical treatment in the hemicornea model is derived from the quantitative analysis of histological sections. In the second approach, tissue viability, as a measure for corneal damage after chemical treatment, is analyzed separately for epithelium and stroma of the hemicornea model. The three independent laboratories that participated in the ring trial produced their own hemicornea models according to the test producer's instructions, thus supporting the open source concept. A total of 9 chemicals with different physicochemical and eye-irritating properties were tested to assess the between-laboratory reproducibility (BLR), the predictive performance, as well as possible limitations of the test systems. The BLR was 62.5% for the first and 100% for the second method. Both methods enabled to discriminate Cat. 1 chemicals from all non-Cat. 1 substances, which qualifies them to be used in a top-down approach. However, the selectivity between No Cat. and Cat. 2 chemicals still needs optimization.


Research and implementation interactions in a social accountability study: utilizing guidance for conducting process evaluations of complex interventions.
In recent years, researchers and evaluators have made efforts to identify and use appropriate and innovative research designs that account for the complexity in studying social accountability. The relationship between the researchers and those implementing the activities and how this impacts the study have received little attention. In this paper, we reflect on how we managed the relationship between researchers and implementers using the United Kingdom Medical Research Council (MRC) guidance on process evaluation of a complex intervention. The MRC guidance focuses on three areas of interaction between researchers and stakeholders involved in developing and delivering the intervention: (i) working with program developers and implementers; (ii) communication of emerging findings between researchers/evaluators and implementers; and (iii) overlapping roles of the intervention and research/evaluation. We summarize how the recommendations for each of the three areas were operationalized in the Community and Provider driven Social Accountability Intervention (CaPSAI) Project and provide reflections based on experience. We co-developed various tools, including standard operating procedures, contact lists, and manuals. Activities such as training sessions, regular calls, and meetings were also conducted to enable a good working relationship between the different partners. Studying social accountability requires the collaboration of multiple partners that need to be planned to ensure a good working relationship while safeguarding both the research and intervention implementation. The MRC guidance is a useful tool for making interaction issues explicit and establishing procedures. Planning procedures for dealing with research and implementers' interactions could be more comprehensive and better adapted to social accountability interventions if both researchers and implementers are involved. There is a need for social accountability research to include clear statements explaining the nature and types of relationships between researchers and implementers involved in the intervention.


[On the neurophysiological basis of color-impulse therapy in pathogenic treatment of eye diseases].
Color-impulse therapy (CIT) is one of the accepted treatment options in current ophthalmology. Many publications deal with the effect of CIT on eye diseases, however its pathogenic basis is poorly studied. An attempt to explain CIT effect from the neurophysiological point is made. Survey of number of publications is performed to prove sanogenic effect of CIT on pathogenesis of different pathological eye conditions.


ATM-ATR-dependent up-regulation of DNAM-1 and NKG2D ligands on multiple myeloma cells by therapeutic agents results in enhanced NK-cell susceptibility and is associated with a senescent phenotype.
There is much evidence to support a role for natural killer (NK) cells in controlling the progression of multiple myeloma (MM), a malignancy characterized by an abnormal plasma cell proliferation in the bone marrow (BM). Induction of DNA damage response has been recently shown capable of enhancing NKG2D ligand (NKG2DL) expression, but nothing is known about DNAM-1 ligand (DNAM-1L) regulation. In this study, we show that myeloma cells treated with low doses of therapeutic agents commonly used in the management of patients with MM, such as doxorubicin, melphalan, and bortezomib, up-regulate DNAM-1 and NKG2D ligands. Accordingly, therapeutic drug treatment of MM cells increases NK-cell degranulation, the NKG2D and DNAM-1 receptors being the major triggering molecules. Similar data were also obtained using ex vivo primary plasma cells derived from MM patients. Drug-induced DNAM-1 and NKG2D ligand expression was abolished after treatment with the ATM (ataxia telangiectasia mutated) and ATR (ATM- and RAD3-related) pharmacologic inhibitors caffeine and KU-55933, and was preferentially associated with senescent cells arrested in the G2 phase of the cell cycle. Altogether, our findings have identified a common pathway that can trigger the up-regulation of different NK cell-activating ligands and suggest that NK cells represent an immunosurveillance mechanism toward cells undergoing stress-induced senescent programs.


Pro-neurotensin depends on renal function and is related to all-cause mortality in chronic kidney disease.
Patients with chronic kidney disease (CKD) have a high risk of premature cardiovascular diseases (CVD) and show increased mortality. Pro-neurotensin (Pro-NT) was associated with metabolic diseases and predicted incident CVD and mortality. However, Pro-NT regulation in CKD and its potential role linking CKD and mortality have not been investigated, so far. In a central lab, circulating Pro-NT was quantified in three independent cohorts comprising 4715 participants (cohort 1: patients with CKD; cohort 2: general population study; and cohort 3: non-diabetic population study). Urinary Pro-NT was assessed in part of the patients from cohort 1. In a 4th independent cohort, serum Pro-NT was further related to mortality in patients with advanced CKD. Tissue-specific Nts expression was further investigated in two mouse models of diabetic CKD and compared to non-diabetic control mice. Pro-NT significantly increased with deteriorating renal function (P < 0.001). In meta-analysis of cohorts 1-3, Pro-NT was significantly and independently associated with estimated glomerular filtration rate (P ≤ 0.002). Patients in the middle/high Pro-NT tertiles at baseline had a higher all-cause mortality compared to the low Pro-NT tertile (Hazard ratio: 2.11, P = 0.046). Mice with severe diabetic CKD did not show increased Nts mRNA expression in different tissues compared to control animals. Circulating Pro-NT is associated with impaired renal function in independent cohorts comprising 4715 subjects and is related to all-cause mortality in patients with end-stage kidney disease. Our human and rodent data are in accordance with the hypotheses that Pro-NT is eliminated by the kidneys and could potentially contribute to increased mortality observed in patients with CKD.


A Comparative Study of Colour Effects on Cognitive Performance in Real-World and VR Environments.
This research explores the influence of colour on cognitive performance and intellectual abilities (i.e., logical and lateral thinking abilities and people's attention to detail) in a conventional laboratory setting and an approximately identical virtual reality (VR) environment. Comparative experiments using psychological methods were carried out in both settings to explore the impact of immersive colour experience. This work builds on earlier studies that suggest that the VR environment enhances user experiences, with results evidencing that a considered approach to colour design can trigger a positive impact on user engagement. The experiments further evaluated the positive effects of immersive colour stimuli in VR by evaluating participants' logical and lateral thinking abilities, as well as their attention to detail. Their response time and error rate when completing each psychometric test were recorded with different hue backgrounds in both environments. The data collected from participants reveal the differential impacts of colour between the reality setting using standard colour imaging displays and in an approximately identical VR environment. Analysis of the psychometric tests shows the differential influence of colours on logical and lateral thinking abilities and people's attention to detail between the physical environment and the VR environment. Our findings add to the data demonstrating that a well-designed immersive colour experience in VR can trigger positive user engagement and, as explored in this study, improve cognitive performance. This again positions immersive colour experience as an important design tool to be fully considered in the creation of effective VR research and applications.


Diversity, biological roles and biosynthetic pathways for sugar-glycerate containing compatible solutes in bacteria and archaea.
A decade ago the compatible solutes mannosylglycerate (MG) and glucosylglycerate (GG) were considered to be rare in nature. Apart from two species of thermophilic bacteria, Thermus thermophilus and Rhodothermus marinus, and a restricted group of hyperthermophilic archaea, the Thermococcales, MG had only been identified in a few red algae. Glucosylglycerate was considered to be even rarer and had only been detected as an insignificant solute in two halophilic microorganisms, a cyanobacterium, as a component of a polysaccharide and of a glycolipid in two actinobacteria. Unlike the hyper/thermophilic MG-accumulating microorganisms, branching close to the root of the Tree of Life, those harbouring GG shared a mesophilic lifestyle. Exceptionally, the thermophilic bacterium Persephonella marina was reported to accumulate GG. However, and especially owing to the identification of the key-genes for MG and GG synthesis and to the escalating numbers of genomes available, a plethora of new organisms with the resources to synthesize these solutes has been recognized. The accumulation of GG as an 'emergency' compatible solute under combined salt stress and nitrogen-deficient conditions now seems to be a disseminated survival strategy from enterobacteria to marine cyanobacteria. In contrast, the thermophilic and extremely radiation-resistant bacterium Rubrobacter xylanophilus is the only actinobacterium known to accumulate MG, and under all growth conditions tested. This review addresses the environmental factors underlying the accumulation of MG, GG and derivatives in bacteria and archaea and their roles during stress adaptation or as precursors for more elaborated macromolecules. The diversity of pathways for MG and GG synthesis as well as those for some of their derivatives is also discussed. The importance of glycerate-derived organic solutes in the microbial world is only now being recognized. Their stress-dependent accumulation and the molecular aspects of their interactions with biomolecules have already fuelled several emerging applications in biotechnology and biomedicine.


[Use of solid-phase immunoenzyme analysis for determining allergen-specific IgE antibodies].
An ELISA system for the detection of allergen-specific IgE antibodies to ragweed allergen has been developed. The system is highly sensitive and specific. Ragweed pollen allergen has been obtained by the dialysis of water-soluble extract through a kidney membrane. The high molecular fraction of ragweed allergen, showing the whole of the allergenic activity detected by skin tests in untreated patients, has been used for coating polystyrene assay plates. To detect IgE antibodies to ragweed allergen, the conjugate of sheep anti-IgE antibodies with horse-radish peroxidase has been used. The level of allergen-specific IgE antibodies has been determined on the basis of the data on the optical density of the samples in comparison with that of the normal sera. The correlation factor of the results obtained in the assay of specific IgE antibodies with the newly developed assay system and with the commercial kit Phadezyme RAST manufactured by Pharmacia AB (Sweden) has proved to be 0.82 at n = 39, p less than 0.01, while the variation factor in the reproduction of the assay results has proved to be 12% at n = 40.


Adipose-derived stem cells promote angiogenesis and tissue formation for in vivo tissue engineering.
Adult mesenchymal stem cells secrete a variety of angiogenic cytokines and growth factors, so we proposed that these paracrine mechanisms may be used to promote vascularization and growth for tissue engineering in vivo. We tested whether or not human adipose-derived stem cells (ASCs) promote tissue formation in rats. ASCs were evaluated in vitro for mRNA expression of angiogenic factors, including the vascular endothelial growth factor, basic fibroblast growth factor, interleukin-8 (IL-8), and stromal cell-derived factor-1 (SDF-1) and proliferative activity on human microvascular endothelial cells. For in vivo analysis, CM-DiI-labeled ASCs were implanted with a rat cardiac extracellular matrix (ECM) extract-derived hydrogel into a chamber with a femoral arteriovenous loop in the groin of male nude rats for 7 days. Vascularization in newly generated tissue was estimated by histomorphometry after endothelial nitric oxide synthase (eNOS) immunostaining. ASCs expressed growth factor mRNA and produced an angiogenic activity in vitro. After implantation, ASCs survived, but remained suspended in the ECM and relatively few were incorporated into the newly formed tissue. The volume of newly generated tissue was significantly higher in chambers containing ASCs and it was enriched with vasculature when compared with the ECM alone. We conclude that human ASCs promote tissue growth and angiogenesis in the rat vascularized chamber, thereby showing promise for tissue-engineering applications for regenerative therapy.


Human phenylethanolamine N-methyltransferase pharmacogenomics: gene re-sequencing and functional genomics.
Phenylethanolamine N-methyltransferase (PNMT, EC22.214.171.124) catalyzes the N-methylation of norepinephrine to form epinephrine. As a step toward understanding the possible contribution of inheritance to individual variation in PNMT-catalyzed epinephrine formation, we 're-sequenced' the entire human PNMT gene, including the three exons, the introns and approximately 1 kb of the 5'-flanking region (5'-FR), using DNA samples from 60 African-American (AA) and 60 Caucasian-American (CA) subjects. Within the 3.5 kb re-sequenced, 18 single nucleotide polymorphisms (SNPs) were observed, including four non-synonymous coding SNPs (cSNPs) that resulted in the following alterations in encoded amino acid sequence: Asn9Ser, Thr98Ala, Arg112Cys and Ala175Thr. When constructs for the non-synonymous cSNPs were transiently expressed in COS-1 cells, the Ala98 allozyme displayed significantly lower levels of both activity and immunoreactive protein (p < 0.002) than did the wild-type (WT) enzyme due, at least in part, to accelerated protein degradation by a proteasome-mediated process. Luciferase reporter gene constructs were also created for the six common PNMT 5'-FR haplotypes observed. Significant differences were observed among haplotypes in their ability to drive transcription. These observations raise the possibility of inherited variation in the ability to form epinephrine from norepinephrine as a result of variant PNMT polymorphisms and haplotypes.


Anorectal melanoma.
Anorectal melanoma is an aggressive mucosal melanoma subtype with a poor prognosis. Although recent advancements have been seen for cutaneous melanoma, the optimal treatment paradigm for management of anorectal melanoma is evolving. In this review, we highlight differences in the pathogenesis of mucosal versus cutaneous melanoma, new concepts of staging for mucosal melanoma, updates to surgical management of anorectal melanoma, and current data for adjuvant radiation and systemic therapy in this unique patient population.


Diabetes induction by subdiabetogenic doses of streptozotocin in BALB/cBOM mice. Noninvolvement of host B-lymphocyte functions.
Multiple subdiabetogenic doses of streptozotocin induce an insulin-dependent progressive hyperglycemia in genetically susceptible strains of mice. We have shown previously that T-cell-dependent autoimmune mechanisms play an obligatory role in this model of diabetogenesis by demonstrating that athymic nude mice and lethally irradiated euthymic mice are selectively resistant to diabetes induction and that the susceptibility can be reconstituted by grafting thymus in nude mice or by giving B-cell-depleted splenic lymphocytes to the irradiated mice. In this report we investigate more directly the possible role of host B-cell functions in the induction of hyperglycemia. Mice were rendered selectively deficient in functional B lymphocytes by repeated injections of a polyclonal antiserum against mouse IgM, starting immediately after birth. These B-cell-suppressed mice had no detectable ability to produce antibodies against a test antigen but appeared to have normal levels of T cells. When treated with multiple low doses of streptozotocin, they developed progressive hyperglycemia in a manner indistinguishable from control mice with normal B-cell functions. These results suggest that host B cells, in contrast to host T cells, are not etiologically involved in the development of diabetes induced by multiple subdiabetogenic doses of streptozotocin.


Dysregulation of ion homeostasis by antifungal agents.
Ion-signaling and transduction networks are central to fungal development and virulence because they regulate gene expression, filamentation, host association, and invasion, pathogen stress response and survival. Dysregulation of ion homeostasis rapidly mediates cell death, forming the mechanistic basis by which a growing number of amphipathic but structurally unrelated compounds elicit antifungal activity. Included in this group is carvacrol, a terpenoid phenol that is a prominent component of oregano and other plant essential oils. Carvacrol triggers an early dose-dependent Ca(2+) burst and long lasting pH changes in the model yeast Saccharomyces cerevisiae. The distinct phases of ionic transients and a robust transcriptional response that overlaps with Ca(2+) stress and nutrient starvation point to specific signaling events elicited by plant terpenoid phenols, rather than a non-specific lesion of the membrane, as was previously considered. We discuss the potential use of plant essential oils and other agents that disrupt ion-signaling pathways as chemosensitizers to augment conventional antifungal therapy, and to convert fungistatic drugs with strong safety profiles into fungicides.


A rare cause of late onset neurological deficit in post tuberculous kyphotic deformity-case report.
Late onset neurological deficit is a rare complication of spinal tuberculosis. Reactivation of the disease and compression by internal gibbus are the common causes for late onset neurological deficit. We report a rare cause of late onset paraplegia in a patient with post tubercular kyphotic deformity. The late onset neurological deficit was due to the adjacent segment degeneration proximal to the kyphotic deformity. Posterior hypertrophied ligamentum flavum and anterior disc osteophyte complex caused the cord compression. The increased stress for prolonged period at the end of the deformity was the reason for the accelerated degeneration. Patient underwent posterior decompression, posterolateral and interbody fusion. Deformity correction was not done. To our best knowledge, this is only the second report of this unusual cause of late onset paraplegia.


Comparative analysis of two complete Corynebacterium ulcerans genomes and detection of candidate virulence factors.
Corynebacterium ulcerans has been detected as a commensal in domestic and wild animals that may serve as reservoirs for zoonotic infections. During the last decade, the frequency and severity of human infections associated with C. ulcerans appear to be increasing in various countries. As the knowledge of genes contributing to the virulence of this bacterium was very limited, the complete genome sequences of two C. ulcerans strains detected in the metropolitan area of Rio de Janeiro were determined and characterized by comparative genomics: C. ulcerans 809 was initially isolated from an elderly woman with fatal pulmonary infection and C. ulcerans BR-AD22 was recovered from a nasal sample of an asymptomatic dog. The circular chromosome of C. ulcerans 809 has a total size of 2,502,095 bp and encodes 2,182 predicted proteins, whereas the genome of C. ulcerans BR-AD22 is 104,279 bp larger and comprises 2,338 protein-coding regions. The minor difference in size of the two genomes is mainly caused by additional prophage-like elements in the C. ulcerans BR-AD22 chromosome. Both genomes show a highly similar order of orthologous coding regions; and both strains share a common set of 2,076 genes, demonstrating their very close relationship. A screening for prominent virulence factors revealed the presence of phospholipase D (Pld), neuraminidase H (NanH), endoglycosidase E (EndoE), and subunits of adhesive pili of the SpaDEF type that are encoded in both C. ulcerans genomes. The rbp gene coding for a putative ribosome-binding protein with striking structural similarity to Shiga-like toxins was additionally detected in the genome of the human isolate C. ulcerans 809. The molecular data deduced from the complete genome sequences provides considerable knowledge of virulence factors in C. ulcerans that is increasingly recognized as an emerging pathogen. This bacterium is apparently equipped with a broad and varying set of virulence factors, including a novel type of a ribosome-binding protein. Whether the respective protein contributes to the severity of human infections (and a fatal outcome) remains to be elucidated by genetic experiments with defined bacterial mutants and host model systems.


A temporoparietal and prefrontal network for retrieving the spatial context of lifelike events.
Virtual reality (VR) and event-related functional magnetic resonance imaging were used to study memory for the spatial context of controlled but lifelike events. Subjects received a set of objects from two different people in two different places within a VR environment. Memory for the objects, and for where and from whom they were received was tested by putting the subject back into a place in the company of a person and giving a paired forced choice of objects. In four conditions objects had to be chosen according to different criteria: which was received in that place, which was received from that person, which object was recognized, and which object was widest. Event-related functional magnetic resonance imaging was performed during testing to identify areas involved in retrieval of the spatial context of an event. A network of areas was identified consisting of a temporoparietal pathway running between the precuneus and parahippocampi via retrosplenial cortex and the parieto-occipital sulcus, left hippocampus, bilateral posterior parietal, dorsolateral, ventrolateral and anterior prefrontal cortices, and the anterior cingulate. Of these areas the parahippocampal, right posterior parietal, and posteriodorsal medial parietal areas were specifically involved in retrieval of spatial context compared to retrieval of nonspatial context. The posterior activations are consistent with a model of long-term storage of allocentric representations in medial temporal regions with translation to body-centered and head-centered representations computed in right posterior parietal cortex and buffered in the temporoparietal pathway so as to provide an imageable representation in the precuneus. Prefrontal activations are consistent with strategic retrieval processes, including those required to overcome the interference between the highly similar events.


Sigma receptors regulate contractions of the guinea pig ileum longitudinal muscle/myenteric plexus preparation elicited by both electrical stimulation and exogenous serotonin.
Sigma receptors are specific, highly localized binding sites in limbic and sensorimotor structures of the brain that interact with many psychotropic drugs. These agents include the psychotomimetic benzomorphan opiates, the psychotomimetic drug phencyclidine and its analogs, as well as numerous typical and atypical antipsychotics such as haloperidol, chlorpromazine, and the novel drugs BMY 14802 and rimcazole. So far, no physiological function has been assigned to these binding sites. We have synthesized a number of novel sigma receptor-active drugs derived from the selective sigma ligand N,N'-di(o-tolyl)guanidine (DTG). DTG and its congeners were found to inhibit contractions of the guinea pig ileal longitudinal muscle/myenteric plexus (LMMP) preparation evoked by electrical stimulation. In addition, the sigma ligands noncompetitively antagonized contractions of the LMMP preparation evoked by serotonin (5-HT). The 5-HT-evoked contractions were found to be largely due to 5-HT's activation of 5-HT3 receptors to release ACh. The activity of DTG congeners in inhibiting electrically or 5-HT-evoked contractions of the LMMP highly correlated with their potency to inhibit binding of both 3H-DTG and (+)3H-3-PPP [3(3-OH-phenyl)-N-(1-propyl)piperidine] to sigma receptors in guinea pig brain homogenates. Two DTG congeners that did not bind to sigma receptors also showed no activity in the bioassay. Many other (but not all) sigma receptor ligands showed a high correlation between their potency to inhibit electrically evoked contractions of the LMMP and their sigma receptor binding affinity. The benzomorphans (+)SKF 10,047 and (+)cyclazocine potentiated electrically evoked contractions of the LMMP. Sigma ligands also inhibited the contractions of the LMMP in the presence of the opiate antagonist naloxone and in preparations in which opioid receptors had been inactivated by treatment with the irreversible opiate antagonist beta-chlornaltrexamine. Control experiments suggested that the sigma ligands act via a neuronal mechanism to inhibit ACh release evoked by electrical stimulation or by stimulation with 5-HT. These results suggest that there are functional sigma receptors on cholinergic nerve terminals or within the myenteric plexus and that these receptors can inhibit stimulated ACh release through an opioid receptor-independent mechanism. However, sigma receptor activation in the ileum has the same effect on ACh release as activation of naloxone-sensitive opioid receptors. The LMMP may be an in vitro bioassay system for characterizing the mechanism of action of sigma receptors and for determining the biological efficacy of drugs known to bind to sigma receptors in radioligand binding assays.


The complete sequences of trout (Salmo gairdneri) thymosin beta 11 and its homologue thymosin beta 12.
Two forms of beta-thymosins, designated thymosin beta 11 and thymosin beta 12, were isolated from trout (Salmo gairdneri) spleen. This suggests that the presence of two beta-thymosins, previously thought to be a property of mammalian tissues only, is a more general phenomenon in vertebrate species. Both trout beta-thymosins were found to be N-terminally blocked by a group identified as acetyl by m.s. Automated protein sequencing of tryptic, thermolytic and Staphylococcus aureus in 41-residue V8 proteinase fragments revealed that one of the two beta-thymosins corresponds to the previously reported 41-residue-long sequence of thymosin beta 11 with two substitutions at positions 5 and 7, i.e. Asn instead of Asp, and Glu instead of Gln, whereas the other beta-thymosin, designated thymosin beta 12, was found to be a 42-residue polypeptide closely similar in sequence to thymosin beta 11, with five substitutions (i.e. at positions 5, 7, 10, 11 and 41, with Asp, Ala, Ser, Asn and Thr instead of Asn, Glu, Ala, Ser and Ser respectively) and one addition at position 42 (Ala). Comparison of the known six sequences of beta-thymosins together with the sequences reported here showed that the sequence similarity of the two beta-thymosins in trout (86%) is greater than that of the two beta-thymosins in mammalian species (74%) and that residues at 28 positions are identical in all beta-thymosins, the longer conserved segments located at positions 16-26 and 31-38.


Enrichment of the β-catenin-TCF complex at the S and G2 phases ensures cell survival and cell cycle progression.
Wnt-β-catenin (β-catenin is also known as CTNNB1 in human) signaling through the β-catenin-TCF complex plays crucial roles in tissue homeostasis. Wnt-stimulated β-catenin-TCF complex accumulation in the nucleus regulates cell survival, proliferation and differentiation through the transcription of target genes. Compared with their levels in G1, activation of the receptor LRP6 and cytosolic β-catenin are both upregulated in G2 cells. However, accumulation of the Wnt pathway negative regulator AXIN2 also occurs in this phase. Therefore, it is unclear whether Wnt signaling is active in G2 phase cells. Here, we established a bimolecular fluorescence complementation (BiFC) biosensor system for the direct visualization of the β-catenin-TCF interaction in living cells. Using the BiFC biosensor and co-immunoprecipitation experiments, we demonstrate that levels of the nucleus-localized β-catenin-TCF complex increase during the S and G2 phases, and declines in the next G1 phase. Accordingly, a subset of Wnt target genes is transcribed by the β-catenin-TCF complex during both the S and G2 phases. By contrast, transient inhibition of this complex disturbs both cell survival and G2/M progression. Our results suggest that in S and G2 phase cells, Wnt-β-catenin signaling is highly active and functions to ensure cell survival and cell cycle progression.


Use of antenatal services and delivery care in Entebbe, Uganda: a community survey.
Disparities in perinatal health care occur worldwide. If the UN Millennium Development Goals in maternal and child health are to be met, this needs to be addressed. This study was conducted to facilitate our understanding of the changing use of maternity care services in a semi-urban community in Entebbe Uganda and to examine the range of antenatal and delivery services received in health care facilities and at home. We conducted a retrospective community survey among women using structured questionnaires to describe the use of antenatal services and delivery care. In total 413 women reported on their most recent pregnancy. Antenatal care attendance was high with 96% attending once, and 69% the recommended four times. Blood pressure monitoring (95%) and tetanus vaccination (91%) were the services most frequently reported and HIV testing (47%), haematinics (58%) and presumptive treatment for malaria (66%) least frequently. Hospital clinics significantly outperformed public clinics in the quality of antenatal service. A significant improvement in the reported quality of antenatal services received was observed by year (p < 0.001). Improvement in the range and consistency of services at Entebbe Hospital over time was associated with an increase in the numbers who sought care there (p = 0.038). Although 63% delivered their newborn at a local hospital, 11% still delivered at home with no skilled assistance and just under half of these women reported financial/transportation difficulties as the primary reason. Less educated, poorer mothers were more likely to have unskilled/no assistance. Simple newborn care practices were commonly neglected. Only 35% of newborns were breastfed within the first hour and delayed wrapping of newborn infants occurred after 27% of deliveries. Although antenatal services were well utilised, the quality of services varied. Women were able and willing to travel to a facility providing a good service. Access to essential skilled birth attendants remains difficult especially for less educated, poorer women, commonly mediated by financial and transport difficulties and several simple post delivery practices were commonly neglected. These factors need to be addressed to ensure that high quality care reaches the most vulnerable women and infants.


Soft biological materials and their impact on cell function.
Most organs and biological tissues are soft viscoelastic materials with elastic moduli ranging from on the order of 100 Pa for the brain to 100 000 Pa for soft cartilage. Biocompatible synthetic materials already have many applications, but combining chemical compatibility with physiologically appropriate mechanical properties will increase their potential for use both as implants and as substrates for tissue engineering. Understanding and controlling mechanical properties, specifically softness, is important for appropriate physiological function in numerous contexts. The mechanical properties of the substrate on which, or within which, cells are placed can have as large an impact as chemical stimuli on cell morphology, differentiation, motility, and commitment to live or die.


[Correlation between agricultural production, clinical and demographic variables and prostate cancer: an ecological study].
Risk factors involved in the etiology of prostate cancer are not well known. The objective of this study was to explore correlations among variables relating to agricultural production, the use of health services, food consumption and socio-demographic characteristics and prostate cancer mortality rates in Brazilian states. Univariate analysis of spatial data for investigation of global spatial autocorrelation in prostate cancer mortality rates in Brazilian states between 2005 and 2009 was conducted. Using bivariate analysis, the correlation between socio-demographic indicators, agricultural production data, variables related to the use of health services dietary intake variables and prostate cancer mortality rates were examined. The production of soybeans and corn were positively correlated with prostate cancer mortality. In multiple linear spatial regression, the variables that showed an association with mortality rates from prostate cancer were tons of soybeans produced (p = 0.030), proportion of the population aged 80 and over (p < 0.001) and consumption of beverages (p < 0.001). A positive correlation between tons of soybeans planted and mortality from prostate cancer was identifed, suggesting the possible existence of an association between exposure to pesticides and prostate cancer.


Impact of sexual harassment victimization by peers on subsequent adolescent victimization and adjustment: a longitudinal study.
To examine gender differences in prevalence and types of sexual harassment victimization experienced in grade 9 and how it contributes to relationship victimization and psychological adjustment 2.5 years later. A total of 1734 students from 23 schools completed self-report surveys at entry to grade 9 and end of grade 11. Self-report data were collected on victimization experiences (sexual harassment, physical dating violence, peer violence, and relational victimization) and adjustment (emotional distress, problem substance use, self-harm, suicidal thoughts, maladaptive dieting, feeling unsafe at school, and perpetration of violent delinquency). Separate analyses by sex were prespecified. Sexual harassment victimization was common among boys (42.4%) and girls (44.1%) in grade 9, with girls reporting more sexual jokes, comments, and unwanted touch than among boys, and with boys reporting more homosexual slurs or receiving unwanted sexual content. For girls, sexual harassment victimization in grade 9 was associated with elevated risk of self-harm, suicidal thoughts, maladaptive dieting, early dating, substance use, and feeling unsafe at school. A similar pattern of risk was found for boys, with the exception of dieting and self-harm behaviors. Adjusted odds ratios (AOR) indicated these students were significantly more likely than nonharassed students to report victimization by peers and dating partners 2.5 years later (AOR for boys and girls, respectively; all p < .01), including sexual harassment (AOR: 2.45; 2.9), physical dating violence (AOR: 2.02; 3.73), and physical peer violence (AOR: 2.75; 2.79). Gr 9 sexual harassment also contributed significantly to emotional distress (AOR: 2.09; 2.24), problem substance use (AOR: 1.79; 2.04), and violent delinquency perpetration (AOR: 2.1; 3.34) 2.5 years later (boys and girls, respectively; all p < .01). Sexual harassment at the beginning of high school is a strong predictor of future victimization by peers and dating partners for both girls and boys, and warrants greater prevention and intervention efforts.


FLUORIDE: ITS EFFECTS ON 2 PARAMETERS OF BONE GROWTH IN ORGAN CULTURE.
Bones of the forepaws of young rats were subjected to varying concentrations of fluoride ions in organ culture. The formation of DNA and protein synthesis were evaluated by measurements of the uptake of tritiated thymidine and C(14)-labeled proline. Fluoride concentrations as high as 10 to 20 parts per million had no demonstrable effect in vitro on these basic parameters of skeletal growth


Effects of perioperative granulocyte colony-stimulating factor on horses with ascending colonic ischemia.
Granulocyte colony-stimulating factor (G-CSF) is a glycoprotein that regulates the proliferation and maturation of hematopoietic progenitor cells and modulates the function of mature neutrophils. The responses to administration of G-CSF alone, and in combination with antimicrobials, were studied in an equine model of ascending colon ischemia. Complete segmental colonic ischemia (3.75 hours) with pelvic flexure enterotomy was created in four treatment groups. Group 1 horses received recombinant canine G-CSF (10 micrograms/kg, every 24 hours, intramuscularly), gentamicin sulfate (2.2 mg/kg, every 8 hours, intravenously), and potassium penicillin G (40,000 IU/kg, every 6 hours, intravenously). Group 2 horses were treated with the G-CSF vehicle and antimicrobials as for group 1. Group 3 horses received G-CSF and the antimicrobial drug vehicles, and group 4 horses served as the untreated control receiving G-CSF vehicle and antimicrobial vehicles. The results for 20 horses, five horses in each group, were compared. Treatment with G-CSF was associated with an increased concentration of white blood cells, band neutrophils, neutrophils, lymphocytes, and monocytes in the peripheral blood after surgery. Antimicrobial administration had no detectable effect on cell concentrations after surgery. Administration of G-CSF was associated with an increased concentration of nucleated cells in the peritoneal fluid including neutrophils, small mononuclear cells and large mononuclear cells. Horses that developed incisional infections had lower neutrophil concentrations in the peripheral blood on postoperative day 2 than horses without infected incisions. These results suggested that the prophylactic administration of G-CSF may be useful in the treatment of patients at risk for developing neutropenia after surgery.


Electro-acupuncture improves epileptic seizures induced by kainic acid in taurine-depletion rats.
Electro-acupuncture (EA) partially inhibited epilepsy with great success. The biological basis underlying EA anti-convulsion remained uncertain, which resulted in limited application and slow improvement of acupuncture. Our previous study indicated that taurine may play an inhibitory role against epilepsy as an inhibitory amino acid in the central nervous system and EA may inhibit epilepsy via up-regulating the expression of taurine transporter to increase the release of taurine. Involvement of taurine in kainic acid (KA)-induced epilepsy and anti-convulsion of EA was further addressed on taurine deficiency animal in the present work. We instituted endogenous taurine-deficiency model by supplementation of beta-alanine (3%) in drinking water for continuous 10 days initially, injected KA into lateral cerebral ventricle to induce epileptic seizure, and performed EA treatment on DU26 "RenZhong" and K "YongQuan" acupoints by an EA apparatus (Model G6805-2) using successive waves with the frequency 64Hz and the current intensity 0.8-1.0 mA for 30 minutes in Sprague-Dawley (SD) rats. Taurine levels markedly decreased in cortex, hippocampus, striatum and cerebellum of rats after beta-alanine administration by fluore-HPLC measurement. EA alleviated epileptic activity in rats at 3.5 h time point after KA injection, whereas beta-alanine-induced taurine depletion rendered rats more susceptible to KA-induced epilepsy. Taurine transporter level increased after EA treatment. These results suggested that taurine participated in epileptogenesis and EA may be related to taurine in controlling epileptic seizure.


Expression of transforming growth factor beta1 in bronchial biopsies in asthma and COPD.
The role of transforming growth factor beta1 (TGF beta1) in airway remodeling in asthma and chronic obstructive pulmonary disease (COPD) has not been fully described. To evaluate the possible pathogenetic role of TGF beta1 in asthma and COPD, immunohistochemical expression of TGF beta1 was described in bronchial biopsies from patients with asthma and COPD compared with healthy individuals. Twelve subjects with asthma, 13 subjects with COPD, and 10 healthy individuals enrolled in the study. Bronchial biopsies were stained with hematoxylin and eosin and anti-TGF beta1 antibody. As a result, immunoreactive TGF beta1 was mainly localized in association with connective tissue in all groups. The staining intensity was not statistically different among the groups in bronchial epithelium, whereas it was significantly higher in the group of asthma in the submucosa. Because there is evidence showing a significant increase of staining intensity in the submucosa from asthmatics but not from subjects with COPD, we may conclude that TGF beta1 may play a significant role in pathogenesis of asthma but not in COPD.


Cloning, nucleotide sequence, and expression in Escherichia coli of the Bacillus stearothermophilus peroxidase gene (perA).
The gene encoding a thermostable peroxidase was cloned from the chromosomal DNA of Bacillus stearothermophilus IAM11001 in Escherichia coli. The nucleotide sequence of the 3.1-kilobase EcoRI fragment containing the peroxidase gene (perA) and its flanking region was determined. A 2,193-base-pair open reading frame encoding a peroxidase of 731 amino acid residues (Mr, 82,963) was observed. A Shine-Dalgarno sequence was found 9 base pairs upstream from the translational starting site. The deduced amino acid sequence coincides with those of the amino terminus and four peptides derived from the purified peroxidase of B. stearothermophilus IAM11001. E. coli harboring a recombinant plasmid containing perA produced a large amount of thermostable peroxidase which comigrated on polyacrylamide gel electrophoresis with the B. stearothermophilus peroxidase. The peroxidase of B. stearothermophilus showed 48% homology in the amino acid sequence to the catalase-peroxidase of E. coli.


MHY1 encodes a C2H2-type zinc finger protein that promotes dimorphic transition in the yeast Yarrowia lipolytica.
The yeast-to-hypha morphological transition (dimorphism) is typical of many pathogenic fungi. Dimorphism has been attributed to changes in temperature and nutritional status and is believed to constitute a mechanism of response to adverse conditions. We have isolated and characterized a gene, MHY1, whose transcription is dramatically increased during the yeast-to-hypha transition in Yarrowia lipolytica. Deletion of MHY1 is viable and has no effect on mating, but it does result in a complete inability of cells to undergo mycelial growth. MHY1 encodes a C2H2-type zinc finger protein, Mhy1p, which can bind putative cis-acting DNA stress response elements, suggesting that Mhy1p may act as a transcription factor. Interestingly, Mhy1p tagged with a hemagglutinin epitope was concentrated in the nuclei of actively growing cells found at the hyphal tip.


Selective association of spectrin with the cytoplasmic surface of human erythrocyte plasma membranes. Quantitative determination with purified (32P)spectrin.
A specific association between spectrin and the inner surface of the human erythrocyte membrane has been examined by measuring the binding of purified [32P]spectrin to inside out, spectrin-depleted vesicles and to right side out ghost vesicles. Spectrin was labeled by incubating erythrocytes with 32Pi, and eluted from the ghost membranes by extraction in 0.3 mM NaPO4, pH 7.6. [32P]Spectrin was separated from actin and other proteins and isolated in a nonaggregated state as a So20,w = 7 S (in 0.3 mM NaPO4) or So20,w = 8 S (in 20 mM KCl, 0.3 mM NaPO4) protein after sedimentation on linear sucrose gradients. Binding of [32P]spectrin to inverted vesicles devoid of spectrin and actin was at least 10-fold greater than to right side out membranes, and exhibited different properties. Association with inside out vesicles was slow, was decreased to the value for right side out vesicles at high pH, or after heating spectrin above 50 degrees prior to assay, and was saturable with increasing levels of spectrin. Binding to everted vesicles was rapid, unaffected by pH or by heating spectrin, and rose linearly with the concentration of spectrin. Scatchard plots of binding to inverted vesicles were linear at pH 7.6, with a KD of 45 microng/ml, while at pH 6.6, plots were curvilinear and consistent with two types of interactions with a KD of 4 and 19 microng/ml, respectively. The maximal binding capacity at both pH values was about 200 microng of spectrin/mg of membrane protein. Unlabeled spectrin competed for binding with 50% displacement at 27 microng/ml. [32P]Spectrin dissociated and associated with inverted vesicles with an identical dependence on ionic strength as observed for elution of native spectrin from ghosts. MgCl2, CaCl2 (1 to 4 mM) and EDTA (0.5 to 1 mM) had little effect on binding in the presence of 20 mM KCl, while at low ionic strength, MgCl2 (1 mM) increased binding and inhibited dissociation to the same extent as 10 to 20 mM KCl. Binding was abolished by pretreatment of vesicles with 0.1 M acetic acid, or with 0.1 microng/ml of trypsin. The periodic acid-Schiff-staining bands were unaffected by trypsin digestion which destroyed binding; mild digestion, which decreased binding only 50%, converted Band 3 almost completely to a membrane-bound 50,000-dalton fragment resistant to further proteolysis. These experiments suggest that attachment of spectrin to the cytoplasmic surface of the membrane results from a selective protein-protein interaction which is independent of erythrocyte actin. A direct role of the major sialoglycoprotein or Band 3 as a membrane binding site appears unlikely.


The combination of thrombophilic genotypes is associated with definite antiphospholipid syndrome.
Thrombosis and pregnancy morbidity are clinical features of the definite antiphospholipid syndrome (APS). These clinical complications are also associated with the presence of inherited thrombophilias. Interactions between acquired and genetic risk factors are becoming increasingly related to a higher thrombotic risk. The aim of our study was to determine the prevalence of four common gene polymorphisms in patients with antiphospholipid antibodies (aPL). A series of 105 consecutive unselected patients with aPL grouped as having APS (n= 69) and not having APS (n= 36) was studied. A control group of 200 healthy subjects was also investigated for the presence of factor V Leiden (FVL), the 20210A allele of the prothrombin (PT-20210A) gene, the thermolabile variant (677TT) of methylenetetrahydrofolate reductase (MTHFR), and the 4G/4G genotype of the plasminogen activator inhibitor (PAI-1) promoter. Two patients who belong to the APS group carried the FVL while PT-20210A was found in 6 patients with APS (8.7%) and in 1 of the non-APS group (2.8%). The prevalence of FVL was similar to that found in the control group whereas PT-20210A was significantly more frequent in APS patients than in normal controls (2.0%, p=0.02). The MTHFR-677TT was found in 22.0%, 15.1% and 13.0%, and the PAI-1 (4G/4G) in 27.5%, 22.8% and 23.5% of APS, non-APS patients and normal controls, respectively. Furthermore, combinations of PT-20210A or FVL with PAI-1 (4G/4G) were significantly more frequent in APS patients (5.8%) than in normal controls (0.5%, p=0.016). This difference was not found between non-APS patients and normal subjects. Present data indicate that testing for heritable thrombophilia would be important to identify aPL subjects with an increased risk of developing APS.


Chronic inhibition of alpha4beta2 nicotinic receptors in the ventral hippocampus of rats: impacts on memory and nicotine response.
Acute and chronic systemic nicotine administration has been shown to cause significant spatial memory improvement. The critical nicotinic receptor subtypes for this effect and their location are still being determined. Nicotinic receptors in the ventral hippocampus have been found to be critically involved in memory. Acute ventral hippocampal infusions of dihydro-beta-erythroidine (DHbetaE), an alpha4beta2 nicotinic receptor antagonist, impaired spatial memory of rats in the radial-arm maze. The current study used chronic ventral hippocampal infusion of DHbetaE as a model of nicotinic receptor loss such as that which occurs in Alzheimer's disease. The therapeutic effect of systemic nicotine treatment in reversing the DHbetaE-induced memory impairment was determined. Rats were pretrained to asymptotic levels of performance on the eight-arm radial maze. Then, they were implanted with bilateral infusion cannulae in the ventral hippocampus, through which 0, 33.3, or 100 microg/side/day of DHbetaE was continuously infused for 4 weeks. The rats were retested on the eight-arm maze throughout infusion period and after withdrawal, and the interaction of acute systemic nicotine injections on memory was tested. The higher (100 microg/side/day) but not the lower (33.3 microg/side/day) DHbetaE dose caused a significant spatial memory impairment. Acute systemic nicotine injections (0, 0.1, 0.2, and 0.4 mg/kg, subcutaneous) attenuated the memory impairing effects of 100 microg/side/day of DHbetaE. There was no significant effect on response latency with the chronic DHbetaE infusion. Acute systemic nicotine infusions did significantly speed responding, an effect which was reversed by chronic hippocampal infusions of DHbetaE. After withdrawal there were no significant lasting effects on choice accuracy or response latency. Wet-dog shakes were significantly elevated during chronic hippocampal DHbetaE administration with no effect during the withdrawal period. These results indicate that chronic inhibition of a subset of nicotinic receptors in the hippocampus results in a significant impairment in the spatial memory choice accuracy. The ability of nicotine to attenuate the impairment supports the development of nicotinic agonist therapy of syndromes, such as Alzheimer's disease, that involve a chronic decrease in the activity of the alpha4beta2 nicotinic receptors and memory impairment.


Update in Lung Cancer 2014.
In the past 2 years, lung cancer research and clinical care have advanced significantly. Advancements in the field have improved outcomes and promise to lead to further reductions in deaths from lung cancer, the leading cause of cancer death worldwide. These advances include identification of new molecular targets for personalized targeted therapy, validation of molecular signatures of lung cancer risk in smokers, progress in lung tumor immunotherapy, and implementation of population-based lung cancer screening with chest computed tomography in the United States. In this review, we highlight recent research in these areas and challenges for the future.


Endovascular Treatment for Acute Stroke Patients With a Pre-stroke Disability: An International Survey.
Background: It is unclear what factors clinicians consider when deciding about endovascular thrombectomy (EVT) in acute ischemic stroke patients with a pre-existing disability. We aimed to explore international practice patterns and preferences for EVT in patients with a pre-stroke disability, defined as a modified Rankin score (mRS) ≥ 2. Methods: Electronic survey link was sent to principal investigators of five major EVT trials and members of a professional interventional neurology society. Results: Of the 81 survey-responding clinicians, 57% were neuro-interventionalists and 33% were non-interventional stroke clinicians. Overall, 64.2% would never or almost never consider EVT for a patient with pre-stroke mRS of 4-5, and 49.3% would always or almost always offer EVT for a patient with pre-stroke mRS 2-3. Perceived benefit of EVT (89%) and severity of baseline disability (83.5%) were identified as the most important clinician-level and patient-level factors that influence EVT decisions in these patients. Conclusion: In this survey of 80 respondents, we found that EVT practice for patients with pre-stroke disability across the world is heterogenous and depends upon patient characteristics. Individual clinician opinions substantially alter EVT decisions in pre-stroke disabled patients.