Changes in the frequencies of human hematopoietic stem and progenitor cells with age and site.
This study enumerated CD45(hi)/CD34(+) and CD45(hi)/CD133(+) human hematopoietic stem cells (HSCs) and progenitor granulocyte-macrophage colony forming cells (GM-CFCs) in blood and trochanteric and femoral bone marrow in 233 individuals. Stem cell frequencies were determined with multiparameter flow cytometry and using an internal control to determine the intrinsic variance of the assays. Progenitor cell frequency was determined using a standard colony assay technique. The frequency of outliers from undetermined methodological causes was highest for blood, but less than 5% for all values. The frequency of CD45(hi)/CD133(+) cells correlated highly with the frequency of CD45(hi)/CD34(+) cells in trochanteric and femoral bone marrow. The frequency of these HSC populations in trochanteric and femoral bone marrow rose significantly with age. In contrast, there was no significant trend of either of these cell populations with age in the blood. Trochanteric marrow progenitor GM-CFCs showed no significant trends with age, but femoral marrow GM-CFCs trended downward with age, potentially because of the reported conversion of red marrow at this site to fat with age. Hematopoietic stem and progenitor cells exhibited changes in frequencies with age that differed between blood and bone marrow. We previously reported that side population (SP) multipotential HSC, which includes the precursors of CD45(hi)/CD133(+) and CD45(hi)/CD34(+), decline with age. Potentially the increases in stem cell frequencies in the intermediate compartment between SP and GM progenitor cells observed in this study represent a compensatory increase for the loss of more potent members of the HSC hierarchy.


Oxidative stress and the regulation of complement activation in human glaucoma.
As part of ongoing studies on proteomic alterations during glaucomatous neurodegeneration, this study focused on the complement system. Human retinal protein samples obtained from donor eyes with (n = 10) or without (n = 10) glaucoma were analyzed by a quantitative proteomic approach using mass spectrometry. Cellular localization of protein expression for different complement components and regulators were also determined by immunohistochemical analysis of an additional group of human donor eyes with glaucoma (n = 34) compared with age-matched control eyes without glaucoma (n = 20). In addition, to determine the regulation of complement factor H (CFH) by oxidative stress, in vitro experiments were performed using rat retinal cell cultures incubated in the presence and absence of an oxidant treatment. Proteomic analysis detected the expression and differential regulation of several complement components in glaucomatous samples, which included proteins involved in the classical and the lectin pathways of complement activation. In addition, several complement regulatory proteins were detected in the human retinal proteome, and glaucomatous samples exhibited a trend toward downregulation of CFH expression. In vitro experiments revealed that oxidative stress, which was also prominently detectable in the glaucomatous human retinas, downregulated CFH expression in retinal cells. These findings expand the current knowledge of complement activation by presenting new evidence in human glaucoma and support that despite important roles in tissue cleaning and healing, a potential deficiency in intrinsic regulation of complement activation, as is evident in the presence of oxidative stress, may lead to uncontrolled complement attack with neurodestructive consequences.


Engagement of L-selectin impairs the actin polymerizing capacity of beta 2-integrins on neutrophils.
A sequential activation of L-selectin and beta 2-integrins on neutrophils is crucial for the rolling, adherence and subsequent migration of these cells on the endothelium. However, little is known about a possible interplay between these adhesion receptors in the final regulation of cell motility. The results presented here show that sulfatides themselves (here used as tools to activate L-selectins), have no major effect on the cellular content of filamentous actin (F-actin), but cause a time-related decrease in the beta 2-integrin-induced formation of F-actin. This effect of sulfatides was abolished in cells lacking L-selectin as a result of pretreatment with chymotrypsin. A similar sulfatide-induced activation of L-selectin also caused a pronounced and time-related decrease of a subsequent chemotactic peptide-induced F-actin response. The effect of sulfatides on both beta 2-integrin- and chemotactic peptide-induced F-actin were abolished if L-selectin were blocked by preincubating the cells with specific antibodies to L-selectin. These effects of L-selectin engagement on cellular F-actin content were neither abolished by blocking the cytosolic free Ca2+ signal with bis-(2-amino-5-methylphenoxy)ethane-N,N,N',N'-tetraaceticacid tetraacetoxymethyly ester (MAPT/AM) nor by blocking a cAMP-induced activation of protein kinase A by pretreating the cells with adenosine-3',5'-cyclic monophos-phorothioate (Rp-cAMPS). Instead we found that L-selectin engagement impaired an early beta 2-integrin-induced tyrosine kinase activation, an event shown to be necessary for a normal beta 2-integrin-mediated F-actin response. The present demonstration of a negative feed-back function of L-selectin on beta 2-integrin-induced modulations of the actin cytoskeleton, suggests that the relative distribution and/or density of the respective L-selectin and beta 2-integrin ligands on endothelial cells might be important factors in determining the final site of firm adhesion and extravasation of neutrophils.


Characterization of food-grade titanium dioxide: the presence of nanosized particles.
Titanium dioxide (TiO2) is widely used in food products, which will eventually enter wastewater treatment plants and terrestrial or aquatic environments, yet little is known about the fraction of this TiO2 that is nanoscale, or the physical and chemical properties of TiO2 that influence its human and environmental fate or toxicity. Instead of analyzing TiO2 properties in complex food or environmental samples, we procured samples of food-grade TiO2 obtained from global food suppliers and then, using spectroscopic and other analytical techniques, quantified several parameters (elemental composition, crystal structure, size, and surface composition) that are reported to influence environmental fate and toxicity. Another sample of nano-TiO2 that is generally sold for catalytic applications (P25) and widely used in toxicity studies, was analyzed for comparison. Food-grade and P25 TiO2 are engineered products, frequently synthesized from purified titanium precursors, and not milled from bulk scale minerals. Nanosized materials were present in all of the food-grade TiO2 samples, and transmission electron microscopy showed that samples 1-5 contained 35, 23, 21, 17, and 19% of nanosized primary particles (<100 nm in diameter) by number, respectively (all primary P25 particles were <100 nm in diameter). Both types of TiO2 aggregated in water with an average hydrodynamic diameter of >100 nm. Food-grade samples contained phosphorus (P), with concentrations ranging from 0.5 to 1.8 mg of P/g of TiO2. The phosphorus content of P25 was below inductively coupled plasma mass spectrometry detection limits. Presumably because of a P-based coating detected by X-ray photoelectron spectroscopy, the ζ potential of the food-grade TiO2 suspension in deionized water ranged from -10 to -45 mV around pH 7, and the iso-electric point for food-grade TiO2 (<pH 4) was significantly lower than that for P25. The presence of other elements in or on the TiO2 (Si content of 0.026-0.062% and Al content of 0.0006-0.810%) was also different from the case for P25 and would influence the environmental fate of TiO2. X-ray diffraction analysis confirmed the presence of anatase and/or rutile in the food-grade materials, and although the presence of amorphous TiO2 could not be ruled out, it is unlikely on the basis of Raman analysis. The food-grade TiO2 was solar photoactive. Cationic dyes adsorbed more readily to food-grade TiO2 than P25, indicating very different potentials for interaction with organics in the environment. This research shows that food-grade TiO2 contains engineered nanomaterials with properties quite different from those of P25, which has previously been used in many ecotoxicity studies, and because food-grade TiO2 is more likely than P25 to enter the environment (i.e., potentially higher exposure levels), there is a need to design environmental (and human) fate and toxicity studies comparing food-grade to catalytic TiO2.


Re-appraisal of the stratigraphy and determination of new U-Pb dates for the Sterkfontein hominin site, South Africa.
Sterkfontein Caves is the single richest early hominin site in the world with deposits yielding one or more species of Australopithecus and possible early Homo, as well as an extensive faunal collection. The inability to date the southern African cave sites accurately or precisely has hindered attempts to integrate the hominin fossil evidence into pan-African scenarios about human evolutionary history, and especially hominin biogeography. We have used U-Pb and U-Th techniques to date sheets of calcium carbonate flowstone inter-bedded between the fossiliferous sediments. For the first time, absolute age ranges can be assigned to the fossil-bearing deposits: Member 2 is between 2.8 +/- 0.28 and 2.6 +/- 0.30 Ma and Member 4 between 2.65 +/- 0.30 and 2.01 +/- 0.05 Ma. The age of 2.01 +/- 0.05 Ma for the top of Member 4 constrains the last appearance of Australopithecus africanus to 2 Ma. In the Silberberg Grotto we have reproduced the U-Pb age of approximately 2.2 Ma of for the flowstones associated with StW573. We believe that these deposits, including the fossil and the flowstones, accumulated rapidly around 2.2 Ma. The stratigraphy of the site is complex as sediments are exposed both in the underground chambers and at surface. We present a new interpretation of the stratigraphy based on surface mapping, boreholes logs and U-Pb ages. Every effort was made to retain the Member system, however, only Members 2 and 4 are recognized in the boreholes. We propose that the deposits formally known as Member 3 are in fact the distal equivalents of Member 4. The sediments of Members 2 and 4 consisted of cone-like deposits and probably never filled up the cave. The U-Th ages show that there are substantial deposits younger than 400 ka in the underground cave, underlying the older deposits, highlighting again that these cave fills are not simple layer-cakes.


The musculoskeletal effects of perioperative smoking.
Although the carcinogenic consequences of smoking are well known, further research is needed on the effects of smoking on musculoskeletal health and surgical outcomes. Orthopaedic perioperative complications of smoking include impaired healing, increased infection, delayed and/or impaired fracture union and arthrodesis, and inferior arthroplasty outcomes. The incorporation of smoking cessation protocols such as transdermal patches, chewing gum, lozenges, inhalers, sprays, bupropion, and varenicline in the perioperative period may result in substantial benefits for patients' musculoskeletal and general health.


Protective effect of treatment with black cumin oil on spatial cognitive functions of rats that suffered global cerebrovascular hypoperfusion.
The fixed oil of black cumin seeds, Nigella sativa L. (NSO), has shown considerable antioxidant and anti-inflammatory activities. Chronic cerebral hypoperfusion has been linked to neurodegenerative disorders including Alzheimer's disease (AD) and its subsequent cognitive impairment in which oxidative stress and neuroinflammation are the principal culprits. Cerebrovascular hypoperfusion was experimentally achieved by bilateral common carotid arteries occlusion (2VO) in rats. Morris water maze (MWM) test was employed to assess the effects of NSO on spatial cognitive function before and after 2VO intervention. Rats were divided into long-term memory (LTM) and short-term memory (STM) groups, each was further subdivided into 3 subgroups: sham control, untreated 2VO and NSO treated 2VO group. All subgroups were tested with MWM at the tenth postoperative week. Working memory test results for both sham control and NSO treated groups showed significantly lower escape latency time and total distance travelled than untreated 2VO group. Similarly, LTM and STM MWM tests for sham control and NSO treated groups revealed significantly better maze test performance as compared to untreated 2VO group. Sham control and NSO treated 2VO groups demonstrated superior probe memory test performance as compared to untreated 2VO group. The fixed oil of Nigella sativa seeds has demonstrated noticeable spatial cognitive preservation in rats challenged with chronic cerebral hypoperfusion which indicates a promising prospective neuroprotective effect.


Glucocorticoid receptor gene mutations in leukemic cells acquired in vitro and in vivo.
Glucocorticoid resistance was investigated in human leukemic CCRF-CEM cells. A mutation (L753F), which renders the human glucocorticoid receptor (hGR) gene functionally hemizygous, was identified in all CEM-derived cell lines analyzed. Allele-specific PCR identified the same mutation in lymph node biopsy material from patient CEM cells. Given the correlation between hGR concentration and glucocorticoid sensitivity, this suggests that loss of functional heterozygosity may result in resistance to glucocorticoid-based chemotherapy. The L753F mutation was probably not responsible for the ontogeny of the disease because it did not appear to be present in all leukemic cells. Thus, it is unlikely that hGR mutations would be detected in leukemic patients at presentation, but they may occur, and be selected for, during treatment. Deletions and point mutations in the hGR gene of cells selected for steroid resistance in vitro were investigated by PCR-single strand conformation polymorphism analysis. Loss of hGR mRNA expression resulted from 5'-deletion of the hGR gene and nonsense mutations in exon 6. These results provide the first evidence for somatic mutation in the hGR gene of a patient with acute lymphoblastic leukemia, offer a potential in vivo mechanism for acquisition of steroid resistance in leukemia, and suggest that screening for additional in vivo mutations will require analysis of genomic DNA.


Integrative analysis of differentially expressed microRNAs of pulmonary alveolar macrophages from piglets during H1N1 swine influenza A virus infection.
H1N1 swine influenza A virus (H1N1 SwIV) is one key subtype of influenza viruses with pandemic potential. MicroRNAs (miRNAs) are endogenous small RNA molecules that regulate gene expression. MiRNAs relevant with H1N1 SwIV have rarely been reported. To understand the biological functions of miRNAs during H1N1 SwIV infection, this study profiled differentially expressed (DE) miRNAs in pulmonary alveolar macrophages from piglets during the H1N1 SwIV infection using a deep sequencing approach, which was validated by quantitative real-time PCR. Compared to control group, 70 and 16 DE miRNAs were respectively identified on post-infection day (PID) 4 and PID 7. 56 DE miRNAs were identified between PID 4 and PID 7. Our results suggest that most host miRNAs are down-regulated to defend the H1N1 SwIV infection during the acute phase of swine influenza whereas their expression levels gradually return to normal during the recovery phase to avoid the occurrence of too severe porcine lung damage. In addition, targets of DE miRNAs were also obtained, for which bioinformatics analyses were performed. Our results would be useful for investigating the functions and regulatory mechanisms of miRNAs in human influenza because pig serves as an excellent animal model to study the pathogenesis of human influenza.


Enantioselective Synthesis of anti-syn-Trihalides and anti-syn-anti-Tetrahalides via Asymmetric β-Elimination.
Structural motifs containing contiguous halide-bearing stereocenters are common in natural products as well as bioactive molecules. A few successful examples have been reported in the area of asymmetric vicinal dihalogenation of alkenes for accessing dihalogenated products; in this report, an alternative generation method of contiguous halide-bearing stereocenters α,β,γ,δ relative to carbonyl group in excellent enantioselectivity is proposed by utilizing a Song's oligoEG catalyst-catalyzed asymmetric β-elimination. According to this methodology, a wide range of anti-syn-trihalides and anti-syn-anti-tetrahalides with high levels of enantioselectivity were synthesized. The synthetic utility of the contiguous halide-bearing stereocenters was demonstrated by several transformations. The results of high-resolution mass spectrometry indicated that the favorable interaction between catalyst and one of the enantiomers of racemic contiguously multihalogenated ketone contributed to the original enantioselectivity of dehydrohalogenation. A deuterium kinetic isotope effect experiment revealed that this β-elimination reaction proceeds by the E2 mechanism. This strategy opens a new pathway for the asymmetric synthesis of contiguous halide-bearing stereocenters of great complexity.


Usefulness of apheresis to extract microfilarias in management of loiasis.
Two patients with loiasis in whom treatment with diethylcarbamazine had to be interrupted because of serious side effects underwent apheresis to extract microfilarias from the blood. After three apheresis sessions the number of microfilarias in the blood had dropped from 8980 to 3412/ml in one patient and from 2950 to 1840/ml in the other. Between 40 and 45 X 10(6) microfilarias were removed from each of the patients. After reduction of the parasitaemia the two patients were given diethylcarbamazine; no appreciable side effects occurred, and the patients were cured.


Housing conditions of urban households with Aboriginal children in NSW Australia: tenure type matters.
Housing is a key determinant of the poor health of Aboriginal Australians. Most Aboriginal people live in cities and large towns, yet research into housing conditions has largely focused on those living in remote areas. This paper measures the prevalence of housing problems amongst participants in a study of urban Aboriginal families in New South Wales, Australia, and examines the relationship between tenure type and exposure to housing problems. Cross-sectional survey data was provided by 600 caregivers of 1406 Aboriginal children aged 0-17 years participating in Phase One of the Study of Environment on Aboriginal Resilience and Child Health (SEARCH). Regression modelling of the associations between tenure type (own/mortgage, private rental or social housing) and housing problems was conducted, adjusting for sociodemographic factors. The majority (60%) of SEARCH households lived in social housing, 21% rented privately and 19% either owned their home outright or were paying a mortgage ("owned"). Housing problems were common, particularly structural problems, damp and mildew, vermin, crowding and unaffordability. Physical dwelling problems were most prevalent for those living in social housing, who were more likely to report three or more physical dwelling problems than those in owned (PR 3.19, 95%CI 1.97, 5.73) or privately rented homes (PR 1.49, 1.11, 2.08). However, those in social housing were the least likely to report affordability problems. Those in private rental moved home most frequently; children in private rental were more than three times as likely to have lived in four or more homes since birth than those in owned homes (PR 3.19, 95%CI 1.97, 5.73). Those in social housing were almost half as likely as those in private rental to have lived in four or more homes since birth (PR 0.56, 95%CI 0.14, 0.77). Crowding did not vary significantly by tenure type. The high prevalence of housing problems amongst study participants suggests that urban Aboriginal housing requires further attention as part of efforts to reduce the social and health disadvantage experienced by Aboriginal Australians. Particular attention should be directed to the needs of those renting in the private and social housing sectors, who are experiencing the poorest dwelling conditions.


Natural history of growth hormone deficiency in a pediatric cohort.
Controversies still exist regarding the evaluation of growth hormone deficiency (GHD) in childhood at the end of growth. The aim of this study was to describe the natural history of GHD in a pediatric cohort. This is a retrospective study of a cohort of pediatric patients with GHD. Cases of acquired GHD were excluded. Univariate logistic regression was used to identify predictors of GHD persisting into adulthood. Among 63 identified patients, 47 (75%) had partial GHD at diagnosis, while 16 (25%) had complete GHD, including 5 with multiple pituitary hormone deficiencies. At final height, 50 patients underwent repeat stimulation testing; 28 (56%) recovered and 22 (44%) remained growth hormone (GH) deficient. Predictors of persisting GHD were: complete GHD at diagnosis (OR 10.1, 95% CI 2.4-42.1), pituitary stalk defect or ectopic pituitary gland on magnetic resonance imaging (OR 6.5, 95% CI 1.1-37.1), greater height gain during GH treatment (OR 1.8, 95% CI 1.0-3.3), and IGF-1 level <-2 standard deviation scores (SDS) following treatment cessation (OR 19.3, 95% CI 3.6-103.1). In the multivariate analysis, only IGF-1 level <-2 SDS (OR 13.3, 95% CI 2.3-77.3) and complete GHD (OR 6.3, 95% CI 1.2-32.8) were associated with the outcome. At final height, 56% of adolescents with GHD had recovered. Complete GHD at diagnosis, low IGF-1 levels following retesting, and pituitary malformation were strong predictors of persistence of GHD.


Therapeutic Effect of Ultrasound-Guided Peripherally Inserted Central Catheter Combined with Predictive Nursing in Patients with Large-Area Severe Burns.
This study was aimed to explore the application value of ultrasound-guided peripherally inserted central catheter (PICC) combined with predictive nursing in the treatment of large-area severe burns. 88 patients with large-area severe burns who visited hospital were chosen as the research objects. They were randomly divided into the observation group and the control group, with 44 cases in each. The patients in the observation group were treated with ultrasound-guided PICC combined with predictive nursing, while those in the control group were treated with traditional PICC and nursing methods. Then, the anxiety of patients was compared between groups by the Self-rating Anxiety Scale (SAS), while the depression was compared by the Self-rating Depression Scale (SDS). The pain of the patients was analyzed by the McGill Pain Questionnaire (MPQ), and a self-made nursing satisfaction questionnaire was adopted to evaluate the nursing satisfaction. The surgery-related indicators of the patients were detected and recorded (the success rate of one-time puncture, the success rate of one-time catheter placement, incidence of complications, heart rate, blood pressure, etc.). The success rates of one-time puncture (93% vs. 86%) and of catheter placement (95% vs. 81%) in the observation group were significantly higher than those in the control group, P < 0.05. The pain scores of the observation group were much lower than those of the control group at each time period, P < 0.05. The number of patients with negative emotions such as anxiety and depression in the observation group was markedly less than that in the control group. The incidence of complications in the observation group was notably lower than that in the control group (4.5% vs 18%), P < 0.05. The nursing satisfaction of the observation group was significantly higher than that of the control group (93% vs 79.5%), P < 0.05. In conclusion, ultrasound-guided PICC and predictive nursing had high clinical application values in the treatment of patients with large-area severe burns.


High-performance surface acoustic wave resonators in the 1 to 3 GHz range using a ScAlN/6H-SiC structure.
This paper describes application of Sc-doped AlN (ScAlN) to wideband SAW devices in the 1 to 3 GHz range. First, it is shown theoretically that large SAW velocity and electromechanical coupling factor are simultaneously achievable when the ScAlN film is combined with a base substrate with extremely high acoustic wave velocities, such as diamond and SiC. Next, SAW delay lines are fabricated on the ScAlN/6H-SiC structure, and reasonable agreement between the theory and experiment is obtained. Finally, one-port SAW resonators are fabricated on the structure, and it is shown that high-performance is achievable in the 1 to 3 GHz range by use of the structure.


A Novel Cellular Model to Study Angiotensin II AT2 Receptor Function in Breast Cancer Cells.
Recent studies have highlighted the AT1 receptor as a potential therapeutic target in breast cancer, while the role of the AT2 subtype in this disease has remained largely neglected. The present study describes the generation and characterization of a new cellular model of human invasive breast cancer cells (D3H2LN-AT2) stably expressing high levels of Flag-tagged human AT2 receptor (Flag-hAT2). These cells exhibit high-affinity binding sites for AngII, and total binding can be displaced by the AT2-selective antagonist PD123319 but not by the AT1-selective antagonist losartan. Of interest, high levels of expression of luciferase and green fluorescent protein make these cells suitable for bioluminescence and fluorescence studies in vitro and in vivo. We provide here a novel tool to investigate the AT2 receptor functions in breast cancer cells, independently of AT1 receptor activation.


A role for chloride transport in lysosomal protein degradation.
Loss of the lysosomal chloride transport protein ClC-7 leads to complex phenotypes in mice and man, including osteopetrosis, accumulation of lysosomal storage material, and neurodegeneration. Using novel tissue-specific ClC-7 knockout mice, we have shown that upon loss of ClC-7, lysosomal degradation of endocytosed protein is slowed down and accumulation of autophagosomes occurs.


Mutational disruption of plasma membrane trafficking of Saccharomyces cerevisiae Yor1p, a homologue of mammalian multidrug resistance protein.
The ATP binding cassette (ABC) transporter protein Yor1p was identified on the basis of its ability to elevate oligomycin resistance when it was overproduced from a high-copy-number plasmid. Analysis of the predicted amino acid sequence of Yor1p indicated that this protein was a new member of a subfamily of ABC transporter proteins defined by the multidrug resistance protein (MRP). In this work, Yor1p is demonstrated to localize to the Saccharomyces cerevisiae plasma membrane by both indirect immunofluorescence and biochemical fractionation studies. Several mutations were generated in the amino-terminal nucleotide binding domain (NBD1) of Yor1p to test if the high degree of sequence conservation in this region of the protein was important for function. Deletion of a phenylalanine residue at Yor1p position 670 led to a mutant protein that appeared to be retained in the endoplasmic reticulum (ER) and that was unstable. As shown by others, deletion of the analogous residue from a second mammalian MRP family member, the cystic fibrosis transmembrane conductance regulator (CFTR), also led to retention of this normally plasma membrane-localized protein in the ER. Changes in the spacing between or the sequences flanking functional motifs of Yor1p NBD1 led to defective trafficking or decreased activity of the mutant proteins. Analyses of the degradation of wild-type and DeltaF670 Yor1p indicated that the half-life of DeltaF670 Yor1p was dramatically shortened. While the vacuole was the primary site for turnover of wild-type Yor1p, degradation of DeltaF670 Yor1p was found to be more complex with both proteasomal and vacuolar contributions.


Effect of lipotoxicity on mitochondrial function and epigenetic programming during bovine in vitro embryo production.
Maternal metabolic disorders may cause lipotoxic effects on the developing oocyte. Understanding the timing at which this might disrupt embryo epigenetic programming and how this is linked with mitochondrial dysfunction is crucial for improving assisted reproductive treatments, but has not been investigated before. Therefore, we used a bovine in vitro model to investigate if pathophysiological palmitic acid (PA) concentrations during in vitro oocyte maturation and in vitro embryo culture alter embryo epigenetic patterns (DNA methylation (5mC) and histone acetylation/methylation (H3K9ac/H3K9me2)) compared to control (CONT) and solvent control (SCONT), at the zygote and morula stage. Secondly, we investigated if these epigenetic alterations are associated with mitochondrial dysfunction and changes in ATP production rate, or altered expression of epigenetic regulatory genes. Compared to SCONT, H3K9ac and H3K9me2 levels were increased in PA-derived zygotes. Also, 5mC and H3K9me2 levels were increased in PA-exposed morulae compared to SCONT. This was associated with complete inhibition of glycolytic ATP production in oocytes, increased mitochondrial membrane potential and complete inhibition of glycolytic ATP production in 4-cell embryos and reduced SOD2 expression in PA-exposed zygotes and morulae. For the first time, epigenetic alterations in metabolically compromised zygotes and morulae have been observed in parallel with mitochondrial dysfunction in the same study.


Expression pattern of alphavbeta3 and alphavbeta5 integrin mRNA in mouse fetal gonads.
The alphavbeta3 and alphavbeta5 integrins are known as transmembrane receptors capable of binding to the RGD amino acid peptide sequence. In mouse early gonadogenesis, some proteins containing the RGD sequence are deposited into extracellular space and participate in morphogenesis. We analyzed the expression patterns of the alphavbeta3 and alphavbeta5 integrins in mouse developing gonads (10.5-13.5 days post coitum) using whole-mount in situ hybridization. The alphav integrin mRNA was homogenously expressed in developing gonadal regions. On the other hand, the beta3 integrin mRNA was found only in large and round cells (presumptive germ cells), whereas beta5 integrin was localized in gonadal somatic cells, with the exception of coelomic epithelial cells. The beta3 integrin-expressed cells were determined to be primordial germ cells because the number of these cells was drastically reduced in busulfan-treated gonads. In this study, we demonstrated that the alphavbeta3 and alphavbeta5 integrins are widely localized in the mouse developing gonads and discussed their presumptive functions on mouse gonadogenesis.


Activation of ethanol metabolism in humans by fructose: importance of experimental design.
Fed human male subjects 18 to 30 years of age received 0.85 g/kg of ethanol p.o. as vodka followed by a second drink (0.3-0.4 g/kg) given 3 to 4 hr later. Blood ethanol levels reached approximately 100 mg% after both doses. Breath samples were taken every 20 to 30 min, and the rate of ethanol elimination was determined. Subjects returned 1 to 2 weeks later and the study was repeated with fructose (1.0 g/kg). When ethanol and fructose were given simultaneously, the peak blood ethanol values reached were reduced significantly. This resulted in calculated increases in the apparent volume of distribution of ethanol; however, the slope (beta) of the ethanol elimination curve was not increased. Thus, under these conditions, fructose appeared to have minimal effects on rates of ethanol elimination (ca. 10%). In another experimental design, subjects received 1 g/kg of fructose 1 hr after ethanol. Under these conditions, fructose did not affect the peak blood ethanol concentration or the apparent volume of distribution; however, the slopes of the ethanol elimination curves (beta) were increased significantly by fructose. Further, the time estimated to eliminate the ethanol completely was reduced by 90 min with fructose. Under these conditions, the stimulation of ethanol elimination by fructose was 20 to 30%. This increase in the rate of ethanol elimination was observed in every subject studied. Similar results were obtained with rats. To understand why blood ethanol values were lower when ethanol and fructose were given simultaneously, female Sprague-Dawley rats were given ethanol alone (1.5 g/kg) or ethanol and fructose (1 g/kg) simultaneously.(ABSTRACT TRUNCATED AT 250 WORDS)


Bone mineral density in newly diagnosed children with neuroblastoma.
Neuroblastoma is the second most common extracranial malignant tumor of childhood and the most common solid tumor of infancy which is characterized by bone metastasis. Previous reports on bone mineral density (BMD) in patients with leukemia and solid malignancies concentrate on long-term survivors and on the effect of chemotherapeutic agents and irradiation. Also, evaluation of BMD in neuroblastoma was reported in few studies which were conducted upon adult survivors of childhood cancer. Previous studies on both acute leukemia and lymphoma patients suggested that the disease process itself played a role in decrease BMD. We evaluated 27 patients with newly diagnosed neuroblastoma for both lumbar (L2-L4) BMD and total BMD using dual energy X-ray absorptiometery (DXA) scan to highlight the effect of neuroblastoma as a disease process on BMD as this disease characterized by bone metastasis. Three out of the 27 patients showed low bone mass in both lumbar and total BMD studies. Low bone mass may occur in early disease process of neuroblastoma and it is important to consider BMD assessment during the early course of the disease as well as the long-term survivors as a part of the patient screening in suspected cases.


[Damage to the structure of DNA and to the metabolism of nucleic acids in the thymus in relation to the dose of tritium oxide].
Single administration of tritium oxide (1.1-44 MBq per 1 kg of body mass) causes a dose-dependent decrease in the level of rat thymus nucleic acids within the range of concentrations of injected tritium 5.5-44 MBq/g. Biosynthesis of nucleic acids turned out to be more sensitive to tritium oxide action. The rate of biosynthesis changed on the 1st day at the lowest dose of 1.1 MBq/g, the absorbed dose on the 1st day was 7.5 cGy. Dose-dependent disorders of DNA structure in the content of 2-strand DNA after the administration of tritium oxide (9-44 MBq/g) were established.


TNF-alpha-induced optic nerve degeneration and nuclear factor-kappaB p65.
To characterize a model of optic nerve axonal degeneration induced by tumor necrosis factor (TNF)-alpha and to determine the role of nuclear factor (NF)-kappaB p65 in axonal degeneration. Groups of rats were euthanatized at 1 day, 1 or 2 weeks, or 1 or 2 months after intravitreal injection of TNF-alpha. Morphometric analyses of neurofilament- or Thy-1-positive cells, retinal ganglion cells (flat preparations stained with cresyl violet or retrograde labeling with a neurotracer), the number of axons, immunostaining for myelin basic protein, and TUNEL assays were performed. Levels of NF-kappaB p65 protein in retina and optic nerve were determined by Western blot analysis and immunohistochemistry. The effects of antisense oligodeoxynucleotide (AS ODN) against NF-kappaB p65 and helenalin, an inhibitor of NF-kappaB p65 activation, on TNF-alpha-induced optic nerve degeneration were determined by counting the number of axons. Intravitreal injections of TNF-alpha induced obvious axonal loss and extensive degeneration of the axons from 2 weeks to 2 months after injection, whereas significant retinal ganglion cell loss was noted only at 2 months after injection. NF-kappaB p65 was increased in the optic nerve but not in the retina and was found to colocalize with ED-1 and Iba1, markers of microglia. Inhibition of NF-kappaB p65 with AS ODN or helenalin significantly ameliorated the effects of TNF-alpha-mediated axonal loss. TNF-alpha causes axonal degeneration with probable delayed loss of retinal ganglion cell bodies. NF-kappaB p65 may play a pivotal role in axonal degeneration, with the possible involvement of microglial cells.


Sensitivity to Enterocins of Biogenic Amine-Producing Faecal Enterococci from Ostriches and Pheasants.
Enterococci are widespread bacteria forming the third largest genus among lactic acid bacteria. Some possess probiotic properties or they can produce beneficial proteinaceous antimicrobial substances called enterocins. On the other hand, some enterococci produce biogenic amines (BAs), so this study is focused on the sensitivity to enterocins of biogenic amine-producing faecal enterococci from ostriches and pheasants. Altogether, 60 enterococci isolated from faeces of ostriches and pheasants were tested for production of BAs. This target of the identified enterococci involved 46 strains selected from 140 ostriches and 17 from 60 pheasants involving the species Enterococcus hirae, E. faecium, E. faecalis, and E. mundtii. Although BAs histamine, cadaverine, putrescine, and tryptamine were not detected in the enterococci tested, in general high BA production by the tested enterococci was noted. The species E. hirae formed the majority of the enterococcal strains from ostrichs faeces (34 strains). High production of tyramine (TYM) was measured with an average amount of 958.16 ± 28.18 mg/ml. Among the enterococci from pheasants, the highest was production of TYM compared to phenylethylamine, spermidine, and spermine. Enterococci featured high BA production; however, they were sensitive to seven enterocins with inhibition activity ranging from 100 up to 25,600 AU/ml.


Two-slab all-optical spring.
It is demonstrated that a waveguide consisting of two dielectric slabs may become an all-optical spring when guiding a superposition of two transverse evanescent modes. Both slabs are transversally trapped in stable equilibrium due to the optical forces developed. A condition for stable equilibrium on the wavenumbers of the two modes is expressed analytically. The spring constant characterizing the system is shown to have a maximal value as a function of the equilibrium distance between the slabs and their width.


Synthesis and characterization of metal-organic frameworks based on 4-hydroxypyridine-2,6-dicarboxylic acid and pyridine-2,6-dicarboxylic acid ligands.
The self-assembly of 4-hydroxypyridine-2,6-dicarboxylic acid (H(3)CAM) and pyridine-2,6-dicarboxylic acid (H2PDA) with Zn(II) salts under hydrothermal conditions gave two novel coordination polymers {[Zn(HCAM)].H2O}n (1) and {[Zn(PDA)(H2O)(1.5)]}n (1a). 1 and 1a comprise of a 2D (4,4) net and a 1D zigzag chain, respectively, in which a new coordination mode of PDA is found. The reactions of H(3)CAM and H2PDA with Nd2O3 in the M/L ratio 2:3 gave {[Nd2(HCAM)3(H2O)4].2H2O}n (2) and {[Nd(2)(PDA)3(H2O)(3)].0.5H2O}n (2a). In 2, a square motif as a building block constructed by four Nd(III) ions was further assembled into a highly ordered 2D (4,4) grid. 2a is a 3D microporous coordination polymer. It is interesting to note that, when Ln(III) salts rather than oxides were employed, the reaction produced {[Ln(CAM)(H2O)3].H2O}n (Ln = Gd, 3; Dy, 4; Er, 5) for H(3)CAM and {[Gd2(PDA)3(H2O)3].H2O}n (3a) for H2PDA. 3-5 are 2D coordination polymers with a 3(3)4(2) uniform net, where hydroxyl groups of H3CAM coordinate with metal ions. The reaction of H3CAM and Er2O3 instead of Er(ClO4)3 produced {[Er2(HCAM)3(H2O)4].2H2O}n (6). The compounds 2a and 3a, 2 and 6 are isomorphous. The stereochemical and supramolecular effects of hydroxyl groups result in the dramatic structural changes from 1D (1a) to 2D (1) and from 2D (2) to 3D (2a). When Ln(III) salts instead of Ln2O3 were employed in the hydrothermal reactions with H(3)CAM, different self-assembly processes gave the products of different metal/ligand ratio with reactants (3-5).


Heterogeneous nuclear ribonucleoprotein A2/B1 regulates the self-renewal and pluripotency of human embryonic stem cells via the control of the G1/S transition.
Self-renewal and pluripotency of human embryonic stem cells (hESCs) are a complex biological process for maintaining hESC stemness. However, the molecular mechanisms underlying these special properties of hESCs are not fully understood. Heterogeneous nuclear ribonucleoprotein A2/B1 (hnRNP A2/B1) is a multifunctional RNA-binding protein whose expression is related to cell proliferation and carcinogenesis. In this study, we found that hnRNP A2/B1 expression was localized to undifferentiated hESCs and decreased upon differentiation of hESCs. hnRNP A2/B1 knockdown reduced the number of alkaline phosphatase-positive colonies in hESCs and led to a decrease in the expression of pluripotency-associated transcription factors OCT4, NANOG, and SOX2, indicating that hnRNP A2/B1 is essential for hESC self-renewal and pluripotency. hnRNP A2/B1 knockdown increased the expression of gene markers associated with the early development of three germ layers, and promoted the process of epithelial-mesenchymal transition, suggesting that hnRNP A2/B1 is required for maintaining the undifferentiated and epithelial phenotypes of hESCs. hnRNP A2/B1 knockdown inhibited hESC proliferation and induced cell cycle arrest in the G0/G1 phase before differentiation via degradation of cyclin D1, cyclin E, and Cdc25A. hnRNP A2/B1 knockdown increased p27 expression and induced phosphorylation of p53 and Chk1, suggesting that hnRNP A2/B1 also regulates the G1/S transition of hESC cell cycle through the control of p27 expression and p53 and Chk1 activity. Analysis of signaling molecules further revealed that hnRNP A2/B1 regulated hESC proliferation in a PI3K/Akt-dependent manner. These findings provide for the first time mechanistic insights into how hnRNP A2/B1 regulates hESC self-renewal and pluripotency.


Generation of an Oocyte-Specific Cas9 Transgenic Mouse for Genome Editing.
The CRISPR/Cas9 system has been developed as an easy-handle and multiplexable approach for engineering eukaryotic genomes by zygote microinjection of Cas9 and sgRNA, while preparing Cas9 for microinjection is laborious and introducing inconsistency into the experiment. Here, we describe a modified strategy for gene targeting through using oocyte-specific Cas9 transgenic mouse. With this mouse line, we successfully achieve precise gene targeting by injection of sgRNAs only into one-cell-stage embryos. Through comprehensive analysis, we also show allele complexity and off-target mutagenesis induced by this strategy is obviously lower than Cas9 mRNA/sgRNA injection. Thus, injection of sgRNAs into oocyte-specific Cas9 transgenic mouse embryo provides a convenient, efficient and reliable approach for mouse genome editing.


Molecular cloning and characterization of the major endothelin receptor subtype in porcine cerebellum.
Endothelin receptors (ETRs) display subtype heterogeneity and are widely distributed throughout the tissues of the periphery and central nervous system. In order to gain further insight into the potential molecular differences of ETRs, we initiated molecular cloning of ETR genes by screening for the appearance of 125I-ET-1 binding activity in COS cells transfected with pools of a porcine cerebellum cDNA expression library. Two independent clones (pPCETR 1.1 and pPCETR 5.6) were identified and isolated by repeated rounds of pool enrichment and COS cell expression. DNA sequence analysis of pPCET 1.1 and pPCET 5.6 indicated that both clones have the same nucleotide sequence; the deduced amino acid sequence indicated that the porcine cerebellum ETR is 443 residues in length and consists of seven potential transmembrane domains, with homology to members of the GTP-binding protein-coupled receptor superfamily. Northern analysis indicated a single mRNA species of about 5 kilobases, which is expressed significantly in cerebellum, lung, kidney, and pituitary. Expression of functional receptor was demonstrated by endothelin-1 (ET-1)-mediated Ca2+ mobilization in COS cells transfected with pPCETR 1.1 (COS/ETR 1.1) and ET-1-mediated electrophysiological responses in Xenopus oocytes injected with RNA derived from pPCETR 1.1. Quantitative comparison of saturation binding of 125I-ET-1 to either porcine cerebellum or COS/ETR 1.1 membranes indicated an identical apparent dissociation constant. The relative efficacy of ET-related peptides to compete for binding of 125I-ET-1 to receptor from porcine cerebellum and COS/ETR 1.1 indicated that both preparations encode a nonselective or ETBR subtype. Chemical cross-linking of 125I-ET-1 to receptor derived from cerebellum or COS/ETR 1 revealed two bands, with apparent molecular masses of 47 and 35 kDa. These data demonstrate that the pPCETR 1.1 encodes the major ETR subtype in the porcine cerebellum.


Associations of Neighborhood Opportunity and Social Vulnerability With Trajectories of Childhood Body Mass Index and Obesity Among US Children.
Physical and social neighborhood attributes may have implications for children's growth and development patterns. The extent to which these attributes are associated with body mass index (BMI) trajectories and obesity risk from childhood to adolescence remains understudied. To examine associations of neighborhood-level measures of opportunity and social vulnerability with trajectories of BMI and obesity risk from birth to adolescence. This cohort study used data from 54 cohorts (20 677 children) participating in the Environmental Influences on Child Health Outcomes (ECHO) program from January 1, 1995, to January 1, 2022. Participant inclusion required at least 1 geocoded residential address and anthropometric measure (taken at the same time or after the address date) from birth through adolescence. Data were analyzed from February 1 to June 30, 2022. Census tract-level Child Opportunity Index (COI) and Social Vulnerability Index (SVI) linked to geocoded residential addresses at birth and in infancy (age range, 0.5-1.5 years), early childhood (age range, 2.0-4.8 years), and mid-childhood (age range, 5.0-9.8 years). BMI (calculated as weight in kilograms divided by length [if aged <2 years] or height in meters squared) and obesity (age- and sex-specific BMI ≥95th percentile). Based on nationwide distributions of the COI and SVI, Census tract rankings were grouped into 5 categories: very low (<20th percentile), low (20th percentile to <40th percentile), moderate (40th percentile to <60th percentile), high (60th percentile to <80th percentile), or very high (≥80th percentile) opportunity (COI) or vulnerability (SVI). Among 20 677 children, 10 747 (52.0%) were male; 12 463 of 20 105 (62.0%) were White, and 16 036 of 20 333 (78.9%) were non-Hispanic. (Some data for race and ethnicity were missing.) Overall, 29.9% of children in the ECHO program resided in areas with the most advantageous characteristics. For example, at birth, 26.7% of children lived in areas with very high COI, and 25.3% lived in areas with very low SVI; in mid-childhood, 30.6% lived in areas with very high COI and 28.4% lived in areas with very low SVI. Linear mixed-effects models revealed that at every life stage, children who resided in areas with higher COI (vs very low COI) had lower mean BMI trajectories and lower risk of obesity from childhood to adolescence, independent of family sociodemographic and prenatal characteristics. For example, among children with obesity at age 10 years, the risk ratio was 0.21 (95% CI, 0.12-0.34) for very high COI at birth, 0.31 (95% CI, 0.20-0.51) for high COI at birth, 0.46 (95% CI, 0.28-0.74) for moderate COI at birth, and 0.53 (95% CI, 0.32-0.86) for low COI at birth. Similar patterns of findings were observed for children who resided in areas with lower SVI (vs very high SVI). For example, among children with obesity at age 10 years, the risk ratio was 0.17 (95% CI, 0.10-0.30) for very low SVI at birth, 0.20 (95% CI, 0.11-0.35) for low SVI at birth, 0.42 (95% CI, 0.24-0.75) for moderate SVI at birth, and 0.43 (95% CI, 0.24-0.76) for high SVI at birth. For both indices, effect estimates for mean BMI difference and obesity risk were larger at an older age of outcome measurement. In addition, exposure to COI or SVI at birth was associated with the most substantial difference in subsequent mean BMI and risk of obesity compared with exposure at later life stages. In this cohort study, residing in higher-opportunity and lower-vulnerability neighborhoods in early life, especially at birth, was associated with a lower mean BMI trajectory and a lower risk of obesity from childhood to adolescence. Future research should clarify whether initiatives or policies that alter specific components of neighborhood environment would be beneficial in preventing excess weight in children.


(3,4-Dihydroxyphenylimino)-2-imidazoline (DPI) and its action at noradrenergic and dopaminergic receptors in the nucleus accumbens of rats: mesolimbic catecholamine receptors and hyperactivity.
Bilateral administration of ergometrine into the nucleus accumbens of rats pretreated 1-3 min earlier with intra-accumbens injections of noradrenaline, phenylephrine, clonidine and phentolamine has been found to produce a normal ergometrine-induced hyperactivity. In contrast, low doses of dopamine and (3,4-dihydroxyphenylimino)-2-imidazoline (DPI) and only high doses of clonidine and phentolamine have been found to attenuate the ergometrine response. These data together with the finding that phentolamine is unable to alter DPI's ability to suppress the ergometrine response provide direct evidence that the latter DPI effect is certainly not due to its ability to act as an agonist at alpha-NE receptors within the nucleus accumbens of rats.


Principles governing oligomer formation in amyloidogenic peptides.
Identifying the principles that describe the formation of protein oligomers and fibrils with distinct morphologies is a daunting problem. Here we summarize general principles of oligomer formation gleaned from molecular dynamics simulations of Abeta-peptides. The spectra of high free energy structures sampled by the monomer provide insights into the plausible fibril structures, providing a rationale for the 'strain phenomenon.' Heterogeneous growth dynamics of small oligomers of Abeta(16-22), whose lowest free energy structures are like nematic droplets, can be broadly described using a two-stage dock-lock mechanism. In the growth process, water is found to play various roles depending on the oligomer size, and peptide length, and sequence. Water may be an explicit element of fibril structure linked to various fibril morphologies.


Interferonogenicity of rabies vaccines as related to their therapeutic and prophylactic activities.
Non-interferonogenic rabies vaccine prepared from a virus grown in sheep brain and devoid of neuroallergenic factor and an interferonogenic vaccine from the same virus strain cultured in Japanese quail embryo cells have been compared. In mouse experiments both preparations appeared to have identical therapeutic and prophylactic efficacy. It seems that interferonogenicity of rabies vaccines cannot be used as a criterion of their prophylactic or therapeutic effects. The effectiveness of exogenous and endogenous interferon (IFN) in mice infected by the highly pathogenic fixed rabies virus has been experimentally demonstrated. Combined application of the vaccine and IFN or its inducer exerted the most marked therapeutic effect. Administration of IFN inducer into mouse brain was much more effective than its extraneural inoculation.


An ultra-short contra-directional coupler utilizing surface plasmon-polaritons at optical frequencies.
A nano-scaled coupled-line coupler based on the guidance of surface plasmon-polaritons (SPPs) is proposed, designed and simulated at optical frequencies. The coupler comprises layered dielectric materials and silver, which serve as two stacked nano-transmission lines to achieve broadside coupling. The key property of this coupler is that it operates based on the principle of contra-directional coupling between a forward and a backward wave giving rise to supermodes that are characterized by complex-conjugate eigenvalues (even when the materials are assumed lossless). The resulting exponential attenuation along the coupler leads to dramatically reduced coupling lengths compared to previously reported co-directional SPP couplers (e.g. from millimeters to submicrons). The effect of material losses and finite coupler width are also analyzed.


Ultrastructure of Meelsvirus: A nuclear virus of arrow worms (phylum Chaetognatha) producing giant "tailed" virions.
Most known giant viruses, i.e., viruses producing giant virions, parasitize amoebae and other unicellular eukaryotes. Although they vary in the level of dependence on host nuclear functions, their virions self-assemble in the host cell's cytoplasm. Here we report the discovery of a new prototype of giant virus infecting epidermal cells of the marine arrow worm Adhesisagitta hispida. Its 1.25 μm-long virions self-assemble and accumulate in the host cell's nucleus. Conventional transmission electron microscopy reveals that the virions have a unique bipartite structure. An ovoid nucleocapsid, situated in a broad "head" end of the virion is surrounded by a thin envelope. The latter extends away from the head to form a voluminous conical "tail" filled with electron-dense extracapsidular material. The 31nm-thick capsid wall has a distinctive substructure resulting from a patterned arrangement of subunits; it bears no ultrastructural resemblance to the virion walls of other known giant viruses. The envelope self-assembles coincident with the capsid and remotely from all host membranes. We postulate that transmission to new hosts occurs by rupture of protruding virion-filled nuclei when infected arrow worms mate. Future genomic work is needed to determine the phylogenetic position of this new virus, which we have provisionally named Meelsvirus.


The radioprotective agent WR1065 protects cells from radiation damage by regulating the activity of the Tip60 acetyltransferase.
The aminothiol WR1065 is a highly effective free radical scavenger which can protect cells from the cytotoxic effects of ionizing radiation. Currently, WR1065 is used clinically to protect patients from radiation injury occurring during radiation therapy protocols. However, it is becoming increasingly clear that WR1065 can alter radiosensitivity through a mechanism which is independent of its ability to function as a free radical scavenger. Here, we examined the ability of WR1065 to directly regulate signaling pathways involved in the DNA damage response. The ability of WR1065 to enhance the survival of irradiated bone marrow cells and primary cultures was established. DNA damage signaling was monitored by measuring activation of the ATM kinase by western blot analysis and activation of Tip60 using an in vitro acetylation assay. Tip60 function was abrogated by expression of a catalytically inactive Tip60, and the effect on radiosensitivity evaluated. Treatment of cells with WR1065 led to a small but significant increase in the kinase activity of ATM. Further, WR1065 robustly activated the Tip60 acetyltransferase, which is a key upstream regulator of the ATM kinase. In addition, WR1065 directly activated the acetyltransferase activity of purified Tip60 in vitro, indicating a direct interaction between WR1065 and Tip60. Finally, cells with reduced levels of Tip60 activity exhibited a significant reduction in radioprotection by WR1065. Direct regulation of Tip60's acetyltransferase activity by WR1065 makes a significant contribution to the radioprotective effects of WR1065. Activators of Tip60 may therefore make effective clinical radioprotectors.


VEGF secretion by neuroendocrine tumor cells is inhibited by octreotide and by inhibitors of the PI3K/AKT/mTOR pathway.
Gastroenteropancreatic (GEP) endocrine tumors are hypervascular tumors able to synthesize and secrete high amounts of VEGF. We aimed to study the regulation of VEGF production in GEP endocrine tumors and to test whether some of the drugs currently used in their treatment, such as somatostatin analogues and mTOR inhibitors, may interfere with VEGF secretion. We therefore analyzed the effects of the somatostatin analogue octreotide, the mTOR inhibitor rapamycin, the PI3K inhibitor LY294002, the MEK1 inhibitor PD98059 and the p38 inhibitor SB203850 on VEGF secretion, assessed by ELISA and Western blotting, in three murine endocrine cell lines, STC-1, INS-r3 and INS-r9. Octreotide and rapamycin induced a significant decrease in VEGF production by all three cell lines; LY294002 significantly inhibited VEGF production by STC-1 and INS-r3 only. We detected no effect of PD98059 whereas SB203850 significantly inhibited VEGF secretion in INS-r3 and INS-r9 cells only. By Western blotting analysis, we observed decreased intracellular levels of VEGF and HIF-1alpha under octreotide, rapamycin and LY294002. For rapamycin and LY294002, this effect was likely mediated by the inhibition of the mTOR/HIF-1/VEGF pathway. In addition to its well-known anti-secretory effects, octreotide may also act through the inhibition of the PI3K/Akt pathway, as suggested by the decrease in Akt phosphorylation detected in all three cell lines. In conclusion, our study points out to the complex regulation of VEGF synthesis and secretion in neoplastic GEP endocrine cells and suggests that the inhibition of VEGF production by octreotide and rapamycin may contribute to their therapeutic effects.


In-house 3D-printed aligners: effect of in vivo ageing on mechanical properties.
To investigate alterations in the mechanical properties of in-house three-dimensional (3D) printed orthodontic aligners after intraoral ageing. Sixteen 3D-printed aligners (TC-85DAC resin, Graphy, Seoul, Korea) were used for the purpose of the study, which were divided into 10 control (not used) aligners and 6 materials retrieved from 4 patients after 1-week service (retrieved group). Samples from the control group were analysed by attenuated total reflectance-Fourier-transform infrared (ATR-FTIR) spectroscopy. Samples from control/retrieved groups were embedded resin and subjected to instrumented indentation testing (IIT) to record force-indentation depth curves, calculating the following (as per ISO 14577-1, 2002 standard): Martens hardness (HM), indentation modulus (EIT), and elastic index (ηIT), and the indentation relaxation index (RIT). Differences between control and retrieved 3D-printed aligners were checked with Mann-Whitney/t-tests at an alpha = 5%. ATR-FTIR analysis showed that aligners were made of a vinyl ester-urethane material. The results of the IIT testing were: HM (control: median 91.5 N/mm2, interquartile range [IQR] 88.0-93.0/as-retrieved: median 90.5 N/mm2, IQR 89.0-93.0); EIT (control, mean 2616.3 MPa, standard deviation [SD] 107.0 MPa/retrieved, mean 2673.2 MPa, SD 149.4 MPa); ηIT (control: median 28.6%, IQR 28.2-30.9%/as-retrieved: median 29.0%, IQR 28.7-29.2%); and RIT (control: median 45.5%, IQR 43.0-47.0%/as-retrieved: median 45.1%, IQR 45.0-45.3%). No differences between as-retrieved and control aligners were found for any of the mechanical properties tested (P > 0.05 in all instances). The mechanical properties of the in-house 3D-printed aligners tested were not affected after 1 week in service period.


The relationship between physician participation in continuing professional development programs and physician in-practice peer assessments.
To investigate the relationship between physicians' performance, as evaluated through in-practice peer assessments, and their participation in continuing professional development (CPD). The authors examined the predictive effects of participating in the CPD programs of the Royal College of Physicians and Surgeons of Canada and the College of Family Physicians of Canada one year before in-practice peer assessments conducted by the medical regulatory authority in Ontario, Canada, in 2008-2009. Two multivariate logistic regression models were used to determine whether physicians who reported participating in any CPD and group-based, assessment-based, and/or self-directed CPD activities were more or less likely to receive satisfactory assessments than physicians who had not. All models were adjusted for the effects of sex, age, specialty certification, practice location, number of patient visits per week, hours worked per week, and international medical graduate status. A total of 617 physicians were included in the study. Analysis revealed that physicians who reported participating in any CPD activities were significantly more likely (odds ratio [OR] = 2.5; P = .021) to have satisfactory assessments than those who had not. In addition, physicians participating in group-based CPD activities were more likely to have satisfactory assessments than those who did not (OR = 2.4; P = .016). There is encouraging evidence supporting a positive predictive association between participating in CPD and performance on in-practice peer assessments. The findings have potential implications for policies which require physicians to participate in programs of lifelong learning.


[Combined idiopathic fibrosing alveolitis and lung cancer].
Data are reported of a rare disease of idiopathic fibrosing alveolitis in association with an adenoma of the thyroid gland in a 68 years old woman. The authors emphasize difficulties in the clinical roentgenological and morphological diagnosis in this kind of combined pathology.


[Clinical experience with a custom-made hip prosthesis developed in Norway].
A custom-made hip prosthesis is developed at St. Olav s Hospital and the Norwegian University of Science and Technology (NTNU) in Trondheim. The purpose was to design an uncemented stem for optimal fit of the proximal femur to achieve secure fixation and optimal strain distribution to the bone. A customized hip prosthesis permits reconstruction of the normal joint mechanism, including reconstruction of leg length, a physiological hip centre and a normal lever arm for the major hip muscles. 6 years of experience with the device at the orthopaedic department at Sørlandet Hospital, Arendal is presented. 43 hips in 37 patients were operated. Median time of observation was 3 years (3 months-6 years). The patients' age was on average 49 years. 70% of the hips had hip dysplasia. The patients were followed prospectively, both clinically and radiologically for up to 6 years (median 3 years). The operations were performed as planned in all patients with an average operation time of 96 minutes. We have had no re-operations, no post-operative dislocations of the operated hips and no infections. No components have loosened and no patient has complained of thigh pain. One patient had a per-operative femoral fissure that was treated with cerclage, and one patient had an affection of nervus ischiadicus. The mean Harris hip score was 49 pre- and 97 post-operation . Our experience with the Norwegian custom-made hip prosthesis is promising and confirms the good results from the Trondheim group.


Purification of Alaskan walleye pollock (Gadus chalcogrammus) and New Zealand hoki (Macruronus novaezelandiae) liver oil using short path distillation.
The beneficial health effects of a diet rich in n-3 long chain polyunsaturated fatty acids (n-3 LC-PUFA) have been extensively researched in recent years. Marine oils are an important dietary source of n-3 LC-PUFA, being especially rich in two of the most important fatty acids of this class, EPA (eicosapentaenoic acid; 20:5n-3) and DHA (docosahexaenoic acid; 22:6n-3). Oils rich in n-3 LC-PUFA are prone to oxidation that leads to loss of product quality. Alaskan pollock (Gadus chalcogrammus Pallas, 1814) and New Zealand's hoki (Macruronus novaezelandiae Hector, 1871) are the highest volume fisheries of their respective countries. Both produce large quantities of fishery byproducts, in particular crude or unrefined n-3 LC-PUFA containing oils. Presently these oils are used as ingredients for animal feed, and only limited quantities are used as human nutritional products. The aim of this research was to investigate the applicability of short path distillation for the purification of pollock and hoki oil to produce purified human-grade fish oil to meet quality specifications. Pollock and hoki oils were subjected to short path distillation and a significant decrease in free fatty acids and lipid oxidation (peroxide and para-anisidine values) products was observed. Purified oils met the Global Organization for EPA and DHA Omega-3 (GOED) standard for edible fish oils.


What's the effect of the implementation of general practitioner cooperatives on caseload? Prospective intervention study on primary and secondary care.
Out-of-hours care in the primary care setting is rapidly changing and evolving towards general practitioner 'cooperatives' (GPC). GPCs already exist in the Netherlands, the United Kingdom and Scandinavia, all countries with strong general practice, including gatekeepers' role. This intervention study reports the use and caseload of out-of-hours care before and after implementation of a GPC in a well subscribed region in a country with an open access health care system and no gatekeepers' role for general practice. We used a prospective before/after interventional study design. The intervention was the implementation of a GPC. One year after the implementation of a GPC, the number of patient contacts in the intervention region significantly increased at the GPC (OR: 1.645; 95% CI: 1.439-1.880), while there were no significant changes in patient contacts at the Emergency Department (ED) or in other regions where a simultaneous registration was performed. Although home visits decreased in all general practitioner registrations, the difference was more pronounced in the intervention region (intervention region: OR: 0.515; 95% CI: 0.411-0.646, other regions: OR: 0.743; 95% CI: 0.608-0.908). At the ED we observed a decrease in the number of trauma cases (OR: 0.789; 95% CI: 0.648-0.960) and of patients who came to hospital by ambulance (OR: 0.687; 95% CI: 0.565-0.836). One year after its implementation more people seek help at the GPC, while the number of contacts at the ED remains the same. The most prominent changes in caseload are found in the trauma cases. Establishing a GPC in an open health care system, might redirect some patients with particular medical problems to primary care. This could lead to a lowering of costs or a more cost-effective out of hours care, but further research should focus on effective usage to divert patient flows and on quality and outcome of care.


Theoretical study of chromophores for biological sensing: Understanding the mechanism of rhodol based multi-chromophoric systems.
Development of two-photon fluorescent probes can aid in visualizing the cellular environment. Multi-chromophore systems display complex manifolds of electronic transitions, enabling their use for optical sensing applications. Time-Dependent Density Functional Theory (TDDFT) methods allow for accurate predictions of the optical properties. These properties are related to the electronic transitions in the molecules, which include two-photon absorption cross-sections. Here we use TDDFT to understand the mechanism of aza-crown based fluorescent probes for metals sensing applications. Our findings suggest changes in local excitation in the rhodol chromophore between unbound form and when bound to the metal analyte. These changes are caused by a charge transfer from the aza-crown group and pyrazol units toward the rhodol unit. Understanding this mechanism leads to an optimized design with higher two-photon excited fluorescence to be used in medical applications.


Inhibition of IRAK1/4 sensitizes T cell acute lymphoblastic leukemia to chemotherapies.
Signaling via the MyD88/IRAK pathway in T cells is indispensable for cell survival; however, it is not known whether this pathway functions in the progression of T acute lymphoblastic leukemia (T-ALL). Here, we determined that compared with thymic and peripheral T cells, T-ALL cells from patients have elevated levels of IRAK1 and IRAK4 mRNA as well as increased total and phosphorylated protein. Targeted inhibition of IRAK1 and IRAK4, either with shRNA or with a pharmacological IRAK1/4 inhibitor, dramatically impeded proliferation of T-ALL cells isolated from patients and T-ALL cells in a murine leukemia model; however, IRAK1/4 inhibition had little effect on cell death. We screened several hundred FDA-approved compounds and identified a set of drugs that had enhanced cytotoxic activity when combined with IRAK inhibition. Administration of an IRAK1/4 inhibitor or IRAK knockdown in combination with either ABT-737 or vincristine markedly reduced leukemia burden in mice and prolonged survival. IRAK1/4 signaling activated the E3 ubiquitin ligase TRAF6, increasing K63-linked ubiquitination and enhancing stability of the antiapoptotic protein MCL1; therefore, IRAK inhibition reduced MCL1 stability and sensitized T-ALL to combination therapy. These studies demonstrate that IRAK1/4 signaling promotes T-ALL progression through stabilization of MCL1 and suggest that impeding this pathway has potential as a therapeutic strategy to enhance chemotherapeutic efficacy.


Interposition of the gallbladder--or the absent common hepatic duct and cystic duct.
Interposition of the gallbladder is a rare anomaly, but its diagnosis is important since it represents a surgically correctable cause of jaundice. The patients present with jaundice, abdominal pain and sometimes an enlarged gallbladder. Radiological diagnosis may be difficult since the condition may be mistaken for a choledochal cyst, hydrops of the gallbladder or Caroli's disease. The ultrasound, cholangiogram and surgical findings of dilated intrahepatic ducts adjacent to a normal or enlarged gallbladder with no dilatation of the common bile duct are presented in two children with this condition.


Prognostic value of serum Krebs von den Lungen-6 (KL-6) levels in COVID-19 pneumonia.
Krebs von den Lungen-6 (KL-6), expressed by damaged type II pneumocytes, is useful in the diagnosis and severity assessment of many diffuse interstitial lung diseases. The objective of our study was to determine the prognostic value of the initial KL-6 plasma level in COVID-19 pneumonia. All patients hospitalized for a suspected COVID-19 pneumonia between March and May 2020 in our Chest department of a French university hospital were included. KL-6 serum concentrations were measured within 72 h of diagnostic suspicion by chemiluminescence enzyme immunoassay Survival analysis was performed using a Cox regression and modeled by a Kaplan-Meier curve. Sixty-six COVID-19 patients (average age = 64 ± 14 years, 71.2 % males) with KL-6 serum measurement were included. Median KL-6 serum concentration was 409 ± 312 U/mL. KL-6 was significantly higher in men (p = 0.003), elders (p = 0.0001) and in patients with greater Charlson's score (p = 0.002). Higher KL-6 concentration was significantly associated with in-hospital mortality (HR: 8.66; 95 % CI:1.1-69.2, p = 0.014), radiological extension of lesions on chest CT scan (p = 0.004) and higher WHO severity score (p = 0.042), but not with admission in intensive care unit. In 9 (14 %) non-surviving COVID-19 patients, KL-6 serum concentration increased whereas it remained stable or decreased in survivors. At 3 months follow-up (n = 48), DLCO was negatively correlated with the initial KL-6 value (r = 0.47, p = 0.001), while FVC, FEV1 and MRC score were not. Initial KL-6 serum concentration is significantly associated with in-hospital mortality, unfavorable outcome, and persistent impairment of DLCO at 3 months. Initial KL-6 plasma determination appears as a prognostic biomarker in COVID-19 pneumonia.


X-linked hypophosphatemic rickets: an Italian experts' opinion survey.
X-linked hypophosphatemic rickets (XLH) is the first cause of inherited hypophosphatemia and is caused by mutation in the PHEX gene, resulting in excessive expression of the phosphaturic factor FGF23. Symptoms are mainly related to rickets in children and osteomalacia in adults and cause several complications that can be highly invalidating. Due to its rarity, XLH is poorly known and diagnosis is frequently delayed. Conventional treatment is based on oral phosphate salts supplementation and activated vitamin D analogs, which however, cannot cure the disease in most cases. Due to the low prevalence of XLH, an experts' opinion survey was conducted across Italian centers to collect data on XLH and on its management. A questionnaire was developed by a group of experts to collect data on XLH epidemiology, diagnosis and treatment in Italy. Data from 10 Italian centers (nine of which pediatric) on 175 patients, followed between 1998 and 2017, were included in the survey. Most patients were followed since childhood and 63 children became adults during the investigated period. The diagnosis was made before the age of 1 and between 1 and 5 years in 11 and 50% of cases, respectively. Clinically apparent bone deformities were present in 95% of patients. These were ranked moderate/severe in 75% of subjects and caused growth stunting in 67% of patients. Other frequent complications included bone pain (40%), dental abscesses (33%), and dental malpositions (53%). Treatment protocols varied substantially among centers. Nephrocalcinosis was observed in 34% of patients. Tertiary hyperparathyroidism developed in 6% of patients. XLH remains a severe condition with significant morbidities.


Effect of the addition of poly(methyl methacrylate) beads on some properties of acrylic resin.
There have been numerous attempts to improve the mechanical properties of acrylic resin. However, the fracture of dentures is still an unresolved problem. The potential advantage of self-reinforcement should be improved mechanical properties over the amorphous random polymer. This study investigated the addition of poly(methyl methacrylate) (PMMA) beads in a variety of percentages on a range of mechanical properties of PMMA denture base acrylic resin. The transverse and impact strengths and hardness of acrylic resin specimens stored in water for periods of 1 week and 6 months prior to testing were investigated. The results showed that for specimens stored for 1 week, the addition of up to 25% beads produced a significant increase in the modulus of elasticity. Addition of 5% beads produced a slight but nonsignificant increase in the modulus of rupture, although additions of increasing percentages produced a decrease in the modulus of rupture. The impact strength was increased for 5% to 25% beads and decreased for 40% beads. The hardness showed only small changes. For specimens stored in water for 6 months, the modulus of rupture decreased with increasing percentages of beads. The addition of beads up to 10% significantly increased the modulus of elasticity. There was no significant increase in impact strength but a significant increase in hardness for 5% beads. The addition of PMMA beads to acrylic resin did not produce a substantial improvement in the mechanical properties and cannot be recommended as a method of reinforcement in its present form.


The horizontal vestibulo-ocular reflex during linear acceleration in the frontal plane of the cat.
Horizontal and vertical eye movements were recorded in alert, restrained cats that were subjected to whole-body rotations with the horizontal semicircular canals in the plane of rotation and the body centered on the axis or 45 cm eccentric from the axis of rotation. Changes in the horizontal vestibulo-ocular reflex (HVOR) due to the resultant of the linear forces (i.e., gravity and linear acceleration) acting on the otolith organs were examined during off-axis rotation when there was a centripetal acceleration along the animal's interaural axis. The HVOR time constant was slightly shortened when the resultant otolith force was not parallel to the animal's vertical axis. This effect was independent of the direction of the otolith force relative to the direction of the slow phase eye velocity. No effect on the HVOR amplitude was observed. In addition to changes in the HVOR dynamics, a significant vertical component of eye velocity was observed during stimulation of the horizontal canals when the resultant otolith force was not parallel with the animal's vertical axis. The effect was greater for larger angles between the resultant otolith force and gravity. An upward or downward component was elicited, depending on the direction of the horizontal component of eye velocity and the direction of the resultant otolith force. The vertical component was always in the direction that would tend to align the eye velocity vector with the resultant otolith force and keep the eye movement in a plane that had been rotated by the angle between the resultant otolith force and gravity.


Hepatic microvasculature in liver injury.
Injury to the hepatic microvasculature, the hepatic sinusoids, manifests in several ways. The sinusoidal endothelial cells (SECs) may lose porosity and scavenger function (capillarization); SECs may loosen from their tetherings to the space of Disse or even detach completely (ischemia-reperfusion injury, early sinusoidal obstruction syndrome, peliosis hepatis, early acetaminophen toxicity); the space of Disse may be completely denuded of sinusoidal lining cells that then embolize and obstruct the sinusoid (early sinusoidal obstruction syndrome); or the sinusoid may be obstructed by fibrosis (hepatic sinusoidal fibrosis, late sinusoidal obstruction syndrome). In many of these microvascular injuries, the change to the sinusoid is a primary event that may lead to hepatocyte hypoxia with liver dysfunction and disruption of the portal circulation. With the exception of hepatic fibrosis, which will be reviewed elsewhere in this issue, each of these types of microvascular injuries will be described in this article.


Pulmonary edema during volume infusion.
The relationship between left ventricular filling pressure and plasma colloid osmotic pressure to pulmonary edema was examined in a group of 37 patients, the majority of whom were hypovolemic. Sixteen patients developed pulmonary edema during fluid infusion. In the 21 patients who did not develop pulmonary edema, the left ventricular filling pressure was slightly elevated but the colloid osmotic pressure was not reduced. The majority of these patients were treated with colloid solutions (group I). In five of the 16 patients who developed pulmonary edema, the left ventricular filling pressure was elevated and there was no reduction in the plasma colloid osmotic pressure. These patients received only colloids (group II). In the other 11 patients who developed pulmonary edema, the left ventricular filling pressure was normal but the plasma colloid osmotic pressure was reduced to 16 +/- 2 torr (group III). The colloid osmotic pressure in this group was significantly less than in the other two groups (P less than 0.01). Most of these patients received large volumes of crystalloid solutions. After administration of furosemide, clearing of pulmonary edema in this group was associated with normalization of the plasma colloid osmotic pressure. Infusion of large volumes of crystalloids in hypovolemic patients can be hazardous, for reduction of the plasma colloid osmotic pressure may predispose to the development of pulmonary edema even when the left ventricular filling pressure remains normal.


Gender associated disparities in atrioventricular nodal reentrant tachycardia: A review article.
Atrioventricular nodal reentrant tachycardia (AVNRT) is the most common sustained supraventricular arrhythmias. An understanding of gender-related differences in AVNRT epidemiology, diagnosis, treatment, outcome, and complications can help guide a more effective diagnosis and treatment of the condition. The study aimed to perform a review of the available literature regarding all aspects of gender-related differences of AVNRT. We focused on all aspects of gender-related differences regarding AVNRT between men and women. A literature search was performed using Google Scholar, PubMed, Springer, Ovid, and Science Direct. Many investigations have demonstrated that the prevalence of AVNRT exhibited a twofold women-to-men predominance. The potential mechanism behind this difference due to sex hormones and autonomic tone. Despite being more common in women, there is a delay in offering and performing the first-line therapy (catheter ablation) compared to men. There were no significant gender-related discrepancies in patients who underwent ablation therapy for AVNRT, regarding the acute success rate of the procedure, long-term success rate, and recurrence of AVNRT. AVNRT is more common in women due to physiological factors such as sex hormones and autonomic tone. Catheter ablation is equally safe and efficacious in men and women; however, the time between the onset of symptoms and ablation is significantly prolonged in women. It is important for the medical community to be aware of this discrepancy and to strive to eliminate such disparities that are not related to patients' choices.


Identification of Drosophila mitotic genes by combining co-expression analysis and RNA interference.
RNAi screens have, to date, identified many genes required for mitotic divisions of Drosophila tissue culture cells. However, the inventory of such genes remains incomplete. We have combined the powers of bioinformatics and RNAi technology to detect novel mitotic genes. We found that Drosophila genes involved in mitosis tend to be transcriptionally co-expressed. We thus constructed a co-expression-based list of 1,000 genes that are highly enriched in mitotic functions, and we performed RNAi for each of these genes. By limiting the number of genes to be examined, we were able to perform a very detailed phenotypic analysis of RNAi cells. We examined dsRNA-treated cells for possible abnormalities in both chromosome structure and spindle organization. This analysis allowed the identification of 142 mitotic genes, which were subdivided into 18 phenoclusters. Seventy of these genes have not previously been associated with mitotic defects; 30 of them are required for spindle assembly and/or chromosome segregation, and 40 are required to prevent spontaneous chromosome breakage. We note that the latter type of genes has never been detected in previous RNAi screens in any system. Finally, we found that RNAi against genes encoding kinetochore components or highly conserved splicing factors results in identical defects in chromosome segregation, highlighting an unanticipated role of splicing factors in centromere function. These findings indicate that our co-expression-based method for the detection of mitotic functions works remarkably well. We can foresee that elaboration of co-expression lists using genes in the same phenocluster will provide many candidate genes for small-scale RNAi screens aimed at completing the inventory of mitotic proteins.


Combined radiocarbon and CO2 flux measurements used to determine in situ chlorinated solvent mineralization rate.
A series of combined measurements was made at the Naval Air Station North Island (NASNI) Installation Restoration Site 5, Unit 2 during July and August 2013. Combined measurements included CO2 respiration rate, CO2 radiocarbon content to estimate chlorinated hydrocarbon (CH) mineralization and a zone of influence (ZOI) model. CO2 was collected continuously over 2 two-week periods by recirculating monitoring well headspace gas through NaOH traps. A series of 12 wells in the main CH plume zone and a background well with no known historical contamination were sampled. The background well CO2 was used to determine radiocarbon content derived from respired natural organic matter. A two end-member mixing model was then used to determine the amount of CH-derived carbon present in the CO2 collected from plume region wells. The ZOI model provided an estimate for the soil volume sampled at each well. CH mineralization rates were highest upgradient and at the plume fringe for areas of high historical contamination and ranged from 0.02 to 5.6 mg CH carbon per day. Using the ZOI model volume estimates, CH-carbon removal ranged from 0.2 to 32 mg CH-carbon m(-3) per day. Because the rate estimates were based on a limited sampling (temporally), they were not further extrapolated to long-term contaminant degradation estimates. However, if the site manager or regulators required them, estimates - subject to long-term variability uncertainties - could be made using volume and rate data determined over short timescales. A more comprehensive seasonal sampling is needed to constrain long-term remediation models for the entire impacted area and identify environmental conditions related to more rapid turnover times amongst the wells.


Tuberculous lumbar spinal epidural abscess in a young adult (case report).
Spinal Epidural abscess (SEA) is an uncommon pathology that needs an urgent intervention to decompress the pressure on the spinal epidural sac, cord, and roots. The authors report a rare case of a young adult with lumbar spinal epidural tuberculous abscess occupying the spinal canal from L2-L5 vertebrae with extesion to the posterior paraspinal muscles and presenting with bilateral progressive lower limb weakness. Case report: A 42 years old male teacher presented with a 15-day history of progressive difficulty to walking and bilateral lower limb weakness associated with fever, malaise and later on urinary incontinence. A magnetic resonance imaging (MRI) scan revealed a paraspinal intermuscular abscess and an abscess occupying the spinal canal compressing the dural sac from L2-L4/5, without any signs of vertebral involvement. Surgery was done by a posterior midline incision. Pus was evacuated from multiple pockets through the paraspinal muscle layers. Laminectomy for L3/4, and hemilaminectomy for L2/3, and L4/5 were performed. Pus and bone specimens were negative for acid-fast bacilli. However, both histopathological studies and Polymerase Chain Reaction (PCR) testing confirmed the presence of tuberculosis (TB). The patient received TB antibiotics, and a follow-up MRI scan at 2 months showed complete evacuation of the abscess. However, signs of L5 spondylitis were evident. No further surgery was needed as there was no vertebral collapse or neural compression and the patient's clinical condition was improving. He had normal right lower limb power and sensation and grade 4+ motor power of the left lower limb. Bowels and bladder function was normal. Isolated tuberculous spinal epidural abscess is a rare disease and should be treated urgently with evacuation and decompression. Signs of spondylitis or spondylodiscitis may appear later and therefore long follow up is recommended in tuberculous cases presenting with an isolated epidural abscess.


SRC inhibitors and angiogenesis.
Angiogenesis is a tightly regulated process that leads to the formation of new blood vessels in limited physiological conditions, and can also occur under pathological situations as retinopathies, arthritis, endometriosis and cancer. Enhanced angiogenesis is present in tumors that need new blood capillaries to grow, remove metabolic waste and transport the cells to locations distal to the primary tumor, facilitating metastasis formation. For these reasons, blockade of angiogenesis is an attractive approach for the treatment of both solid and haematological malignancies. Antiangiogenic therapy should be less toxic in comparison with conventional treatments such as chemotherapy, being angiogenesis a process relatively restricted to the growing tumor. The Src family of tyrosine kinases has been implicated in the intracellular signaling cascade that acts downstream of cell surface receptors to elicit different cellular functions, including growth, proliferation, adhesion and motility. Src kinases are frequently activated in human malignancies, causing tumor progression, metastasis formation and deregulating expression of proangiogenic molecules. This review reports several studies performed by different authors demonstrating the involvement of Src tyrosine kinases in angiogenesis by regulating different signalling pathways. Moreover, we report selective Src inhibitors for which a direct involvement with angiogenesis has been demonstrated, even if every Src inhibitor could potentially possesses also antiangiogenic properties. Biological data, structures and mechanisms of action of selected molecules, in terms of Src protein-inhibitor interactions, are also reported.


Regulation of the expression on mouse T lymphocytes of the epitope identified by monoclonal antibody 3A35.
Monoclonal antibody 3A35 (MA 3A35) has previously been shown to be an activation marker of macrophages and T lymphocytes. It immunoprecipitated from macrophages a 200-kDa molecule belonging to the T200 family and from T cells a 85-kDa antigen. In the present work, the factors controlling the expression of the epitope identified by MA 3A35 on polyclonal activated T cells and T-cell clones, as well as the ability of 3A35 alone or together with complement to interfere with T-cell functions, were investigated. Corticoresistant thymocytes unreactive with MA 3A35 became fully reactive after 2 days of in vitro stimulation by PMA and IL-2 and the level of reactivity per cell declined to a low level thereafter. In helper and cytolytic T-cell clones, the expression of the epitope defined by MA 3A35 was also maximal soon after antigenic stimulation then declined. In helper-T-cell clones, the epitope remained detectable during the entire culture period, whereas in cytolytic clones its expression was markedly reduced at the end of the culture. The lineage of cytotoxic T lymphocytes (CTL) as studied in a bulk culture of spleen cells primed in vivo against a syngeneic tumor exhibited similar regulation by antigenic stimulation. The CTL precursors were resistant to lysis by MA 3A35 plus complement; after 3 days of culture with the stimulatory antigen, they became highly sensitive but their sensitivity then diminished and mature CTL were completely resistant. MA 3A35 plus complement also killed the activated T cells which responded to macrophage-presented antigens and were thought to be mainly Lyt-1+. Therefore, the epitope identified by MA 3A35 was expressed predominantly at an early stage of T-cell activation. At a late stage, it persisted almost exclusively on helper and Lyt-1+ cells. In addition, MA 3A35 plus complement lysed NK cells, AK cells, and their precursors present in normal spleen. In the absence of complement, MA 3A35 had no detectable effect on T-cell functions.


Development of a food frequency questionnaire for children from 7 to 10 years old.
To present and discuss the process of the development of a Quantitative Food Frequency Questionnaire (QFFQ) aimed to 7-10 year-old children. A methodological study was performed based on the application of a Food Diary (FD) fulfilled by 85 children. Based on registered foods, the diet composition in relation to energy and macronutrients was calculated. Foods were grouped into items and the percentage of the contribution of each item to the diet in terms of calories and macronutrients were calculated. The QFFQ was constructed with 92 food items, 76 were selected by Block's method and 16 were identified from other instruments from literature developed for children and adolescents. The food items were grouped in 17 food groups with 3 serving sizes and 7 categories to measure food frequency. Among the food items, the rice was the one that showed the biggest percentage of contribution for energy and carbohydrates. For other nutrients, the meat and the milk were the ones that contributed most to the consumption of proteins and lipids, respectively.


Surface enhanced Raman spectroscopy (SERS) sensors for gas analysis.
This paper presents work towards a gas sensor based on surface enhanced Raman spectroscopy (SERS). Sensitive SERS analysis of CO and N(2)O was performed using a mixed AgPd nanoparticle substrate that facilitated effective gas adsorption and strong SERS generation. The adsorption, desorption and displacement of both gases from the AgPd substrate were studied. We find that heating/cooling of the AgPd is necessary for CO adsorption but after this step a maximum SERS enhancement factor (EF) of 4 x 10(5) was measured and subsequent desorption and displacement occurred readily. For N(2)O, adsorption occurred at ambient temperature and the maximum SERS EF was 1 x 10(5), but desorption required heating and CO would not displace N(2)O from the surface but was rather co-adsorbed. We conclude that it is necessary to ensure analytes adsorb relatively weakly to the SERS substrate to enable continuous, real-time analysis of multiple analytes and to avoid substrate contamination.


Ridge preservation with the use of Bio-Oss collagen: A 6-month study in the dog.
In previous short-term studies, it was observed that while the placement of biomaterial in alveolar sockets may promote bone formation and ridge preservation, the graft may in fact also delay healing. The objective of the present experiment was to evaluate the more long-term effect on hard tissue formation and the amount of ridge augmentation that can occur by the placement of a xenogeneic graft in extraction sockets of dogs. Five beagle dogs were used. The third mandibular premolars were hemi-sected. The distal roots were carefully removed. A graft consisting of Bio-Oss collagen was placed in one socket while the contra-lateral site was left without grafting. After 6 months of healing, the dogs were euthanized and biopsies were sampled. From each experimental site, four ground sections - two from the mesial root and two from the healed socket - were prepared, stained and examined under a microscope. The placement of Bio-Oss collagen in the fresh extraction socket served as a scaffold for tissue modeling but did not enhance new bone formation. In comparison with the non-grafted sites, the dimension of the alveolar process as well as the profile of the ridge was better preserved in Bio-Oss-grafted sites. The placement of a biomaterial in an extraction socket may modify modeling and counteract marginal ridge contraction that occurs following tooth removal.


Rhabdochona (Rhabdochona) keralaensis sp. nov. (Rhabdochonidae) and some other nematodes in the Indian mottled eel Anguilla bengalensis bengalensis from India.
A new species of parasitic nematode, Rhabdochona (Rhabdochona) keralaensis sp. nov., is described based on specimens recovered from the intestine of Anguilla bengalensis bengalensis (Gray) collected from the freshwater bodies of Kerala State, southern India. It is characterized mainly by the presence of ten anterior prostomal teeth, absence of basal teeth, simple deirids, length of spicules, number and arrangement of preanal papillae and non-filamented eggs. Two species of camallanid nematodes, adults of Camallanus cotti Fujita, 1927 and a single Procamallanus (Spirocamallanus) sp. fourth-stage larva, were also recorded from A. bengalensis. Camallanus cotti, a pathogenic parasite of many species of wild and aquarium-reared fishes in many countries of South and East Asia, Europe, South and North America, West Indies and Australia, is reported for the first time from a representative of the fish family Anguillidae. Descriptions of R. (R.) keralaensis and Indian specimens of C. cotti, based on light and scanning electron microscopical examinations, are provided. Camallanus oxygasterae Gupta et Bakshi, 1983 is considered a junior synonym of C. cotti.


Role of cathepsin A and lysosomes in the intracellular activation of novel antipapillomavirus agent GS-9191.
GS-9191, a bis-amidate prodrug of the nucleotide analog 9-(2-phosphonylmethoxyethyl)-N6-cyclopropyl-2,6-diaminopurine (cPrPMEDAP), was designed as a topical agent for the treatment of papillomavirus-associated proliferative disorders, such as genital warts. In this study, we investigated the mechanism of conversion of GS-9191 to cPrPMEDAP. We observed that GS-9191 is hydrolyzed in the presence of the lysosomal carboxypeptidase cathepsin A (CatA) in vitro and is less efficiently metabolized in CatA-deficient fibroblasts than in control cells. In addition, knockdown of CatA by small interfering RNA (siRNA) reduced the intracellular accumulation of GS-9191 metabolites. However, intracellular CatA levels did not correlate with the susceptibility of tested cell lines to GS-9191, indicating that the CatA step is unlikely to be rate limiting for the activation of GS-9191. Further analysis showed that upon the hydrolysis of the carboxylester bond in one of the GS-9191 amidate moieties, the unmasked carboxyl group displaces L-phenylalanine 2-methylpropyl ester from the other amidate moiety. The cPrPMEDAP-L-phenylalanine conjugate (cPrPMEDAP-Phe) formed is not metabolized by Hint1 (histidine triad nucleotide binding protein 1) phosphoramidase but undergoes spontaneous degradation to cPrPMEDAP in acidic pH that can be significantly enhanced by the addition of SiHa cell extract. Pretreatment of SiHa cells with bafilomycin A or chloroquine resulted in an 8-fold increase in the intracellular concentration of cPrPMEDAP-Phe metabolite and the accumulation of GS-9191 metabolites in the lysosomal/endosomal fraction. Together, these observations indicate that the conversion of GS-9191 to cPrPMEDAP occurs in lysosomes via CatA-mediated ester cleavage, followed by the release of cPrPMEDAP, most likely through the combination of enzyme-driven and spontaneous pH-driven hydrolysis of a cPrPMEDAP-Phe intermediate.