Correlation of Copper and Zinc in Spontaneous Abortion.
Humans require minute amounts of trace metals to maintain body's normal growth and physiological functions; such elements may also play a vital role in pregnancy and pregnancy outcome. The present study was conducted to assess the role of two trace metals, zinc (Zn) and copper (Cu) in women with history of spontaneous abortion (SAb cases) in comparison to women without such history (controls). In this retrospective study, a total of 277 subjects were enrolled from the Obstetrics and Gynecology Department, Civil Hospital, Ahmedabad, India. Personal demographic information, medical history, reproductive history especially details of number of SAb, duration of last SAb, number of children, etc. were recorded using predesigned and pre-tested proforma. Serum Zn and Cu levels were measured by an atomic absorption spectrophotometer. The data indicated that the serum level of Cu (P<0.01) and Zn was lower in SAb cases as compared to controls. Correlation between the number of SAbs and trace metals levels showed a significant negative correlation between Cu and Cu/Zn and the number of SAbs. Cu/Zn was higher in controls and women having at least one child as compared cases and women without child, respectively. Pregnant women had higher levels of trace elements as compared to non-pregnant women at the time of enrollment. The data revealed that trace metals such as Zn and Cu have a positive role in pregnancy outcome and optimum levels of Zn and Cu might be able to decline the chances of SAb occurrence in addition to other factors. The ratio of Cu/Zn has a positive role in reproductive outcomes.


Facial nerve palsy and Kawasaki disease.
We report on a case of facial nerve palsy associated with Kawasaki disease in a 2-year-old boy. Facial nerve palsy is one of the rare neurological manifestations of Kawasaki disease. Twenty-seven other cases that have been reported in the literature are reviewed. There is a high incidence of coronary artery aneurysm (52%) and a female predilection in patients with Kawasaki disease. The facial palsy associated with the disease is self-limiting. Recovery is spontaneous in surviving patients, although the use of intravenous immunoglobulin may be able to hasten the recovery.


Homeobox gene HLX is a regulator of HGF/c-met-mediated migration of human trophoblast-derived cell lines.
Homeobox gene transcription factors play a critical role in normal placental development and are expressed in specialized trophoblast cells. Abnormal trophoblast function is associated with clinically significant pregnancy disorders, including fetal growth restriction (FGR). Our previous studies demonstrated that homeobox gene HLX is expressed in proliferating and migrating (but not invading) human trophoblast cells and that HLX expression is significantly decreased in human FGR. We have also shown that HLX is a regulator of colony-stimulating-factor-1-dependent trophoblast proliferation. Hepatocyte growth factor (HGF) activates trophoblast cell migration in a paracrine fashion, and its receptor, c-met, is expressed on trophoblast cells. Given that HGF is a regulator of trophoblast migration, we hypothesize that HLX is a mediator of HGF/c-met-dependent trophoblast migration but not invasion. Here we investigated the potential role of HLX in HGF/c-met-mediated trophoblast migration and invasion in two human trophoblast-derived cell lines, SGHPL-4 and HTR-8/SVneo. Results showed that in cultured trophoblast cells, HGF stimulation significantly increased HLX mRNA and protein expression. HLX inactivation significantly decreased trophoblast migration but not invasion. When HLX was inactivated in the presence of HGF stimulation, migration remained significantly decreased. SU11274-mediated inhibition of the receptor c-met significantly decreased HLX mRNA and protein expression. In the presence of HGF stimulation, HLX expression remained significantly decreased with c-met inhibition. This is the first study to show that homeobox gene HLX is a downstream effector gene of HGF, that HLX regulates human trophoblast-derived cell migration, and that HGF, via receptor c-met, acts through HLX to control cell migration.


Rapid Bench to Bedside Therapeutic Bacteriophage Production.
It has been over 100 years since bacteriophages (phages) were used as a human therapeutic. Since then, phage production has dramatically evolved. Current phage preparations have fewer adverse effects due to their low bacterial toxin content. As a result, therapeutic phages have become a predominant class of new antimicrobials and are being widely used for compassionate treatment of multidrug-resistant (MDR) infections. We describe herein a protocol for the production and ultrapurification of phages. By this technique, it is possible for a lab experienced with the process to produce >109 plaque-forming units (PFU) per mL of Gram-negative phages that meet FDA endotoxins limits for intravenous infusions in as little as 48 hours. We provide illustrations of the process and tips on how to safely remove bacterial toxins from phage lysates. Although dependent on the phage strain, the approach described can rapidly generate and purify phages for a variety of applications.


New oxybromide cobaltites with layered perovskite-related structures: 18R-Ba(6)Co(5)BrO(14) and 14H-Ba(7)Co(6)BrO(17).
Single crystals of the title compounds were prepared by solid-solid reaction using BaBr(2) flux at 1373 K. The structures of these two new cobaltites were solved and refined. The two compounds are built from a close-packing of [BaO(3)] and [BaOBr] layers with stacking sequences (c'chhcc')(3) and (c'chhhcc')(2) for the 18R and 14H structures, respectively, which create Co(3)O(12) trimers or Co(4)O(15) tetramers of face-sharing octahedra connected at their extremities to isolated tetrahedra by corner-sharing. These new materials are strongly related to the 5H-Ba(5)Co(5)O(14)/12H-Ba(0.9)CoO(2.6) and 10H-Ba(5)Co(5)ClO(13)/6H-Ba(6)Co(6)ClO(16) materials, with the existence of common blocks. In Ba(6)Co(5)BrO(14) and Ba(7)Co(6)BrO(17), all the atoms in the vicinity of the [BaOBr] layers are disordered, whereas the rest of the structure is perfectly ordered.


Progressive dopaminergic degeneration in the chronic MPTPp mouse model of Parkinson's disease.
Parkinson's disease (PD) is characterized by a progressive degeneration of dopamine (DA) neurons and gradual worsening of motor symptoms. The investigation of progressive degenerative mechanisms and potential neuroprotective strategies relies on experimental models of the chronic neuropathology observed in human. The present study investigated the progressive nature of neurodegeneration in the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine/probenecid (MPTPp) chronic mouse model of PD. MPTP (25 mg/kg) plus probenecid (250 mg/kg) were administered twice a week for 5 weeks. We evaluated behavioral deficits (olfactory and motor impairment), neurodegeneration (loss of tyrosine hydroxylase (TH)-positive cells in the substantia nigra pars compacta, SNc), biochemical markers of functional impairment in the caudate-putamen (CPu) (striatal enkephalin mRNA, DA and DOPAC levels), and glial reactivity (CD11b and GFAP immunoreactivity in the SNc and CPu) at progressive time-points (after 1, 3, 7, and 10 administrations of MPTPp). Olfactory dysfunction already appeared after the 1st MPTPp injection, whereas motor dysfunction appeared after the 3rd and worsened upon subsequent administrations. Moreover, starting after three MPTPp injections, we observed a gradual decline of TH-positive cells in the SNc, and a gradual raise of enkephalin mRNA in the CPu. Striatal DA levels reduction was not different among all time-points evaluated, whereas DOPAC levels were similarly reduced after 1-7 MPTP injections, but were further decreased after the 10th injection. Reactive microglia and astroglia were observed in both the SNc and CPu from the 1st MPTPp administration. In the SNc, gliosis displayed a gradual increase over the treatment. After 2 months, TH, DA, DOPAC, and reactive glia in the SNc were still altered in MPTPp-treated mice as compared to controls. By showing a progressive development of behavioral deficits and nigral neurodegeneration, together with impairment of biochemical parameters and gradual increase of glial response, results suggest that the chronic MPTPp protocol is a model of progressive PD, which may be suitable to investigate chronic pathological processes and neuroprotective strategies in PD.


The evolutionary differentiation of two histone H2A.Z variants in chordates (H2A.Z-1 and H2A.Z-2) is mediated by a stepwise mutation process that affects three amino acid residues.
The histone H2A family encompasses the greatest number of core histone variants of which the replacement variant H2A.Z is currently one of the most heavily studied. No clear mechanism for the functional variability that H2A.Z imparts to chromatin has yet been proposed. While most of the past studies have referred to H2A.Z generically as a single protein, in vertebrates it is a mixture of two protein forms H2A.Z-1 (previously H2A.Z) and H2A.Z-2 (previously H2A.F/Z or H2A.V) that differ by three amino acids. We have performed an extensive study on the long-term evolution of H2A.Z across metazoans with special emphasis on the possible selective mechanisms responsible for the differentiation between H2A.Z-1 and H2A.Z-2. Our results reveal a common origin of both forms early in chordate evolution. The evolutionary process responsible for the differentiation involves refined stepwise mutation change within the codons of the three differential residues. This eventually led to differences in the intensity of the selective constraints acting upon the different H2A.Z forms in vertebrates. The results presented in this work definitively reveal that the existence of H2A.Z-1 and H2A.Z-2 is not a whim of random genetic drift. Our analyses demonstrate that H2A.Z-2 is not only subject to a strong purifying selection but it is significantly more evolutionarily constrained than H2A.Z-1. Whether or not the evolutionary drift between H2A.Z-1 and H2A.Z-2 has resulted in a functional diversification of these proteins awaits further research. Nevertheless, the present work suggests that in the process of their differently constrained evolutionary pathways, these two forms may have acquired new or complementary functions.


Pyrimidine-2,4,6-trione derivatives and their inhibition of mutant SOD1-dependent protein aggregation. Toward a treatment for amyotrophic lateral sclerosis.
Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by the progressive loss of motor neurons, leading to muscle weakness, paralysis, and death, most often from respiratory failure. The only FDA-approved drug for the treatment of ALS, riluzole, only extends the median survival in patients by 2-3 months. There is an urgent need for novel therapeutic strategies for this devastating disease. Using a high-throughput screening assay targeting an ALS cultured cell model (PC12-G93A-YFP cell line), we previously identified three chemotypes that were neuroprotective. We present a further detailed analysis of one promising scaffold from that group, pyrimidine-2,4,6-triones (PYTs), characterizing a number of PYT analogues using SAR and ADME. The PYT compounds show good potency, superior ADME data, low toxicity, brain penetration, and excellent oral bioavailability. Compounds from this series show 100% efficacy in the protection assay with a good correlation in activity between the protection and protein aggregation assays. The modifications of the PYT scaffold presented here suggest that this chemical structure may be a novel drug candidate scaffold for use in clinical trials in ALS.


Mitotic spindle asymmetry in rodents and primates: 2D vs. 3D measurement methodologies.
Recent data have uncovered that spindle size asymmetry (SSA) is a key component of asymmetric cell division (ACD) in the mouse cerebral cortex (Delaunay et al., 2014). In the present study we show that SSA is independent of spindle orientation and also occurs during cortical progenitor divisions in the ventricular zone (VZ) of the macaque cerebral cortex, pointing to a conserved mechanism in the mammalian lineage. Because SSA magnitude is smaller in cortical precursors than in invertebrate neuroblasts, the unambiguous demonstration of volume differences between the two half spindles is considered to require 3D reconstruction of the mitotic spindle (Delaunay et al., 2014). Although straightforward, the 3D analysis of SSA is time consuming, which is likely to hinder SSA identification and prevent further explorations of SSA related mechanisms in generating ACD. We therefore set out to develop an alternative method for accurately measuring spindle asymmetry. Based on the mathematically demonstrated linear relationship between 2D and 3D analysis, we show that 2D assessment of spindle size in metaphase cells is as accurate and reliable as 3D reconstruction provided a specific procedure is applied. We have examined the experimental accuracy of the two methods by applying them to different sets of in vivo and in vitro biological data, including mouse and primate cortical precursors. Linear regression analysis demonstrates that the results from 2D and 3D reconstructions are equally powerful. We therefore provide a reliable and efficient technique to measure SSA in mammalian cells.


Comparison of the canine and human olfactory receptor gene repertoires.
Olfactory receptors (ORs), the first dedicated molecules with which odorants physically interact to arouse an olfactory sensation, constitute the largest gene family in vertebrates, including around 900 genes in human and 1,500 in the mouse. Whereas dogs, like many other mammals, have a much keener olfactory potential than humans, only 21 canine OR genes have been described to date. In this study, 817 novel canine OR sequences were identified, and 640 have been characterized. Of the 661 characterized OR sequences, representing half of the canine repertoire, 18% are predicted to be pseudogenes, compared with 63% in human and 20% in mouse. Phylogenetic analysis of 403 canine OR sequences identified 51 families, and radiation-hybrid mapping of 562 showed that they are distributed on 24 dog chromosomes, in 37 distinct regions. Most of these regions constitute clusters of 2 to 124 closely linked genes. The two largest clusters (124 and 109 OR genes) are located on canine chromosomes 18 and 21. They are orthologous to human clusters located on human chromosomes 11q11-q13 and HSA11p15, containing 174 and 115 ORs respectively. This study shows a strongly conserved genomic distribution of OR genes between dog and human, suggesting that OR genes evolved from a common mammalian ancestral repertoire by successive duplications. In addition, the dog repertoire appears to have expanded relative to that of humans, leading to the emergence of specific canine OR genes.


Hypothermia: role of alpha 1- and alpha 2-noradrenergic receptors in the hypothalamus of the cat.
The purpose of this study was to characterize the alpha 1- and alpha 2-noradrenergic receptor sub-types which could mediate the hypothermic response produced by norepinephrine (NE) and other alpha-noradrenergic agonists applied to the thermosensitive zone of the hypothalamus. An array of four guide tubes was implanted stereotaxically so that their tips rested just above the anterior hypothalamic, preoptic area (AH/POA) of the cat. Following post-operative recovery, a micro-injection of an agonist or antagonist of NE receptors or control CSF vehicle was given in a volume of 1.0-2.0 microliter in the AH/POA in each of the unrestrained cats. The alpha 1-noradrenergic receptor agonist, phenylephrine, but not methoxamine, applied to the AH/POA produced a dose-dependent hypothermia of up to 2.0 degrees C. When applied similarly, the alpha 2-noradrenergic agonist clonidine, as well as norepinephrine, which acts on both alpha 1- and alpha 2-noradrenergic receptors, also induced a decline in the cat's core temperature of up to 1.5 degrees C. The hypothermic response of clonidine was inhibited by pre-treatment of the AH/POA with a micro-injection of the selective alpha 2-noradrenergic blocking agent, yohimbine. However, yohimbine given similarly in the cat's AH/POA potentiated significantly both the phenylephrine and norepinephrine-induced hypothermia. The combined alpha 1-, alpha 2-noradrenergic receptor antagonist, phentolamine, also injected into AH/POA inhibited the thermolytic response evoked by both phenylephrine and norepinephrine, whereas it was virtually ineffective against the clonidine-induced hypothermia. These results, therefore, strongly suggest that both alpha 1- and alpha 2-noradrenergic receptors subserve the coordinated thermoregulatory mechanisms in AH/POA which are required for the functional dissipation of body heat and the consequent evocation of hypothermia.


Clinical presentation and outcomes of pediatric anaplastic large cell lymphoma: A tertiary center experience from India.
Anaplastic large cell lymphoma (ALCL) is an uncommon T-cell lymphoma, accounting for 10-15% of pediatric non-Hodgkin lymphoma (NHL). Outcomes of ALCL have not improved in recent years despite better chemotherapy and supportive care. This study aimed to analyze the clinical profile and outcomes of pediatric ALCL patients at our center. Medical records of children with ALCL from July 2017 to December 2019 were reviewed. Their clinical and laboratory details were noted. The outcomes of all patients were recorded. Ten patients were diagnosed with ALCL during this period. The median age at diagnosis was 8 years, with 7 patients being boys. Lymph nodes were the primary site of involvement in four children. Extranodal involvement was seen in 60%. One patient had the underlying chronic granulomatous disease (CGD). Systemic (B) symptoms were seen in 70%. Three children had hemophagocytic lymphohistiocytosis (HLH) at presentation. All of them had stage-III disease. Seven patients were treated on the Berlin-Frankfurt-Münster-non-Hodgkin lymphomas (BFM-NHL) 90 protocol and three children are currently in remission. Four patients had disease relapse/progressive disease while on therapy. Three children had HLH at presentation and all had poor outcomes. Pediatric ALCL has diverse and atypical clinical presentation and a high index of suspicion is needed for diagnosis. ALCL is an aggressive malignancy and is responsive to multiagent chemotherapy, though survival has not improved over the past decade. The integration of novel agents may improve the outcome.


Axillary versus femoral arterial cannulation in type A acute aortic dissection: evidence from a meta-analysis of comparative studies and adjusted risk estimates.
There is a growing perception that femoral arterial cannulation (FAC), by reversing the flow in the thoracoabdominal aorta, may increase the risk of retrograde brain embolization, dissection and organ malperfusion in type A aortic dissection. Axillary artery cannulation (AXC) has been reported to improve operative outcomes by allowing antegrade blood flow. However, FAC still remains largely utilized as a consensus for the routine use of AXC has not yet been reached. A meta-analysis on comparative studies reporting operative outcomes using AXC versus FAC was performed. Pooled weighted incidence rates for end points of interest (both adjusted and unadjusted) have been computed using an inverse variance model. Overall, a total of 8 studies including 793 patients were analysed (AXC = 396, FAC = 397). AXC was associated with reduced risk for in-hospital mortality [risk ratio (RR): 0.41; 95% confidence interval (CI): 0.29-0.58; P < 0.001] and permanent neurological deficit (PND) (RR: 0.59; 95% CI: 37.-0.93; P = 0.02) when compared with FAC. Pooled adjusted estimates confirmed that AXC was independently associated with a significantly reduced incidence of in-hospital mortality (adjusted OR: 0.54; 95% CI: 0.36-0.82; P = 0.004; I(2) = 57%) and PND (adjusted OR: 0.19; 95%CI: 0.07-0.54; P = 0.002; I(2) = 0%). The present meta-analysis demonstrated that AXC is superior to FAC in reducing in-hospital mortality and the incidence of PND in patients operated on for type A acute aortic dissection.


Protective immunity against Streptococcus mutans infection in mice after intranasal immunization with the glucan-binding region of S. mutans glucosyltransferase.
Here we present the construction and characterization of a chimeric vaccine protein combining the glucan-binding domain (GLU) of the gtfB-encoded water-insoluble glucan-synthesizing glucosyltransferase enzyme (GTF-I) from Streptococcus mutans and thioredoxin from Escherichia coli, which increases the solubility of coexpressed recombinant proteins and stimulates proliferation of murine T cells. The protective potential of intranasal (i.n.) immunization with this chimeric immunogen was compared to that of the GLU polypeptide alone in a mouse infection model. Both immunogens were able to induce statistically significant mucosal (salivary and vaginal) and serum responses (P < 0.01) which were sustained to the end of the study (experimental day 100). Following infection with S. mutans, sham-immunized mice maintained high levels of this cariogenic organism ( approximately 60% of the total oral streptococci) for at least 5 weeks. In contrast, animals immunized with the thioredoxin-GLU chimeric protein (Thio-GLU) showed significant reduction (>85%) in S. mutans colonization after 3 weeks (P < 0.05). The animals immunized with GLU alone required 5 weeks to demonstrate significant reduction (>50%) of S. mutans infection (P < 0.05). Evaluation of dental caries activity at the end of the study showed that mice immunized with either Thio-GLU or GLU had significantly fewer carious lesions in the buccal enamel or dentinal surfaces than the sham-immunized animals (P < 0.01). The protective effects against S. mutans colonization and caries activity following i.n. immunization with GLU or Thio-GLU are attributed to the induced salivary immunoglobulin A (IgA) anti-GLU responses. Although in general Thio-GLU was not significantly better than GLU alone in stimulating salivary IgA responses and in protection against dental caries, the finding that the GLU polypeptide alone, in the absence of any immunoenhancing agents, is protective against disease offers a promising and safe strategy for the development of a vaccine against caries.


The effects of subthalamic and pallidal deep brain stimulation on postural responses in patients with Parkinson disease.
The effect of deep brain stimulation (DBS) for Parkinson disease (PD) on balance is unclear. The goal of this study was to investigate how automatic postural responses (APRs) were affected in patients randomized to either subthalamic nucleus (STN) or globus pallidus internus (GPi) surgery. The authors tested 24 patients with PD who underwent bilateral DBS, 9 control patients with PD who did not undergo DBS, and 17 age-matched control volunteers. The electrode placement site was randomized and blinded to the patients and to the experimenters. Kinematic, kinetic, and electromyographic recordings of postural responses to backward disequilibrium via forward translations of the standing surface were recorded in the week prior to surgery while the patients were off (OFF) and on (ON) antiparkinsonian medication (levodopa), and then 6 months after surgery in 4 conditions: 1) off medication with DBS switched off (OFF/OFF); 2) off medication with DBS on (DBS); 3) on medication with DBS off (DOPA); and 4) with both medication and DBS on (DBS+DOPA). Stability of the automatic postural response (APR) was measured as the difference between the displacement of the center of pressure and the projected location of the center of body mass. Patients with PD had worse APR stability than controls. Turning the DBS on at either site improved APR stability compared with the postoperative OFF condition by lengthening the tibialis response, whereas medication did not show an appreciable effect. The STN group had worse APR stability in their best functional state (DBS+DOPA) 6 months after the DBS procedure compared with their best functional state (ON levodopa) before the DBS procedure. In contrast, the GPi group and the PD control group showed no change over 6 months. The APR stability impairment in the STN group was associated with smaller tibialis response amplitudes, but there was no change in response latency or coactivation with gastrocnemius. Turning the DBS current on improved APR stability for both STN and GPi sites. However, there was a detrimental DBS procedural effect for the STN group, and this effect was greater than the benefit of the stimulating current, making overall APR stability functionally worse after surgery for the STN group.


Anorexia nervosa and body dysmorphic disorder are associated with abnormalities in processing visual information.
Anorexia nervosa (AN) and body dysmorphic disorder (BDD) are characterized by distorted body image and are frequently co-morbid with each other, although their relationship remains little studied. While there is evidence of abnormalities in visual and visuospatial processing in both disorders, no study has directly compared the two. We used two complementary modalities--event-related potentials (ERPs) and functional magnetic resonance imaging (fMRI)--to test for abnormal activity associated with early visual signaling. We acquired fMRI and ERP data in separate sessions from 15 unmedicated individuals in each of three groups (weight-restored AN, BDD, and healthy controls) while they viewed images of faces and houses of different spatial frequencies. We used joint independent component analyses to compare activity in visual systems. AN and BDD groups demonstrated similar hypoactivity in early secondary visual processing regions and the dorsal visual stream when viewing low spatial frequency faces, linked to the N170 component, as well as in early secondary visual processing regions when viewing low spatial frequency houses, linked to the P100 component. Additionally, the BDD group exhibited hyperactivity in fusiform cortex when viewing high spatial frequency houses, linked to the N170 component. Greater activity in this component was associated with lower attractiveness ratings of faces. Results provide preliminary evidence of similar abnormal spatiotemporal activation in AN and BDD for configural/holistic information for appearance- and non-appearance-related stimuli. This suggests a common phenotype of abnormal early visual system functioning, which may contribute to perceptual distortions.


Forced cesareans.
This review attempts primarily to provide guidance to physicians and others who may face the dilemma raised by a patient who refuses a recommended cesarean. This is, therefore, not a traditional literature review. The ethical-legal nature of the topic necessitates a different approach. Accordingly, this review briefly describes two recent cases in which courts have reversed orders to override a woman's refusal of consent, suggests different ways in which medical practice trends and emerging medical data may be implicated in forced cesarean cases, and then considers the implications of recent cases in the light of social-political factors.


Comment on "Widespread RNA and DNA sequence differences in the human transcriptome".
Li et al. (Research Articles, 1 July 2011, p. 53; published online 19 May 2011) reported widespread differences between the RNA and DNA sequences of the same human cells, including all 12 possible mismatch types. Before accepting such a fundamental claim, a deeper analysis of the sequencing data is required to discern true differences between RNA and DNA from potential artifacts.


Relationships between demographic variables and lead toxicosis in cattle evaluated at North American veterinary teaching hospitals.
To determine associations between age, sex, breed, and month and year of admission and the diagnosis of lead toxicosis in cattle. Retrospective case-control study. Sample Population-Records of all cattle evaluated at North American veterinary teaching hospitals during the years 1963 to 2002, which were available through the Veterinary Medical Database. Logistic regression was used to evaluate the associations between postulated risk factors and the occurrence of lead toxicosis in cattle and predict the occurrence of the diagnosis of lead toxicosis in cattle. 413 cases of lead intoxication and 202,363 control cattle were identified and met the inclusion criteria. Cattle < 4 years of age were at increased risk for the diagnosis of lead intoxication relative to cattle > or = 4 years of age. Cattle > or = 2 months and < 6 months of age had the greatest risk for lead intoxication (odds ratio, 12.3). Angus cattle were at greater risk for toxicosis (odds ratio, 1.95), compared with other breeds. The risk of lead toxicosis was greater before 1985 (odds ratio, 1.94) than the risk thereafter. The risk of lead toxicosis diagnosis was greatest in the months of May, June, July, and August. Lead toxicosis in cattle was associated with age < 4 years and the Angus breed. A seasonal pattern existed with peak occurrence in the late spring and summer. The occurrence of lead toxicosis has declined over time.


Eosinophil chemotactic factor (ECF). I. Release from polymorphonuclear leukocytes by the calcium ionophore A23187.
Eosinophil chemotactic factor (ECF), previously thought to be primarily associated with human basophils and mast cells, could be released from human neutrophils (PMN) and eosinophils but not lymphocytes by the calcium ionophore A23187. Release of ECF from PMN was time and dose-dependent. Like antigen-induced, basophil-derived ECF, PMN-derived ECF had a high selectivity for eosinophils as determined by differential counts of migrating cells and by deactivation studies. Chromatographic analysis of PMN-derived ECF on Sephadex G-25 showed an elution pattern very similar to that of basophil-derived ECF. With rat peritoneal cells, it was possible to show that mast cells as well as mast cell-depleted cell preparations could be induced to release an ECF that appears to be similar or identical to human ECF. These findings suggest that ECF may play a role in inflammatory processes involving cells other than basophils and mast cells.


Long non-coding RNA MIR17HG impedes FOSL2-mediated transcription activation of HIC1 to maintain a pro-inflammatory phenotype of microglia during intracerebral haemorrhage.
Activation and polarization of microglia play decisive roles in the progression of intracerebral haemorrhage (ICH), and lactate exposure correlates with microglia polarization. This study explores molecules influencing lactate production and microglia phenotype alteration following ICH. A murine model of ICH was induced by intracerebral injection of collagenase. The mice experienced autonomous neurological function recovery, haematoma resolution and rapid lactate production, along with a gradual increase in angiogenesis activity, neuronal recovery and an M1-to-M2 phenotype change of microglia. Galloflavin, a lactate dehydrogenase antagonist, suppressed this phenotype change and the functional recovery in mice. FOS like 2 (FOSL2) was significantly upregulated in the brain tissues from day 7 post-ICH. Overexpression of FOSL2 induced an M1-to-M2 phenotype shift in microglia and accelerated lactate production in vivo and in haemoglobin-treated microglia in vitro. Long non-coding RNA MIR17HG impeded FOSL2-mediated transcription activation of hypermethylated in cancer 1 (HIC1). MIR17HG overexpression induced pro-inflammatory activation of microglia in mice, which was blocked by further HIC1 overexpression. Overall, this study demonstrates that MIR17HG maintains a pro-inflammatory phenotype of microglia during ICH progression by negating FOSL2-mediated transcription activation of HIC1. Specific inhibition of MIR17HG or upregulation of FOSL2 or HIC1 may favour inflammation inhibition and haematoma resolution in ICH.


N-Acetyltransferase 2 genetic polymorphisms and risk of colorectal cancer.
To investigate the possible association between meat intake, cigarette smoking and N-acetyltransferase 2 (NAT2) genetic polymorphisms on colorectal cancer (CRC) risk. Patients with CRC were matched for gender and age to healthy controls. Meat intake and cigarette smoking were assessed using a specific frequency questionnaire. DNA was extracted from peripheral blood and the genotypes of the polymorphism were assessed by polymerase chain reaction-restriction fragment length polymorphism. Five NAT2 alleles were studied (WT, M1, M2, M3 and M4) using specific digestion enzymes. A total of 147 patients with colorectal cancer (76 women and 90 men with colon cancer) and 212 controls were studied. The mean age of the two groups was 62 years. More than half the subjects (59.8% in the case group and 51.9% in the control group) were NAT2 slow acetylators. The odds ratio for colorectal cancer was 1.38 (95% CI: 0.90-2.12) in slow acetylators. Although the number of women was small (n = 76 in the case group), the cancer risk was found to be lower in intermediate (W/Mx) acetylators [odds ratio (OR): 0.55, 95% confidence interval (95% CI): 0.29-1.02]. This difference was not observed in men (OR: 0.56, 95% CI: 0.16-2.00). Among NAT2 fast acetylators (W/W or W/Mx), meat consumption more than 3 times a week increased the risk of colorectal cancer (OR: 2.05, 95% CI: 1.01-4.16). In contrast, cigarette smoking increased the risk of CRC among slow acetylators (OR: 1.97, 95% CI: 1.02-3.79). The risk of CRC was higher among fast acetylators who reported a higher meat intake. Slow NAT2 acetylation was associated with an increased risk of CRC.


CRISPR/Cas9: a molecular Swiss army knife for simultaneous introduction of multiple genetic modifications in Saccharomyces cerevisiae.
A variety of techniques for strain engineering in Saccharomyces cerevisiae have recently been developed. However, especially when multiple genetic manipulations are required, strain construction is still a time-consuming process. This study describes new CRISPR/Cas9-based approaches for easy, fast strain construction in yeast and explores their potential for simultaneous introduction of multiple genetic modifications. An open-source tool (http://yeastriction.tnw.tudelft.nl) is presented for identification of suitable Cas9 target sites in S. cerevisiae strains. A transformation strategy, using in vivo assembly of a guideRNA plasmid and subsequent genetic modification, was successfully implemented with high accuracies. An alternative strategy, using in vitro assembled plasmids containing two gRNAs, was used to simultaneously introduce up to six genetic modifications in a single transformation step with high efficiencies. Where previous studies mainly focused on the use of CRISPR/Cas9 for gene inactivation, we demonstrate the versatility of CRISPR/Cas9-based engineering of yeast by achieving simultaneous integration of a multigene construct combined with gene deletion and the simultaneous introduction of two single-nucleotide mutations at different loci. Sets of standardized plasmids, as well as the web-based Yeastriction target-sequence identifier and primer-design tool, are made available to the yeast research community to facilitate fast, standardized and efficient application of the CRISPR/Cas9 system.


Artificially long legs directly enhance long sprint running performance.
This comment addresses the incomplete presentation and incorrect conclusion offered in the recent manuscript of Beck et al. (R. Soc. Open Sci. 9, 211799 (doi:10.1098/rsos.211799)). The manuscript introduces biomechanical and performance data on the fastest-ever, bilateral amputee 400 m runner. Using an advantage standard of not faster than the fastest non-amputee runner ever (i.e. performance superior to that of the intact-limb world record-holder), the Beck et al. manuscript concludes that sprint running performance on bilateral, lower-limb prostheses is not unequivocally advantageous compared to the biological limb condition. The manuscript acknowledges the long-standing support of the authors for the numerous eligibility applications of the bilateral-amputee athlete. However, it does not acknowledge that the athlete's anatomically disproportionate prosthetic limb lengths (+15 cm versus the World Para Athletics maximum) are ineligible in both Olympic and Paralympic track competition due to their performance-enhancing properties. Also not acknowledged are the slower sprint performances of the bilateral-amputee athlete on limbs of shorter length that directly refute their manuscript's primary conclusion. Our contribution here provides essential background information and data not included in the Beck et al. manuscript that make the correct empirical conclusion clear: artificially long legs artificially enhance long sprint running performance.


Hepatic pedicle clamping does not worsen survival after hepatic resection for colorectal liver metastasis: results from a systematic review and meta-analysis.
Hepatic pedicle clamping (HPC) has been demonstrated to be effective for short-term outcomes during hepatic resection. However, HPC-induced hepatic ischemia/reperfusion injury can accelerate the outgrowth of hepatic micrometastases in experimental studies. The conclusive evidence regarding effects of HPC on long-term patient outcomes after hepatic resection for colorectal liver metastasis (CRLM) has not been determined. A comprehensive electronic literature search was performed to identify studies evaluating the oncological effects of HPC after hepatic resection for CRLM. The main outcome measures were intrahepatic recurrence (IHR), disease-free survival (DFS), and overall survival (OS). A meta-analysis was performed using the random-effects models to compute odds ratio (OR) along with 95% confidence intervals (CI). Four studies, with a total of 2,114 patients (73.7% HPC, 26.3% non-HPC), matched the inclusion criteria. Meta-analyses revealed that IHR (OR 0.88; 95% CI 0.69-1.11; P = 0.27), DFS (OR 0.88; 95% CI 0.70-1.10; P = 0.27) and OS (OR 0.99; 95% CI 0.79-1.24; P = 0.90) did not differ significantly between the HPC and non-HPC groups. This meta-analysis provides persuasive evidence that HPC during hepatic resection for CRLM has no significant adverse oncological outcomes. HPC should be considered an option during parenchymal liver resection from current available evidence.


Successful Management of a Patient with Autoimmune Hemorrhaphilia due to Anti-Factor XIII/13 Antibodies Complicated by Pulmonary Thromboembolism.
Autoimmune hemophilia-like disease (hemorrhaphilia) due to anti-factor XIII (FXIII) antibodies (AH13) is a very rare, life-threatening bleeding disorder. A 77-year-old woman developed macrohematuria and a right renal pelvic hematoma. The coagulation times were not prolonged, but FXIII activity and antigen levels were severely and moderately reduced to 9 and 29% of normal values, respectively. Accordingly, the FXIII-specific activity turned out to be low. FXIII inhibitor and anti-FXIII-A subunit autoantibodies were detected by a 1:1 crossmixing test and immunoblot and immunochromatographic assays. She was therefore diagnosed with "definite AH13" and treated with plasma-derived FXIII concentrates to arrest the hemorrhage. In addition to a highly compressed inferior vena cava by a huge renal pelvic hematoma, deep vein thrombosis (DVT) and pulmonary thromboembolism (PE) were identified by systemic computed tomography. The patient was immediately started on anticoagulation therapy with low-dose heparin. Emboli disappeared quickly, probably because under-crosslinked thrombi caused by severe FXIII deficiency are vulnerable to fibrinolysis. After about 1.5 years, anti-FXIII-A subunit autoantibodies still remained despite the use of rituximab, steroid pulse therapy, oral prednisolone, and oral cyclophosphamide treatments. In conclusion, an extremely rare AH13 case complicated by DVT and PE was successfully managed by balancing anticoagulation therapy with hemostatic therapy.


Luminescent amino-functionalized or erbium-doped silica spheres for biological applications.
This work presents the morphological and optical properties of luminescent silica spheres, discussing applications in bioimaging and biosensing. The spheres are obtained by the hydrolysis and condensation of tetraethylorthosilicate (TEOS) and can be synthesized by following either a basic or an acidic route. Luminescence emission is induced after incorporation of aminopropyltriethoxysilane (APTES) during synthesis or by introducing an optically active element, such as erbium, or other rare-earth elements. The luminescence properties of APTES-functionalized silica spheres have been investigated and optimized by varying the annealing temperature. On the other hand, erbium incorporation in silica spheres was also studied and the corresponding Er(3+) luminescence emission at 1.54 microm was evaluated for intensity and lifetime. The basic pH environment in the synthesis allows good control of the size of the spheres (approximately 200 nm in diameter), whereas the acidic route produces a wide dispersion in particle size (200-5000 nm). Both these approaches, however, can be followed to obtain an efficient photoluminescence (PL) emission for the APTES-functionalized silica spheres after 400-600 degrees C thermal treatment. If Er(NO(3))(3) is introduced in the basic solution, a rapid precipitation of Er(OH)(3) occurs, but erbium can be easily and efficiently incorporated in the acid-synthesized spheres, showing high PL intensity at 1.54 microm with lifetime of 3.9 ms. Finally, I discuss perspectives for the applications of these luminescent silica spheres, in particular as biological markers for bioimaging and biosensing.


Different Cold-Signaling Pathways Function in the Responses to Rapid and Gradual Decreases in Temperature.
In plants, cold temperatures trigger stress responses and long-term responses that result in cold tolerance. In Arabidopsis thaliana, three dehydration-responsive element (DRE) binding protein 1/C-repeat binding factors (DREB1/CBFs) act as master switches in cold-responsive gene expression. Induction of DREB1 genes triggers the cold stress-inducible transcriptional cascade, followed by the induction of numerous genes that function in the cold stress response and cold tolerance. Many regulatory factors involved in DREB1 induction have been identified, but how these factors orchestrate the cold stress-specific expression of DREB1s has not yet been clarified. Here, we revealed that plants recognize cold stress as two different signals, rapid and gradual temperature decreases, and induce expression of the DREB1 genes. CALMODULIN BINDING TRANSCRIPTION ACTIVATOR3 (CAMTA3) and CAMTA5 respond to a rapid decrease in temperature and induce the expression of DREB1s, but these proteins do not respond to a gradual decrease in temperature. Moreover, they function during the day and night, in contrast to some key circadian components, including CIRCADIAN CLOCK ASSOCIATED1 and LATE ELONGATED HYPOCOTYL, which regulate cold-responsive DREB1 expression as transcriptional activators only during the day. Thus, plants efficiently control the acquisition of freezing tolerance using two different signaling pathways in response to a gradual temperature decrease during seasonal changes and a sudden temperature drop during the night.


Lung cancer in pregnancy: report of nine cases from an international collaborative study.
Lung cancer is an uncommon diagnosis during pregnancy. The combination of smoking in young women, increased maternal age during pregnancy, and increasing incidence of lung cancer worldwide may cause an increase of pregnancy associated lung cancer. The aim of this study was to describe all cases of lung cancer during pregnancy, registered in the international Cancer in Pregnancy registration study (CIP study; www.cancerinpregnancy.org). We present nine cases, all advanced lung cancer during the course of pregnancy. Collected data included demographic features of the study patients, cancer treatment, pregnancy outcome as well as maternal and fetal outcomes. Nine pregnant patients from 4 European centres with a median age of 33 years old (range, 26-42) were included. The median gestational age at diagnosis was 17 weeks (range, 6-28). All patients presented with metastatic disease including bone, lung, brain, spinal cord, pleura, lymph nodes, adrenal and liver. Histopathology was compatible with adenocarcinoma in 4 patients, non-small cell lung cancer with unidentified subtype in 2 patients and squamous-cell, large-cell and a poorly differentiated carcinoma in 3 patients, respectively. Eight patients were treated with systemic therapy, five of them during gestation. No responses were seen. The maternal postpartum outcome was poor with less than one year survival following delivery. One patient experienced a spontaneous abortion and three pregnancies were terminated. Five infants were all born premature due to poor maternal status by cesarean section, with a median gestational age of 30 weeks (range 26-33). To summarize, lung cancer in pregnancy has a dismal maternal outcome in our series. We add nine new cases and discuss both therapeutic and prognostic results.


Perturbed mechanotransduction by endothelial surface glycocalyx modification greatly impairs the arteriogenic process.
The mechanisms that trigger initiation of arteriogenesis in response to pathogenic obstruction of arterial flow are not fully understood. Our objective is to determine whether glycocalyx mediated mechanotransduction of fluid shear stress to the endothelial layer is an essential first step in inducing arteriogenesis. Mice were implanted with an osmotic minipump containing saline or hyaluronan synthase inhibitor 4-methylesculetin (4ME) 2 wk before femoral artery ligation. 4ME was effective in modifying the endothelial glycocalyx as measured by dextran exclusion and perfused boundary region changes. Glycocalyx modification resulted in a 52% (P = 0.002) reduction in perfusion restoration through the 21-day follow-up [area under the curve, 4.9 ± 1.1 (n = 11) vs. 10.2 ± 3.2 (n = 10), 4ME vs. control (Ctrl)]. Upon femoral artery ligation, no change in collateral vessel diameter in 4ME treated mice (49.8 ± 26.3 vs. 47.1 ± 14.0 μm, ligated vs unligated) was observed (Ctrl, 88.5 ± 18.8 vs. 35.1 ± 3.0 μm, ligated vs unligated, P < 0.05). This impaired arteriogenic process was accompanied by lack of local induction of both endothelial and smooth muscle cell activation (Ki67, endothelial nitric oxide synthase, and ICAM-1), as well as a failure to recruit CD11b-positive cells in 4ME-treated collateral vessels (0.012 ± 0.003 vs. 0.010 ± 0.003 cells/μm vessel perimeter, ligated vs. unligated), whereas in Ctrls, the number of CD11b cells was increased (0.024 ± 0.002 vs. 0.010 ± 0.004 cells/μm vessel perimeter, P < 0.05). Modification of the glycocalyx by inhibition of hyaluronan synthesis renders the endothelium unresponsive to altered hemodynamic conditions resulting from femoral artery ligation, which results in a hampered restoration of distal perfusion.


Decoding distress among pandemic pregnancies: Examining pregnancy distress and COVID-19 stress as concurrent predictors of current and anticipated postpartum sleep during the COVID-19 pandemic.
The goal of the present study was to examine pregnancy distress, COVID-19 stress, COVID-19 compliance, and life satisfaction as concurrent predictors of perceived current and anticipated postpartum sleep duration (SD) and quality among a U.S. sample of pregnant individuals. The sample comprised 544 pregnant individuals (62.3% non-Hispanic White; mean age = 28.0 and SD = 6.2), most of whom were in their third trimester (42.3%). Participants were recruited through an online panel (Qualtrics) between October and November 2020. The primary analysis was based on a concurrent path model, which examined pregnancy distress, COVID-19 stress, COVID-19 compliance, and life satisfaction as predictors of current and anticipated postpartum SD and sleep quality, controlling for several demographic (eg, age and income), familial (eg, number of children and adults in the home), sleep-related (eg, chronotype and use of sleeping medication), and psychosocial functioning (eg, COVID-19 impact) variables. Results indicated that higher COVID-19 compliance was significantly associated with poorer current sleep quality. Furthermore, both pregnancy distress and COVID-19 stress predicted worse anticipated postpartum sleep quality (but not duration). Life satisfaction, however, was consistently significantly associated with current and anticipated postpartum SD and quality. Findings are critical for understanding how various forms of stress correlate with sleep health behaviors during pregnancy, as well as expectations for postpartum sleep among a vulnerable population at risk for both psychological and sleep challenges during a period of unprecedented global stress.


Bleustein-Gulyaev-Shimizu surface acoustic waves in two-dimensional piezoelectric phononic crystals.
In this paper, we present a study on the existence of Bleustein-Gulyaev-Shimizu piezoelectric surface acoustic waves in a two-dimensional piezoelectric phononic crystal (zinc oxide, ZnO, and cadmium-sulfide, CdS) using the plane wave expansion method. In the configuration of ZnO (100)/CdS(100) phononic crystal, the calculated results show that this type of surface waves has higher acoustic wave velocities, high electromechanical coupling coefficients, and larger band gap width than those of the Rayleigh surface waves and pseudosurface waves. In addition, we find that the folded modes of the Bleustein-Gulyaev-Shimizu surface waves have higher coupling coefficients.


Association between glycated hemoglobin and diet and other lifestyle factors in a nondiabetic population: cross-sectional evaluation of data from the Potsdam cohort of the European Prospective Investigation into Cancer and Nutrition Study.
Glycation reactions of proteins and other compounds, depending on blood glucose concentrations, have a detrimental effect on health. The association of diet and other lifestyle factors with glycated hemoglobin (Hb A(1c)) values was examined in a nondiabetic population. This was a cross-sectional study of 1773 middle-aged men and women. Mean Hb A(1c) values were calculated for categories of diet and lifestyle factors, and odds ratios (ORs) for the highest versus lowest tertiles of Hb A(1c) were determined and compared. The OR of being in the highest Hb A(1c) tertile compared with the lowest increased with greater age [age 40-44 y compared with >60 y: men (OR: 2.86; 95% CI: 1.60, 5.20) and women: (6.11; 3.15, 12.30)] and greater obesity [body mass index (in kg/m(2)) >25 and waist-hip ratio >1.0 in men and >0.8 in women): men (2.80; 1.48, 5.45) and women (1.73; 1.15, 2.61)]. High energy and energy-adjusted saturated fat intakes were associated with increased risk of being in the highest tertile of Hb A(1c) [highest compared with lowest quintile: (1.53; 1.04, 2.26; P for trend = 0.013) and (1. 98; 1.33, 2.95; P for trend = 0.003), respectively]. No significant associations were observed for intakes of carbohydrates, protein, dietary fiber, or beta-carotene; however, some of the associations were nearly significant. Alcohol, vitamin C, and vitamin E intakes were inversely related to risk [highest compared with lowest quintile: (0.56; 0.38, 0.83; P for trend = 0.001), (0.50; 0.33, 0. 74; P for trend = 0.003), and (0.65; 0.43, 0.96; P for trend = 0. 036), respectively]. Hb A(1c) values might be modifiable by diet and other lifestyle factors.


MiR-33a-5p inhibits the growth and metastasis of melanoma cells by targeting SNAI2.
MicroRNAs have been verified as critical regulators in the development of melanoma. miR-33a-5p was significantly downregulated in melanoma, however, the specific role and regulatory mechanism of miR-33a-5p in melanoma were still unclear. The present study identified that miR-33a-5p was downregulated in melanoma tissues and cells, while SNAI2 was upregulated. miR-33a-5p directly targeted SNAI2 and negatively regulated its expression in melanoma cells. Overexpression of miR-33a-5p repressed proliferation, migration, invasion, EMT and promoted apoptosis of melanoma cell in vitro, these effects were partially reversed by SNAI2 overexpression. In addition, miR-33a-5p impaired melanoma growth in vivo by inhibiting SNAI2. Mechanistically, miR-33a-5p repressed activation of the PI3K/AKT/mTOR pathway by targeting SNAI2. In conclusion, miR-33a-5p repressed the progression of melanoma by targeting SNAI2 via inactivation of the PI3K/AKT/mTOR signaling pathway, providing a potential molecular mechanism for the treatment of melanoma.


Lentivirus expressing shRNAs inhibit the replication of contagious ecthyma virus by targeting DNA polymerase gene.
Contagious ecthyma or Orf is known as a zoonotic disease remains prevalently worldwide despite the application of some control strategies against it. RNAi particularly shRNA provides us with the chance to tackle this obstacle by an encouraging new approach. The current study indicates the design and experiment of third-generation lentivirus packaging systems delivering shRNAs to inhibit Orf virus (ORFV) replication and infection. Given the importance of DNA-pol gene in virus replication, in this study, three shRNAs against this gene were designed and cloned into lentiviral vectors to stabilize the expression of shRNAs. After producing lentivectors expressing ORFV-DNA- pol in HEK293T cells, the synthesized shRNAs were applied to downregulate viral replication and gene expression. The reduction in viral titer and RNA was evaluated by TCID50 test as well as real-time RT-PCR. The results were then analyzed in comparison with the control group. Designed shRNAs significantly reduced virus yield approximately 90 to 97% and 96.8 to 99.4%, respectively compared to the control groups (cells infected with ORFV and infected with ORFV and scrambled vector) by TCID50 test. Real-time RT-PCR revealed a dramatic reduction in the expression of viral RNA approximately 99% compared to cells infected with ORFV and from 92.6 to 99%, respectively compared to cells infected with ORFV and scrambled vector. Therefore, it can be stated that RNAi is capable of being used as a potent therapeutically option against viruses like ORFV.


[Amino acid composition of the rat quadriceps femoris muscle after a flight on the Kosmos-936 biosatellite].
The amino acid composition of the quadriceps muscle of rats flown onboard the biosatellite Cosmos-936 and exposed to the ground-based synchronous control experiment was studied. The weightless rats showed changes in the amino acid concentration in the quadriceps muscle. The centrifuged flight and synchronous rats displayed an accumulation of free amino acids in the above muscle.


Dielectric and gravimetric studies of water binding to lysozyme.
Time domain dielectric spectroscopy and hydration isotherm measurements as a function of temperature have been applied to hydrated lysozyme powder. Two dielectric dispersions were identified, the first centred at approximately 8 MHz and a second above 1 GHz. The higher dispersion is considered to be the result of rotational relaxation of water molecules bound to the enzyme. In this case the results indicate the existence of a population of 32 water molecules per lysozyme molecule which are irrotationally bound to the lysozyme structure. A larger population of water molecules is relatively free to respond to the electric field and exhibits a dipole moment close to that of vapour phase water molecules. Multi-temperature hydration isotherm measurements are used to calculate enthalpies and entropies associated with the binding of water to lysozyme. Discontinuities both in dielectric and in thermodynamic characteristics in the range 10-14% hydration are interpreted as a re-ordering of the water structure on the enzyme surface.


The selective removal of phosphorus from soil: is event size important?
Data from the Woburn Erosion Reference Experiment (Bedfordshire, UK) were used to test the hypothesis that losses of phosphorus (P) in small erosion events are as great as those in infrequent large events, and to examine the effect of storm characteristics on the selective enrichment of P in eroded sediment. For almost every plot event in the period 1988 to 1994, the clay-sized fraction of the sediment was enriched compared with the soil of the plots. There was more variation in clay enrichment for smaller erosion events than for larger ones. The clay and P contents of the sediment were strongly correlated (p < 0.01), and there was a wider range of P concentrations in the sediment derived from small events than in that from large events. However, individual events resulting in small soil losses (< 100 kg) did not account for greater P losses than larger events (> 100 kg). The greater frequency of smaller events, combined with the likelihood of higher P concentrations in the sediment, therefore accounted for a greater proportion of the P lost over the 6-yr period than the infrequent large events. Phosphorus concentrations generally increased with increasing peak discharge and decreased with increasing event duration. For the same return period, P losses were generally greater from plots cultivated up and down the slope than from those cultivated across the slope. Overall, our results suggest that small erosion events should be controlled to prevent P contamination of surface waters and that the most effective means of doing this are by the introduction of minimal tillage techniques and across-slope cultivations.


STRait Razor: a length-based forensic STR allele-calling tool for use with second generation sequencing data.
Recent studies have demonstrated the capability of second generation sequencing (SGS) to provide coverage of short tandem repeats (STRs) found within the human genome. However, there are relatively few bioinformatic software packages capable of detecting these markers in the raw sequence data. The extant STR-calling tools are sophisticated, but are not always applicable to the analysis of the STR loci commonly used in forensic analyses. STRait Razor is a newly developed Perl-based software tool that runs on the Linux/Unix operating system and is designed to detect forensically-relevant STR alleles in FASTQ sequence data, based on allelic length. It is capable of analyzing STR loci with repeat motifs ranging from simple to complex without the need for extensive allelic sequence data. STRait Razor is designed to interpret both single-end and paired-end data and relies on intelligent parallel processing to reduce analysis time. Users are presented with a number of customization options, including variable mismatch detection parameters, as well as the ability to easily allow for the detection of alleles at new loci. In its current state, the software detects alleles for 44 autosomal and Y-chromosome STR loci. The study described herein demonstrates that STRait Razor is capable of detecting STR alleles in data generated by multiple library preparation methods and two Illumina(®) sequencing instruments, with 100% concordance. The data also reveal noteworthy concepts related to the effect of different preparation chemistries and sequencing parameters on the bioinformatic detection of STR alleles.


Electroacupuncture induced spinal plasticity is linked to multiple gene expressions in dorsal root deafferented rats.
The underlying mechanism for electroacupuncture (EA) associated functional improvement in patients suffering from spinal cord injury (SCI) is largely unknown. Collateral sprouting is one plausible factor, where the cord microenvironment may contribute greatly. The present study evaluated the effects of EA on collateral sprouting from spared dorsal root ganglion (DRG), sensory functional restorations, and differential gene expressions in spinal cord after partial DRG removal in the rat. Following EA, N1 waveform latencies for cortical somatosensory evoked potential significantly shortened. The densities of terminal sprouting from the spared DRG significantly increased on the EA versus the non-EA side. Microarray analysis revealed that several genes were upregulated on the acupunctured side at different time points; they were ciliary neurotrophic factor (CNTF) at 1 day postoperation (dpo), fibroblast growth factor (FGF)-1, insulin-like growth factor (IGF) 1 receptor, neuropeptide Y, and FGF-13 at 7 dpo, and CNTF and calcitonin gene-related polypeptide-alpha at 14 dpo, respectively. Meanwhile, five genes (CNTF, p75-like apoptosis-inducing death domain protein, IGF-1, transforming growth factor-beta 2, and FGF-4) were downregulated at 7 dpo. Furthermore, reverse transcriptase polymerase chain reaction results supported the gene chip analysis. It was concluded that the EA induced sensory functional restorations following partial DRG ganglionectomies could be brought about by intraspinal sprouting from the spared DRG, as well as multiple differential gene expressions in the spinal cord. The results could have clinical application in EA treatment of patients after spinal injury.


Load capacity improvements in nucleic acid based systems using partially open feedback control.
Synthetic biology is facilitating novel methods and components to build in vivo and in vitro circuits to better understand and re-engineer biological networks. Recently, Kim and Winfree have synthesized a remarkably elegant network of transcriptional oscillators in vitro using a modular architecture of synthetic gene analogues and a few enzymes that, in turn, could be used to drive a variety of downstream circuits and nanodevices. However, these oscillators are sensitive to initial conditions and downstream load processes. Furthermore, the oscillations are not sustained since the inherently closed design suffers from enzyme deactivation, NTP fuel exhaustion, and waste product build up. In this paper, we show that a partially open architecture in which an [Symbol: see text]1 adaptive controller, implemented inside an in silico computer that resides outside the wet-lab apparatus, can ensure sustained tunable oscillations in two specific designs of the Kim-Winfree oscillator networks. We consider two broad cases of operation: (1) the oscillator network operating in isolation and (2) the oscillator network driving a DNA tweezer subject to a variable load. In both scenarios, our simulation results show a significant improvement in the tunability and robustness of these oscillator networks. Our approach can be easily adopted to improve the loading capacity of a wide range of synthetic biological devices.


[Significance of hereditary factors in multiple organ dysfunction syndrome in children with infections].
In order to elucidate the contribution of hereditary factor (dose of ribosome genes) to the realization of organ dysfunction syndromes in children with surgical infection, the total size of silver nitrate-stained nucleoli-forming regions (Ag-NFR) of acrocentric chromosomes in karyotypes was assessed in 22 patients aged from 14 months to 12 years, 17 of these with appendicular peritonitis, 4 with destructive pneumonia, and 1 with acute hematogenic osteomyelitis. Pyoseptic diseases involved no organ dysfunction in 10 patients, and in 12, multiple organ dysfunction was diagnosed. Our findings indicate that the carriers of low-copy variants of ribosome genes are characterized by hereditary predisposition to lowered individual resistance of the organism. Therefore, children with surgical infection with a decreased size of Ag-NFR (low dose of ribosome genes) in the karyotype potentially represent a group at risk of multiple organ dysfunction realization.


Mutational analysis of cis-acting RNA signals in segment 7 of influenza A virus.
The genomic viral RNA (vRNA) segments of influenza A virus contain specific packaging signals at their termini that overlap the coding regions. To further characterize cis-acting signals in segment 7, we introduced synonymous mutations into the terminal coding regions. Mutation of codons that are normally highly conserved reduced virus growth in embryonated eggs and MDCK cells between 10- and 1,000-fold compared to that of the wild-type virus, whereas similar alterations to nonconserved codons had little effect. In all cases, the growth-impaired viruses showed defects in virion assembly and genome packaging. In eggs, nearly normal numbers of virus particles that in aggregate contained apparently equimolar quantities of the eight segments were formed, but with about fourfold less overall vRNA content than wild-type virions, suggesting that, on average, fewer than eight segments per particle were packaged. Concomitantly, the particle/PFU and segment/PFU ratios of the mutant viruses showed relative increases of up to 300-fold, with the behavior of the most defective viruses approaching that predicted for random segment packaging. Fluorescent staining of infected cells for the nucleoprotein and specific vRNAs confirmed that most mutant virus particles did not contain a full genome complement. The specific infectivity of the mutant viruses produced by MDCK cells was also reduced, but in this system, the mutations also dramatically reduced virion production. Overall, we conclude that segment 7 plays a key role in the influenza A virus genome packaging process, since mutation of as few as 4 nucleotides can dramatically inhibit infectious virus production through disruption of vRNA packaging.


Matrix-assisted laser desorption/ionization mass spectrometric analysis of aliphatic biodegradable photoluminescent polymers using new ionic liquid matrices.
In this study, two novel ionic liquid matrices (ILMs), N,N-diisopropylethylammonium 3-oxocoumarate and N,N-diisopropylethylammonium dihydroxymonooxoacetophenoate, were tested for the structural elucidation of recently developed aliphatic biodegradable polymers by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). The polymers, formed by a condensation reaction of three components, citric acid, octane diol, and an amino acid, are fluorescent, but the exact mechanism behind their luminescent properties has not been fully elucidated. In the original studies, which introduced the polymer class (J. Yang et al., Proc. Natl. Acad. Sci. USA 2009, 106, 10086-10091), a hyper-conjugated cyclic structure was proposed as the source for the photoluminescent behavior. With the use of the two new ILMs, we present evidence that supports the presence of the proposed cyclization product. In addition, the new ILMs, when compared with a previously established ILM, N,N-diisopropylethylammonium α-cyano-3-hydroxycinnimate, provided similar signal intensities and maintained similar spectral profiles. This research also established that the new ILMs provided good spot-to-spot reproducibility and high ionization efficiency compared with corresponding crystalline matrix preparations. Many polymer features revealed through the use of the ILMs could not be observed with crystalline matrices. Ultimately, the new ILMs highlighted the composition of the synthetic polymers, as well as the loss of water that was expected for the formation of the proposed cyclic structure on the polymer backbone.


Efficacy of glove combinations in reducing cell culture infection after glove puncture with needles contaminated with human immunodeficiency virus type 1.
To study the effect of various latex and treated glove combinations in reducing the frequency of human immunodeficiency virus (HIV) infection of tissue culture cells after puncture by surgical needles contaminated with infectious human immunodeficiency virus type 1 (HIV-1). One, two, or three layers of sterile latex glove material, or two latex layers with intermediate cotton or Kevlar (with or without the virucidal compound nonoxynol-9) were used to cover 24-well cell culture dishes containing MT2 cells in cell culture medium. Surgical needles wet with cell culture medium containing HIV-1 (HTLV IIIA strain) were passed through the glove materials into the culture medium in the wells of the culture dishes. The culture medium in each well was then assayed biweekly for HIV-1 p24 antigen as a test for infection of cells in the well. The rate of HIV-1 infection of cell cultures after glove puncture was greater than 90% with a single latex surgical glove barrier, 23% to 60% with double or triple layers of latex gloves, less than 8% with an intermediate cotton glove impregnated with 4% nonoxynol-9, 6% with an intermediate Kevlar glove, and 0% with an intermediate Kevlar glove impregnated with nonoxynol-9. An intermediate glove of Kevlar or of Kevlar or cotton impregnated with virucidal compound nonoxynol-9 between standard latex gloves may improve surgical glove safety, compared with latex gloves alone with respect to needlestick transmission of HIV-1. The experimental model used may permit rapid investigation of other glove systems as barriers to the transfer of infectious agents through gloves by needlestick.


Early induction of uncoupling protein-2 in pulmonary macrophages in hyperoxia-associated lung injury.
High concentrations of inspired oxygen contribute to the pathogenesis of neonatal bronchopulmonary dysplasia and adult acute respiratory distress syndrome. Animal models of hyperoxia-associated lung injury (HALI) are characterized by enhanced generation of reactive oxygen species (ROS) and an adaptive antioxidant response. ROS contribute to pathogenesis, partly through enhancing pro-inflammatory activity in macrophages. Uncoupling protein-2 (UCP2) is an inner mitochondrial membrane protein whose expression lowers mitochondrial superoxide (O₂ⁱ⁻) production. UCP2, therefore, has potential to contribute to antioxidant response. It is inducible in macrophages. We hypothesized that induction of UCP2 occurred in response to pulmonary hyperoxia in vivo and that expression localized to pulmonary macrophages. We then investigated mechanisms of UCP2 regulation in hyperoxia-exposed macrophages in vitro and correlated changing UCP2 expression with mitochondrial membrane potential (Δψm) and O₂ⁱ⁻ production. UCP2 is induced in lungs of mice within 1 h of hyperoxia exposure. Induction occurs in pulmonary alveolar macrophages in vivo, and can be replicated in vitro in isolated macrophages. UCP2 mRNA does not change. UCP2 increases quickly after the first hyperoxia-induced burst of mitochondrial O₂ⁱ⁻ generation. Suppression of Δψm and mitochondrial O₂ⁱ⁻ production follow and persist while UCP2 is elevated. Induction of UCP2 is an early response to hyperoxia in pulmonary macrophages. The mechanism is post-transcriptional. UCP2 induction follows a transient rise in mitochondrial ROS generation. The subsequent falls in Δψm and mitochondrial O₂ⁱ⁻ support the notion that regulable UCP2 expression in macrophages acts to contain mitochondrial ROS generation. That, in turn, may limit inappropriate pro-inflammatory activation in HALI.


Sex-Disparity in the Association Between Birthweight and Cardiovascular Parameters in 4-Year-Old Children: A Chinese Cohort Study.
Background: Sex-related differences in cardiovascular parameters have been well documented in adults, and the impact of birthweight on cardiovascular health in later life has been acknowledged. However, data was limited regarding the association between birthweight and cardiovascular outcomes at an early age, and the sex-disparity in the association remained unclear. Objective: To investigate the association between birthweight and cardiovascular parameters in 4-year-old children. Furthermore, to explore whether sex-disparity exist in this association or in cardiovascular risk. Methods: Follow-up data from the Shanghai Birth Cohort (SBC) was analyzed. Detailed perinatal information including both maternal and offspring datum were recorded. Blood pressure, echocardiography, and anthropometry assessment were conducted during the follow-up of 4-year-old children. Linear regression models were used to analyze the association between birthweight and left ventricle (LV) structure and function changes in each sex and birthweight category. Multivariable logistic regression models were used to compare risk of left ventricular hypertrophy (LVH) in different birthweight subgroups. Results: Overall, macrosomia was significantly associated with thickened LV posterior wall thickness in systole [LVPWs, (β = 0.26, 95% CI: 0.06, 0.45)] and diastole [LVPWd, (β = 0.18, 95% CI: 0.06, 0.30)], and thickened interventricular septal thickness in diastole [IVSd, (β = 0.16, 95% CI: 0.05, 0.28)]. Boys with macrosomia showed a higher left ventricle mass index [LVMI, (β = 1.29, 95% CI: 0.14, 2.43)], thickened LVPWs (β = 0.30, 95% CI: 0.05, 0.56) and LVPWd (β = 0.21, 95% CI: 0.06, 0.36), and thickened IVSd (β = 0.23, 95% CI: 0.09, 0.36). However, no significant association of structural changes was found in girls. Furthermore, an increased risk of LVH was found solely in macrosomic boys (OR = 2.79, 95% CI: 1.17, 6.63). Conclusion: Children with macrosomia developed cardiovascular changes as early as 4 years of age. Macrosomia was associated with LV structural changes and higher LVH risk in pre-school-aged boys, while no association was found in girls.


Universal method for synthesis of artificial gel antibodies by the imprinting approach combined with a unique electrophoresis technique for detection of minute structural differences of proteins, viruses, and cells (bacteria): II. Gel antibodies against virus (Semliki Forest Virus).
Artificial and highly selective antibodies (in the form of gel granules) against proteins can easily be synthesized by a simple, cost-effective imprinting technique [Liao, J.-L. et al., Chromatographia 1996, 42, 259-262]. Using the same method for synthesis of gel antibodies against viruses in combination with analysis by free zone electrophoresis in a rotating narrow bore tube we have shown that artificial gel antibodies against Semliki Forest Virus (wild type) can sense the difference between this virus and a mutant, although they differ in their chemical composition only by three amino acids in one of the three proteins on the surface of the virus particle. The reason for this extremely high resolution is explained by the fact that we use three types of selectivity: (i) shape selectivity (created by the close fit between the antigen and its imprint in the gel), (ii) bond selectivity in the contact area between the antigen and its imprint in the gel antibody, and (iii) charge selectivity, originating from slightly different structures or/and conformations of the antigens.


Antikeratin antibodies in sera of patients with connective tissue disease and liver disease.
Sera from 93 patients with connective tissue disease, 36 rheumatoid arthritis, 41 systemic lupus erythematosus, 12 polymyositis/dermatomyositis and 4 systemic sclerosis and sera from 12 patients with liver disease, along with sera of 10 healthy subjects, were tested for antikeratin antibodies using enzyme-linked immunosorbent assay in which the wells were coated with human epidermal keratin. Values above the mean+SD of the 10 healthy subjects were found in 8.3% of rheumatoid arthritis, 29.3% of systemic lupus erythematosus, 33.3% of polymyositis/dermatomyositis, 50% of systemic sclerosis, 16.7% of liver disease patients, and 20% of healthy subjects. The results indicated a character of naturally occurring antibody of antikeratin antibodies in human sera.


Stringent or nonstringent complete remission and prognosis in acute myeloid leukemia: a Danish population-based study.
Stringent complete remission (sCR) of acute myeloid leukemia is defined as normal hematopoiesis after therapy. Less sCR, including non-sCR, was introduced as insufficient blood platelet, neutrophil, or erythrocyte recovery. These latter characteristics were defined retrospectively as postremission transfusion dependency and were suggested to be of prognostic value. In the present report, we evaluated the prognostic impact of achieving sCR and non-sCR in the Danish National Acute Leukaemia Registry, including 769 patients registered with classical CR (ie, <5% blasts in the postinduction bone marrow analysis). Individual patients were classified as having sCR (n = 360; 46.8%) or non-sCR (n = 409; 53.2%) based on data from our national laboratory and transfusion databases. Survival analysis revealed that patients achieving sCR had superior overall survival (hazard ratio [HR], 1.34; 95% confidence interval [CI], 1.10-1.64) as well as relapse-free survival (HR, 1.25; 95% CI, 1.03-1.51) compared with those with non-sCR after adjusting for covariates. Cox regression analysis regarding the impact of the stringent criteria for blood cell recovery identified these as significant and independent variables. In conclusion, this real-life register study supports the international criteria for response evaluation on prognosis and, most importantly, documents each of the 3 lineage recovery criteria as contributing independently.


Antagonistic effects of selenium on lead-induced autophagy by influencing mitochondrial dynamics in the spleen of chickens.
Lead (Pb) may damage the immune function in human and animal. Selenium (Se) has antagonistic effects on Pb. In our study, brown layer chickens were randomly allocated to control group, Se group (1 mg/kg Se), Se+Pb group (1 mg/kg Se and 350 mg/kg Pb), and Pb group (350 mg/kg Pb). The chickens were sacrificed on the 90th day; spleen tissues were subjected to observation of ultrastructure and detection of spleen-related indexes. The results revealed that in the Pb group, expression levels of the cytokines IL-1 and TNF-α significantly increased, and expression levels of IL-2 and INF-γ significantly decreased; activities of antioxidant enzyme GPX, SOD and CAT significantly decreased, and expression level of malondialdehyde (MDA) significantly increased; expression levels of mitochondrial fission-related genes (Mff and Drp1) significantly increased, and expression levels of mitochondrial fusion-related genes (Opa1, Mfn1 and Mfn2) significantly decreased; expression of autophagy-related genes (Beclin 1, Dynein, Atg 5, LC3-I and LC-II) was upregulated, while expression of mammalian target of rapamycin (mTOR) was downregulated. The results of transmission electron microscopy indicated that Pb induced mitochondrial fragmentation, and triggered autophagy in the spleen of chickens. The Se and Pb co-treatment remarkably alleviated these injuries induced by Pb in the spleen of chickens. In conclusion, Pb can induce the oxidative stress to influence the mitochondrial dynamics balance and lead to autophagy, which triggers the immune dysfunction in the spleen of chickens; the Se exhibits the antagonistic effects on lead-induced autophagy by influencing mitochondrial dynamics in the spleen of chickens.


Effects of HMW-GS Ax1 or Dx2 absence on the glutenin polymerization and gluten micro structure of wheat (Triticum aestivum L.).
Wheat (Triticum aestivum L.) dough strength and extensibility are mainly determined by the polymerization of glutenin. The number of high-molecular-weight glutenin subunits (HMW-GS) differs in various wheat varieties due to the silencing of some genes. The effects of Ax1 or Dx2 subunit absence on glutenin polymerization, dough mixing properties and gluten micro structure were investigated with two groups of near-isogenic lines. The results showed that Ax1 or Dx2 absence decreased the accumulation rate of glutenin polymers and thus delayed the rapid increase period for glutenin polymerization by at least ten days, which led to lower percentage of polymeric protein in mature grain. Ax1 or Dx2 absence significantly decreased the dough development time and dough stability, but increased the uniformity of micro structure. Lacunarity, derived from quantitative analysis of gluten network, is suggested as a new indicator for wheat quality.


Combination of adoptive immunotherapy with Herceptin for patients with HER2-expressing breast cancer.
Clinical use of Herceptin (trastuzumab), which is a humanized monoclonal antibody against HER2, started for patients with HER2-overexpressing breast cancer. To potentiate the efficacy of the Herceptin therapy, this study focused on the combination of Herceptin with activated immune lymphocytes. We used peripheral blood mononuclear cells (PBMCs) as effector cells and used HER2-unexpressing K562 cells, HER2-weakly-expressing breast carcinoma cells (Breast-M), or HER2-strongly-expressing breast carcinoma cells (BT-474) as target cells. Interleukin-2 (IL-2)-activated PBMCs, IL-2/OKT-3-activated PBMCs and a streptococcal preparation OK-432-activated PBMCs were generated and used as effector cells. Cytotoxic activity was determined with 4-hour 51Cr release assay. Both fresh PBMCs and activated PBMCs exhibited Herceptin-dependent cytotoxicity. Importantly, Herceptin-dependent cytotoxicity was found even at a lower effector to target cell ratio (E/T ratio) than that of Herceptin-independent cytotoxicity. In addition, Herceptin-dependent cytotoxicity by these activated PBMCs was observed even in HER2-weakly-expressing Breast-M cells. Since gamma-globulin or anti-CD16 antibody abrogated Herceptin-dependent cytotoxicity, it seems likely that antibody-dependent cellular cytotoxicity (ADCC) plays an important role in the Herceptin-dependent cytotoxicity. We present a recurrent breast cancer patient with malignant pleural effusion, in which HER2-strongly-expressing tumor cells were present, who was undergoing Herceptin therapy. Cluster formation between tumor cells and intrapleural mononuclear cells was induced 24 hours after intravenous injection of Herceptin (4 mg/kg). Mononuclear cells bound specifically to HER2-strongly-expressing tumor cells but not to other cells, such as mesothelial cells, suggested a Herceptin-mediated binding like ADCC in vivo. Taken together, these findings suggest that the combination of Herceptin with various types of activated lymphocytes may be a new therapeutic strategy, not only for HER2-strongly-expressing breast cancer but also for HER2-weakly-expressing cancer.


Prevalence of Periodontal Destruction and Putative Periodontal Pathogens in the Same Lebanese Family.
Periodontal diseases are associated with microorganisms rich in Gram-negative species. Several studies have indicated the presence of few a periodontopathic microorganisms in the same family. A parent with severe adult periodontitis, who is infected with bacteria associated with periodontal disease, may function as a source of infection. Their children may be at a greater risk to become colonized with bacteria. The purpose of this investigation was (1) to explore the presence of three bacteria, such as Porphyromonas gingivalis (PG), Prevotella intermedia (PI), and Aggregatibacter actinomycetemcomitans (AA) in the same Lebanese family and (2) to study the clinical destruction in the same family and their relations as members of this family due to the presence of PG. A total of 10 families were screened; only 5 (13 females and 5 males) were selected for this study, and at least one member of the family had untreated periodontal disease, chronic or aggressive. Every participant signed an informed consent form. A total of 18 available deoxyribonucleic acid (DNA) samples were taken to analyze the presence of three periodontal bacteria. Multiple logistic regression was used for the exact methods. All 18 patients showed a positive result for PI. Also, PG. was recognized in 15 patients while AA was not detected in any of the subjects. All couples suffered from periodontitis, chronic or aggressive forms, five children suffered from gingivitis, three children had no clinical manifestation, and only one suffered from localized aggressive periodontitis. The statistical analysis showed with each 1 year of increase in age, the odds of having periodontal disease multiply by 1.39, i.e., age as a risk factor for periodontal disease due to the presence of PG and sharing the same plate. This investigation demonstrates a high prevalence of periodontal microorganisms in children and young adults of Lebanese periodontitis parents and a microbiological similarity between the children and their mothers. All these factors could be a high risk of developing periodontal disease in the future. This article shows that vertical transmission of microorganisms is a possible risk factor for developing periodontal disease in the offspring.


p66Shc Mediates Mitochondrial Dysfunction Dependent on PKC Activation in Airway Epithelial Cells Induced by Cigarette Smoke.
Airway epithelial mitochondrial injury plays a critical role in the pathogenesis of chronic obstructive pulmonary disease (COPD). The p66Shc adaptor protein is a newly recognized mediator of mitochondrial dysfunction. However, little is known about the effect of p66Shc on airway epithelial damage in the development of COPD. The aim of the present study is to investigate the roles of p66Shc and its upstream regulators in the mitochondrial injury of airway epithelial cells (Beas-2b) induced by cigarette smoke extract (CSE). Our present study revealed that CSE increased p66Shc expression and its mitochondrial translocation in concentration and time-dependent manners in airway epithelial cells. And p66Shc siRNA significantly attenuated mitochondrial dysfunction and cell injury when airway epithelial cells were stimulated with 7.5% CSE. The total and phosphorylated expression of PKCβ and PKCδ was significantly increased associated with mitochondrial dysfunction and cell injury when airway epithelial cells were exposed to 7.5% CSE. The pretreatments with pharmacological inhibitors of PKCβ and PKCδ could notably suppress p66Shc phosphorylation and its mitochondrial translocation and protect the mitochondria and cells against oxidative damage when airway epithelial cells were incubated with 7.5% CSE. These data suggest that a novel PKCβ/δ-p66Shc signaling pathway may be involved in the pathogenesis of COPD and other oxidative stress-associated pulmonary diseases and provide a potential therapeutic target for these diseases.


[LC-MS/MS analysis of herbicide residues in waters from Navarre].
The aim of the study was to obtain information on the residues of herbicides (imazametabenz, simazine, isoproturon, clortoluron, metribuzine, atrazine, cianazine, terbutrin, propanil, terbutilazine, alachlor and pendimetalin) present in water proceeding from Navarre. A total of 465 samples of water were taken: 378 were taken from 141 water supplies; the remaining 87 proceeded from fountains, springs and borings in agricultural areas in the south of Navarre. The technique employed was LC-MS/MS. Herbicide residues were detected in 33% of the water supplies, and their concentration exceeded the normative limit in 8% of them. Herbicide residues were found in 86% of the samples from wells and fountains of the agricultural areas of the south of Navarre. The number of water supplies with herbicide residues increased from north to south. However, the percentage of water supplies in which the concentration of herbicides exceeded the limits was greater in the centre of Navarre than in the north and south. Imazametabenz was the herbicide most detected in the samples from the northern, central and southern areas. Atrazine, terbutilazine and imazametabenz were most detected in the samples of the southern area. The differences amongst the three areas of Navarre can be explained as being due to the different agricultural practices of each area. Concentrations of herbicides exceeding the normative limits were not detected in any of the water samples proceeding from the larger towns of Navarre.


Independent 5' and 3'-end determination of multiple dihydrofolate reductase transcripts.
Multiple dihydrofolate reductase (dhfr) mRNAs, differing substantially in abundance, are produced as a result of the utilization of multiple transcription initiation sites and multiple polyadenylation sites. We have shown that dhfr mRNAs initiating from an upstream promoter region utilize the same collection of six polyadenylation sites and generate multiple dhfr mRNAs at the same relative abundance as do the mRNAs initiating from the major transcription promoter region. These results indicate that the 5' and 3' ends of dhfr mRNAs are independently determined. We show that the relative abundance of steady-state dhfr mRNAs was the same in nuclear and cytoplasmic RNA fractions. This finding makes it unlikely that differences in mRNA stability account for differences in the relative abundance of the multiple dhfr mRNAs in the cytoplasm. Our analysis of the dhfr promoter region revealed the existence of stable cytoplasmic polyadenylated transcripts complementary to the first 300 nucleotides of the dhfr transcripts initiating from the upstream promoter region. Therefore, the dhfr locus hosts two divergent and partially overlapping genes which share the same promoter region.


Diagnostic approach in cases with suspected work-related asthma.
Work-related asthma (WRA) is a major cause of respiratory disease in modern societies. The diagnosis and consequently an opportunity for prevention are often missed in practice. Based on recent studies and systematic reviews of the literature methods for detection of WRA and identification of specific causes of allergic WRA are discussed. All workers should be asked whether symptoms improve on days away from work or on holidays. Positive answers should lead to further investigation. Spirometry and non-specific bronchial responsiveness should be measured, but carefully performed and validly analysed serial peak expiratory flow or forced expiratory volume in one second (FEV1) measurements are more specific and confirm occupational asthma in about 82% of those still exposed to the causative agent. Skin prick testing or specific immunoglobulin E assays are useful to document allergy to high molecular weight allergens. Specific inhalational challenge tests come closest to a gold standard test, but lack standardisation, availability and sensitivity. Supervised workplace challenges can be used when specific challenges are unavailable or the results non-diagnostic, but methodology lacks standardisation. Finally, if the diagnosis remains unclear a follow-up with serial measurements of FEV1 and non-specific bronchial hyperresponsiveness should detect those likely to develop permanent impairment from their occupational exposures.


High Prevalence of Diabetes Distress in a University Population.
Diabetes distress is an affective condition that addresses an individual's frustrations, worries, and concerns about living with diabetes. It is associated with fewer self-care behaviors, suboptimal glycemic control, and lower quality of life (QOL). For these reasons, diabetes care guidelines recommend routine assessment of diabetes distress. To assess diabetes distress in a university population. This study was conducted using a descriptive, cross-sectional design. Researchers assessed diabetes distress and other psychosocial factors via an electronic anonymous survey among students, faculty, and staff at a large university in the Midwest. A total of 173 participants completed the survey (mean [SD] age, 35.1 [16.7] years), with 108 [62.4%] female and 142 [82.1%] white participants). Eighty-five participants had type 1 diabetes mellitus (T1DM), and 88 had type 2 diabetes mellitus (T2DM). Of the 85 T1DM participants, 23 (27.4%) reported high diabetes distress, and 27 (30.7%) T2DM participants reported high diabetes distress. Sixteen T1DM (18.8%) and 15 T2DM (17.0%) participants screened positive for severe depression. Severe depression was associated with high distress for both T1DM and T2DM participants (T1DM: χ2=28.845, P<.001; T2DM: χ2=20.679, P<.001). Participants with T1DM reported more frequent self-care behaviors (mean [SD], 62.3 [17.1] vs 52.2 [19.2]; P<.001), but lower diabetes QOL (63.3 [14.1] vs 68.5 [15.5]; P=.021) compared with T2DM participants. No differences were observed in depressive symptoms, diabetes self-efficacy, and coping styles. Linear regression models showed that high diabetes distress scores (standardized β=.323, P=.025; standardized β=.604, P<.001) were independently associated with higher hemoglobin A1C levels and lower diabetes QOL after controlling for depressive symptoms, age, and gender in T1DM participants. Similarly, high diabetes distress scores (standardized β=.434, P<.001) were associated with lower diabetes QOL in T2DM participants after controlling for the same variables. High diabetes distress levels were associated with lower diabetes QOL for both T1DM and T2DM participants. These findings suggest that attending or working at a university may be associated with high diabetes distress scores and lower diabetes QOL. Additional research with a larger, more diverse sample from multiple universities is needed to confirm these findings.


Preoperative identification of facial nerve in vestibular schwannomas surgery using diffusion tensor tractography.
Facial nerve palsy is a common complication of treatment for vestibular schwannoma (VS), so preserving facial nerve function is important. The preoperative visualization of the course of facial nerve in relation to VS could help prevent injury to the nerve during the surgery. In this study, we evaluate the accuracy of diffusion tensor tractography (DTT) for preoperative identification of facial nerve. We prospectively collected data from 11 patients with VS, who underwent preoperative DTT for facial nerve. Imaging results were correlated with intraoperative findings. Postoperative DTT was performed at postoperative 3 month. Facial nerve function was clinically evaluated according to the House-Brackmann (HB) facial nerve grading system. Facial nerve courses on preoperative tractography were entirely correlated with intraoperative findings in all patients. Facial nerve was located on the anterior of the tumor surface in 5 cases, on anteroinferior in 3 cases, on anterosuperior in 2 cases, and on posteroinferior in 1 case. In postoperative facial nerve tractography, preservation of facial nerve was confirmed in all patients. No patient had severe facial paralysis at postoperative one year. This study shows that DTT for preoperative identification of facial nerve in VS surgery could be a very accurate and useful radiological method and could help to improve facial nerve preservation.


Seven Ventilators Challenged With Leaks During Neonatal Nasal CPAP: An Experimental Pilot Study.
Nasal CPAP is the most common respiratory support for neonates. Several factors are considered important for effective treatment, including leaks at the patient interface and the delivery of pressure-stable CPAP. Investigations of pressure stability during leaks should include both the change in the mean delivered CPAP and the pressure variation during each breath. The aim of this study was to examine the response of ventilators delivering nasal CPAP when challenged with leaks at the patient interface. Seven ventilators providing nasal CPAP at 4 cm H2O were challenged with leaks during simulated neonatal breathing. Leak was applied for 15 consecutive breaths at a constant level (1-4 L/min). The 2 aspects of pressure stability were evaluated by measuring the mean delivered CPAP and the amplitude of pressure swings before, during, and after leaks. The ability to maintain the delivered CPAP and the amplitude of pressure swings varied greatly among the 7 ventilators before, during, and after leaks. Four of the ventilators tested have built-in leak compensation. There was no simple relationship between maintaining delivered CPAP during leaks and providing CPAP with low pressure swing amplitude. Maintaining the delivered CPAP and providing this without pressure swings are 2 separate aspects of pressure stability, and investigations concerning the clinical importance of pressure stability should address both aspects. This study also shows that compensation for leaks does not necessarily provide pressure-stable CPAP.


EEG measures index neural and cognitive recovery from sleep deprivation.
Sleep deprivation impairs many cognitive abilities, but these impairments can be reversed after a certain quantity and quality of sleep. The ability to inhibit responding is particularly susceptible to disruption after prolonged wakefulness. How recovery sleep (RS) alters brain activity, leading to improved performance on a variety of cognitive tasks, remains unclear. This issue was examined in the current study using spectral analysis of electroencephalogram (EEG) data during sleep. These measures of sleep physiology were acquired after both normal sleep (NS) and RS, and were related to measures of inhibitory control and concurrent brain activity. Subjects were nine young adults who underwent functional magnetic resonance imaging twice, after 9 h of NS and after 10 h of RS that followed 38 h of being awake. A multiple regression model was used to examine differences between conditions in (1) EEG spectral power during sleep, (2) probability of successful inhibition in a go/no-go task, and (3) activation within a region of right prefrontal cortex during the task. Performance recovery, as indexed by reduced performance differences between conditions, was predicted by increased delta power and decreased sigma power in RS compared with NS. These EEG variables predicted most of the variance in inhibitory performance difference between conditions. Regressions also suggested that RS improved performance because of changes in brain function including prefrontal regions that resulted from delta rebound. We thus propose that slow waves, reflected in delta power during RS, act to restore brain function, thereby improving cognitive performance that entails response inhibition.