Embryogenesis and Plant Regeneration from Isolated Wheat Zygotes.
Wheat zygotes can be mechanically isolated and cultivated to continue their development in vitro. Since each zygote needs to be individually isolated, only relatively few of these cells are available per experiment. To facilitate embryonic growth despite of this limitation, the zygotes are kept within a culture insert placed in a larger dish which itself contains embryogenic pollen cocultivated for continuous medium conditioning. This setup ensures that the two cultures, while being physically separated from one another, can exchange essential intercellular signal molecules passing through the bottom of the insert which is made of a permeable membrane. Thanks to the natural fate of zygotes, which is to form an embryo followed by the generation of a plant, embryogenesis and plant regeneration are achieved at much higher efficiency as compared to other single-cell systems. While the method is largely independent of the genotype, it allows for the nondestructive observation, manipulation, and individual analysis of zygotes and very young embryos.


Acquisition of extended-spectrum cephalosporin-resistant Gram-negative bacteria: epidemiology and risk factors in a 6-year cohort of 507 severe trauma patients.
Severe trauma patients are at higher risk of infection and often exposed to antibiotics, which could favor acquisition of antimicrobial resistance. In this study, we aimed to assess prevalence, acquisition, and factors associated with acquisition of extended-spectrum cephalosporin-resistant Gram-negative bacteria (ESCR-GNB) in severe trauma patients. We conducted a retrospective monocentric cohort study in a French level one Regional Trauma Centre between 01 January 2010and 31 December 2015. Patients admitted for ≥ 7 days, with an Injury Severity Score ≥ 15, and ≥ 1 microbiological sample were included in the analysis. Prevalence and acquisition rate of ESCR-GNB were determined then, factors associated with ESCR-GNB acquisition were assessed using a Cox model. Of 1873 patients admitted during the study period, 507 were included (median Injury Severity Score = 29 [22-34] and median intensive care unit length of stay = 16 days [10-28]). Most of them (450; 89%) had an antimicrobial therapy. Prevalence of ESCR-GNB increased from 13% to 33% during intensive care unit stay, bringing the ESCR-GNB acquisition rate to 29%. Acquisition of ESCR-GNB was mainly related to AmpC beta-lactamase Enterobacterales and was independently associated with mechanical ventilation needs (hazard ratio [HR] = 6.39; 95% confidence interval [CI] [1.51-27.17]; P = 0.01), renal replacement therapy needs (HR = 2.44; 95% CI [1.24-4.79]; P = 0.01), exposure to cephalosporins (HR = 1.06; 95% CI [1.01-1.12]; P = 0.02), and/or combination therapy with non-beta-lactam antibiotics such as vancomycin, linezolid, clindamycin, or metronidazole (HR = 1.03; 95% CI [1.01-1.06]; P = 0.02). Acquisition of ESCR-GNB was prevalent in severe trauma patients. Our results suggest selecting antibiotics with caution, particularly in the most severely ill.


Zinc binding and its trapping by allosteric transition in glucosamine-6-phosphate deaminase from Escherichia coli.
Glucosamine-6-phosphate isomerase deaminase from Escherichia coli, a typical allosteric enzyme, becomes less cooperative and 50% inhibited when treated with zinc. This metal cation behaving as a tight-bound and slow partial inhibitor. Modification of a pair of vicinal reactive thiols with some sulfhydryl reagents mimics this effect. On the other hand, sulfhydryl reactivity disappears in the presence of saturating concentrations of Zn2+, which does not modify the kinetics of S-methylated enzyme, a finding that indicates that vicinal thiols are an essential part of the zinc-binding site. Allosteric activation of the deaminase causes trapping of the metal, which cannot be released by dialysis against a buffer containing EDTA. Cadmium and nickel(II) cations also produce a similar effect.


NR2A and NR2B subunits differentially mediate MAP kinase signaling and mitochondrial morphology following excitotoxic insult.
NMDA receptors are essential for neurotransmission and key mediators of synaptic signaling, but they can also trigger deleterious degenerative processes that lead to cell death. Growing evidence suggests that selective blockade of the heterogeneous subunits that comprise the NMDA receptor may enable better control of pharmacotherapies for treating neurological diseases and injuries. We investigated the relationship between NMDAR activation, MAPK signaling, and mitochondrial shape following an excitotoxic insult. NR2A- and NR2B-containing NMDARs differentially mediated acute changes in cytosolic calcium, alterations in mitochondrial morphology, and phosphorylation of the MAPKs ERK and JNK. Activation of NR2A-containing NMDARs was associated with JNK phosphorylation that was neuroprotective in neuronal cultures subjected to excitotoxicity. In contrast, activation of NR2B-containing NMDARs triggered calcium accumulation in mitochondria that was strongly associated with mitochondrial swelling and neuronal cell death. Indeed, while blockade of NR2B-containing receptors was neuroprotective, this protection was lost when NR2A-initiated JNK phosphorylation was inhibited. Given the modest selectivity of the NR2A inhibitor, NVP-AAM077, the results highlight the significance of the relative, rather than absolute, activation of these two NMDA subtypes in modulating cell death pathways. Therefore, the balance between concurrent activation of NR2B-containing and NR2A-containing NMDARs dictates neuronal fate following excitotoxicity.


Improved Outcomes With an Enhanced Recovery Approach to Cesarean Delivery.
To examine the results of a quality-improvement study that implemented an enhanced recovery after surgery (ERAS) program for cesarean delivery. A pre-post design was used to assess changes in opioid use, length of stay, and costs among all patients undergoing cesarean delivery before and after implementation of an evidence-based ERAS pathway for the preoperative, intraoperative, and postoperative management of patients beginning December 2018. A total of 3,679 cesarean deliveries (scheduled and emergent) were included from January 1, 2018, through August 31, 2019, of which 2,171 occurred before implementation on December 17, 2018, and 1,508 occurred postimplementation. Eighty-four percent of patients received opioids as inpatients after cesarean delivery during the preimplementation period, as compared with 24% in the postimplementation period (odds ratio [OR] 16.8, 95% CI 14.3-19.9). Among patients who required any opioids, the total morphine milligram equivalents also significantly decreased (median 56.5 vs 15.0, mean relative change 0.32, 95% CI 0.28-0.35). Compared with the preimplementation period, those in the postimplementation period had a shorter postcesarean length of stay (3.2 vs 2.7 days, mean relative change 0.82, 95% CI 0.80-0.83, median 3 days in both periods), lower median direct costs by $349 (mean relative change 0.93, 95% CI 0.91-0.95), and no change in the 30-day readmission rate (1.4% vs 1.7%, OR 0.83, 95% CI 0.49-1.41). An ERAS approach for the cesarean delivery population is associated with improved outcomes including decreases in opioid use, length of stay, and costs.


Ethnicity, gender, and utilization of mental health services in a Medicaid population.
This study examines gender and ethnic differences in mental health utilization and expenditures in a fee-for-service Medicaid eligible population in Monroe County, New York. The analyses demonstrate that Medicaid poor are not a homogeneous population in terms of mental health utilization, and that their patterns of care use are quite unlike those generally attributed to the middle class patients. For example, Medicaid eligible males are more likely to be mental health users than females, although they are less likely to use medical services. Ethnicity appears to be a strong determinant of ambulatory mental health utilization, but not of inpatient care. Whites experience significantly higher levels of psychiatric ambulatory use than do the nonwhites. Nonwhites, on the other hand experience greater utilization of alcohol ambulatory services than do the whites. The data indicate that although mental health care cost represents at least 20% of total expenditures in this population, this cost is predictable and stable from year to year. Other analyses dealing with the cost of medical care for mental health users, and with the impact of a 'gate keeper' on mental health utilization patterns, are presented. Both clinical and public policy implications are discussed.


Preliminary molecular characterization of a human immunodeficiency virus (HIV-I) associated with neuropathology.
A human immunodeficiency virus (HIV-I) was isolated from the brain of a patient with progressive dementia but no obvious immunosuppression. This isolate, designated as HIV-IBR, was molecularly cloned and sequenced, and its long terminal repeat (LTR) and envelope sequences were compared with those of other HIV isolates not uniquely associated with dementia. The HIV-IBR LTR showed marked homology with the LTR sequences of the other HIV-I isolates. The predicted amino acid sequence of the external glycoprotein (gp120) of HIV-IBR revealed a pattern of conserved and variable regions similar to that of other HIV isolates. The sequence of the transmembrane portion of envelope, gp 41, was highly homologous to the counterpart region of other isolates. Further analysis is required to determine whether specific sequence variation can account for neurological manifestations of HIV-IBR infection.


Active moss biomonitoring applied to an industrial site in Romania: relative accumulation of 36 elements in moss-bags.
Active moss biomonitoring using the species Sphagnum girgensohnii was tested at a strongly polluted site in Romania (Baia Mare) according to a novel sampling design. Nine moss transplants from each of the two background areas (Dubna, Russia and Vitosha Mountain, Bulgaria) were deployed in parallel on balconies about 24 m above street level for 4 months. The samples were analyzed for 36 elements using instrumental neutron activation analysis (INAA). Based on the results obtained the sampling variability is discussed in relation to the analytical variability, and the relative uptake of the different elements is assessed. The moss-bags using Sphagnum girgensohnii demonstrate a high or a very high relative uptake for a majority of the 36 investigated elements, but the values depend on the initial element concentration in the moss. Moss leaves analyzed separately showed somewhat higher levels than stems for many elements. Practical considerations however still speak in favor of using the whole moss for transplants.


[Effects and mechanism of emodin and sandostatin on pancreatic ischemia in acute haemorrhagic necrotizing pancreatitis].
To investigate pancreatic ischemia and abnormal metabolism of eicosanoids in acute haemorrhagic-necrotizing pancreatits (AHNP) and the effects of emodin or sandostatin on them. Rats with AHNP were triggered with sodium taurocholate; the pancreatic blood flow (PBF) was detected with computerized tissue blood flowmeter, and plasma prostaglandin E2 (PGE2), 6-keto-prostaglandin F1 alpha (6-keto-PGF1 alpha) and thromboxane (TXB2) were determined with radioimmunoassay. There was a significant decrease of PBF in the early stage of AHNP. Compared with that in the untreated group, significant improvement of PBF was demonstrated in emodin as well as in sandostatin group which showed reduced PBF following infusion of sandostatin before triggering AHNP. In untreated group plasma TXB was significantly higher, with an increase of 4.5 times, than that in sham-operated group while 6-keto-PGF1 alpha or PGE2 tended to decrease. The above mentioned abnormal synthesis of eicosanoids was blocked either in emodin or in sandostatin group in which lessened damage of acini cells was shown by pathologic scoring or transmission electron microscope. Both of the two groups shared significantly lower mortalities than the untreated group. Either emodion or sandostatin could partly reverse the decrease of PBF in the early stage of AHNP, which may be ascribed at least in part to inhibition of abnormal synthesis of eicosanoids and improvement of cytoprotection of acini cells, and combined application of the two drugs might promise positively synergetic action as well.


Disruption of basement membrane, extracellular matrix metalloproteinases and E-cadherin in renal-cell carcinoma.
A retrospective study was performed to determine the prognostic value of Basement Membrane (BM) integrity, Matrix Metalloproteinases (MMPs) and E-Cadherin expression in renal cell carcinoma (RCC). An immunohistochemical study on laminin and collagen IV, MMPs 1 and 2, and E-Cadherin was carried out on 71 RCCs. BM fragmentation was considered taking 75% as a cut-off. MMP 1 and MMP2 immunostaining, as well as E-Cadherin was considered taking 25% as a cut-off. An inverse relationship was seen between E-Cadherin with laminin, collagen IV and MMPs. More than 75% loss of laminin, collagen IV and E-Cadherin, as well as higher expression of MMPs, were associated with symptoms, tumoral size and worse grade. Loss of collagen IV and E-Cadherin were of prognostic value. Both BM and E-Cadherin are good prognostic markers. MMPs patterns show a relationship between BM proteins and E-Cadherin, but evaluation is more time-consuming and provide no better prognostication; consequently they are not useful in routine clinical applications.


The kidney sodium-phosphate co-transporter alters bone quality in an age and gender specific manner.
Mutations in the kidney NaPiIIa co-transporter are clinically associated with hypophosphatemia, hyperphosphaturia (phosphate wasting), hypercalcemia, nephrolithiasis and bone demineralization. The mouse lacking this co-transporter system was reported to recover its skeletal defects with age, but the "quality" of the bones was not considered. To assess changes in bone quality we examined both male and female NaPiIIa knockout (KO) mice at 1 and 7months of age using micro-computed tomography (micro-CT) and Fourier transform infrared imaging (FTIRI). KO cancellous bones at both ages had greater bone volume fraction, trabecular thickness and lesser structure model index based on micro-CT values relative to age- and sex-matched wildtype animals. There was a sexual-dimorphism in the micro-CT parameters, with differences at 7months seen principally in males. Cortical bone at 1month showed an increase in bone volume fraction, but this was not seen at 7months. Cortical thickness which was elevated in the male and female KO at 1month was lower in the male KO at 7months. FTIRI showed a reduced mineral and acid phosphate content in the male and female KO's bones at 1month with no change in acid phosphate content at 7months. Collagen maturity was reduced in KO cancellous bone at 1month. The observed sexual dimorphism in the micro-CT data may be related to altered phosphate homeostasis, differences in animal growth rates and other factors. These data indicate that the bone quality of the KO mice at both ages differs from the normal and suggests that these bone quality differences may contribute to skeletal phenotype in humans with mutations in this co-transporter.


Hyperhomocysteinemia Is Associated with Vitamin B-12 Deficiency: A Cross-sectional Study in a Rural, Elderly Population of Shanxi China.
To report the association of lifestyle factors and plasma vitamin B-12 with hyperhomocysteinemia in a large sample of men and women living in a region of China where there is an increased risk of NTDs. Community-based, cross-sectional study of Lvliang City, Shanxi Province, China. Hyperhomocysteinemia is an independent risk factor for cardiovascular disease (CVD) and a sensitive marker of vitamin B-12 and folate deficiency. A total of 2355 (1044 men and 1311 women) participants born before 1 January 1958 (≥55 years of age) and living in Lvliang City for at least 2 months a year were included. The participants were assessed regarding demographic characteristics, height, weight, as well as having a physical examination and blood sampling for serum cholesterol, total homocysteine (tHcy), folate, and vitamin B12 levels. The median (25th-75th percentile) tHcy concentration was 21.5 (15.8-33.6) µmol/L in men and 18.0 (13.4-24.8) µmol/L in women. The overall prevalence of hyperhomocysteinemia (tHcy ≥15 µmol/L) was 72.6% (84.3% in men and 63.2% in women), inversely correlated with folate (r=-0.230, P=0.006) and vitamin B-12 (r=-0.540, P<0.001), and positively correlated with uric acid (r=0.054, P<0.001). Vitamin B-12 and folate deficiency, older age, and male gender were associated with elevated tHcy; with vitamin B-12 deficiency being the strongest. Plasma tHcy concentration and hyperhomocysteinemia were significantly higher in this population than in previously studied populations. Vitamin B-12 and folate supplementation, concomitant lifestyle changes such as smoking cessation, and lipid-lowering treatments may help to decrease plasma tHcy concentrations and reduce the CVD risk in this population.


The role of physical activity in cancer survivors' quality of life.
As a result of a cancer diagnosis and treatment, many cancer survivors experience persistent physical, mental, and emotional symptoms that affect their quality of life. Physical activity has been identified as an intervention that may help to manage the side effects of a cancer diagnosis and its treatment. The purpose of this study was to investigate the role of physical activity on overall quality of life in adult cancer survivors. One-on-one semi-structured interviews were conducted in person or via telephone with 13 adult (≥18 yrs) cancer survivors who had completed cancer treatment. These cancer survivors described their physical activity as improving their physical functioning and mental health, as a means of positive social engagement, and adding positivity to their daily life. Even though the cancer survivors in this study experienced diverse types of cancer and for different lengths of time as well as varying methods of treatment(s), these results support the role of physical activity in enhancing cancer survivors' quality of life. Further research is warranted to (a) expand this research with a larger sample and quantitative methods, (b) examine healthcare providers' knowledge and application of exercise guidelines to cancer survivors in cancer care, and (c) explore implementation strategies for greater advocacy for healthcare providers to share the exercise recommendations with cancer survivors.


Sweep frequency impedance measures in Australian Aboriginal and Caucasian neonates.
Despite high prevalence of otitis media in Aboriginal children, the acoustic-mechanical properties of their outer and middle ear during the neonatal period remain obscured. The objective of this study was to compare the acoustic-mechanical properties of outer and middle ear using Sweep Frequency Impedance (SFI) measures between Australian Aboriginal and Caucasian neonates. SFI data from 40 ears of 24 Aboriginal neonates (16 males, 8 females) with mean gestational age of 39.57 weeks (SD = 1.25) and 160 ears of 119 Caucasian neonates (57 males, 62 females) with mean gestational age of 39.28 weeks (SD = 1.25) serving as controls were analysed. SFI data in terms of resonance frequency (RF) and mobility of the outer and middle ear (ΔSPL) were collected from neonates who passed a test battery that included automated auditory brainstem response, distortion product otoacoustic emissions test and 1000-Hz tympanometry. SFI data were analysed using descriptive statistics and analysis of variance. There was no significant difference in mean gestational age, age of testing and birth weight between the Aboriginal and Caucasian neonates. The mean resonance frequencies for the outer ear (mean RF1 = 264.9 Hz, SD = 58.6 Hz) and middle ear (mean RF2 = 1144 Hz, SD = 228.8 Hz) for Aboriginal neonates were significantly lower than that of Caucasian neonates (mean RF1 = 295.3 Hz, SD = 78.4 Hz and mean RF2 = 1241.8 Hz, SD = 216.6 Hz). However, no significant difference in the mobility of outer ear (ΔSPL1) and middle ear (ΔSPL2) between the two groups was found. Middle ear resonance was absent in 22.5% (9 ears) of Aboriginal ears but present in all Caucasian ears. This study provided evidence that despite passing the test battery, Aboriginal neonates had significantly lower resonance frequencies of the outer and middle ear than Caucasian neonates. Furthermore, 22.5% of Aboriginal neonates showed no middle ear resonance, indicating the possibility of subtle middle ear issues not detected by the test battery. Reasons for the different acoustic-mechanical properties between the two ethnic groups remain unclear and require further investigation.


Association of Molecular Genetic Markers of TP53, MDM2, and CDKN1A Genes with Progression-Free Survival of Patients with Ovarian Cancer after Platinum-Based Chemotherapy.
We studied the association of polymorphic markers of cell cycle control genes (Arg72Pro of the TP53 gene, T(-410)G of the MDM2 gene, and Ser31Arg of the CDKN1A gene) in ovarian cancer and progression-free survival following platinum-based chemotherapy. Tumor tissue samples obtained from 49 patients who had undergone chemotherapy were examined. Patients received standard platinum-based chemotherapy and were observed until disease progression. Polymorphic markers of genes were evaluated by PCR-RFLP and real-time PCR. In patients carrying the G allele of the T(-410)G marker of the MDM2 gene, a decreasing trend was observed in median progression-free survival. An increase in the median progression-free survival was observed in carriers of the Pro allele of the TP53 gene (p=0.045). Furthermore, a stronger association was noted with carriers of the minor Pro/Pro homozygous genotype relative to the Arg/Arg genotype (p=0.007). In the subgroup of patients who underwent optimal or complete cytoreductive surgery, carriage of the minor Arg allele of the Ser31Arg marker (CDN1A gene) was associated with a decrease in the median progression-free survival time (p=0.004).


A screening assay for monoclonal antibodies based on the availability of a polyclonal antiserum.
A double-sandwich enzyme-linked immunospecific assay (ELISA) has been developed for screening hybridoma clones for monoclonal antibodies against human alpha-fetoprotein (AFP). The assay uses commercial polyclonal antiserum to bind alpha-fetoprotein from biological fluids, and the complex then binds the specific monoclonal antibodies in hybridoma culture supernatants. The sandwich is completed with commercial peroxidase-linked anti-mouse immunoglobulin. In exploring the variables of this system, it was found that the quality of the polyclonal anti-alpha-fetoprotein antiserum was the most critical factor. The assay functioned with as little as 10 ng alpha-fetoprotein from cord serum or amniotic fluid. A variety of commercial peroxidase-linked second antibodies was satisfactory, and both IgG and IgA class monoclonals were detected. Comparison of the ELISA with a standard radioimmunoassay showed the former to perform very adequately as a general hybridoma screening assay.


Endogenous recovery after brain damage: molecular mechanisms that balance neuronal life/death fate.
Neuronal survival depends on multiple factors that comprise a well-fueled energy metabolism, trophic input, clearance of toxic substances, appropriate redox environment, integrity of blood-brain barrier, suppression of programmed cell death pathways and cell cycle arrest. Disturbances of brain homeostasis lead to acute or chronic alterations that might ultimately cause neuronal death with consequent impairment of neurological function. Although we understand most of these processes well when they occur independently from one another, we still lack a clear grasp of the concerted cellular and molecular mechanisms activated upon neuronal damage that intervene in protecting damaged neurons from death. In this review, we summarize a handful of endogenously activated mechanisms that balance molecular cues so as to determine whether neurons recover from injury or die. We center our discussion on mechanisms that have been identified to participate in stroke, although we consider different scenarios of chronic neurodegeneration as well. We discuss two central processes that are involved in endogenous repair and that, when not regulated, could lead to tissue damage, namely, trophic support and neuroinflammation. We emphasize the need to construct integrated models of neuronal degeneration and survival that, in the end, converge in neuronal fate after injury. Under neurodegenerative conditions, endogenously activated mechanisms balance out molecular cues that determine whether neurons contend toxicity or die. Many processes involved in endogenous repair may as well lead to tissue damage depending on the strength of stimuli. Signaling mediated by trophic factors and neuroinflammation are examples of these processes as they regulate different mechanisms that mediate neuronal demise including necrosis, apoptosis, necroptosis, pyroptosis and autophagy. In this review, we discuss recent findings on balanced regulation and their involvement in neuronal death.


The kinetics of assembly of normal and variant human oxyhemoglobins.
The kinetics of assembly have been monitored spectrophotometrically for normal and variant human oxyhemoglobins in 0.1 M Tris, 0.1 M NaCl, 1 mM Na2EDTA, pH 7.4, at 21.5 degrees C. Oxyhemoglobin versus oxy chain static difference spectra were performed and revealed subtle but significant absorption changes in both the visible and Soret regions. Kinetic experiments were performed by rapidly mixing equivalent (in heme) concentrations of alpha and beta A chains and following the change in absorbance at 583 nm with time. Over a protein concentration range of 10-100 microM in heme prior to mixing, these time courses were homogeneous and followed first-order kinetics, yielding a value of 0.069 s-1 for the apparent rate constant of dissociation of oxygenated beta A chain tetramers. Under these conditions, the overall assembly of oxyhemoglobins S (beta 6Glu----Val) and N-Baltimore (beta 95Lys----Glu) were also governed by the rates of dissociation of their respective oxygenated beta S and beta N-Baltimore chain tetramers with the apparent first-order rate constants of 0.044 and 0.15 s-1, respectively. In the Soret region, the alpha, beta monomer combination reaction could be observed if the protein concentration (heme basis) was lowered and if protein nonequivalency (beta chain exceeded alpha chain concentration) mixing experiments were performed. A kinetic oxyhemoglobin A, oxy-alpha, oxy-beta A monomer difference spectrum could be generated, and simple second-order kinetics were observed (415 nm) yielding rate constants of 2.3, 3.3, and 4.8 X 10(5) M-1 s-1 for the assembly of oxyhemoglobins S, A, and N-Baltimore, respectively. To our knowledge, this is the first kinetic study to reveal significant differences between the rate of association of alpha and beta monomers of hemoglobin A and those of two distinctly charged hemoglobin variants.


DNA integrity is maintained after freeze-drying of human spermatozoa.
To evaluate the effects on human spermatozoa of freeze-drying, also known as lyophilization, and of cryopreservation in liquid nitrogen. Prospective experimental study. Reproductive medicine unit and a private IVF center. Thirty healthy male donors. Sperm samples from 30 donors divided as two aliquots, one to be lyophilized and the other to be cryopreserved in liquid nitrogen. Assessment of count, motility, morphology, viability, DNA integrity, chromosomal status, and birefringence properties of lyophilized and cryopreserved human spermatozoa compared with the same parameters in the fresh sample. Although sperm viability and motility were totally compromised after freeze-drying, the sperm chromatin structure was not altered in comparison with fresh samples, which demonstrated that the procedure did not affect DNA integrity. The sperm-head inner protoplasmic structures were also preserved, which was estimated by assessing the corresponding birefringence characteristics. After cryopreservation with liquid nitrogen, the motility, viability, and DNA integrity of spermatozoa were statistically significantly reduced compared with the fresh samples; the proportion of sperm cells with abnormal head birefringence increased meaningfully. The process of freeze-drying deeply damages cell membranes; however, unlike with liquid nitrogen preservation, it does not affect DNA integrity.


Local myotoxicity of bupivacaine in rabbits after continuous supraclavicular brachial plexus block.
Local anesthetics including bupivacaine have been known to be myotoxic. Continuous brachial plexus blocks with bupivacaine provide good postoperative pain relief, but their effects on the surrounding muscles have not been studied. Eight rabbits were given continuous bilateral brachial plexus blocks with 0.25% bupivacaine (BUPI) and another eight (SALINE) equivalent volumes of saline infused into the plexus region under pentobarbital-ketamine sedation. At the end of the 24-hour blocks, muscle biopsies were taken from one side of the neck in both groups. A week later, the biopsy procedure was repeated on the other side. Plasma concentrations of bupivacaine were measured before and at the end of the 24-hour blocks. Inflammatory cell infiltration and muscle fiber necrosis were significantly more common in the BUPI than in the SALINE group (p < 0.001). At the end of the continuous block, predominantly neutrophils were found, but in the BUPI group large amounts of eosinophilic granulocytes also were found. A week later lymphocytes, plasma cells, macrophages, and fibroblasts were seen as a sign of beginning scar formation. Plasma concentrations of bupivacaine at 24 hours were at a nontoxic level (0.28-0.71 microgram/ml). Brachial plexus blocks with bupivacaine caused reversible myotoxicity and inflammation in the rabbit. A week after the continuous blocks, muscle fiber regeneration had already started.


Reaction and strain engineering for improved stereo-selective whole-cell reduction of a bicyclic diketone.
Reduction of bicyclo[2.2.2]octane-2,6-dione to (1R, 4S, 6S)-6-hydroxy-bicyclo[2.2.2]octane-2-one by whole cells of Saccharomyces cerevisiae was improved using an engineered recombinant strain and process design. The substrate inhibition followed a Han-Levenspiel model showing an effective concentration window between 12 and 22 g/l, in which the activity was kept above 95%. Yeast growth stage, substrate concentration and a stable pH were shown to be important parameters for effective conversion. The over-expression of the reductase gene YDR368w significantly improved diastereoselectivity compared to previously reported results. Using strain TMB4110 expressing YDR368w in batch reduction with pH control, complete conversion of 40 g/l (290 mM) substrate was achieved with 97% diastereomeric excess (de) and >99 enantiomeric excess (ee), allowing isolation of the optically pure ketoalcohol in 84% yield.


A selective fluorescence probe based on naphthalene for the detection of barium(II).
A designed naphthalene derivative (potassium 2,2'-(naphthalen-1-ylazanediyl)diacetate, ND) was synthesized successfully which characterized by spectroscopic techniques (NMR, IR and HRMS-ESI-TOF). The sensitivity and selectivity were high toward Ba2+ in aqueous solution (100.0 mM HEPES buffer, 100.0 mM KNO3, pH 7.4) over a wide variation of metal ions such as Li+, Na+, K+, Cs+, Mg2+, Ca2+, Al3+, Pb2+, Fe2+, Fe3+, Co2+, Ni2+, Cu+, Cu2+, Ag+, Zn2+, Cd2+ and Ce3+. The limit of detection (LOD) was found to be 9.56×10-8 M. Density functional theory (DFT) calculations at the B3LYP level employing the 6-31G(d,p) and LANL2DZ basis set were performed for optimization of ND and ND-Ba2+ complex. The calculated results were consistent with a favorable complexation corresponding to the proposed coordination.


Endovascular treatment of traumatic internal carotid artery pseudoaneurysm.
Traumatic internal carotid artery pseudoaneurysm (TICAP) is the most common cause of stroke in young adults. The treatment of TICAP with open surgery poses excess risk, thus during last decade endovascular treatment strategies have been applied. To assess the efficacy and the existing experience of endovascular treatment of TICAP. A systematic review of the literature was undertaken to identify all reported cases of endovascular treatment of TICAP from 1998 to 2015 in MEDLINE, EMBASE and Cochrane Central Register of Controlled Trials. A total of 193 patients (139 males, 75%) with mean age of 30.8±2.2 years in 23 case studies, were treated for their TICAP with endovascular treatment. The main causes of TICAP were road traffic accidents 51%, assaults 12%, fall from height 8% and other miscellaneous causes were 29%. In 8/23 studies, the patients were operated emergently, in 9/23 at least 1 month after the carotid injury, and in 6/23 the time between the injury and the operation was not reported. The total success rate of pseudoaneurysm occlusion was 84% (162/193). The reported peri-procedural morbidity rate was 6% (11/185; 3 TIA, 7 strokes and 1 subclavian artery dissection), and the peri-operative mortality rate was 1.2% (2/162). Most patients received post-operatively antiplatelet therapy (either single or dual) and the duration of the administration ranged from 3 months to long term. During their follow up (ranging from 4 days to 13 years) only 6 patients required re-intervention, and this was undertaken with endovascular approach. Endovascular therapy tends to be an effective option for the treatment of TICAP with low morbidity and mortality rates.


[The assessment of newly qualified nurses in competitive exams for hospital employment: a descriptive study].
In the public sector the selection of the health personnel is based on qualifications and exams with a written, an oral and practical test, on contents selected by the committee. The level of complexity and contents of the tests is very variable across committees. To analyze the tests for the selection of nurses, the level of complexity and contents assessed. A sample of tests was analysed and for each item, the cognitive level tested, according to Bloom's taxonomy, areas and health problems investigated. One-hundred-twenty-eight selection tests of 14 hospitals and districts were analysed. Open questions are used in the majority of tests to assess mainly knowledge and not complex cognitive functions or problem solving skills. The contents vary widely and are not focused on priority health problems. Questions on prevention, health education and symptoms monitoring are missing. Written, oral and practical tests focus on the same contents and test mainly knowledge.


Abi1 regulates the activity of N-WASP and WAVE in distinct actin-based processes.
Neural Wiskott-Aldrich syndrome protein (N-WASP) and WAVE are members of a family of proteins that use the Arp2/3 complex to stimulate actin assembly in actin-based motile processes. By entering into distinct macromolecular complexes, they act as convergent nodes of different signalling pathways. The role of WAVE in generating lamellipodial protrusion during cell migration is well established. Conversely, the precise cellular functions of N-WASP have remained elusive. Here, we report that Abi1, an essential component of the WAVE protein complex, also has a critical role in regulating N-WASP-dependent function. Consistently, Abi1 binds to N-WASP with nanomolar affinity and, cooperating with Cdc42, potently induces N-WASP activity in vitro. Molecular genetic approaches demonstrate that Abi1 and WAVE, but not N-WASP, are essential for Rac-dependent membrane protrusion and macropinocytosis. Conversely, Abi1 and N-WASP, but not WAVE, regulate actin-based vesicular transport, epidermal growth factor receptor (EGFR) endocytosis, and EGFR and transferrin receptor (TfR) cell-surface distribution. Thus, Abi1 is a dual regulator of WAVE and N-WASP activities in specific processes that are dependent on actin dynamics.


Recent trends in traffic injury.
Data in the Registrar-General's statistical reviews for 1958-75 and in the reports of the Hospital In-Patient Enquiry for 1964-73 have been examined to ascertain the light they shed on the changes in the patterns of injury from road accidents over this period. Causes of death, primary injury for hospital inpatients and average length of hospital stay have been examined. It seems that an increasing proportion of deaths are being ascribed to internal injuries and a decreasing proportion to fracture of the skull.


Sense of community, unfairness, and psychosomatic symptoms: a multilevel analysis of Italian schools.
This study examines the role of school sense of community and school unfairness in predicting psychosomatic symptoms among Italian early adolescents. The study used hierarchical linear modeling to examine the relations between individual and school characteristics, and students' psychosomatic symptoms. Data were analyzed using a two-level model based on 22,552 students (49.8% boys; M age = 15.70, SD = .69) nested within 1,111 schools. A questionnaire devised by the Health Behavior in School-aged Children (HBSC) international group was used. Sense of community and procedural unfairness in the school at the individual level showed a significant association with students' psychosomatic symptoms. School characteristics predicted between-school variation in psychosomatic symptoms, in that procedural unfairness at the school level increased the risk for psychosomatic symptoms and a sense of community in the school protected students. The present study suggests that the quality-of-school context may have a key role in predicting students' psychosomatic symptoms.


Integrative proteomics and n-glycoproteomics reveal the synergistic anti-tumor effects of aspirin- and gemcitabine-based chemotherapy on pancreatic cancer cells.
Pancreatic cancer is a highly malignant tumor that is well known for its poor prognosis. Based on glycosylation, we performed integrated quantitative N-glycoproteomics to investigate the synergistic anti-tumor effects of aspirin and gemcitabine on pancreatic cancer cells and explore the potential molecular mechanisms of chemotherapy in pancreatic cancer. Two pancreatic cancer cell lines (PANC-1 and BxPC-3) were treated with gemcitabine, aspirin, and a combination (gemcitabine + aspirin). We found that the addition of aspirin enhanced the inhibitory effect of gemcitabine on the activity of PANC-1 and BxPC-3 cells. Quantitative N-glycoproteome, proteome, phosphorylation, and transcriptome data were obtained from integrated multi-omics analysis to evaluate the anti-tumor effects of aspirin and gemcitabine on pancreatic cancer cells. Mfuzz analysis of intact N-glycopeptide profiles revealed two consistent trends associated with the addition of aspirin, which showed a strong relationship between N-glycosylation and the synergistic effect of aspirin. Further analysis demonstrated that the dynamic regulation of sialylation and high-mannose glycoforms on ECM-related proteins (LAMP1, LAMP2, ITGA3, etc.) was a significant factor for the ability of aspirin to promote the anti-tumor activity of gemcitabine and the drug resistance of pancreatic cancer cells. In-depth analysis of N-glycosylation-related processes and pathways in pancreatic cancer cells can provide new insight for future studies regarding pancreatic cancer therapeutic targets and drug resistance mechanisms.


A novel injectable local hydrophobic drug delivery system: Biodegradable nanoparticles in thermo-sensitive hydrogel.
In this article, a novel local hydrophobic drug delivery system: nanoparticles in thermo-sensitive hydrogel, was demonstrated. First, honokiol, as a model hydrophobic drug, loaded poly(epsilon-caprolactone)-poly(ethylene glycol)-poly(epsilon-caprolactone) (PCEC) nanoparticles were prepared by emulsion solvent evaporation method, and then were incorporated into thermo-sensitive F127 hydrous matrix. The obtained injectable hydrophobic drug delivery system can act as a depot for sustained release of honokiol in situ. The lower critical solution temperature (LCST) of the composite matrix increases with increase in the mass of incorporated nanoparticles, or with decrease in the amount of residual organic solvent in the system. Honokiol release profile in vitro was studied, and the results showed that honokiol could be sustained released from the system. The described injectable drug delivery system might have great potential application for local delivery of hydrophobic drugs such as honokiol.


Interstrand DNA crosslinks due to AP (apurinic/apyrimidinic) sites.
Storage of a solution of DNA containing apurinic sites, even at 4 degrees C leads to the appearance of interstrand crosslinks. Possible biological consequences of these crosslinks, when they appear in cell DNA, are briefly discussed. Formation of interstrand crosslinks in DNA containing tritium-labelled thymine and kept in an aqueous solution might be due, at least partly, to the loss of bases by the autoirradiated DNA.


Impact of temperature on immediate and secondary compression of MSW with high and low food contents.
Landfills in developing countries usually show municipal solid wastes (MSW) with large amount of food wastes, extraordinarily high moisture contents, and high internal temperatures. Because of these specific characteristics, significant post-closure settlements in landfills with high food waste are expected over time. This paper focused on the assessment of temperature impact on immediate and secondary compression behaviors of MSW with large contents of food, water, and plastic, comparatively to an aged low food content waste. A compression test was developed having a temperature-control system. The immediate compression index (C'c) of HFWC samples was found to be higher than those of low food waste content (LFWC) owing to the soft behavior of food content, although immediate compression was 15-30% of total strains for HFWC wastes, while for LFWC samples it was 80% of total strains. Mechanical creep was also greater in HFWC owing to the soft behavior of the wet food components intensified by temperature increases. Mechanical creep of LFWC samples was attributed to the deformation of large parcel of soft plastics, also accelerated by temperature. The HFWC waste showed a first dominant phase of biocompression with an intense and rapid biological degradation, and a second phase characterized by reduced biological activity. Temperatures higher and high compression stresses are required to provide significant impact on biocompression index magnitudes. Overall, the compressibility of high food content wastes has shown to be significantly higher and the temperature impact led to twice total settlements of the MSW with low organic content.


Cortical beta-band power modulates with uncertainty in effector selection during motor planning.
Although beta-band activity during motor planning is known to be modulated by uncertainty about where to act, less is known about its modulations to uncertainty about how to act. To investigate this issue, we recorded oscillatory brain activity with EEG while human participants (n = 17) performed a hand choice reaching task. The reaching hand was either predetermined or of participants' choice, and the target was close to one of the two hands or at about equal distance from both. To measure neural activity in a motion artifact-free time window, the location of the upcoming target was cued 1,000-1,500 ms before the presentation of the target, whereby the cue was valid in 50% of trials. As evidence for motor planning during the cuing phase, behavioral observations showed that the cue affected later hand choice. Furthermore, reaction times were longer in the choice trials than in the predetermined trials, supporting the notion of a competitive process for hand selection. Modulations of beta-band power over central cortical regions, but not alpha-band or theta-band power, were in line with these observations. During the cuing period, reaches in predetermined trials were preceded by larger decreases in beta-band power than reaches in choice trials. Cue direction did not affect reaction times or beta-band power, which may be due to the cue being invalid in 50% of trials, retaining effector uncertainty during motor planning. Our findings suggest that effector uncertainty modulates beta-band power during motor planning.NEW & NOTEWORTHY Although reach-related beta-band power in central cortical areas is known to modulate with the number of potential targets, here we show, using a cuing paradigm, that the power in this frequency band, but not in the alpha or theta band, is also modulated by the uncertainty of which hand to use. This finding supports the notion that multiple possible effector-specific actions can be specified in parallel up to the level of motor preparation.


Association of Backpack Weight with Musculoskeletal Status among Adolescents.
Adolescence period is a critical stage of musculoskeletal development and carrying heavy backpack to school daily puts them at risk of musculoskeletal discomfort. This study aims to find out the association of backpack weight with musculoskeletal status among adolescents. A descriptive cross-sectional study was conducted among 291 grade 4 to 8 students from three government schools of Lalitpur. Data was analyzed using descriptive and inferential statistics. Around 68% of the adolescents carried schoolbag of weight greater than 10% of their body weight. Similarly, 66.7% perceived musculoskeletal pain of which 29.9% had ever missed school due to pain, 70.1% felt tired while carrying their schoolbag and 23.4% adopted forward/sideways leaning posture while carrying their schoolbag. Age (p< 0.001), backpack percentage to body weight (p< 0.001) and perceived weight of schoolbag (p= 0.006) were significantly associated with musculoskeletal pain. Similarly, age (p= 0.023), sex (p= 0.005), grade (p= 0.030) and perceived weight of schoolbag (p= 0.007) were significantly associated with tiredness while carrying schoolbag. Also, backpack percentage to body weight (p= 0.008), duration of carrying schoolbag (p= 0.010) and perceived weight of schoolbag (p= 0.001) were significantly associated with posture assumed while carrying schoolbag. Students carrying backpack weight more than recommended limit are more likely to develop musculoskeletal pain and change in posture while carrying their schoolbag. Thus, necessary steps must be taken to further analyze the situation and develop management strategies on reducing backpack weight and its possible effect on adolescents.


Severity of post-ERCP pancreatitis directly proportional to the invasiveness of endoscopic intervention: a pilot study in a canine model.
Pancreatitis complicates 1% - 22% of endoscopic retrograde cholangiopancreatography procedures. The study aims were to develop a reproducible animal model of post-ERCP pancreatitis (PEP), and investigate the impact of endoscopic technique on severity of PEP. ERCP was carried out in six male hound dogs. Pancreatitis was induced by one of three escalating methods: 1) pancreatic acinarization with 20 - 30 mL of contrast; 2) acinarization + ductal balloon occlusion + sphincterotomy; 3) acinarization + intraductal synthetic bile injection + ductal balloon occlusion + sphincterotomy. Dogs 5 and 6 received a pancreatic stent. Necropsy was performed on postoperative day 5. All pancreatic specimens were graded by two blinded pathologists according to a validated scoring system. All dogs were compared with three control dogs. Dogs 1 - 4 developed clinical pancreatitis and hyperamylasemia (11 736 vs. 722 U/L, P = 0.02). Total injury scores were significantly elevated compared with controls (6.85 vs. 1.06, P = 0.004). There was significant increase in acinar cell necrosis (0.86 vs. 0.06, P = < 0.001), and all other categories (except fibrosis) demonstrated elevated injury scores . Dogs 5 and 6 developed clinical pancreatitis without significant hyperamylasemia; total injury scores were elevated compared with controls (4.83 vs. 1.06, P = 0.01), but lower than in Dogs 1 - 4 (4.83 vs. 6.85, P = 0.25). There was escalating severity of pancreatic injury from Dogs 1 to 4 correlating with the method of endoscopic injury used. Severity of PEP is directly proportional to invasiveness of endoscopic intervention. Pancreatic acinarization, even without balloon occlusion and sphincterotomy, can be used as a reliable animal model for future studies investigating therapy and prevention of disease.


Phase II trial of bevacizumab with dose-dense paclitaxel as first-line treatment in patients with advanced ovarian cancer.
To assess the tolerability and efficacy of bevacizumab with carboplatin and weekly paclitaxel as first-line adjuvant therapy for advanced stage ovarian cancer. After IRB approval, this single-institution, phase II study enrolled patients with stage III or IV epithelial ovarian cancer after primary cytoreductive surgery to treatment with carboplatin (AUC 5), weekly paclitaxel (80mg/m2), and bevacizumab (15mg/kg) every 3weeks for at least 6cycles. The primary endpoint was tolerability of at least 4cycles of therapy, with a target treatment success rate of >60%. Secondary endpoints included progression-free survival (PFS) and response rate. Plasma biomarkers were analyzed by the multiplex ELISA assays. Thirty-three patients were enrolled with 30 evaluable patients receiving at least one cycle of combination treatment. Twenty-three patients (77%) were able to complete at least 4cycles of therapy per protocol, and the posterior probability that the treatment success rate is >60% is 0.77. Twenty-one patients (70%) were able to complete ≥6cycles of therapy. Median PFS was 22.4months for patients with optimal (R0) compared to 16.9months for optimal≤1cm (HR 1.71, 95% CI 0.58-4.98, p=0.33), and 16.9months for suboptimal>1cm (HR 3.75, 95% CI 1.05-13.34, p=0.04) disease. Increases in mean Flt-3L was significantly higher in responders versus non-responders (83.4 vs. 28pg/mL, p=0.05). Adjuvant bevacizumab with dose-dense chemotherapy is associated with acceptable toxicity and a high likelihood of completing 4cycles of therapy. Dynamic changes in Flt-3L may represent a predictive marker to treatment response.


The mechanism of neuronal resistance and adaptation to hypoxia.
In this work we provide a theoretical explanation for the observations that: (i) young animals are more resistant to hypoxia than adult ones and (ii) repeated exposure to a hypoxic insult increases the tolerance of young animals and isolated brain tissue to that insult. Considered here is the role of taurine, a putative Ca2+ transport modulator, in attenuating Ca2+ influx and overload in brain tissue upon hypoxia. It is proposed that the higher resistance of young animals to hypoxia stems from their higher brain content of taurine as compared with adults. The increased resistance to lack of oxygen upon re-exposure to hypoxia may occur as a result of protein and coenzyme A (CoA) breakdown which leads to the accumulation of products like cystine, cysteine, cysteamine and other sulfur-containing compounds. Upon reoxygenation, these compounds are oxidized to form taurine, which in turn attenuates neuronal Ca2+ accumulation. The sulfur-containing compounds are considered to be natural scavengers of oxygen-derived free radicals which are formed upon reoxygenation and have been implicated as a major component in the process leading to ischemic/hypoxic brain damage. Repeated hypoxic insults bring about the formation of higher levels of taurine and hence the observed adaptation to oxygen lack. The hypothesis presented here is supported by experimental observations in our laboratory and those of others.


Familial dilated cardiomyopathy: clinical features in French families.
The aims of the study were to analyze the clinical features, the penetrance and the mode of inheritance of 13 French families with dilated cardiomyopathy using diagnostic criteria recently established by a European collaboration. Screening consisted of physical examination, ECG and Echo of all the probands first degree relatives (n = 118). Using major Echo criteria [ejection fraction (EF) < 45% or FS < 25% and left ventricular diameter (LVD) > 117% of the predictive value], or combined minor Echo/ECG criteria, relatives were classified as affected, unknown or healthy. (1) Adult affected relatives (n = 31) were identified with major Echo criteria in 74% of cases, and with combined minor Echo/ECG criteria in 26% of cases. (2) In the unknown relatives (n = 21), the most common abnormality was an isolated left ventricular dilation (67%). (3) Mode of inheritance was autosomal dominant (AD) in 11 families and possibly autosomal recessive in two. (4) In AD families, the penetrance was incomplete in adults (72%), age-related (O.R.: 1.3 per 10 years; 95% CI 1.03-1.56) and sex-related [greater in men (87%) than in women (61%), actuarial survival curve: P<0.002]. (5) Mortality related to end stage heart failure was 2.2 times as high as mortality related to sudden death (11% vs. 5%). (1) In the absence of a specific phenotype of FDC, the characterization of relatives appears more accurate when minor criteria were added. (2) Since high mortality (16%) and incomplete penetrance frequently give rise to small nuclei of clinically affected and alive relatives per family, the accurate model of penetrance that we proposed might be helpful in the future to enhance the statistical power of linkage analysis in this disease.


Heme-associated subunit complex of cytochrome c oxidase identified by a new two-dimensional gel electrophoresis.
A Novel two-dimensional system (isoelectric focusing in the first dimension and sodium dodecyl sulfate-urea electrophoresis in the second) was used to determine the pI values of beef heart subunits of cytochrome c oxidase. The results identified a heme-associated complex of Subunits I, II, and IV. Conditions mitigate against migration of free heme.


Head to head comparison of nomograms predicting probability of lymph node invasion of prostate cancer in patients undergoing extended pelvic lymph node dissection.
To validate the Briganti nomogram and compare it with 2 current lymph node invasion (LNI) nomograms (the Cagiannos nomogram and the updated 2007 Partin tables). The Briganti nomogram predicts the probability of LNI in patients undergoing extended pelvic lymph node dissection (EPLND) during radical prostatectomy for prostate cancer. Irrespective of the risk of LNI, 173 consecutive patients were treated for localized prostate cancer with radical laparoscopic prostatectomy and EPLND. The area under the receiver operating characteristics curve was used to estimate the predictive accuracy of the nomograms, and calibration plots were used for comparisons between the predicted and observed probabilities of LNI. The median number of nodes removed was 15 (range 10-34). Of the 173 patients, 12 (6.9%) had LNI. The Briganti nomogram achieved a receiver operating characteristic curve of 0.88 versus 0.83 with the Cagiannos nomogram and 0.84 with the 2007 Partin tables. The difference in predictive accuracy was not statistically significant (P < .2). The Briganti nomogram showed only minor departures from the ideal predictions in the low-risk range and the Cagiannos nomogram showed major departures from the ideal predictions for the entire risk range. The Briganti nomogram provides highly accurate predictions of the risk of LNI after EPLND. Its performance tended to be increased without being significantly better. The other tools also performed reasonably well but underestimate the true risk of LNI. We recommend the use of these tools to identify patients at low risk of LNI for whom EPLND can be safely spared.


Effect of obesity on the relationship between very low density lipoprotein production rate and plasma triglyceride concentration in normal and hypertriglyceridemic subjects.
The effect of obesity on the relationship between very low density lipoprotein (VLDL)-triglyceride (TG) production rate and concentration was studied in 80 subjects whose TG concentrations ranged from 41-1315 mg/100 ml and whose relative weight varied from 0.74-1.46. There was a positive correlation between VLDL-TG production rate and plasma TG concentration in all 80 patients (r = 0.78), which was highly statistically significant (p less than 0.001). Both male and female subjects were subdivided into quartiles on the basis of relative weight; the relationship between VLDL-TG production rate and plasma TG concentration was found to be comparable in all subgroups. These observations suggest that moderate degrees of obesity do not affect VLDL-TG kinetics in patients whose plasma TG levels vary over an extremely wide range.


A protocol for imaging microvilli biogenesis on the surface of cultured porcine kidney epithelial cell monolayers.
A key facet of epithelial differentiation is the assembly of actin-based protrusions known as microvilli, which amplify apical membrane surface area for various cell functions. To probe mechanisms of microvillus assembly, we developed a protocol using spinning disk confocal microscopy to directly visualize microvillus biogenesis on the surface of cultured porcine kidney epithelial cell monolayers engineered to express fluorescent proteins. This protocol offers access to the molecular details of individual protrusion growth events at high spatiotemporal resolution. For complete details on the use and execution of this protocol, please refer to Gaeta et al. (2021).


Microplastics in the surgical environment.
Atmospheric microplastics (MPs) have been consistently detected within indoor and outdoor air samples. Locations with high human activity are reported to have high MP levels. The aim was to quantify and characterise the MPs present within the surgical environment over a one-week sampling period. MPs were collected in samplers placed around an operating theatre and adjoining anaesthetic room at 12 h intervals. Particles were filtered onto 0.02 µm membranes and analysed using micro-Fourier-transform infrared spectroscopy. The number of MPs identified during the working day sampling period varied, with a mean of 1,924 ± 3,105 MP m-2 day-1 and a range of 0 - 9,258 MP m-2 day-1 observed in the theatre, compared with a mean of 541 ± 969 MP m-2 day-1 and a range of 0 - 3,368 MP m-2 day-1 for the anaesthetic room. Across both rooms and at all sampling points, an increase in levels with a decrease in MP size was observed. Identified particles consisted of mainly fragment shaped MPs (78 %) with polyethylene terephthalate (37 %), polypropylene (25 %), polyethylene (7 %) and nylon (13 %) representing the most abundant polymer types. MPs were not detected in the theatre during non-working hours. The results provide novel information on defining polymer levels and types, in a room environment where the use of single plastics has been regarded as beneficial to practice. These results can inform cellular toxicity studies, investigating the consequences of human MP exposure as well as represent a potentially novel route of exposure for humans for this emerging contaminant of concern, via surgery.


Encapsulating lithium at the microscale: selective deposition in carbon-doped graphitic carbon nitride spheres.
For stable lithium deposition without dendrites, three-dimensional (3D) porous structure has been intensively investigated. Here, we report the use of carbon-doped graphitic carbon nitride (C-doped g-C3N4) microspheres as a 3D host for lithium to suppress dendrite formation, which is crucial for stable lithium deposition. The C-doped g-C3N4microspheres have a high surface area and porosity, allowing for efficient lithium accommodation with high accessibility. The carbon-doping of the g-C3N4microspheres confers lithiophilic properties, which facilitate the regulation of Li+flux and dense filling of cavities with nucleated lithium, thereby preventing volume expansion and promoting dendrite-free Li deposition. The electrochemical performance was improved with cyclic stability and high Coulombic efficiency over 260 cycles at 1.0 mA cm-2for 1.0 mAh cm-2, and even over 70 cycles at 5.0 mA cm-2for 3.0 mAh cm-2. The use of C-doped g-C3N4microspheres as a 3D Li host shows promising results for stable lithium deposition without dendrite formation.


Nitric oxide induces the apoptosis of human BCR-ABL-positive myeloid leukemia cells: evidence for the chelation of intracellular iron.
Anti-leukemia activity of human macrophages involves the generation of nitric oxide (NO) derivatives. However, leukemic transformation may involve mechanisms that rescue cells from NO-mediated apoptosis. In the present work, we analyzed the effects of exogenous NO on the proliferation of BCR-ABL(+) chronic myelogenous leukemia (CML) cells. As normal leukocytes, the proliferation of leukemia cells was inhibited by SNAP (S-nitroso-N-acetyl-penicillamine), GEA (Oxatriazolium amino-chloride), and SIN-1 (Morpholino-sydnonimine), whereas SNP (sodium nitroprusside) had no effect on leukemia cell growth. SIN-1 induced higher anti-proliferation activity in BCR-ABL(+) cells, compared to normal hemopoietic cells. Inhibition of leukemia cell proliferation correlated with increased apoptosis and DEVDase activity. The simultaneous addition of exogenous iron reversed NO-mediated inhibition of cell growth, caspase activation and apoptosis in all BCR-ABL(+) cells tested. The quantification of intracellular iron levels in leukemia cells indicated that NO induced an early, dose-dependent decrease in ferric iron levels. Accordingly, elevation of intracellular iron protected leukemia cells from NO-mediated apoptosis. Together, the present work reveals the presence of an iron-dependant mechanism for leukemia cell rescue from NO-induced growth inhibition and apoptosis.


Radiation dose reduction in perfusion CT imaging of the brain: A review of the literature.
Perfusion CT (PCT) of the brain is widely used in the settings of acute ischemic stroke and vasospasm monitoring. The high radiation dose associated with PCT is a central topic and has been a focus of interest for many researchers. Many studies have examined the effect of radiation dose reduction in PCT using different approaches. Reduction of tube current and tube voltage can be efficient and lead to a remarkable reduction of effective radiation dose while preserving acceptable image quality. The use of novel noise reduction techniques such as iterative reconstruction or spatiotemporal smoothing can produce sufficient image quality from low-dose perfusion protocols. Reduction of sampling frequency of perfusion images has only little potential to reduce radiation dose. In the present article we aimed to summarize the available data on radiation dose reduction in PCT imaging of the brain.


Visual Sensor Networks and Related Applications.
The use of sensing devices to perform monitoring tasks has continuously evolved in the past decades [...].


Skeletal staging in Langerhans cell histiocytosis: a multimodality imaging review.
Evaluating the extent of skeletal disease in Langerhans cell histiocytosis (LCH) is a major predictor of patient outcome. Traditionally, whole-body skeletal staging consists of plain radiography and bone scintigraphy. However, more recently whole-body MRI has been shown to be accurate in detecting osseous and extra-osseous lesions, but no large-scale studies analysing its role within the diagnostic algorithm of LCH skeletal staging currently exist. In addition, FDG PET-CT provides useful information regarding disease activity and treatment response, but has an inherent radiation dose which is not ideal in children. Currently, radiographic skeletal survey remains the gold standard with cross-sectional imaging only performed for further characterisation. However, radiographs have shown a wide sensitivity range for skeletal staging and have clear limitations in detecting extra-skeletal disease, a crucial component of stratification in identifying 'at risk' organs. We aim to highlight the various appearances of bony LCH across all the imaging modalities for primary skeletal staging. We will also review the advantages, disadvantages, sensitivity and specificity of each, and establish their role in staging skeletal LCH. Recent studies using whole-body MRI have shown promising results, with radiographs and other modalities playing a more complementary role.


Divergent chondro/osteogenic transduction laws of fibrocartilage stem cell drive temporomandibular joint osteoarthritis in growing mice.
The anterior disc displacement (ADD) leads to temporomandibular joint osteoarthritis (TMJOA) and mandibular growth retardation in adolescents. To investigate the potential functional role of fibrocartilage stem cells (FCSCs) during the process, a surgical ADD-TMJOA mouse model was established. From 1 week after model generation, ADD mice exhibited aggravated mandibular growth retardation with osteoarthritis (OA)-like joint cartilage degeneration, manifesting with impaired chondrogenic differentiation and loss of subchondral bone homeostasis. Lineage tracing using Gli1-CreER+; Tmfl/-mice and Sox9-CreER+;Tmfl/-mice showed that ADD interfered with the chondrogenic capacity of Gli1+ FCSCs as well as osteogenic differentiation of Sox9+ lineage, mainly in the middle zone of TMJ cartilage. Then, a surgically induced disc reposition (DR) mouse model was generated. The inhibited FCSCs capacity was significantly alleviated by DR treatment in ADD mice. And both the ADD mice and adolescent ADD patients had significantly relieved OA phenotype and improved condylar growth after DR treatment. In conclusion, ADD-TMJOA leads to impaired chondrogenic progenitor capacity and osteogenesis differentiation of FCSCs lineage, resulting in cartilage degeneration and loss of subchondral bone homeostasis, finally causing TMJ growth retardation. DR at an early stage could significantly alleviate cartilage degeneration and restore TMJ cartilage growth potential.


State patients who committed violent crimes and were admitted to Weskoppies Hospital from 2005 to 2014: Profiles and trends.
Most of the available literature focus on characteristics of violent offenders and trends of crime patterns in the general population. The crime trends in the population of those who may be mentally ill have not been well described. To describe the profiles and trends of mentally ill persons who committed violent crimes prior to admission as state patients. The study was conducted at Weskoppies Psychiatric Hospital. A retrospective record review of state patients admitted between 2005 and 2014 was conducted to describe demographic, clinical, forensic and victim profiles of state patients admitted from 2005 to 2014. Trends over time were also assessed. Hundred and seventy state patient files were reviewed. The majority were males (91.43%), with a history of substance use (55.0%) %), previous psychiatric treatment (46.4%), and diagnosed with a psychotic disorder (82.1%). The 10 year trend showed that murder was the leading charge during 2006 and 2007. It was surpassed by sexual assault crimes as the most common charge after 2007, except for 2009 when murder was again the most common. From 2010 onward, sexual assault remained the most common offence leading to admission as a state patient. This study found changes over time in crime patterns of state patients who committed murder and sexual assault. State patients may have different criminal patterns than the general public. This together with the high rates of substance use and previous psychiatric treatment can be important focus areas for future research.


Oxidative changes of lipids monitored by MALDI MS.
Oxidation processes of lipids are of paramount interest from many viewpoints. For instance, oxidation processes are highly important under in vivo conditions because molecules with regulatory functions are generated by oxidation of lipids or free fatty acids. Additionally, many inflammatory diseases are accompanied by lipid oxidation and, therefore, oxidation products are also useful disease (bio)markers. Thus, there is also considerable interest in methods of (oxidized) lipid analysis. Nowadays, soft ionization mass spectrometric (MS) methods are regularly used to study oxidative lipid modifications due to their high sensitivities and the extreme mass resolution. Although electrospray ionization (ESI) MS is so far most popular, applications of matrix-assisted laser desorption and ionization (MALDI) MS are increasing. This review aims to summarize the so far available data on MALDI analyses of oxidized lipids. In addition to model systems, special attention will be paid to the monitoring of oxidized lipids under in vivo conditions, particularly the oxidation of (human) lipoproteins. It is not the aim of this review to praise MALDI as the "best" method but to provide a critical survey of the advantages and drawbacks of this method.


Differential response of normal and cirrhotic liver to vasoactive agents. A study in the isolated perfused rat liver.
In cirrhosis, endogenous vasoactive agents could act as modulators of intrahepatic resistance and thus portal pressure. The aim of this work was to study the effects of norepinephrine, angiotensin II and arg8-vasopressin on intrahepatic portal resistance in isolated perfused livers from normal rats and rats with carbon tetrachloride-induced cirrhosis. Livers were perfused at a constant pressure and the measured variable was portal blood flow. Dose-response curves were obtained by cumulative addition of agonists to the perfusate. The three vasoactive agents increased resistance in normal and cirrhotic livers. The maximal amplitude of response was similar in normal and cirrhotic livers. The cirrhotic livers exhibited an increased sensitivity to norepinephrine, a decreased sensitivity to angiotensin II but an unchanged sensitivity to arg8-vasopressin. The shape of the dose-response curve for norepinephrine and arg8-vasopressin, but not for angiotensin II, was modified in cirrhotic livers. We conclude that the cirrhotic liver retains a strong vascular reactivity to vasoactive agents and particularly to norepinephrine.


Integrin-alpha3 mediates binding of Chordin to the cell surface and promotes its endocytosis.
Dorsoventral patterning in animal development is regulated by a morphogenetic gradient of Bone morphogenetic protein signalling, which is established by a set of proteins that are conserved from Drosophila to vertebrates. These include Chordin (Chd)/Short gastrulation, Xolloid/Tolloid and Twisted gastrulation. Here, we report the identification of a cell-surface component of this morphogenetic pathway. Prompted by the observation that Chd protein bound to the surface of certain cell lines with subnanomolar affinity, we identified two cell-surface proteins that bind to Chd, one of which corresponds to Integrin-alpha3. Integrin-alpha3 and Chd are co-expressed in the Xenopus embryo. Transfection of Integrin-alpha3 increased the binding of Chd to the cell surface, which was competed by an excess of soluble Integrin-alpha3. After binding to the cell surface, Chd was translocated into intracellular endocytic compartments in a temperature-dependent manner. We propose that Integrin-alpha3 may regulate the concentration of Chd protein in the extracellular space by endocytosis.


Secreted proteases from dermatophytes.
Dermatophytes are highly specialized pathogenic fungi that exclusively infect the stratum corneum, nails or hair, and it is evident that secreted proteolytic activity is important for their virulence. Endo- and exoproteases-secreted by dermatophytes are similar to those of species of the genus Aspergillus. However, in contrast to Aspergillus spp., dermatophyte-secreted endoproteases are multiple and are members of two large protein families, the subtilisins (serine proteases) and the fungalysins (metalloproteases). In addition, dermatophytes excrete sulphite as a reducing agent. In the presence of sulphite, disulphide bounds of the keratin substrate are directly cleaved to cysteine and S-sulphocysteine, and reduced proteins become accessible for further digestion by various endo- and exoproteases secreted by the fungi. Sulphitolysis is likely to be an essential step in the digestion of compact keratinized tissues which precedes the action of all proteases.


Electrophysiologic properties differ in the ventricular endocardium and epicardium of the Japanese monkey.
We measured action potential duration (APD) from the endocardium (Endo) and epicardium (Epi) of the left ventricular free wall in Japanese monkey hearts and found that the APD of Endo is significantly longer than that of Epi at a stimulus cycle length of 1500 msec in normal Tyrode solution (control condition). We then hypothesized that shorter APD of Epi results from greater outward pump current and that the difference in the current may be due to a difference in membrane Na,K-ATPase activity between Endo and Epi. If this were the case, interventions which alter the Na,K-pump activity should alter electrophysiologic characteristics in the endocardium and the epicardium to different degrees. An application of ouabain (10(-6) M), an inhibitor of Na,K-ATPase, produced greater shortening of APD and greater depolarization of the resting potential in Endo as compared with Epi. Shortening of stimulus cycle length shortened the APD more markedly in Endo than Epi, resulting in a significantly longer APD in Epi than Endo at a stimulus cycle length of 200 msec. The hyperpolarization and the APD shortening produced when the tissues were returned to 5.4 mM K+ Tyrode solution from K+-free medium were also more marked in Endo than in Epi. Such findings suggest that endocardial cells are more liable to accumulate Na ions intracellularly and K ions extracellularly when the Na, K-pump is suppressed by ouabain or K+-free perfusion, presumably due to lower activity of Na, K-ATPase in Endo compared to Epi.(ABSTRACT TRUNCATED AT 250 WORDS)


[Intravenously stimulated insulin reserve of the pancreas following experimental pancreatic duct ligation in the swine--studies 2, 4 and about 60 days following ligation].
After pancreatic duct ligation (PDL) intravenous glucose tolerance tests (i.v. GTT) were performed in 12 pigs. The release of insulin and the corresponding blood sugar levels were controlled, the pancreata were examined histologically and immunohistochemically. In spite of a reduced number of beta-cells no reduction of glucose tolerance could be shown 60 days after PDL. Two days after the i.v. GTT showed an increased release of insulin corresponding with lowered blood sugar levels. This phenomenon is explained as a stimulated state of the beta-cell in the phase of interstitial edema.


Virtual screening of potentially endocrine-disrupting chemicals against nuclear receptors and its application to identify PPARγ-bound fatty acids.
Nuclear receptors (NRs) are key regulators of energy homeostasis, body development, and sexual reproduction. Xenobiotics binding to NRs may disrupt natural hormonal systems and induce undesired adverse effects in the body. However, many chemicals of concerns have limited or no experimental data on their potential or lack-of-potential endocrine-disrupting effects. Here, we propose a virtual screening method based on molecular docking for predicting potential endocrine-disrupting chemicals (EDCs) that bind to NRs. For 12 NRs, we systematically analyzed how multiple crystal structures can be used to distinguish actives and inactives found in previous high-throughput experiments. Our method is based on (i) consensus docking scores from multiple structures at a single functional state (agonist-bound or antagonist-bound), (ii) multiple functional states (agonist-bound and antagonist-bound), and (iii) multiple pockets (orthosteric site and alternative sites) of these NRs. We found that the consensus enrichment from multiple structures is better than or comparable to the best enrichment from a single structure. The discriminating power of this consensus strategy was further enhanced by a chemical similarity-weighted scoring scheme, yielding better or comparable enrichment for all studied NRs. Applying this optimized method, we screened 252 fatty acids against peroxisome proliferator-activated receptor gamma (PPARγ) and successfully identified 3 previously unknown fatty acids with Kd = 100-250 μM including two furan fatty acids: furannonanoic acid (FNA) and furanundecanoic acid (FUA), and one cyclopropane fatty acid: phytomonic acid (PTA). These results suggested that the proposed method can be used to rapidly screen and prioritize potential EDCs for further experimental evaluations.


Polyethylene glycol as an alternative polymer solvent for nanoparticle preparation.
Solvent toxicity is one of the major drawbacks in the preparation of polymeric nanoparticles today. Here, polyethylene glycols (PEGs) are proposed as non-toxic solvents for the preparation of polymeric nanoparticles. Based on a preparation process similar to the solvent displacement technique, several process parameters were examined for their effects on the properties of the prepared nanoparticles by this method to achieve the optimum preparation conditions. The investigated parameters included polymer type and concentration, volume and temperature of the dispersing phase, methods of dispersing the solvent phase into the non-solvent phase, duration and speed of stirring and washing by dialysis. Ammonio methacrylate copolymer (Eudragit RL), poly-lactide-co-glycolide (PLGA), and PEG-PLGA were found to be successful polymer candidates for the preparation of nanoparticles by this method. Nanoparticles with diameters ranging from 80 to 400 nm can be obtained. The encapsulation efficiencies of bovine serum albumin, and lysozyme as model proteins were ranging from 7.3±2.2% to 69.3±1.8% depending on the strength of polymer-protein interaction. Biological assays confirmed a full lysozyme activity after the preparation process. PEG proved to be a suitable non-toxic solvent for the preparation of polymeric protein-loaded nanoparticles, maintaining the integrity of protein.


Multiple forms of inducible drug-metabolizing enzymes: a reasonable mechanism by which any organism can cope with adversity.
All organisms possess a number of genetically regulated mechanisms in order to cope with rapid adverse changes in the environment. The two systems which appear to respond to a seemingly endless array of chemical specificities are the immune response and the induction of drug-metabolizing enzymes. Similarities and differences between the immunoglobulin and the cytochrome P-450-mediated monooxygenase systems are described. DNA insertion sequences, plasmid "transposons," maize "controlling elements," gene duplication, intervening sequences, and high-frequency intergenic recombination are all discussed as possible methods by which organisms can "adapt" quickly to a new selective pressure. If the regulation of P-450 induction resembles in any way the other methods by which pro- and eukaryotes cope genetically with numerous forms of environmental adversity, therefore, it is very likely that mammalian tissues contain hundreds, if not thousands, of inducible forms of P-450.


1H NMR study on the interaction of cupric ion with ribonuclease A.
The reassignment of the 1H NMR C-2 histidine signals of the bovine pancreatic ribonuclease A has required a revision of the 1H NMR data on the role of the different histidines in their interaction with the Cu2+. The results of our measurements carried out at p2H 5.5 and 7.0 reduce the importance of His-12 as main site of interaction. At p2H 5.5 a very strong binding site involves His-119, while a weaker one contains certainly His-105. On the contrary, at p2H 7.0 the histidines 105 and 119 seem to possess binding constants of the same order of magnitude and in addition they provide stronger ligands for the Cu2+ than His-12. The comparison with X-ray data in the crystal shows numerous analogies. Finally, preliminary results on the competitive inhibition effect between the Cu2+ and 2',3'-cytidine monophosphoric acid are discussed.


11beta-hydroxysteroid dehydrogenase and the pre-receptor regulation of corticosteroid hormone action.
Two isozymes of 11beta-hydroxysteroid dehydrogenase (11beta-HSD1 and 11beta-HSD2) catalyse the interconversion of hormonally active cortisol and inactive cortisone. The enzyme evolved from a metabolic pathway to a novel mechanism underpinning human disease with the elucidation of the role of the type 2 or 'kidney' isozyme and an inherited form of hypertension, 'apparent mineralocorticoid excess'. 'Cushing's disease of the kidney' arises because of a failure of 11beta-HSD2 to inactivate cortisol to cortisone resulting in cortisol-induced mineralocorticoid excess.Conversely, 11beta-HSD1 has been linked to human obesity and insulin resistance, but also to other diseases in which glucocorticoids have historically been implicated (osteoporosis, glaucoma). Here, the activation of cortisol from cortisone facilitates glucocorticoid hormone action at an autocrine level. The molecular basis for the putative human 11beta-HSD1 'knockout'--'cortisone reductase deficiency'--has recently been described, an observation that also answers a long standing conundrum relating to the set-point of 11beta-HSD1 activity. In each case, these clinical studies have been underpinned by studies in vitro and the manipulation of enzyme expression in vivo using recombinant mouse models.


Factor analysis in predominantly severe COPD: identification of disease heterogeneity by easily measurable characteristics.
The clinical and demographic variables defining the heterogeneity of chronic obstructive pulmonary disease (COPD) are unclear. A post-hoc analysis of five randomised studies in patients with a history of previous exacerbations examined the clinical and demographic characteristics describing moderate-to-very-severe COPD. Factor analysis was performed on all continuous baseline demographic and clinical data, without variable selection. Analyses were based on the full cohort and on stratifications by pack-years smoked, smoking status, gender, and comorbidities; patient exacerbation history was analysed in two of the five studies. 6162 COPD patients were evaluated (70% male; 40% current smokers; mean pre-bronchodilator forced expiratory volume in 1 s [FEV1] 35.2% predicted). Baseline clinical and demographic variables loaded differentially on six factors with minimal overlap, explaining 60.4% of the heterogeneity: 1) symptoms (cough, dyspnoea, sleep disturbance), health status, reliever use; 2) pre-bronchodilator FEV1, FEV1/forced vital capacity, morning peak expiratory flow (PEF), body mass index (BMI); 3) blood pressure; 4) age, months since first COPD symptoms; 5) PEF variability; 6) pulse, FEV1 reversibility. Most factors loaded similarly in stratified and exacerbation analyses. BMI loaded with reversibility in females, and with age and months since first COPD symptoms in ex-smokers. Exacerbations loaded to factor 6. Readily available data can explain ≈ 60% of COPD heterogeneity in a large dataset of predominantly severe COPD patients. Factors were robust over determinants of disease outcome; gender, smoking status, pack-years smoked, and comorbidities. The main factors were largely unchanged by adding exacerbations. Only BMI loaded to other factors.


A novel smoothed (SMO) point mutation in congenital tibial hemimelia: a case report.
Congenital tibial hemimelia (CTH [MIM: 275220]) is a rare congenital limb deficiency that manifests as a shortened, curved, dysplastic or absent tibia with polydactyly. In previous studies, mutations of a distant sonic hedgehog (SHH) cis-regulator (ZRS) and a Shh repressor (GLI3) were identified. Here, we admitted a 20-month-old boy who manifested with right tibial deformity, varus foot, ankle dislocation, and ipsilateral preaxial polydactyly. After genetic sequencing and data analysis, the results revealed a 443 A > G mutation in the father and a 536 C > T mutation in the mother in exon 2 of the Smoothed (SMO) gene at 7q32.1, with the coexistence of both mutant alleles in the proband/patient. Our report suggests that even though not previously reported, SMO mutations may be associated with limb anomalies such as tibial hemimelia via Hh signaling in humans and has implications for genetic counseling.


Dynamics of the response to variations in dietary choline content in rats.
A mathematical model was developed based on the analysis of body response to variations in choline content in the ration of growing rats. The peculiarities of choline metabolism and the expression of α7-nicotinic receptors in the brain were studied. The information obtained can be useful for comparative analysis of choline administration for correction of certain cerebral pathologies.


Cholangiocyte senescence caused by lysophosphatidylcholine as a potential implication in carcinogenesis.
The incidence of biliary tract cancer in patients with pancreaticobiliary maljunction or intrahepatic cholelithiasis is markedly high with undefined mechanism. In these diseases, biliary lysophosphatidylcholine (LPC) level is reportedly increased. This study investigated the influence of LPC on cholangiocytes focusing on cellular senescence and its potential contribution to carcinogenesis. Cultured MMNK-1, an immortalized human cholangiocyte was treated with LPC in vitro and its effect was evaluated. Lysophosphatidylcholine demonstrated cytotoxicity with generation of intracellular reactive oxygen species. Accordingly, LPC provoked oxidative DNA injury, whereas the gene expressions of DNA repair enzyme (OGG1, MUTYH, MTH1) remained unchanged. Interestingly, LPC caused global DNA hypomethylation, which is frequently observed in cancer tissues. Microarray analysis identified differentially regulated genes in response to LPC, which included the components of senescence-associated secretory phenotype (SASP) including interleukin-8 (IL-8), IL-6, transforming growth factor-β and plasminogen activator inhibitor-1. Significant induction of these genes was further confirmed by quantitative real-time polymerase chain reaction. In addition to upregulation of p21 gene expression, senescence-associated beta-galactosidase activity, a widely used marker of cellular senescence was significantly induced by the treatment of LPC. Based on these data, cholangiocyte senescence and SASP caused by LPC are potential pathogenic mechanisms in the development of biliary tract cancer.


Constructing Effective Hole Transport Channels in Cross-Linked Hole Transport Layer by Stacking Discotic Molecules for High Performance Deep Blue QLEDs.
The inadequate hole injection limits the efficiency and lifetime of the blue quantum dot light-emitting diodes (QLEDs), which severely hampers their commercial applications. Here a new discotic molecule of 3,6,10,11-tetrakis(pentyloxy)triphenylene-2,7-diyl bis(2,2-dimethylpropanoate) (T5DP-2,7) is introduced, in which the hole transport channels with superior hole mobility (2.6 × 10-2 cm2 V-1 s-1 ) is formed by stacking. The composite hole transport material (HTM) is prepared by blending T5DP-2,7 with the cross-linked 4,4'- bis(3-vinyl-9H-carbazol-9-yl)-1,1'biphenyl (CBP-V) which shows the deep highest occupied molecular orbital energy level. The increased hole mobility of the target composite HTM from 10-4 to 10-3 cm2 V-1 s-1 as well as the stepwise energy levels facilitates the hole transport, which would be beneficial for more balanced carrier injection. This composite hole transport layer (HTL) has improved the deep-blue-emission performances of Commission International de I'Eclairage of (0.14, 0.04), luminance of 44080 cd m-2 , and external quantum efficiency of 18.59%. Furthermore, when L0 is 100 cd m-2 , the device lifetime T50 is extended from 139 to 502 h. The state-of-the-art performance shows the successful promotion of the high-efficiency for deep blue QLEDs, and indicates that the optimizing HTL by discotic molecule stacking can serve as an excellent alternative for the development of HTL in the future.


Evaluation for tetanus antibodies in Korean-Americans living in the New York area: a pilot study.
Public health initiatives to immunize children and adults have effectively reduced the number of tetanus cases in the USA. However, in the third National Health and Nutrition Examination Survey (NHANES III), immigrants from Mexico had a 67% non-protective anti-tetanus antibody (ATA) level. Less work has been conducted among other immigrant populations to determine the extent of this observation. Objective To measure ATA levels among the Korean-American immigrant population. Methods A convenience sample of 50 Korean Americans born outside the USA was recruited to determine the levels of ATA. A non-protective level of ATA was defined as below 0.15 IU/ml. Results The mean age was 59.5 years and 82% were female. There were 43/50 (86% (95% confidence limits 76, 96)) patients with a non-protective ATA level. Those between the ages of 50-59 years (94% were seronegative) and 60 years-highest age (92% were seronegative) were among the least likely to be protected. Neither gender nor a self-reported history of past tetanus immunization or military service predicted protection to tetanus. Discussion In this pilot study we found that 86% of Korean immigrants did not have protective ATA levels, with patients in the 50-59 year age range as unlikely to be protected as the older subjects. Patient reported history was unreliable in determining whether an individual had protective levels. Conclusion The vast majority of sampled Korean American immigrants lack protective ATA levels and are in need of immunization. Additional study is needed to determine the risk of other immigrant groups to tetanus.


Incidence and associated factors of pediatric post-intensive care syndrome using the VSCAREMD model.
The VSCAREMD model is used for evaluating vaccination, sleep, and parental care burden, which includes daily activity and social interaction, rehabilitation requirements, hearing, mood, and development. It has been proposed to detect post-intensive care syndrome (PICS) in children. This study aimed to outline the incidence of PICS in children using the VSCAREMD model and to describe the associated factors. All children ages 1 month to 15 years and admitted to the intensive care unit for at least 48 hours were evaluated using the VSCAREMD model within 1 week of intensive care discharge. Abnormal findings were assorted into four domains: physical, cognitive, mental, and social. Descriptive statistics were performed using chi-square, univariate, and multivariate analyses. A total of 78 of 95 children (82.1%) had at least one abnormal domain. Physical, cognitive, mental, and social morbidity were found in 64.2%, 26.3%, 13.7%, and 38.9% of the children, respectively. Prolonged intensive care unit stay greater than 7 days was associated with dysfunction in physical (adjusted odds ratio [aOR], 3.80; 95% confidence interval [CI], 1.31-11.00), cognitive (aOR, 10.11; 95% CI, 3.01-33.89), and social domains (aOR, 5.01; 95% CI, 2.01-12.73). Underlying medical conditions were associated with cognitive (aOR, 13.63; 95% CI, 2.64- 70.26) and social morbidity (aOR, 2.81; 95% CI, 1.06-7.47). The incidence of PICS using the VSCAREMD model was substantially high and associated with prolonged intensive care. This model could help evaluate PICS in children.


Integrin-dependent control of translation: engagement of integrin alphaIIbbeta3 regulates synthesis of proteins in activated human platelets.
Integrins are widely expressed plasma membrane adhesion molecules that tether cells to matrix proteins and to one another in cell-cell interactions. Integrins also transmit outside-in signals that regulate functional responses of cells, and are known to influence gene expression by regulating transcription. In previous studies we found that platelets, which are naturally occurring anucleate cytoplasts, translate preformed mRNA transcripts when they are activated by outside-in signals. Using strategies that interrupt engagement of integrin alphaIIbbeta3 by fibrinogen and platelets deficient in this integrin, we found that alphaIIbbeta3 regulates the synthesis of B cell lymphoma 3 (Bcl-3) when platelet aggregation is induced by thrombin. We also found that synthesis of Bcl-3, which occurs via a specialized translation control pathway regulated by mammalian target of rapamycin (mTOR), is induced when platelets adhere to immobilized fibrinogen in the absence of thrombin and when integrin alphaIIbbeta3 is engaged by a conformation-altering antibody against integrin alphaIIbbeta3. Thus, outside-in signals delivered by integrin alphaIIbbeta3 are required for translation of Bcl-3 in thrombin-stimulated aggregated platelets and are sufficient to induce translation of this marker protein in the absence of thrombin. Engagement of integrin alpha2beta1 by collagen also triggered synthesis of Bcl-3. Thus, control of translation may be a general mechanism by which surface adhesion molecules regulate gene expression.


The effect of doxorubicin (adriamycin) on cytoplasmic microtubule system in cardiac cells.
The objective of this study was to determine whether the antineoplastic agent doxorubicin (adriamycin) affects the microtubule system in isolated cardiac cells. Embryonic chick heart cultures were exposed to adriamycin concentrations of 40, 80, or 200 microM for 3, 12, or 24 h before fixation prior to immunofluorescence. Adriamycin produced marked distortions in the normal microtubule system. This was a function of adriamycin concentration and the duration of the exposure to adriamycin. Even when adriamycin was removed from the medium for a 12 h period, cells treated with adriamycin showed abnormalities of the microtubule system. The effect of adriamycin on the microtubule system extends to its reassembly after colchicine. Cells allowed to recover after colchicine in the presence of adriamycin (40 microM) showed impairment in microtubule regeneration. These data indicate that the microtubule system in cardiac myocytes is affected by adriamycin both by damaging microtubule structure and by impairing microtubule reassembly.


Kinetic analysis of urea-inactivation of beta-galactosidase in the presence of galactose.
The effect of galactose on the inactivation of purified beta-galactosidase from the black bean, Kestingiella geocarpa, in 5 M urea at 50 degrees C and at pH 4.5, was determined. Lineweaver-Burk plots of initial velocity data in the presence and absence of urea and galactose were used to determine the relevant K(m) and V(max) values, with p-nitrophenyl beta-D-galactopyranoside (PNPG) as substrate, S. The inactivation data were analysed using the Tsou equation and plots. Plots of ln([P](infinity) - [P](t) ) against time in the presence of urea yielded the inactivation rate constant, A. Plots of A vs [S] at different galactose concentrations were zero order showing that A was independent of [S]. Plots of [P](infinity) vs [S] were used to determine the mode of inhibition of the enzyme by galactose, and slopes and intercepts of the 1/[P](infinity) vs. 1/[S] yielded k(+0) and k '(+0), the microscopic rate constants for the free enzyme and the enzyme-substrate complex, respectively. Plots of k(+0) and k '(+0) vs. galactose concentrations showed that galactose protected the free enzyme and not the enzyme-substrate complex against urea inactivation via a noncompetitive mechanism at low galactose concentrations and a competitive pattern of inhibition at high galactose concentrations. The implication of the different modes of inhibition in protecting the free enzyme was discussed.


A simplified method for freezing and storage of human semen.
The use of 10-cm-long plastic tuberculin syringes for freezing and storage of human semen is introduced. This new method reduces the number of prefreezing and postthawing manipulations to a minimum. Postthawing motility, penetration of spermatozoa into cervical mucus, and results of the hamster ovum test were similar for spermatozoa stored-frozen in tuberculin syringes and for spermatozoa stored-frozen in french straws. The results of the postinsemination test performed in six women were slightly better with fresh semen than with semen stored-frozen in tuberculin syringes. Donor insemination with fresh semen resulted in eight pregnancies per 24 insemination cycles; with semen stored-frozen in tuberculin syringes five pregnancies were achieved in 22 insemination cycles.


Decreased levels of testosterone and gonadotrophins in men with long-standing tetraplegia.
Blood samples were frequently collected during a 24-h period from six tetraplegic men. The results were compared with those of eight able-bodied controls. Previous studies have reported conflicting results regarding the plasma concentrations of testosterone, luteinizing hormone (LH) and follicle-stimulating hormone (FSH) in tetraplegia. The objective of this study was to examine the pituitary-gonadal axis by determining the plasma concentrations and circadian variations of these hormones in men with long-standing tetraplegia. Sunnaas Hospital, Norway. The plasma concentrations of hormones were measured with standardized assays. All three hormones and free testosterone index were decreased in the tetraplegic subjects compared with the able-bodied controls (P<0.05). We also determined the morning levels of hormones with regulatory effects on testosterone, LH and FSH. Whereas plasma leptin was significantly higher in the tetraplegic group, no significant differences in the morning plasma values for insulin, SHBG, GH or IGF-1, or in the 24-h urine concentrations of cortisol were detected between the two groups. The plasma concentration of LH displayed a circadian variation (P<0.05) in the tetraplegic group, but not among the able-bodied. No circadian variation was noted for the plasma concentrations of testosterone and FSH in either group. Our data indicate that, over time, tetraplegic male subjects might be at risk of developing hypogonadism.