Effect of liquid surface area on hydrogen sulfide oxidation during micro-aeration in dairy manure digesters.
Although there are a variety of commercially available biological and chemical treatments for removal of hydrogen sulfide (H2S) from biogas, managing biogas H2S remains a significant challenge for agricultural digesters where labor and operational funds are very limited compared to municipal and industrial digesters. The objectives of this study were to evaluate headspace aeration for reducing H2S levels in low cost plug flow digesters and to characterize the relationship between the liquid surface area and H2S oxidation rates. Experiments with replicate field scale plug flow digesters showed that H2S levels decreased from 3500 ppmv to <100 ppmv when headspace oxygen levels were 0.5 to 1%. Methane production was not affected by aeration rates that resulted in headspace oxygen levels of up to 1%. Pilot scale experiments using 65 to 104 L desulfurization units showed that H2S oxidation rates increased with increases in liquid surface area. These results support the hypothesis that H2S oxidation rates are limited, in part, by the surface area available for oxygen transfer, and can be increased by growth of biofilms containing H2S oxidizing bacteria. Maximum removal rates corresponded to 40 to 100 g S m-2 d-1 of liquid surface area at biogas retention times of 30 to 40 min.


An αII Spectrin-Based Cytoskeleton Protects Large-Diameter Myelinated Axons from Degeneration.
Axons must withstand mechanical forces, including tension, torsion, and compression. Spectrins and actin form a periodic cytoskeleton proposed to protect axons against these forces. However, because spectrins also participate in assembly of axon initial segments (AISs) and nodes of Ranvier, it is difficult to uncouple their roles in maintaining axon integrity from their functions at AIS and nodes. To overcome this problem and to determine the importance of spectrin cytoskeletons for axon integrity, we generated mice with αII spectrin-deficient peripheral sensory neurons. The axons of these neurons are very long and exposed to the mechanical forces associated with limb movement; most lack an AIS, and some are unmyelinated and have no nodes. We analyzed αII spectrin-deficient mice of both sexes and found that, in myelinated axons, αII spectrin forms a periodic cytoskeleton with βIV and βII spectrin at nodes of Ranvier and paranodes, respectively, but that loss of αII spectrin disrupts this organization. Avil-cre;Sptan1f/f mice have reduced numbers of nodes, disrupted paranodal junctions, and mislocalized Kv1 K+ channels. We show that the density of nodal βIV spectrin is constant among axons, but the density of nodal αII spectrin increases with axon diameter. Remarkably, Avil-cre;Sptan1f/f mice have intact nociception and small-diameter axons, but severe ataxia due to preferential degeneration of large-diameter myelinated axons. Our results suggest that nodal αII spectrin helps resist the mechanical forces experienced by large-diameter axons, and that αII spectrin-dependent cytoskeletons are also required for assembly of nodes of Ranvier.SIGNIFICANCE STATEMENT A periodic axonal cytoskeleton consisting of actin and spectrin has been proposed to help axons resist the mechanical forces to which they are exposed (e.g., compression, torsion, and stretch). However, until now, no vertebrate animal model has tested the requirement of the spectrin cytoskeleton in maintenance of axon integrity. We demonstrate the role of the periodic spectrin-dependent cytoskeleton in axons and show that loss of αII spectrin from PNS axons causes preferential degeneration of large-diameter myelinated axons. We show that nodal αII spectrin is found at greater densities in large-diameter myelinated axons, suggesting that nodes are particularly vulnerable domains requiring a specialized cytoskeleton to protect against axon degeneration.


Pretraining NMDA receptor blockade in the basolateral complex, but not the central nucleus, of the amygdala prevents savings of conditional fear.
The acquisition of conditional freezing is abolished by N-methyl-D-aspartate (NMDA) receptor antagonism in the basolateral complex of the amygdala (BLA) during fear conditioning, suggesting that memory formation is prevented. The present study examined whether there is residual memory, or "savings," for fear conditioning in rats trained under amygdaloid NMDA receptor blockade. Rats infused with D,L-2-amino-5-phosphonovalerate (APV) into the BLA or central nucleus of the amygdala (CEA) during fear conditioning did not acquire either auditory or contextual fear conditioning. However, savings of conditional fear was exhibited by rats infused with APV into the CEA but not the BLA. These results suggest that both the BLA and CEA play a critical role in the acquisition of conditional fear but that the BLA is able to process and retain some aspects of aversive memories in the absence of the CEA.


Government spending, recession, and suicide: evidence from Japan.
Austerity has been shown to have an adverse influence on people's mental health and suicide rates. Most existing studies have focused on the governments' reactions to a single event, for example, the Great Recession of 2008. This study focused on significant changes in fiscal policy between 2001 and 2014 in Japan. The size of expenditures by national and local governments decreased dramatically between 2001 and 2006 under the neoliberal reform and then increased after the global economic crisis and the Great East Japan Earthquake. Using the data from 47 prefectures between 2001 and 2014, we tested whether more spending by the local governments was associated with a lower suicide rate in their jurisdiction. We also investigated whether this relationship was particularly salient during a more severe recession. Our analysis revealed that an increase of 1% in the per capita local government expenditures was associated with a decrease of 0.2% in the suicide rates among males and females aged between 40 and 64 and that this correlation was strengthened as the unemployment rate increased, particularly among males. Government's reaction to economic crises can either exacerbate or mitigate the negative impact of the economic recession on people's mental health and suicide rates.


Preoperative somatostatin analogues versus direct transsphenoidal surgery for newly-diagnosed acromegaly patients: a systematic review and meta-analysis using the GRADE system.
Whether the preoperative use of somatostatin analogues (SA) improves surgical outcomes in acromegaly is still a matter of debate. We conducted a systematic review of randomized, controlled trials that compared the short-term outcomes of preoperative use of SA (Pre-SA) with direct TSS (No-SA) for the treatment of newly diagnosed acromegaly. Embase, Pubmed, Lilacs, and Central Cochrane were used as our data sources. The primary outcomes were no need for any adjuvant treatment 3 months after surgery, based on biochemical results (GH nadir after OGTT <1 μg/L and normal IGF-1 for age and gender), quality of life and mortality. The included trials were analyzed using the Grading of Recommendations Assessment, Development, and Evaluation approach. A total of 2.099 references were identified and two reviewers independently screened the titles and abstracts. From the 14 potentially eligible studies, four were included and ten were excluded due to lack of randomization or different outcomes. A pool of 261 patients was randomly assigned to Pre-SA or No-SA. Meta-analysis of IGF1 normalization showed a significant difference in favor of Pre-SA (RR 2.47; 95% CI 1.66, 3.77). Adding a GH nadir on OGTT ≤1 μg/L, we found a RR of 2.15 (95% CI 1.39, 3.33). Quality of evidence for no need of adjuvant postoperative treatment was moderate, but for improving quality of life was very low and for mortality was absent. Pre-SA increases the chance of biochemical control of acromegaly 3 months after TSS in patients harboring GH-secreting pituitary macroadenomas.


Boron Nitride Nanoparticles with a Petal-Like Surface as Anticancer Drug-Delivery Systems.
Nanoparticles (NPs) have a great potential as nanosized drug-delivery carriers. Such systems must safely deliver the drug to the site of the tumor without drug leakage, effectively penetrate inside cancer cells, and provide intracellular drug release. Herein we developed an original and simple method aimed at the fabrication of spherical boron nitride NPs (BNNPs), 100-200 nm in diameter, with peculiar petal-like surfaces via chemical vapor deposition. Such structures were found to be able to absorb a large amount of antitumor drug-killing tumor cells. They revealed low cytotoxicity and rapid cellular uptake. BNNPs were saturated with doxorubicin (DOX) and then dispersed. The BNNPs loaded with DOX (BNNPs-DOX) were stable at neutral pH but effectively released DOX at pH 4.5-5.5. MTT assay and cell growth testing showed that the BNNPs-DOX nanocarriers had been toxic for IAR-6-1 cells. BNNPs loaded with DOX penetrated into the neoplastic IAR-6-1 cells using endocytic pathways, and then DOX released into the cytoplasm and cell nuclei and resulted in cell death.


Identifying the Recognition Site for Selective Trapping of 99TcO4- in a Hydrolytically Stable and Radiation Resistant Cationic Metal-Organic Framework.
Effective and selective removal of 99TcO4- from aqueous solution is highly desirable for both waste partitioning and contamination remediation purposes in the modern nuclear fuel cycle, but is of significant challenge. We report here a hydrolytically stable and radiation-resistant cationic metal-organic framework (MOF), SCU-101, exhibiting extremely fast removal kinetics, exceptional distribution coefficient, and high sorption capacity toward TcO4-. More importantly, this material can selectively remove TcO4- in the presence of large excesses of NO3- and SO42-, as even 6000 times of SO42- in excess does not significantly affect the sorption of TcO4-. These superior features endow that SCU-101 is capable of effectively separating TcO4- from Hanford low-level waste melter off-gas scrubber simulant stream. The sorption mechanism is directly unraveled by the single crystal structure of TcO4--incorporated SCU-101, as the first reported crystal structure to display TcO4- trapped in a sorbent material. A recognition site for the accommodation of TcO4- is visualized and is consistent with the DFT analysis results, while no such site can be resolved for other anions.


Incidence and risk factors of massive subretinal hemorrhage in retinal angiomatous proliferation.
To evaluate the incidence and associated risk factors of massive subretinal hemorrhage (SRH) in patients with retinal angiomatous proliferation (RAP). A total of 187 eyes of 135 treatment-naıve patients diagnosed with RAP were evaluated retrospectively. Clinical records including the time between the initial visit, last anti-vascular endothelial growth factor (VEGF) treatment, last stable examination, and the date of massive SRH were reviewed. Imaging findings including indocyanine green angiography (ICGA) and optical coherence tomography (OCT) were analyzed. Massive SRH developed in 18 eyes (9.6%) a median of 20 months after the initial presentation. Kaplan-Meier survival analysis revealed that the incidence (2.8, 5.8, 13.1, and 21.0% after 1,2,5 and 10 years, respectively) continuously increased. Among 14 eyes with discernable vascular anastomosis on baseline ICGA, 13 (92.8%) showed retinal arteriole involvement. On spectral-domain OCT imaging of the last visit prior to the massive SRH, a layered lamellar tissue complex was noted under the retinal pigment epithelium in 9 of 13 eyes, which was significantly associated with massive SRH[hazard ratio(HR),5.883;P = .010]. The average time between the last stable examination/last injection and the massive SRH was 2 and 5 months, respectively. The patients were treated with anti-VEGF, gas and recombinant tissue plasminogen activator injection; however, all except one eye had visual acuity worse than 20/1000 at the final visit. Massive SRH can occur in RAP in the course of anti-VEGF treatment, resulting in severe vision loss. A proactive dosing regimen may be more appropriate for these RAP eyes.


Iridium Ziegler-type hydrogenation catalysts made from [(1,5-COD)Ir(mu-O2C8H15)](2) and AlEt3: spectroscopic and kinetic evidence for the Ir(n) species present and for nanoparticles as the fastest catalyst.
Ziegler-type hydrogenation catalysts, those made from a group 8-10 transition metal precatalyst and an AlR(3) cocatalyst, are often used for large scale industrial polymer hydrogenation; note that Ziegler-type hydrogenation catalysts are not the same as Ziegler-Natta polymerization catalysts. A review of prior studies of Ziegler-type hydrogenation catalysts (Alley et al. J. Mol. Catal. A: Chem. 2010, 315, 1-27) reveals that a approximately 50 year old problem is identifying the metal species present before, during, and after Ziegler-type hydrogenation catalysis, and which species are the kinetically best, fastest catalysts--that is, which species are the true hydrogenation catalysts. Also of significant interest is whether what we have termed "Ziegler nanoclusters" are present and what their relative catalytic activity is. Reported herein is the characterization of an Ir Ziegler-type hydrogenation catalyst, a valuable model (vide infra) for the Co-based industrial Ziegler-type hydrogenation catalyst, made from the crystallographically characterized [(1,5-COD)Ir(mu-O(2)C(8)H(15))](2) precatalyst plus AlEt(3). Characterization of this Ir model system is accomplished before and after catalysis using a battery of physical methods including Z-contrast scanning transmission electron microscopy (STEM), high resolution (HR)TEM, and X-ray absorption fine structure (XAFS) spectroscopy. Kinetic studies plus Hg(0) poisoning experiments are then employed to probe which species are the fastest catalysts. The main findings herein are that (i) a combination of the catalyst precursors [(1,5-COD)Ir(mu-O(2)C(8)H(15))](2) and AlEt(3) gives catalytically active solutions containing a broad distribution of Ir(n) species ranging from monometallic Ir complexes to nanometer scale, noncrystalline Ir(n) nanoclusters (up to Ir(approximately 100) by Z-contrast STEM) with the estimated mean Ir species being 0.5-0.7 nm, Ir(approximately 4-15) clusters considering the similar, but not identical results from the different analytical methods; furthermore, (ii) the mean Ir(n) species are practically the same regardless of the Al/Ir ratio employed, suggesting that the observed changes in catalytic activity at different Al/Ir ratios are primarily the result of changes in the form or function of the Al-derived component (and not due to significant AlEt(3)-induced changes in initial Ir(n) nuclearity). However (iii), during hydrogenation, a shift in the population of Ir species toward roughly 1.0-1.6 nm, fcc Ir(0)(approximately 40-150), Ziegler nanoclusters occurs with, significantly, (iv) a concomitant increase in catalytic activity. Importantly, and although catalysis by discrete subnanometer Ir species is not ruled out by this study, (v) the increases in activity with increased nanocluster size, plus Hg(0) poisoning studies, provide the best evidence to date that the approximately 1.0-1.6 nm, fcc Ir(0)(approximately 40-150), heterogeneous Ziegler nanoclusters are the fastest catalysts in this industrially related catalytic hydrogenation system (and in the simplest, Ockham's Razor interpretation of the data). In addition, (vi) Ziegler nanoclusters are confirmed to be an unusual, hydrocarbon-soluble, highly coordinatively unsaturated, Lewis-acid containing, and highly catalytically active type of nanocluster for use in other catalytic applications and other areas.


Increases in intracellular calcium triggered by channelrhodopsin-2 potentiate the response of metabotropic glutamate receptor mGluR7.
The metabotropic glutamate receptor 7a (mGluR7a), a heptahelical Galphai/o-coupled protein, has been shown to be important for presynaptic feedback inhibition at central synapses and certain forms of long term potentiation and long term depression. The intracellular C terminus of mGluR7a interacts with calmodulin in a Ca2+-dependent manner, and calmodulin antagonists have been found to abolish presynaptic inhibition of glutamate release in neurons and mGluR7a-induced activation of G-protein-activated inwardly rectifying K+ channel (GIRK) channels in HEK293 cells. Here, we characterized the Ca2+ dependence of mGluR7a signaling in Xenopus oocytes by using channelrhodopsin-2 (ChR2), a Ca2+-permeable, light-activated ion channel for triggering Ca2+ influx, and a GIRK3.1/3.2 concatemer to monitor mGluR7a responses. Application of the agonist (S)-2-amino-4-phosphonobutanoic acid (l-AP4) (1-100 microm) caused a dose-dependent inward current in high K+ solutions due to activation of GIRK channels by G-protein betagamma subunits released from mGluR7a. Elevation of intracellular free Ca2+ by light stimulation of ChR2 markedly increased the amplitude of L-AP4 responses, and this effect was attenuated by the calcium chelator 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid tetrakis (acetoxymethyl ester). l-AP4 responses were potentiated by submembranous [Ca2+] levels within physiological ranges and with a threshold close to resting [Ca2+]i values, as determined by recording the endogenous Xenopus Ca2+-activated chloride conductance. Together, these results show that L-AP4-dependent mGluR7a signaling is potentiated by physiological levels of [Ca2+]i, consistent with a model in which presynaptic mGluR7a acts as a coincidence detector of Ca2+ influx and glutamate release.


A novel lysozyme mutation Phe57Ile associated with hereditary renal amyloidosis.
Variant forms of lysozyme, a ubiquitous bacteriolytic enzyme, are known to lead to hereditary non-neuropathic renal amyloidosis and, so far, three different mutations of the lysozyme gene have been reported. In this study, we report a novel lysozyme variant, Phe57Ile, associated with renal amyloidosis in three patients in one Italian Canadian family. The proband was a 52-year-old woman who developed renal failure at the age of 42 years. Renal biopsy demonstrated replacement of glomeruli by amyloid. Her younger sister and her younger daughter who underwent renal transplantation also had renal amyloidosis. The proband's older daughter and her niece were in good health. To elucidate pathogenesis of this hereditary renal amyloidosis, DNA analyses of the lysozyme gene, including single strand confirmation polymorphism, direct DNA sequence, and restriction fragment length polymorphism analyses, were performed. DNA analyses of the lysozyme gene revealed a T to A transversion at the first position of codon 57 of the lysozyme gene in the proband, her sister, and her affected and unaffected daughters, indicating a replacement of Phe by Ile at residue 57. In addition, DNA sequencing demonstrated a C to A transversion at the second position of codon 70, denoting a replacement of Thr by Asn at residue 70, in the proband's sister and her niece. Thus, the proband's sister is compound heterozygous for the Phe57Ile and Thr70Asn alleles. Distinctive clinical features in patients of this family are nephropathy due to renal amyloidosis. Our results indicate that the novel lysozyme variant Phe57Ile is associated with renal amyloidosis in this family. From our results, a clear relation between the Thr70Asn polymorphism and renal amyloidosis could not be demonstrated.


Mild hypothermia ameliorates ubiquitin synthesis and prevents delayed neuronal death in the gerbil hippocampus.
The purpose of the present study is to determine the effect of mild hypothermia on the synthesis of ubiquitin, an important protein for maintenance of cell viability, in the hippocampal neurons following transient cerebral ischemia. Transient ischemia was induced by occluding both common carotid arteries for 5 minutes. In experiment 1, the animals were divided into four groups according to the rectal and scalp temperatures during ischemia: the normothermia group and the graded hypothermia A, B, and C groups (n = 9 per group). CA1 neuronal density was assessed at 7 days after ischemia. In experiment 2, the animals were divided into two groups designated the normothermia and the hypothermia groups (n = 6 per group). The presence of ubiquitin was examined by immunohistochemistry at 6, 24, and 48 hours after transient ischemia in various regions of the hippocampus. In experiment 1, the mean +/- SEM neuronal density per millimeter was 12 +/- 1 in the normothermia group and 126 +/- 25, 225 +/- 10, and 214 +/- 9 in hypothermia groups A, B, and C, respectively. Mild hypothermia in groups B and C, in which the brain temperature was below 33 degrees C, ameliorated markedly the extent of ischemic neuronal damage in the CA1 sector (p less than 0.01). In experiment 2, ubiquitin immunoreactivity had disappeared in all regions of the hippocampus at 6 hours after ischemia and showed no subsequent recovery in the CA1 pyramidal neurons under normothermic conditions. Under hypothermic conditions, however, it had recovered significantly in the CA1 pyramidal neurons at 24 and 48 hours after ischemia (p less than 0.01). We conclude that mild hypothermia, in which the brain temperature is below 33 degrees C, markedly improves the ischemic delayed neuronal damage in the CA1 sector, and that increased ubiquitin synthesis and protein ubiquitination could be one essential part of the protective mechanism afforded by mild hypothermia against delayed neuronal death.


Genetic factors in drug metabolism.
Patients vary widely in their response to drugs. Having an understanding of the pharmacokinetic and pharmacodynamic properties of various medications is importantwhen assessing ethnic differences in drug response. Genetic factors can account for 20 to 95 percent of patient variability. Genetic polymorphisms for many drug-metabolizing enzymes and drug targets (e.g., receptors) have been identified. Although currently limited to a few pathways, pharmacogenetic testing may enable physicians to understand why patients react differently to various drugs and to make better decisions about therapy. Ultimately, this understanding may shift the medical paradigm to highly individualized therapeutic regimens.


Prevalence and risk factors of arthritis in a middle-aged and older Chinese population: the China health and retirement longitudinal study.
The aims of this study were to estimate the prevalence of arthritis and to identify risk factors of arthritis in a middle-aged and older Chinese adult population. The China Health and Retirement Longitudinal Study (CHARLS) national survey data were used to estimate overall arthritis prevalence and prevalence by age and gender groups taking into account the complex survey design and response rate. The PROC SURVEYLOGISTIC procedure (SAS 9.3; SAS Institute, Cary, NC, USA) was applied to identify factors associated with arthritis using the CHARLS national survey data. Significant factors were further evaluated in the longitudinal CHARLS pilot study. The overall prevalence of arthritis among middle-aged and older Chinese adults was 31.4% (95% CI 30.3, 32.4). Prevalence increased with age. Females had a higher prevalence of arthritis than males in each age group. In the cross-sectional analysis, age, gender, education, BMI, sleep duration, vigorous physical activity and self-reported doctor-diagnosed chronic lung disease, hypertension, chronic liver disease, cardiovascular disease, stroke, chronic kidney disease and chronic digestive disease were associated with arthritis. Age, gender, vigorous physical activity and cardiovascular disease were confirmed to be risk factors of arthritis in the longitudinal analysis. Participants with cardiovascular disease were 1.67 times (95% CI 1.02, 2.74) more likely to have self-reported arthritis in a 4-year period of follow-up in the CHARLS pilot study. Middle-aged and older Chinese adults had a high prevalence of arthritis. Cardiovascular disease is a novel risk factor for arthritis in this population.


Accumulation of advanced glycation end products potentiate human retinal capillary endothelial cells mediated diabetic retinopathy.
The major ophthalmic complication in patients with diabetes is diabetic retinopathy (DR), which is one of the major eye diseases that causes blindness. It is well established that the occurrence and duration of DR is positively correlated with duration of diabetes. Advanced glycation end product (AGE) accumulation in patients with diabetes is one factor that leads to the development of DR. However, the underlying mechanisms remain unclear. In the present study, the role of phosphoinositide 3‑kinase/protein kinase B (Akt) signaling in AGE‑induced DR development was investigated. An in vitro experimental system was used to study the effects of AGEs on human retinal capillary endothelial cells (HRCECs) and Müller cells. Flow cytometry, MTT, western blotting and BrdU incorporation assays were performed. Reverse transcription‑quantitative polymerase chain reaction was used to measure the expression of angiogenesis‑associated genes. Functional assays of angiogenesis, including HRCEC invasion and tube formation assays. It was demonstrated that the expression of receptor for AGEs was upregulated in HRCECs and Müller cells following treatment with AGEs. AGE treatment did not affect Müller cell viability, but enhanced HRCEC viability. Akt inhibition increased cell apoptosis and death in HRCECs. AGE treatment upregulated the expression of pro‑angiogenic genes, which was suppressed by Akt inhibitor treatment. In addition, Akt inhibitor treatment suppressed HRCEC invasion and tube formation ability. The present study suggested that Akt‑mediated signaling may serve critical roles in the development of DR due to the accumulation of AGEs. Akt may be a potential therapeutic target in DR.


Subtle deficits in instrumental activities of daily living in subtypes of mild cognitive impairment.
Greater cognitive and functional deficits in mild cognitive impairment (MCI) are associated with higher rates of dementia. We explored the relationship between these factors by comparing instrumental activities of daily living (IADLs) among cognitive subtypes of MCI and examining associations between IADL and neuropsychological indices. We analyzed data from 1,108 MCI and 3,036 normal control subjects included in the National Alzheimer's Coordinating Center Uniform Data Set who were assessed with the Functional Activities Questionnaire (FAQ). IADL deficits were greater in amnestic than nonamnestic MCI, but within these subgroups, did not differ between those with single or multiple domains of cognitive impairment. FAQ indices correlated significantly with memory and processing speed/executive function. IADL deficits are present in both amnestic MCI and nonamnestic MCI but are not related to the number of impaired cognitive domains. These cross-sectional findings support previous longitudinal reports suggesting that cognitive and functional impairments in MCI may be independently associated with dementia risk.


Construction and validation of a low-cost surgical trainer based on iPhone technology for training laparoscopic skills.
In this article, we describe the construction and validation of a laparoscopic trainer using an iPhone 5 and a plastic document holder case. The abdominal cavity was simulated with a clear plastic document holder case. On 1 side of the case, 2 holes for entry of laparoscopic instruments were drilled. We added a window to place the camera of the iPhone, which works as our camera of the trainer. Twenty residents carried out 4 tasks using the iPhone Trainer and a physical laparoscopic trainer. The time of all tasks were analyzed with a simple paired t test. The construction of the trainer took 1 hour, with a cost of <US$90. Results showed no significant differences in time for the 4 tasks performed in both the trainers. iPhone Trainer is a reusable and fully functional device that allows surgeons to practice their skills anywhere and at their own pace.


Energy determinants GAPDH and NDPK act as genetic modifiers for hepatocyte inclusion formation.
Genetic factors impact liver injury susceptibility and disease progression. Prominent histological features of some chronic human liver diseases are hepatocyte ballooning and Mallory-Denk bodies. In mice, these features are induced by 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC) in a strain-dependent manner, with the C57BL and C3H strains showing high and low susceptibility, respectively. To identify modifiers of DDC-induced liver injury, we compared C57BL and C3H mice using proteomic, biochemical, and cell biological tools. DDC elevated reactive oxygen species (ROS) and oxidative stress enzymes preferentially in C57BL livers and isolated hepatocytes. C57BL livers and hepatocytes also manifested significant down-regulation, aggregation, and nuclear translocation of glyceraldehyde 3-phosphate dehydrogenase (GAPDH). GAPDH knockdown depleted bioenergetic and antioxidant enzymes and elevated hepatocyte ROS, whereas GAPDH overexpression decreased hepatocyte ROS. On the other hand, C3H livers had higher expression and activity of the energy-generating nucleoside-diphosphate kinase (NDPK), and knockdown of hepatocyte NDPK augmented DDC-induced ROS formation. Consistent with these findings, cirrhotic, but not normal, human livers contained GAPDH aggregates and NDPK complexes. We propose that GAPDH and NDPK are genetic modifiers of murine DDC-induced liver injury and potentially human liver disease.


A distance-performance trade-off in the phenotypic basis of dispersal.
Across taxa, individuals vary in how far they disperse, with most individuals staying close to their origin and fewer dispersing long distances. Costs associated with dispersal (e.g., energy, risk) are widely believed to trade off with benefits (e.g., reduced competition, increased reproductive success) to influence dispersal propensity. However, this framework has not been applied to understand variation in dispersal distance, which is instead generally attributed to extrinsic environmental factors. We alternatively hypothesized that variation in dispersal distances results from trade-offs associated with other aspects of locomotor performance. We tested this hypothesis in the stream salamander Gyrinophilus porphyriticus and found that salamanders that dispersed farther in the field had longer forelimbs but swam at slower velocities under experimental conditions. The reduced swimming performance of long-distance dispersers likely results from drag imposed by longer forelimbs. Longer forelimbs may facilitate moving longer distances, but the proximate costs associated with reduced swimming performance may help to explain the rarity of long-distance dispersal. The historical focus on environmental drivers of dispersal distances misses the importance of individual traits and associated trade-offs among traits affecting locomotion.


Click chemistry approach for imaging intracellular and intratissue distribution of curcumin and its nanoscale carrier.
This study was aimed at developing a fluorescence imaging approach to simultaneously characterize the delivery and distribution of a bioactive molecule, curcumin, and its micelle based nanoscale carrier in cells and tissue models. To enable imaging of curcumin, a monoalkyne derivative of curcumin was synthesized and purified using LC-MS. Intracellular uptake of curcumin was characterized using a click chemistry reaction between a monoalkyne modified curcumin and Alexa-488 azide fluorescent dye in cells and tissues. Fluorescence images of cells and tissues incubated with monoalkyne curcumin showed specific detection of intracellular delivered monoalkyne curcumin using the click chemistry reaction. The fluorescence imaging results also demonstrated significant improvement in detection sensitivity of intracellular delivered curcumin as compared to measurements based on native fluorescence of unmodified curcumin. Intracellular uptake of monoalkyne curcumin was characterized as a function of incubation time and concentration. The results show a rapid uptake of monoalkyne curcumin during the first 4 h of incubation. Modification of curcumin to its monoalkyne derivative did not impact its apoptotic activity in cancer cells. DSPE-PEG micelles labeled with Alexa-647 were selected as a representative nanoscale carrier to enhance the solubility and delivery of monoalkyne curcumin. Fluorescence images of cells and tissues incubated with fluorescently labeled micelles containing monoalkyne curcumin clearly illustrate significant differences in intracellular and intratissue localization of DSPE-PEG and encapsulated monoalkyne curcumin. The imaging approach developed in this study can be used to understand delivery and distribution of diverse bioactive compounds and their nanocarrier systems as well as in situ measurement of interactions of bioactives with cellular and tissue targets.


Careful with understudied phyla: the case of chaetognath.
A recent study by Barthélémy et al. described a set of ribosomal protein (RP) genes extracted from a collection of expressed sequence tags (ESTs) of the chaetognath (arrow worm) Spadella cephaloptera. Three main conclusions were drawn in this paper. First, the authors stated that RP genes present paralogous copies, which have arisen through allopolyploidization. Second, they reported two alternate nucleotide stretches conserved within the 5' untranslated regions (UTR) of multiple ribosomal cDNAs and they suggested that these motifs are involved in the differential transcriptional regulation of paralogous RP genes. Third, they claimed that the phylogenetic position of chaetognaths could not be accurately inferred from a RP dataset because of the persistence of two problems: a long branch attraction (LBA) artefact and a compositional bias. We reconsider here the results described in Barthélémy et al. and question the evidence on which they are based. We find that their evidence for paralogous copies relies on faulty PCR experiments since they attempted to amplify DNA fragments absent from the genomic template. Our PCR experiments proved that the conserved motifs in 5'UTRs that they targeted in their amplifications are added post-transcriptionally by a trans-splicing mechanism. Then, we showed that the lack of phylogenetic resolution observed by these authors is due to limited taxon sampling and not to LBA or to compositional bias. A ribosomal protein dataset thus fully supports the position of chaetognaths as sister group of all other protostomes. This reinterpretation demonstrates that the statements of Barthélémy et al. should be taken with caution because they rely on inaccurate evidence. The genomic study of an unconventional model organism is a meaningful approach to understand the evolution of animals. However, the previous study came to incorrect conclusions on the basis of experiments that omitted validation procedures.


[Clinical Analysis of Autologous Peripheral Blood Hematopoietic Stem Cells Mobilization and Collection in Lymphoma Patients].
To analyze the factors influencing collection of autologous peripheral blood hematopoietic stem cells in lymphoma patients. Clinical data of 74 patients who received autologous peripheral blood hematopoietic stem cells mobilization and collection in the 940th Hospital of Joint Logistic Support Force of PLA from April 2009 to April 2021 were collected. The effects of gender, age, disease type, stage, course of disease, chemotherapy cycle number, relapse, radiotherapy, disease status and blood routine indexes on the day of collection on peripheral blood hematopoietic stem cell collection were analyzed. The success rate of collection was 95.9%(71/74), and the excellent rate of collection was 71.6%(53/74). There was a significantly statistical differentce in the number of CD34+ cells in grafts collected from patients with chemotherapy cycle ≤6 and >6 [(9.1±5.2)×106/kg vs (6.4±3.7)×106/kg, P=0.031]. The number of CD34+ cells in the first collection was positively correlated with WBC count, hemoglobin, platelet count, neutrophil count, lymphocyte count, monocyte count and hematocrit value on the day of collection ( r value was 0.424,0.486,0.306,0.289,0.353,0.428,0.528, respectively). WBC count, hemoglobin, monocyte count and hematocrit value have higher predictive value for the first collection of CD34+ cells. The area under the receiver operating characteristic was 0.7061,0.7845,0.7319,0.7848, respectively. Low dose CTX and VP16 chemotherapy combined with G-CSF can effectively mobilize autologous peripheral blood stem cells. The cycle number of chemotherapy relates to the collection of autologous peripheral blood stem cells. After mobilization, the success of the first collection can be better predicted by the blood routine indexes.


Pannexin 1 channels play essential roles in urothelial mechanotransduction and intercellular signaling.
Urothelial cells respond to bladder distension with ATP release, and ATP signaling within the bladder and from the bladder to the CNS is essential for proper bladder function. In other cell types, pannexin 1 (Panx1) channels provide a pathway for mechanically-induced ATP efflux and for ATP-induced ATP release through interaction with P2X7 receptors (P2X7Rs). We report that Panx1 and P2X7R are functionally expressed in the bladder mucosa and in immortalized human urothelial cells (TRT-HU1), and participate in urothelial ATP release and signaling. ATP release from isolated rat bladders induced by distention was reduced by the Panx1 channel blocker mefloquine (MFQ) and was blunted in mice lacking Panx1 or P2X7R expression. Hypoosmotic shock induced YoPro dye uptake was inhibited by MFQ and the P2X7R blocker A438079 in TRT-HU1 cells, and was also blunted in primary urothelial cells derived from mice lacking Panx1 or P2X7R expression. Rinsing-induced mechanical stimulation of TRT-HU1 cells triggered ATP release, which was reduced by MFQ and potentiated in low divalent cation solution (LDPBS), a condition known to enhance P2X7R activation. ATP signaling evaluated as intercellular Ca2+ wave radius was significantly larger in LDPBS, reduced by MFQ and by apyrase (ATP scavenger). These findings indicate that Panx1 participates in urothelial mechanotransduction and signaling by providing a direct pathway for mechanically-induced ATP release and by functionally interacting with P2X7Rs.


Psychotherapy in consultation-liaison psychiatry.
Psychotherapy in the Consultation-Liaison (C-L) setting is shaped by the realities of the patient's situation, since all patients referred are dealing with physical illness. The patient's state of physical and mental health will determine both the type of therapeutic work possible and the focus of such work. Tailoring the therapeutic intervention to the patient's specific needs and flexibility in altering and adapting therapeutic strategies over time in line with the patient's changing needs are essential. Although periods of treatment may range from single session to long term, supportive, insight oriented, group, family, cognitive and behavioral techniques have all been used successfully in a C-L setting, with measurable impact on well-being. Psychotherapeutic work in C-L is unique in that the focus of the therapist extends beyond the patient and family to include all caregivers, including other health care professionals, in line with the biopsychosocial model.


Offline modeling for product quality prediction of mineral processing using modeling error PDF shaping and entropy minimization.
This paper presents a novel offline modeling for product quality prediction of mineral processing which consists of a number of unit processes in series. The prediction of the product quality of the whole mineral process (i.e., the mixed concentrate grade) plays an important role and the establishment of its predictive model is a key issue for the plantwide optimization. For this purpose, a hybrid modeling approach of the mixed concentrate grade prediction is proposed, which consists of a linear model and a nonlinear model. The least-squares support vector machine is adopted to establish the nonlinear model. The inputs of the predictive model are the performance indices of each unit process, while the output is the mixed concentrate grade. In this paper, the model parameter selection is transformed into the shape control of the probability density function (PDF) of the modeling error. In this context, both the PDF-control-based and minimum-entropy-based model parameter selection approaches are proposed. Indeed, this is the first time that the PDF shape control idea is used to deal with system modeling, where the key idea is to turn model parameters so that either the modeling error PDF is controlled to follow a target PDF or the modeling error entropy is minimized. The experimental results using the real plant data and the comparison of the two approaches are discussed. The results show the effectiveness of the proposed approaches.


A continuous professional development process for first-year pharmacy students.
To develop, pilot test, and evaluate a continuous professional development (CPD) process for first-year pharmacy (P1) students. Students and faculty members were introduced to the important elements of the CPD process via a live training program. Students completed the year-long 4-step CPD cycle by identifying a learning objective, creating a plan, completing the learning activity, evaluating their learning outcome, documenting each step, and meeting with their faculty advisor for feedback and advice. Seventy-five first-year students (100%) successfully completed the CPD process during the 2009-2010 academic year. The students spent an average of 7 hours (range 2 to 20 hours) on the CPD process. The majority of faculty members (83%) completing the survey instrument found the process valuable for the students and would like to see the program continued. Integrating a CPD requirement for students in a college or school of pharmacy is feasible and valuable to students' developing life-long learning skills. Effective and frequent training of faculty members and students is a key element in the CPD process.


Temporal frequency tuning of cortical face-sensitive areas for individual face perception.
In a highly dynamic visual environment the human brain needs to rapidly differentiate complex visual patterns, such as faces. Here, we defined the temporal frequency tuning of cortical face-sensitive areas for face discrimination. Six observers were tested with functional magnetic resonance imaging (fMRI) when the same or different faces were presented in blocks at 11 frequency rates (ranging from 1 to 12 Hz). We observed a larger fMRI response for different than same faces - the repetition suppression/adaptation effect - across all stimulation frequency rates. Most importantly, the magnitude of the repetition suppression effect showed a typical Gaussian-shaped tuning function, peaking on average at 6 Hz for all face-sensitive areas of the ventral occipito-temporal cortex, including the fusiform and occipital "face areas" (FFA and OFA), as well as the superior temporal sulcus. This effect was due both to a maximal response to different faces in a range of 3 to 6 Hz and to a sharp drop of the blood oxygen level dependent (BOLD) signal from 6 Hz onward when the same face was repeated during a block. These observations complement recent scalp EEG observations (Alonso-Prieto et al., 2013), indicating that the cortical face network can discriminate each individual face when these successive faces are presented every 160-170 ms. They also suggest that a relatively fast 6 Hz rate may be needed to isolate the contribution of high-level face perception processes during behavioral discrimination tasks. Finally, these findings carry important practical implications, allowing investigators to optimize the stimulation frequency rates for observing the largest repetition suppression effects to faces and other visual forms in the occipito-temporal cortex.


Introduction of Large Sequence Inserts by CRISPR-Cas9 To Create Pathogenicity Mutants in the Multinucleate Filamentous Pathogen Sclerotinia sclerotiorum.
The necrotrophic fungal plant pathogen Sclerotinia sclerotiorum is responsible for substantial global crop losses annually resulting in localized food insecurity and loss of livelihood. Understanding the basis of this broad-host-range and aggressive pathogenicity is hampered by the quantitative nature of both host resistance and pathogen virulence. To improve this understanding, methods for efficient functional gene characterization that build upon the existing complete S. sclerotiorum genome sequence are needed. Here, we report on the development of a clustered regularly interspaced short palindromic repeat (CRISPR)-CRISPR-associated protein 9 (CRISPR-Cas9)-mediated strategy for creating gene disruption mutants and the application of this technique for exploring roles of known and hypothesized virulence factors. A key finding of this research is that transformation with a circular plasmid encoding Cas9, target single guide RNA (sgRNA), and a selectable marker resulted in a high frequency of targeted, insertional gene mutation. We observed that 100% of the mutants integrated large rearranged segments of the transforming plasmid at the target site facilitated by the nonhomologous end joining (NHEJ) repair pathway. This result was confirmed in multiple target sites within the same gene in three independent wild-type isolates of S. sclerotiorum and in a second independent gene. Targeting the previously characterized Ssoah1 gene allowed us to confirm the loss-of-function nature of the CRISPR-Cas9-mediated mutants and explore new aspects of the mutant phenotype. Applying this technology to create mutations in a second previously uncharacterized gene allowed us to determine the requirement for melanin accumulation in infection structure development and function.IMPORTANCE Fungi that cause plant diseases by rotting or blighting host tissue with limited specificity remain among the most difficult to control. This is largely due to the quantitative nature of host resistance and a limited understanding of fungal pathogenicity. A mechanistic understanding of pathogenicity requires the ability to manipulate candidate virulence genes to test hypotheses regarding their roles in disease development. Sclerotinia sclerotiorum is among the most notorious of these so-called broad-host-range necrotrophic plant pathogens. The work described here provides a new method for rapidly constructing gene disruption vectors to create gene mutations with high efficiency compared with existing methods. Applying this method to characterize gene functions in S. sclerotiorum, we confirm the requirement for oxalic acid production as a virulence factor in multiple isolates of the fungus and demonstrate that melanin accumulation is not required for infection. Using this approach, the pace of functional gene characterization and the understanding of pathogenicity and related disease resistance will increase.


DOVIS: an implementation for high-throughput virtual screening using AutoDock.
Molecular-docking-based virtual screening is an important tool in drug discovery that is used to significantly reduce the number of possible chemical compounds to be investigated. In addition to the selection of a sound docking strategy with appropriate scoring functions, another technical challenge is to in silico screen millions of compounds in a reasonable time. To meet this challenge, it is necessary to use high performance computing (HPC) platforms and techniques. However, the development of an integrated HPC system that makes efficient use of its elements is not trivial. We have developed an application termed DOVIS that uses AutoDock (version 3) as the docking engine and runs in parallel on a Linux cluster. DOVIS can efficiently dock large numbers (millions) of small molecules (ligands) to a receptor, screening 500 to 1,000 compounds per processor per day. Furthermore, in DOVIS, the docking session is fully integrated and automated in that the inputs are specified via a graphical user interface, the calculations are fully integrated with a Linux cluster queuing system for parallel processing, and the results can be visualized and queried. DOVIS removes most of the complexities and organizational problems associated with large-scale high-throughput virtual screening, and provides a convenient and efficient solution for AutoDock users to use this software in a Linux cluster platform.


Expression of Cystatin C in human stomach neoplasms.
Cystatin C is a member of the cysteine protease inhibitor family and functions to decrease protease production. A recent study showed that it is aberrantly expressed in many malignant tumors in association with tumor invasion and metastasis. Our study aimed to detect Cystatin C expression in stomach neoplasm tissues and adjacent reparative normal tissues. Samples of cancerous and non-cancerous stomach tissue were obtained via surgery as matched pairs from 12 patients with stomach neoplasms and preserved in paraffin. The expression of Cystatin C in the samples was investigated by immunohistochemistry. Fisher's exact test was used to analyze the relationship between stomach neoplasms and adjacent normal tissues. Additionally, mRNA was extracted and analysed by reverse transcriptase-polymerase chain reaction. The intensity of Cystatin C immunostaining was increased in the tumor tissues compared to the adjacent normal tissues. Cystatin C mRNA expression was increased in stomach neoplasms compared to the normal tissues (p<0.05). The results indicate that the expression of Cystatin C may serve as a marker for stomach neoplasms.


In vivo ethanol decreases phosphorylated MAPK and p70S6 kinase in the developing rat brain.
Exposure to ethanol during pregnancy is detrimental to fetal development, and individuals affected by the fetal alcohol syndrome present a number of CN system dysfunctions including microencephaly and mental retardation. Recently, it has been suggested that ethanol-induced inhibition of glial cell proliferation may be relevant in the causation of microencephaly. In this study, we measured the developmental changes of MAPK (ERKl/2) and p70S6 kinase, which are considered to play a prominent role in cell proliferation, and their phosphorylated proteins in rat brain, and examined the effects of in vivo ethanol administration. MAPK and phospho-MAPK increased gradually after birth, and reached adult levels on postnatal day 21. In contrast, levels of both p70S6 kinase and phospho-p70S6 kinase decreased after birth. Exposure to ethanol (2-6 g/kg, from postnatal day 4 to 7) had no effects on MAPK or p70S6 kinase levels, but caused a dose-dependent decrease of both phosphoproteins. These results suggest that phosphorylation of MAPK and p70S6 kinase may represent relevant targets for the developmental neurotoxicity of ethanol, and may be involved in microencephaly.


Link between cell apical morphology and H+ secretion in salamander small intestine.
To determine the morphological basis for the cellular secretion of H+ by in vitro salamander jejunum the transepithelial proton flux (JH) was measured under different conditions and compared with the ultrastructure of the villus absorptive cells using transmission electron microscopy in conjunction with morphometric analysis. Norepinephrine (NE) stimulated JH and significantly increased the density of small cytoplasmic vesicular and tubular profiles in the region immediately below the apical plasma membrane of the absorptive cells. Cytoplasmic tubular elements were often observed in direct contact with the plasma membrane at the base of the microvilli, suggesting that expansion of the plasma membrane had occurred. The alpha 2-antagonist, yohimbine, blocked NE-stimulated H+ secretion and reduced the number of tubulovesicles. Theophylline reduced the rate of spontaneous acid secretion and the density of apical tubules and vesicles. Exposure to horseradish peroxidase revealed a physical connection between the extracellular space and the apical tubulovesicular compartment. The direct relationship between the number of apical vesicular and tubular profiles and JH suggests that the luminal membrane plays a fundamental role in the cellular events in H+ secretion. The membranes of the tubules and vesicles may be elements of a dynamic pool of transporting membranes under the control of exogenous effectors such as catecholamines.


Characterization of the Hypothalamic-Pituitary-Adrenal-Axis in Familial Longevity under Resting Conditions.
The hypothalamic-pituitary-adrenal (HPA)-axis is the most important neuro-endocrine stress response system of our body which is of critical importance for survival. Disturbances in HPA-axis activity have been associated with adverse metabolic and cognitive changes. Humans enriched for longevity have less metabolic and cognitive disturbances and therefore diminished activity of the HPA axis may be a potential candidate mechanism underlying healthy familial longevity. Here, we compared 24-h plasma ACTH and serum cortisol concentration profiles and different aspects of the regulation of the HPA-axis in offspring from long-lived siblings, who are enriched for familial longevity and age-matched controls. Case-control study within the Leiden Longevity study cohort consisting of 20 middle-aged offspring of nonagenarian siblings (offspring) together with 18 partners (controls). During 24 h, venous blood was sampled every 10 minutes for determination of circulatory ACTH and cortisol concentrations. Deconvolution analysis, cross approximate entropy analysis and ACTH-cortisol-dose response modeling were used to assess, respectively, ACTH and cortisol secretion parameters, feedforward and feedback synchrony and adrenal gland ACTH responsivity. Mean (95% Confidence Interval) basal ACTH secretion was higher in male offspring compared to male controls (645 (324-1286) ngl/L/24 h versus 240 (120-477) ng/L/24 h, P = 0.05). Other ACTH and cortisol secretion parameters did not differ between offspring and controls. In addition, no significant differences in feedforward and feedback synchrony and adrenal gland ACTH responsivity were observed between groups. These results suggest that familial longevity is not associated with major differences in HPA-axis activity under resting conditions, although modest, sex-specific differences may exist between groups that might be clinically relevant.


To update or not to update the ESCMID guidelines for the diagnosis and treatment of biofilm infections - That is the question! The opinion of the ESGB board.
The work on the ESGB guidelines for diagnosis and treatment of biofilm infections began in 2012 and the result was published in 2014. The guidelines have been and still are frequently cited in the literature proving its usefulness for people working with biofilm infections. At the ESGB Biofilm conference in Mallorca 2022 (Eurobiofilms2022) the board of the ESGB decided to evaluate the 2014-guidelines and relevant publications since 2014 based on a lecture given at the Eurobiofilms2022. The Delphi method for working on production of guidelines and the current ESCMID rules for guidelines are presented. The criteria for evaluation of relevant literature are very strict and especially for treatment, most clinicians and regulatory authorities require convincing results from Level I (randomized controlled trials) publications to justify changes of treatments. The relevant new biofilm literature and the relevant biofilm presentations from the Eurobiofilms meetings and ECCMID conferences was used for evaluating the contemporary relevance of the ESGB 2014 guidelines. Several infectious diseases have been recognized as biofilm infections since 2014, but the diagnostic methods and therapeutic strategies are still the same as recommended in the 2014 ESGB guidelines which are summarized in this opinion paper. Some promising new in vitro and in vivo (animal experiments) observations and reports for therapy of biofilm infections are mentioned, but they still await clinical trials. The interim opinion at the present time (2022) is therefore, that the guidelines do not need revision now, but there is a need for survey articles discussing new methods of diagnosis and treatment of biofilm infections in order - hopefully - to give inspiration to conduct clinical trials which may lead to progress in diagnosis and treatment of patients with biofilm infections.


Lack of understanding of acute HIV infection among newly-infected persons-implications for prevention and public health: The NIMH Multisite Acute HIV Infection Study: II.
Acute/early HIV infection is a period of high HIV transmission. Consequently, early detection of HIV infection and targeted HIV prevention could prevent a significant proportion of new transmissions. As part of an NIMH-funded multisite study, we used in-depth interviews to explore understandings of acute HIV infection (AHI) among 34 individuals diagnosed with acute/early HIV infection in six US cities. We found a marked lack of awareness of AHI-related acute retroviral symptoms and a lack of clarity about AHI testing methods. Most participants knew little about the meaning and/or consequences of AHI, particularly that it is a period of elevated infectiousness. Over time and after the acute stage of infection, many participants acquired understanding of AHI from varied sources, including the Internet, HIV-infected friends, and health clinic employees. There is a need to promote targeted education about AHI to reduce the rapid spread of HIV associated with acute/early infection within communities at risk for HIV.


Design of a robust fuzzy controller for the arc stability of CO(2) welding process using the Taguchi method.
CO(2) welding is a complex process. Weld quality is dependent on arc stability and minimizing the effects of disturbances or changes in the operating condition commonly occurring during the welding process. In order to minimize these effects, a controller can be used. In this study, a fuzzy controller was used in order to stabilize the arc during CO(2) welding. The input variable of the controller was the Mita index. This index estimates quantitatively the arc stability that is influenced by many welding process parameters. Because the welding process is complex, a mathematical model of the Mita index was difficult to derive. Therefore, the parameter settings of the fuzzy controller were determined by performing actual control experiments without using a mathematical model of the controlled process. The solution, the Taguchi method was used to determine the optimal control parameter settings of the fuzzy controller to make the control performance robust and insensitive to the changes in the operating conditions.


Raising the speed limits for 4D fluorescence microscopy.
Three-dimensional time-lapse (4D) fluorescence microscopy is becoming a routine experimental tool. This article summarizes current technologies, and describes a new method for speeding image acquisition during 4D confocal microscopy.


Galanin-like immunoreactivity within the primate basal forebrain: differential staining patterns between humans and monkeys.
Galanin-like immunoreactivity (GAL-ir) was examined within the basal forebrain and adjacent regions of eight young adult New World monkeys (Cebus apella), one aged Old World monkey (Macaca mulatta), and eight humans without clinical or pathological evidence of neurological disease. All monkeys demonstrated similar patterns of immunoreactive profiles characterized by a continuum of GAL-ir magnocellular neurons located within the medial septum, diagonal band nuclei, and nucleus basalis. Colocalization experiments revealed that most (greater than 90%) of GAL-ir basal forebrain neurons also expressed the receptor for nerve growth factor (NGFR), an excellent marker for primate cholinergic basal forebrain neurons. A few smaller parvicellular GAL-ir neurons were also observed within the monkey basal forebrain. In contrast, identical cytochemical experiments revealed that virtually none of the magnocellular neurons within the basal forebrain of humans were GAL-ir. Rather, a network of GAL-containing fibers and terminal-like profiles were observed encompassing the magnocellular cholinergic neurons in humans. This immunohistochemical species difference does not appear to be mediated by procedural or technical factors since human brains contained numerous GAL-ir perikarya and fibers within adjacent regions including the bed nucleus of the stria terminalis and medial hypothalamus. These data demonstrate that there is a prominent phylogenetic transformation in primates with respect to the processing of GAL-mediated information. This species difference potentially relates to the severe basal forebrain degeneration reported in human dementias and illustrates the possible need for a reevaluation of the use of monkeys as an animal model of human basal forebrain-related cognitive dysfunction.


Genomic rearrangements in trypanosomatids: an alternative to the "one gene" evolutionary hypotheses?
Most molecular trees of trypanosomatids are based on point mutations within DNA sequences. In contrast, there are very few evolutionary studies considering DNA (re) arrangement as genetic characters. Waiting for the completion of the various parasite genome projects, first information may already be obtained from chromosome size-polymorphism, using the appropriate algorithms for data processing. Three illustrative models are presented here. First, the case of Leishmania (Viannia) braziliensis/L. (V.) peruviana is described. Thanks to a fast evolution rate (due essentially to amplification/deletion of tandemly repeated genes), molecular karyotyping seems particularly appropriate for studying recent evolutionary divergence, including eco-geographical diversification. Secondly, karyotype evolution is considered at the level of whole genus Leishmania. Despite the fast chromosome evolution rate, there is qualitative congruence with MLEE- and RAPD-based evolutionary hypotheses. Significant differences may be observed between major lineages, likely corresponding to major and less frequent rearrangements (fusion/fission, translocation). Thirdly, comparison is made with Trypanosoma cruzi. Again congruence is observed with other hypotheses and major lineages are delineated by significant chromosome rearrangements. The level of karyotype polymorphism within that "species" is similar to the one observed in "genus" Leishmania. The relativity of the species concept among these two groups of parasites is discussed.


Increased EID1 nuclear translocation impairs synaptic plasticity and memory function associated with pathogenesis of Alzheimer's disease.
Though loss of function in CBP/p300, a family of CREB-binding proteins, has been causally associated with a variety of human neurological disorders, such as Rubinstein-Taybi syndrome, Huntington's disease and drug addiction, the role of EP300 interacting inhibitor of differentiation 1 (EID1), a CBP/p300 inhibitory protein, in modulating neurological functions remains completely unknown. Through the examination of EID1 expression and cellular distribution, we discovered that there is a significant increase of EID1 nuclear translocation in the cortical neurons of Alzheimer's disease (AD) patient brains compared to that of control brains. To study the potential effects of EID1 on neurological functions associated with learning and memory, we generated a transgenic mouse model with a neuron-specific expression of human EID1 gene in the brain. Overexpression of EID1 led to an increase in its nuclear localization in neurons mimicking that seen in human AD brains. The transgenic mice had a disrupted neurofilament organization and increase of astrogliosis in the cortex and hippocampus. Furthermore, we demonstrated that overexpression of EID1 reduced hippocampal long-term potentiation and impaired spatial learning and memory function in the transgenic mice. Our results indicated that the negative effects of extra nuclear EID1 in transgenic mouse brains are likely due to its inhibitory function on CBP/p300 mediated histone and p53 acetylation, thus affecting the expression of downstream genes involved in the maintenance of neuronal structure and function. Together, our data raise the possibility that alteration of EID1 expression, particularly the increase of EID1 nuclear localization that inhibits CBP/p300 activity in neuronal cells, may play an important role in AD pathogenesis.


MLC-inhibiting serum in mice after transfusion with 3 M KCl-extracted soluble antigen.
Balb/c (H-2d) mice were transfused weekly with 3 M KCl-extracted soluble antigen prepared from splenocytes of C3H/HeJ (C3H)(H-3k) mice. One week after each transfusion, spleen and serum samples were collected from transfused mice and pooled. The serum was absorbed with erythrocytes and spleen cells from C3H mice and heat inactivated. Spleen cells from transfused mice were tested for proliferative responses in mixed lymphocyte culture (MLC) against stimulator cells from the antigen donor C3H or from third party SJL (H-2s) mice. The proliferative responses of lymphocytes from soluble-antigen-transfused Balb/c mice to stimulator cells from C3H and SJL mice were not suppressed. Furthermore, suppressor cells could not be demonstrated in spleens of transfused mice in in vitro coculture experiments. The MLC inhibition test was utilized to investigate the presence of MLC-inhibiting serum from transfused mice. The results demonstrate that serum capable of inhibiting responses of Balb/c mice were induced after three weekly injections of soluble antigen and that this inhibition in MLC was specific for the stimulator cells from the antigen donor C3H mice. These findings differ from our studies using whole blood transfusions where (1) MLC inhibiting antibodies developed in Balb/c mice after only one transfusion of C3H whole blood, and (2) serum from blood transfused mice achieve greater inhibition than soluble-antigen-induced serum. These results suggest that although soluble antigen is capable of inducing MLC inhibiting serum, the kinetics of this induction may be different from transfusion with whole blood.


A new simple /spl infin/OH neuron model as a biologically plausible principal component analyzer.
A new approach to unsupervised learning in a single-layer neural network is discussed. An algorithm for unsupervised learning based upon the Hebbian learning rule is presented. A simple neuron model is analyzed. A dynamic neural model, which contains both feed-forward and feedback connections between the input and the output, has been adopted. The, proposed learning algorithm could be more correctly named self-supervised rather than unsupervised. The solution proposed here is a modified Hebbian rule, in which the modification of the synaptic strength is proportional not to pre- and postsynaptic activity, but instead to the presynaptic and averaged value of postsynaptic activity. It is shown that the model neuron tends to extract the principal component from a stationary input vector sequence. Usually accepted additional decaying terms for the stabilization of the original Hebbian rule are avoided. Implementation of the basic Hebbian scheme would not lead to unrealistic growth of the synaptic strengths, thanks to the adopted network structure.


Permeability characteristics and osmotic sensitivity of rhesus monkey (Macaca mulatta) oocytes.
Permeability characteristics and sensitivity to osmotic shock are principal parameters that are important to derive procedures for the successful cryopreservation of mammalian oocytes. The osmotically inactive volume of rhesus monkey oocytes was determined by measuring their volumes in the presence of hypertonic solutions of sucrose from 0.2 to 1.5 mol/l, compared with their volume in isotonic TALP-HEPES solution. Boyle-van't Hoff plots at infinite osmolality indicated that the non-osmotic volumes of immature and mature oocytes were 20 and 17% respectively. Osmotic responses of oocytes exposed to 1.0 mol/l solutions of glycerol, dimethylsulphoxide (DMSO) and ethylene glycol (EG) were determined. Rhesus monkey oocytes appeared to be less permeable to glycerol than to DMSO or to EG. Sensitivity of oocytes to osmotic shock was determined by exposing them to various solutions of EG (0.1 to 5.0 mol/l) and then abruptly diluting them into isotonic medium. Morphological survival, as measured by membrane integrity, of oocytes diluted out of EG depended significantly on the concentration of EG (P < 0.01). Determination of permeability characteristics and sensitivity to osmotic shock of rhesus oocytes will aid in the derivation of procedures for their cryopreservation.


[The diagnostic value of enzyme-linked immunosorbent assay technique in detection of anti-tuberculosis antibodies in groups of patients with HIV-positive and HIV-negative reaction].
The article analyzes the results of evaluation of diagnostic value of enzyme-linked immunosorbent assay for detection of antituberculosis antibodies with purpose to diagnose tuberculosis of lungs separately in groups of both HIV-positive and HIV-negative persons. A moderate diagnostic value of enzyme-linked immunosorbent assay is established under screening for tuberculosis of lungs in HIV-negative patients. The evidence of reliability of results depended mainly of actual form of tuberculosis of lungs. In view of many positive results in detection of anti-tuberculosis antibodies in control group of patients with HIV-infection the data testified the low diagnostic significance of enzyme-linked immunosorbent assay in group of patients with HIV-infection. The drawbacks of enzyme-linked immunosorbent assay technique of diagnostic of tuberculosis of lungs in common group of HIV-infected patients partially compensated by possibility of its application at late stages of development of HIV-infection.


Safety evaluation of allogeneic umbilical cord blood mononuclear cell therapy for degenerative conditions.
The current paradigm for cord blood transplantation is that HLA matching and immune suppression are strictly required to prevent graft versus host disease (GVHD). Immunological arguments and historical examples have been made that the use of cord blood for non-hematopoietic activities such as growth factor production, stimulation of angiogenesis, and immune modulation may not require matching or immune suppression. 114 patients suffering from non-hematopoietic degenerative conditions were treated with non-matched, allogeneic cord blood. Doses of 1-3 x 10(7) cord blood mononuclear cells per treatment, with 4-5 treatments both intrathecal and intravenously were performed. Adverse events and hematological, immunological, and biochemical parameters were analyzed for safety evaluation. No serious adverse effects were reported. Hematological, immunological, and biochemical parameters did not deviate from normal ranges as a result of therapy. The current hematology-based paradigm of need for matching and immune suppression needs to be revisited when cord blood is used for non-hematopoietic regenerative purposes in immune competent recipients.


Flap dynamics of plasmepsin proteases: insight into proposed parameters and molecular dynamics.
Herein, for the first time, we report the flap opening and closing in Plasmepsin proteases - plasmepsin II (PlmII) was used as a prototype model. We proposed different combined parameters to define the asymmetric flap motion; the distance, d1, between the flap tip residues (Val78 and Leu292); the dihedral angle, ϕ; in addition to TriCα angles Val78-Asp34-Leu292, θ1, and Val78-Asp214-Leu292, θ2. Only three combined parameters, the distance, d1, the dihedral angle, ϕ, and the TriCα angle, θ1, were found to appropriately define the observed "twisting' motion during the flap opening and closing. The coordinated motions of the proline-rich loop adjacent to the binding cavity rim appeared to exert steric hindrance on the flap residues, driving the flap away from the active site cavity. This loop may also have increased movements around the catalytic dyad residue, Asp214, making TriCα, θ2, unreliable in describing the flap motion. The full flap opening at d1, 23.6 Å, corresponded to the largest TriCα angle, θ1, at 78.6° on a ∼46 ns time scale. Overall the average θ1 and θ2 for the bound was ∼46° and ∼53°, respectively, compared to ∼50° and ∼59° for the Apo PlmII, indicating a drastic increase in TriCα as the active site cavity opens. Similar trends in the distance, d1, and the dihedral angle, ϕ, were observed during the simulation. The asymmetrical opening of the binding cavity was best described by the large shift in ϕ from -33.91° to +21.00° corresponding to the partial opening of the flap in the range of 22-31 ns. Though, the dihedral angle described the twisting of the flap, the extent of flap opening can appropriately be defined by combining d1 and θ1. The results presented here, on the combined parameters, will certainly augment current efforts in designing potent structure-based inhibitors against plasmepsins.


[Organization and ways of extending the scope of surgical services in a polyclinic in relation to total dispensary examination].
The authors analyze the first experience with widening the volume of surgical aid aimed at sanitation of surgical patients under conditions of polyclinics. Advantages of this progressive method are shown. It will improve qualification of polyclinic surgeons, contribute to rational use of surgical places in hospitals, rapid sanitation of population in the course of total dispensary examination which gives a considerable economic effect.


Clinical Availability of ATRA for Patients With Suspected Acute Promyelocytic Leukemia: Why Guidelines May Not Be Followed.
All-trans retinoic acid (ATRA) serves as the backbone of the management of patients with acute promyelocytic leukemia (APL), with guidelines recommending the initiation of ATRA as soon as APL is suspected. As a regional referral center for patients with acute leukemia, those who are suspected of having APL are often transferred to our facility. However, many referring centers are unable to initiate treatment using ATRA. We conducted an exploratory analysis of the clinical availability of ATRA and the factors limiting access to this critical drug. The United States was divided into 6 geographic regions: Northwest, Southwest, Central, Southeast, Northeast, and the Great Lakes. Twenty hospitals were randomly selected from states within each of these regions and were surveyed as to whether they typically treated patients with acute leukemia, the availability of ATRA at their institution, and reported reasons for not stocking ATRA (if not available). Less than one-third of hospitals queried (31%) had ATRA in stock. Neither the size of the hospital nor the hospital's status as academic versus nonacademic (53% vs 31%; P=.08) influenced ATRA availability. Of the hospitals that referred patients with APL, only 14% (7/49) had ATRA readily available. Hospitals that treated patients with APL were more likely to have ATRA available than referring centers (58% vs 14%; P=.000002). Nearly two-thirds of the hospitals surveyed that cared for patients with acute leukemia do not have ATRA immediately available. Moreover, the vast majority of hospitals that refer patients to other centers do not have ATRA. These findings should spur investigation into the impact of immediate ATRA availability on the morbidity and mortality of patients with APL. A call by hematologists nationwide to their formulary committees is warranted to ensure that this lifesaving medication is available to patients suspected of having APL.


Presence of pathogens and indicator microbes at a non-point source subtropical recreational marine beach.
Swimming in ocean water, including ocean water at beaches not impacted by known point sources of pollution, is an increasing health concern. This study was an initial evaluation of the presence of indicator microbes and pathogens and the association among the indicator microbes, pathogens, and environmental conditions at a subtropical, recreational marine beach in south Florida impacted by non-point sources of pollution. Twelve water and eight sand samples were collected during four sampling events at high or low tide under elevated or reduced solar insolation conditions. The analyses performed included analyses of fecal indicator bacteria (FIB) (fecal coliforms, Escherichia coli, enterococci, and Clostridium perfringens), human-associated microbial source tracking (MST) markers (human polyomaviruses [HPyVs] and Enterococcus faecium esp gene), and pathogens (Vibrio vulnificus, Staphylococcus aureus, enterovirus, norovirus, hepatitis A virus, Cryptosporidium spp., and Giardia spp.). The enterococcus concentrations in water and sand determined by quantitative PCR were greater than the concentrations determined by membrane filtration measurement. The FIB concentrations in water were below the recreational water quality standards for three of the four sampling events, when pathogens and MST markers were also generally undetectable. The FIB levels exceeded regulatory guidelines during one event, and this was accompanied by detection of HPyVs and pathogens, including detection of the autochthonous bacterium V. vulnificus in sand and water, detection of the allochthonous protozoans Giardia spp. in water, and detection of Cryptosporidium spp. in sand samples. The elevated microbial levels were detected at high tide and under low-solar-insolation conditions. Additional sampling should be conducted to further explore the relationships between tidal and solar insolation conditions and between indicator microbes and pathogens in subtropical recreational marine waters impacted by non-point source pollution.


At the crossroads of conspicuous and concealable: what race categories communicate about sexual orientation.
We found that judgments of a perceptually ambiguous social category, sexual orientation, varied as a function of a perceptually obvious social category, race. Sexual orientation judgments tend to exploit a heuristic of gender inversion that often promotes accuracy. We predicted that an orthogonal social category that is itself gendered, race, would impact both sexual orientation categorizations and their accuracy. Importantly, overlaps in both the phenotypes and stereotypes associated with specific race and sex categories (e.g., the categories Black and Men and the categories Asian and Women) lead race categories to be decidedly gendered. Therefore, we reasoned that race categories would bias judgments of sexual orientation and their accuracy because of the inherent gendered nature. Indeed, both gay and straight perceivers in the United States were more likely to judge targets to be gay when target race was associated with gender-atypical stereotypes or phenotypes (e.g., Asian Men). Perceivers were also most accurate when judging the sexual orientation of the most strongly gender-stereotyped groups (i.e., Asian Women and Black Men), but least accurate when judging the sexual orientation of counter-stereotypical groups (i.e., Asian men and Black Women). Signal detection analyses confirmed that this pattern of accuracy was achieved because of heightened sensitivity to cues in groups who more naturally conform to gendered stereotypes (Asian Women and Black Men). Implications for social perception are discussed.


DNA helicase HIM-6/BLM both promotes MutSγ-dependent crossovers and antagonizes MutSγ-independent interhomolog associations during caenorhabditis elegans meiosis.
Meiotic recombination is initiated by the programmed induction of double-strand DNA breaks (DSBs), lesions that pose a potential threat to the genome. A subset of the DSBs induced during meiotic prophase become designated to be repaired by a pathway that specifically yields interhomolog crossovers (COs), which mature into chiasmata that temporarily connect the homologs to ensure their proper segregation at meiosis I. The remaining DSBs must be repaired by other mechanisms to restore genomic integrity prior to the meiotic divisions. Here we show that HIM-6, the Caenorhabditis elegans ortholog of the RecQ family DNA helicase BLM, functions in both of these processes. We show that him-6 mutants are competent to load the MutSγ complex at multiple potential CO sites, to generate intermediates that fulfill the requirements of monitoring mechanisms that enable meiotic progression, and to accomplish and robustly regulate CO designation. However, recombination events at a subset of CO-designated sites fail to mature into COs and chiasmata, indicating a pro-CO role for HIM-6/BLM that manifests itself late in the CO pathway. Moreover, we find that in addition to promoting COs, HIM-6 plays a role in eliminating and/or preventing the formation of persistent MutSγ-independent associations between homologous chromosomes. We propose that HIM-6/BLM enforces biased outcomes of recombination events to ensure that both (a) CO-designated recombination intermediates are reliably resolved as COs and (b) other recombination intermediates reliably mature into noncrossovers in a timely manner.


Double-blind analysis of the relation between adult periodontitis and systemic host response to suspected periodontal pathogens.
By using a sensitive enzyme-linked immunosorbent assay, 200 randomly selected sera from Red Cross blood donors were screened for immunoglobulin G (IgG), IgA, and IgM levels against Actinobacillus actinomycetemcomitans, Bacteroides gingivalis, and Bacteroides intermedius. A subgroup of 79 blood donors was clinically examined for type and extent of periodontal destruction, and serological and clinical data were subjected in all possible dual combinations to correlation analyses. The results revealed that the majority of the blood donors suffered from moderate to severe adult periodontitis, often coupled with severe gingival inflammation. No cases of localized juvenile periodontitis or rapidly progressive periodontitis were observed. The extent of periodontal destruction proved to be significantly correlated only to the IgG response levels against B. gingivalis. Corresponding correlation tests assessing the relationships of loss of attachment, bone loss, pocket depth, and papillary bleeding index with the IgG responses to A. actinomycetemcomitans were of marginal significance, while the IgG responses to B. intermedius revealed no relationship to the periodontal health status. The specific IgM responses proved to be unrelated to the clinical parameters, but interestingly, they were found to be highly correlated with each other. Specific IgA levels were frequently too low for enzyme-linked immunosorbent assay testing and, therefore, had to be exempted from statistical analyses. Assessments of the serotype specificity of strongly elevated IgG responses to A. actinomycetemcomitans disclosed no evidence for an association of a particular serotype-specific IgG response with the occurrence of adult periodontal destruction. In contrast to results of earlier studies, a number of sera were found to contain strongly elevated IgG levels against two or even all three serotypes. Although derived by an alternative approach, the reported results largely corroborate earlier observations linking only the occurrence of elevated anti-B. gingivalis IgG responses to the presence of marked periodontal lesions in adults.


Excretion of pantoprazole in human breast.
To measure the amount of pantoprazole in human milk following its oral administration to a breast-feeding mother and to estimate human exposure. One woman was studied over a 24-hour interval after oral administration of 40-mg pantoprazole. Serial plasma and milk samples were collected over 24 hours, and pantoprazole concentrations were measured by high-performance liquid chromatography. A milk/plasma ratio of 0.022 was observed at tmax, 2 hours after dose administration. Infant exposure was measured as maximum concentration in milk multiplied by an estimated maximum consumption of 200 mL at this time. The relative infant dose was estimated to be 7.3 microg of pantoprazole, which is equivalent to 0.14% of the weight-normalized dose received by the mother. Because pantoprazole is unstable in acidic pH, the systemic dose received by the infant is expected to be lower if the ingested pantoprazole is exposed to acid in the infant's stomach. The mother detected no adverse events in the infant. These limited data show that pantoprazole is minimally excreted into breast milk. While it is not known if pantoprazole affects breast milk production, women who are breast-feeding do not have to stop breastfeeding when taking pantoprazole chronically.


IGF-I activation of the AKT pathway is impaired in visceral but not subcutaneous preadipocytes from obese subjects.
Obesity morbidity is associated with excess visceral adiposity, whereas sc adipose tissue is much less metabolically hazardous. Human abdominal sc preadipocytes have greater capacity for proliferation, differentiation, and survival than omental preadipocytes. IGF-I is a critical mediator of preadipocyte proliferation, differentiation, and survival through multiple signaling pathways. We investigated IGF-I action in primary cultures of human preadipocytes isolated from sc and omental adipose tissue of obese subjects. IGF-I-stimulated DNA synthesis was significantly lower in omental compared with sc preadipocytes. IGF-I phosphorylation of the IGF-I receptor and the ERK pathway was comparable in sc and omental cells. However, omental preadipocytes had decreased insulin receptor substrate (IRS)-1 protein associated with increased IRS-1-serine(636/639) phosphorylation and degradation. IGF-I-stimulated phosphorylation of AKT on serine(473) but not threonine(308) was decreased in omental cells, and activation of downstream targets, including S6Kinase, glycogen synthase kinase-3, and Forkhead box O1 was also impaired. CyclinD1 abundance was decreased in omental cells due to increased degradation. Over-expression of IRS-1 by lentivirus in omental preadipocytes increased IGF-I-stimulated AKT-serine(473) phosphorylation. The mammalian target of rapamycin (mTOR)-Rictor complex regulates phosphorylation of AKT-serine(473) in 3T3-L1 adipocytes, but knockdown of Rictor by lentivirus-delivered short hairpin RNA in sc preadipocytes did not affect AKT-serine(473) phosphorylation by IGF-I. These data reveal an intrinsic defect in IGF-I activation of the AKT pathway in omental preadipocytes from obese subjects that involves IRS-1 but probably not mTOR-Rictor complex. We conclude that impaired cell cycle regulation by AKT contributes to the distinct growth phenotype of preadipocytes in visceral fat of obese subjects.


MPK-1/ERK is required for the full activity of resveratrol in extended lifespan and reproduction.
Resveratrol (RSV) extends the lifespan of various organisms through activation of sirtuin. However, whether RSV-mediated longevity is entirely dependent upon sirtuin is still controversial. Thus, understanding additional mechanisms concerning the genetic requirements for the biological activity of RSV needs to be clarified to utilize the beneficial effects of RSV. In this study using Caenorhabditis elegans as a model system, we found that MPK-1 (an ERK homolog) signaling is necessarily required for RSV-mediated longevity of sir-2.1/sirtuin mutants as well as for wild-type worms. We demonstrated that MPK-1 contributes to RSV-mediated longevity through nuclear accumulation of SKN-1 in a SIR-2.1/DAF-16 pathway-independent manner. The positive effect of RSV in regulating lifespan was completely abolished by RNA interference against mpk-1 in the sir-2.1 and daf-16 mutants, strongly indicating that the MPK-1/SKN-1 pathway is involved in RSV-mediated longevity, independently of SIR-2.1/DAF-16. We additionally found that RSV protected worms from oxidative stress via MPK-1. In addition to organismal aging, RSV prevented the age-associated loss of mitotic germ cells, brood size, and reproductive span through MPK-1 in C. elegans germline. Therefore, our findings not only provide new mechanistic insight into the controversial effects of RSV on organismal longevity, but additionally have important implications in utilizing RSV to improve the outcome of aging-related diseases.


Precise Preparation of Triarylboron-Based Graphdiyne Analogues for Gas Separation.
A series of triarylboron-based graphdiyne analogues (TAB-GDYs) with tunable pore size were prepared through copper mediated coupling reaction. The elemental composition, chemical bond, morphology of TAB-GDYs were well characterized. The crystallinity was confirmed by selected area electron diffraction (SAED) and stacking modes were studied in combination with high resolution transmission electron microscope (HRTEM) and structure simulation. The absorption and desorption isotherm revealed relatively high specific surface area of these TAB-GDYs up to 788 m2g-1 for TMTAB-GDY, which decreased as pore size enlarged. TAB-GDYs exhibit certain selectivity for CO2/N2 (21.9), CO2/CH4 (5.3), CO2/H2 (41.8) and C2H2/CO2 (2.3). This work has developed a series of boron containing two-dimensional frameworks with clear structures and good stability, and their tunable pore sizes have laid the foundation for future applications in the gas separation field.


Pellino3, a novel member of the Pellino protein family, promotes activation of c-Jun and Elk-1 and may act as a scaffolding protein.
Toll-like receptors and the IL-1R are part of the innate immune response aimed at mobilizing defense mechanisms in response to infections or injury. These receptors can initiate common intracellular signaling cascades. One intermediate component in these signaling cascades is Pellino, which was first identified in Drosophila and shown to interact with IL-1R-associated kinase. Two homologues, Pellino1 and Pellino2, have been identified in mammals. A novel member of the Pellino protein family has been identified and named Pellino3. Pellino3 shares 84 and 85% amino acid identity with Pellino1 and Pellino2, respectively. Two alternatively spliced Pellino3 mRNAs, Pellino3a and Pellino3b, are widely expressed. Pellino3 physically interacts with IL-1R-associated kinase-1, TNF receptor-associated factor-6, TGF-beta-activated kinase-1, and NF-kappaB-inducing kinase in an IL-1-dependent manner, suggesting that it plays a role as a scaffolding protein. In reporter assays Pellino3 leads to activation of c-Jun and Elk-1, but not NF-kappaB. Pellino3 also leads to activation of c-Jun N-terminal kinase. These data suggest that Pellino3 plays an important role in the innate immune response.


Cross-resistance to methotrexate and metals in human cisplatin-resistant cell lines results from a pleiotropic defect in accumulation of these compounds associated with reduced plasma membrane binding proteins.
Cross-resistance to a wide array of toxic chemicals is a common phenomenon in cisplatin-resistant cell lines. In this study, two independently isolated cisplatin-resistant cell lines derived from a human hepatoma and a cervical adenocarcinoma were shown to be cross-resistant to methotrexate (MTX) and several metal salts, such as sodium arsenite, sodium arsenate, antimony potassium tartrate, and cadmium chloride. A pleiotropic defect resulting in reduced accumulation of cisplatin, 3[H]MTX, 73As3+, and 73As5+ was found in both cisplatin-resistant cell lines. Analysis by immunoblot, indirect immunofluorescence, and Northern hybridization showed dramatically reduced expression of the folate binding protein that mediates MTX uptake in both human cisplatin-resistant cell lines. By photoaffinity labeling with UV irradiation, specific binding proteins of Mr 230,000 and Mr 48,000 for 73As3+ and Mr 190,000 for 73As5+ were found in enriched plasma membrane of both human cisplatin-sensitive parental cell lines. Expression of these specific binding proteins was decreased in cells selected for cisplatin resistance. A protein band at Mr 36,000 that binds to 73As3+ was overexpressed in both human cisplatin-resistant cell lines. The finding of loss of distinct binding proteins for MTX, arsenate, and arsenite in association with decreased accumulation of these agents in cisplatin-resistant cells suggests a pleiotropic, possibly regulatory, alteration in these cells.


Using sequential e-mail messages to promote health behaviors: evidence of feasibility and reach in a worksite sample.
US adults report suboptimal physical activity and fruit and vegetable intake. Innovative strategies to promote healthy behaviors are needed. Employee health promotion programs have been associated with reductions in health risks but are labor-intensive and costly to implement. E-mail and Web-based worksite programs have the potential to reach a broad adult population and to provide a cost-effective approach to employee wellness programming. To assess the feasibility of using sequential e-mail messages to promote physical activity and increase fruit and vegetable intake among employed adults. Employees at one worksite of a large insurance company in New York State were invited to participate. Interested workers provided written consent. After completing a baseline survey, participants received daily e-mails, Monday through Friday, for 26 weeks. The e-mails provided (a) succinct strategies to encourage physical activity or increase fruit and vegetable intake and (b) links to detailed Web-based information and tools. Program reach was assessed by the number of e-mails opened, measures of sustained participation over 6 months, and the number of health-related Web-links clicked. Of 960 employees, 388 (40%) consented to participate; of these, 345 (89%) completed the baseline health survey. After 6 months, 70% of the 345 participants had opened 50% or more of the daily e-mails. In addition, 75% of participants continued to open at least one e-mail a week through week 26 of the study. E-mail opening rates did not vary by gender, age, income, education, ethnicity, or baseline health behavior. The rate of enrollment and sustained participation document the feasibility, broad reach, employee acceptance, and potential value of using electronic communications for health promotion in the workplace.


A large-animal model to evaluate the clinical potential of fetal pig pancreas fragment transplantation.
The long-term goal of this study is to assess the feasibility of using fetal pig pancreas fragment (FPPF) transplantation to treat patients with type I diabetes. Using the highly inbred Westran Pigs, our initial aim was to establish a rejection-free transplant model of FPPF grafted into sibling recipient pigs without immunosuppression. FPPFs were isolated from 80-100-day-old fetuses of either Westran Pigs or outbred pigs and transplanted into the thymus, spleen, liver, or kidney of the recipient Westran pig. Biopsies were taken from each transplant site at set time points and assessed histologically for islet viability, rejection, and endocrine function. Fifty-eight fetal donors were used to transplant 16 recipient pigs. A nonspecific inflammation was seen for both outbred and inbred FPPF donor tissue at day 3 and was considered a response to ischemic necrosis. However, all the transplanted outbred FPPF donor tissue was acutely rejected and lost by day 10-14. In contrast, inbred FPPF tissue showed little evidence of graft necrosis after 3 days, and growth and formation of epithelial islet cell nest-like structures were seen to 28 days after transplantation. With time after transplantation, increasing amounts of insulin immunoperoxidase staining was seen together with chromogranin and somatostatin staining. In summary, this study confirms the potential of the Westran pig to answer the unproven ability of fetal pancreatic tissue to reverse type I diabetes in a large animal model.


MDRSA: A Web Based-Tool for Rapid Identification of Multidrug Resistant Staphylococcus aureus Based on Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry.
As antibiotics resistance on superbugs has risen, more and more studies have focused on developing rapid antibiotics susceptibility tests (AST). Meanwhile, identification of multiple antibiotics resistance on Staphylococcus aureus provides instant information which can assist clinicians in administrating the appropriate prescriptions. In recent years, matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has emerged as a powerful tool in clinical microbiology laboratories for the rapid identification of bacterial species. Yet, lack of study devoted on providing efficient methods to deal with the MS shifting problem, not to mention to providing tools incorporating the MALDI-TOF MS for the clinical use which deliver the instant administration of antibiotics to the clinicians. In this study, we developed a web tool, MDRSA, for the rapid identification of oxacillin-, clindamycin-, and erythromycin-resistant Staphylococcus aureus. Specifically, the kernel density estimation (KDE) was adopted to deal with the peak shifting problem, which is critical to analyze mass spectra data, and machine learning methods, including decision trees, random forests, and support vector machines, which were used to construct the classifiers to identify the antibiotic resistance. The areas under the receiver operating the characteristic curve attained 0.8 on the internal (10-fold cross validation) and external (independent testing) validation. The promising results can provide more confidence to apply these prediction models in the real world. Briefly, this study provides a web-based tool to provide rapid predictions for the resistance of antibiotics on Staphylococcus aureus based on the MALDI-TOF MS data. The web tool is available at: http://fdblab.csie.ncu.edu.tw/mdrsa/.