Single low-dose targeted bevacizumab infusion in adult patients with steroid-refractory radiation necrosis of the brain: a phase II open-label prospective clinical trial.
There is an unmet need for safe and rapidly effective therapies for refractory brain radiation necrosis (RN). The aim of this prospective single-arm phase II trial was to evaluate the safety and efficacy of a single low-dose targeted bevacizumab infusion after blood-brain barrier disruption (BBBD) in adult patients with steroid-refractory brain RN. Ten adults with steroid-refractory, imaging-confirmed brain RN were enrolled between November 2016 and January 2018 and followed for 12 months after treatment. Bevacizumab 2.5 mg/kg was administered as a one-time targeted intra-arterial infusion immediately after BBBD. Primary outcomes included safety and > 25% decrease in lesion volume. Images were analyzed by a board-certified neuroradiologist blinded to pretrial diagnosis and treatment status. Secondary outcomes included changes in headache, steroid use, and functional status and absence of neurocognitive sequelae. Comparisons were analyzed using the Fisher exact test, Mann-Whitney U-test, linear mixed models, Wilcoxon signed-rank test, and repeated-measures 1-way ANOVA. Ten adults (mean ± SD [range] age 35 ± 15 [22-62] years) participated in this study. No patients died or exhibited serious adverse effects of systemic bevacizumab. At 3 months, 80% (95% CI 44%-98%) and 90% (95% CI 56%-100%) of patients demonstrated > 25% decrease in RN and vasogenic edema volume, respectively. At 12 months, RN volume decreased by 74% (median [range] 76% [53%-96%], p = 0.012), edema volume decreased by 50% (median [range] 70% [-11% to 83%], p = 0.086), and headache decreased by 84% (median [range] 92% [58%-100%], p = 0.022) among the 8 patients without RN recurrence. Only 1 (10%) patient was steroid dependent at the end of the trial. Scores on 12 of 16 (75%) neurocognitive indices increased, thereby supporting a pattern of cerebral white matter recovery. Two (20%) patients exhibited RN recurrence that required further treatment at 10 and 11 months, respectively, after bevacizumab infusion. For the first time, to the authors' knowledge, the authors demonstrated that a single low-dose targeted bevacizumab infusion resulted in durable clinical and imaging improvements in 80% of patients at 12 months after treatment without adverse events attributed to bevacizumab alone. These findings highlight that targeted bevacizumab may be an efficient one-time treatment for adults with brain RN. Further confirmation with a randomized controlled trial is needed to compare the intra-arterial approach with the conventional multicycle intravenous regimen. Clinical trial registration no.: NCT02819479 (ClinicalTrials.gov).


Amino acid substitutions in mutant forms of histidinol dehydrogenase from Neurospora crassa.
Amino acid changes in the enzyme l-histidinol dehydrogenase (l-histidinol-NAD oxidoreductase, EC 184.108.40.206) have been determined between the wild-type Neurospora crassa and two temperature-sensitive mutants. Comparison was made between amino acid analyses of peptides of differing electrophoretic and chromatographic mobilities resulting from tryptic and chymotryptic digestion of protein from wild-type and mutant K26, and wild-type and mutant K445 strains, respectively. The analyses demonstrate the substitution of aspartic acid for alanine in mutant K26, and leucine for histidine in mutant K445. The effects of the resulting changes in polarity and charge are discussed in relation to the catalytic functioning of the proteins.


Amine-Functionalized Covalent Organic Framework for Efficient SO2 Capture with High Reversibility.
Removing sulfur dioxide (SO2) from exhaust flue gases of fossil fuel power plants is an important issue given the toxicity of SO2 and subsequent environmental problems. To address this issue, we successfully developed a new series of imide-linked covalent organic frameworks (COFs) that have high mesoporosity with large surface areas to support gas flowing through channels; furthermore, we incorporated 4-[(dimethylamino)methyl]aniline (DMMA) as the modulator to the imide-linked COF. We observed that the functionalized COFs serving as SO2 adsorbents exhibit outstanding molar SO2 sorption capacity, i.e., PI-COF-m10 record 6.30 mmol SO2 g-1 (40 wt%). To our knowledge, it is firstly reported COF as SO2 sorbent to date. We also observed that the adsorbed SO2 is completely desorbed in a short time period with remarkable reversibility. These results suggest that channel-wall functional engineering could be a facile and powerful strategy for developing mesoporous COFs for high-performance reproducible gas storage and separation.


The Rapid Apple Decline Phenomenon: Current Status and Expected Associated Factors in Korea.
Rapid apple decline (RAD) is a complex phenomenon affecting cultivated apple trees and particularly dwarf rootstocks on grafted young apple trees. Since its first appearance in the United States, RAD has been reported worldwide, for example in Canada, South America, Africa, and Asia. The phenomenon has also been observed in apple orchards in Korea, and it presented similar symptoms regardless of apple cultivar and cultivation period. Most previous reports have suggested that RAD may be associated with multiple factors, including plant pathogenic infections, abiotic stresses, environmental conditions, and the susceptibility of trees to cold injury during winter. However, RAD was observed to be more severe and affect more frequently apple trees on the Malling series dwarf rootstock. In this study, we reviewed the current status of RAD worldwide and surveyed biotic and abiotic factors that are potentially closely related to it in Korea.


FOXO1 mediates RANKL-induced osteoclast formation and activity.
We have previously shown that the transcription factor FOXO1 is elevated in conditions with high levels of bone resorption. To investigate the role of FOXO1 in the formation of osteoclasts, we examined mice with lineage-specific deletion of FOXO1 in osteoclast precursors and by knockdown of FOXO1 with small interfering RNA. The receptor activator for NF-κB ligand (RANKL), a principal bone-resorbing factor, induced FOXO1 expression and nuclear localization 2 d after stimulation in bone marrow macrophages and RAW264.7 osteoclast precursors. RANKL-induced osteoclast formation and osteoclast activity was reduced in half in vivo and in vitro with lineage-specific FOXO1 deletion (LyzM.Cre(+)FOXO1(L/L)) compared with matched controls (LyzM.Cre(-)FOXO1(L/L)). Similar results were obtained by knockdown of FOXO1 in RAW264.7 cells. Moreover, FOXO1-mediated osteoclast formation was linked to regulation of NFATc1 nuclear localization and expression as well as a number of downstream factors, including dendritic cell-specific transmembrane protein, ATP6vod2, cathepsin K, and integrin αv. Lastly, FOXO1 deletion reduced M-CSF-induced RANK expression and migration of osteoclast precursors. In the present study, we provide evidence that FOXO1 plays a direct role in osteoclast formation by mediating the effect of RANKL on NFATc1 and several downstream effectors. This is likely to be significant because FOXO1 and RANKL are elevated in osteolytic conditions.


The complete mitochondrial genomes of two rice planthoppers, Nilaparvata lugens and Laodelphax striatellus: conserved genome rearrangement in Delphacidae and discovery of new characteristics of atp8 and tRNA genes.
Nilaparvata lugens (the brown planthopper, BPH) and Laodelphax striatellus (the small brown planthopper, SBPH) are two of the most important pests of rice. Up to now, there was only one mitochondrial genome of rice planthopper has been sequenced and very few dependable information of mitochondria could be used for research on population genetics, phylogeographics and phylogenetic evolution of these pests. To get more valuable information from the mitochondria, we sequenced the complete mitochondrial genomes of BPH and SBPH. These two planthoppers were infected with two different functional Wolbachia (intracellular endosymbiont) strains (wLug and wStri). Since both mitochondria and Wolbachia are transmitted by cytoplasmic inheritance and it was difficult to separate them when purified the Wolbachia particles, concomitantly sequencing the genome of Wolbachia using next generation sequencing method, we also got nearly complete mitochondrial genome sequences of these two rice planthoppers. After gap closing, we present high quality and reliable complete mitochondrial genomes of these two planthoppers. The mitogenomes of N. lugens (BPH) and L. striatellus (SBPH) are 17, 619 bp and 16, 431 bp long with A + T contents of 76.95% and 77.17%, respectively. Both species have typical circular mitochondrial genomes that encode the complete set of 37 genes which are usually found in metazoans. However, the BPH mitogenome also possesses two additional copies of the trnC gene. In both mitochondrial genomes, the lengths of the atp8 gene were conspicuously shorter than that of all other known insect mitochondrial genomes (99 bp for BPH, 102 bp for SBPH). That two rearrangement regions (trnC-trnW and nad6-trnP-trnT) of mitochondrial genomes differing from other known insect were found in these two distantly related planthoppers revealed that the gene order of mitochondria might be conservative in Delphacidae. The large non-coding fragment (the A+T-rich region) putatively corresponding responsible for the control of replication and transcription of mitochondria contained a variable number of tandem repeats (VNTRs) block in different natural individuals of these two planthoppers. Comparison with a previously sequenced individual of SBPH revealed that the mitochondrial genetic variation within a species exists not only in the sequence and secondary structure of genes, but also in the gene order (the different location of trnH gene). The mitochondrial genome arrangement pattern found in planthoppers was involved in rearrangements of both tRNA genes and protein-coding genes (PCGs). Different species from different genera of Delphacidae possessing the same mitochondrial gene rearrangement suggests that gene rearrangements of mitochondrial genome probably occurred before the differentiation of this family. After comparatively analyzing the gene order of different species of Hemiptera, we propose that except for some specific taxonomical group (e.g. the whiteflies) the gene order might have diversified in family level of this order. The VNTRs detected in the control region might provide additional genetic markers for studying population genetics, individual difference and phylogeographics of planthoppers.


Sensing of enantiomeric excess in chiral carboxylic acids.
Cinchona alkaloids (quinine, quinidine, cinchonine, cinchonidine) alkylated at N(1) with chloromethyl anthracene can serve as fluorescent sensors for chiral carboxylic acids. These cinchona ammonium salts are shown to bind chiral carboxylic acids while displaying an increase in fluorescence intensity that can be utilized in determination of enantiomeric excess (ee). Sensor arrays composed of four cinchona ammonium salts are used for quantitative analysis of ee in several non-steroidal anti-inflammatory drugs (NSAIDs), such as enantiomers of ibuprofen, ketoprofen, and naproxen.


Evaluating the efficiency of the 2020 ban of BPA and BPS in thermal papers in Switzerland.
Thermal printing technology requires a color developer to activate the dye under the action of heat. Bisphenol A (BPA) has traditionally been used for this purpose, although it has increasingly been replaced by bisphenol S (BPS) in recent years. Due to concerns regarding their toxicity, the Swiss authorities have banned both BPA and BPS from thermal papers since 2020. The impact of this regulatory decision was evaluated during 3 monitoring campaigns: in 2013-2014, 2019 and 2021. They were used to describe the starting point, the transition phase, and the status after entry into force of the ban, respectively. Whereas the use of BPA as color developer dropped from 82.2% in 2013/14 to 10.8% in 2021, the fraction of BPS-based thermal paper rose from 3.1% to 19.1% during the same period, despite being banned. However, Pergafast® 201 (PF201) is now the main color developer in thermal paper in Switzerland, with an occurrence of 60.3%. Other alternatives such as D-8, TGSA, PPSMU, NKK-1304, BPS-MAE, D-90 and Blue4est® have only been marginally detected. This study demonstrates the efficiency of the regulatory measure and the feasibility to substitute BPA in thermal papers with less-toxic alternatives.


The impact of comprehensive blood conservation program on major complications after total aortic arch replacement.
Patients undergoing total aortic arch replacement (TAAR) usually require blood products perioperatively. This cohort study aimed to investigate the impact of a comprehensive blood conservation program on the major complications in these patients. Patients with traditional or comprehensive blood management intraoperatively from January 2017 to December 2018 were included. We compared the rates of major complications (cerebral vascular accident, acute kidney injury, or mortality) between the two groups after propensity score matching (PSM). The association between blood management and outcomes was assessed by logistic regression. Restricted cubic splines (RCS) were built to evaluate the impact of fresh frozen plasma (FFP) on complications. Patients were stratified by the ratio of FFP/RBC (red blood cell) to investigate the effect of the ratio on complications. After 1:1 PSM, 200 patients were selected. 35% (35/100) of patients suffered major complications in the traditional group, while it decreased to 22% (22/100) in the comprehensive management group (OR = 0.524, p = 0.043). Multivariable logistic regression showed that FFP was a risk factor (OR = 1.186, p = 0.014). RCS results indicated that with the increase of FFP, the risk of complications gradually increases. The cut-off value was 402 mL. Patients in the group of ratio = 0 ∼ 0.5 had a higher chance than those without transfusion (OR = 7.487, p < 0.001). Comprehensive blood conservation program in patients undergoing TAAR is safe and can reduce the incidence of major complications, which are associated with FFP volume and the ratio of FFP/RBC.


Influencing Activity of Bats by Dimly Lighting Wind Turbine Surfaces with Ultraviolet Light.
Wind energy producers need deployable devices for wind turbines that prevent bat fatalities. Based on the speculation that bats approach turbines after visually mistaking them for trees, we tested a potential light-based deterrence method. It is likely that the affected bats see ultraviolet (UV) light at low intensities. Here, we present the results of a multi-month experiment to cast dim, flickering UV light across wind turbine surfaces at night. Our objectives were to refine and test a practical system for dimly UV-illuminating turbines while testing whether the experimental UV treatment influenced the activity of bats, birds, and insects. We mounted upward-facing UV light arrays on turbines and used thermal-imaging cameras to quantify the presence and activity of night-flying animals. The results demonstrated that the turbines can be lit to the highest reaches of the blades with "invisible" UV light, and the animal responses to such experimental treatment can be concurrently monitored. The UV treatment did not significantly change nighttime bat, insect, or bird activity at the wind turbine. Our findings show how observing flying animals with thermal cameras at night can help test emerging technologies intended to variably affect their behaviors around wind turbines.


[The interface between hydroxyapatite ceramic and newly formed bone in scanning electron microscopy].
Bone samples, which have been embedded in methylmethacrylate in an undecalcified state can be evaluated by a scanning electron microscopy after removal of the methylmethacrylate. Hydroxy apatite (HA) has been implanted in 6 mm bore holes of the rabbits' femoral condyle. The animals were sacrificed after 2, 4, 6, 8 weeks and 8 months. The interface between HA and the newly built bone was observed. A direct contact has been found regularly through high magnification (up to 16,000 times). The three dimensional embedding of the implanted HA-particles into the newly built bone can be shown. Collagen filaments are spreading out from bone to HA-particles. We think that there is not only a mechanical but also a chemical connection between both.


Rapid kinetic studies link tetrahydrobiopterin radical formation to heme-dioxy reduction and arginine hydroxylation in inducible nitric-oxide synthase.
To understand how heme and (6R)-5,6,7,8-tetrahydro-l-biopterin (H(4)B) participate in nitric-oxide synthesis, we followed ferrous-dioxy heme (Fe(II)O(2)) formation and disappearance, H(4)B radical formation, and Arg hydroxylation during a single catalytic turnover by the inducible nitric-oxide synthase oxygenase domain (iNOSoxy). In all cases, prereduced (ferrous) enzyme was rapidly mixed with an O(2)-containing buffer to start the reaction. A ferrous-dioxy intermediate formed quickly (53 s(-1)) and then decayed with concurrent buildup of ferric iNOSoxy. The buildup of the ferrous-dioxy intermediate preceded both H(4)B radical formation and Arg hydroxylation. However, the rate of ferrous-dioxy decay (12 s(-1)) was equivalent to the rate of H(4)B radical formation (11 s(-1)) and the rate of Arg hydroxylation (9 s(-1)). Practically all bound H(4)B was oxidized to a radical during the reaction and was associated with hydroxylation of 0.6 mol of Arg/mol of heme. In dihydrobiopterin-containing iNOSoxy, ferrous-dioxy decay was much slower and was not associated with Arg hydroxylation. These results establish kinetic and quantitative links among ferrous-dioxy disappearance, H(4)B oxidation, and Arg hydroxylation and suggest a mechanism whereby H(4)B transfers an electron to the ferrous-dioxy intermediate to enable the formation of a heme-based oxidant that rapidly hydroxylates Arg.


Exact mapping of Illumina blind spots in the Mycobacterium tuberculosis genome reveals platform-wide and workflow-specific biases.
Whole-genome sequencing (WGS) is fundamental to Mycobacterium tuberculosis basic research and many clinical applications. Coverage across Illumina-sequenced M. tuberculosis genomes is known to vary with sequence context, but this bias is poorly characterized. Here, through a novel application of phylogenomics that distinguishes genuine coverage bias from deletions, we discern Illumina 'blind spots' in the M. tuberculosis reference genome for seven sequencing workflows. We find blind spots to be widespread, affecting 529 genes, and provide their exact coordinates, enabling salvage of unaffected regions. Fifty-seven pe/ppe genes (the primary families assumed to exhibit Illumina bias) lack blind spots entirely, while the remaining pe/ppe genes account for 55.1 % of blind spots. Surprisingly, we find coverage bias persists in homopolymers as short as 6 bp, shorter tracts than previously reported. While G+C-rich regions challenge all Illumina sequencing workflows, a modified Nextera library preparation that amplifies DNA with a high-fidelity polymerase markedly attenuates coverage bias in G+C-rich and homopolymeric sequences, expanding the 'Illumina-sequenceable' genome. Through these findings, and by defining workflow-specific exclusion criteria, we spotlight effective strategies for handling bias in M. tuberculosis Illumina WGS. This empirical analysis framework may be used to systematically evaluate coverage bias in other species using existing sequencing data.


Pharmacokinetics, safety, and endocrine and appetite effects of ghrelin administration in young healthy subjects.
It has been demonstrated that ghrelin plays a major role in the regulation of GH secretion and food intake. These actions make ghrelin a strong candidate for the treatment of GH deficiency, anorexia and cachexia. However, only preliminary studies have been performed to assess ghrelin administration in humans. In this study, we have conducted a double-blind, randomized, placebo-controlled trial to investigate the pharmacokinetics, safety, and endocrine and appetite effects of ghrelin in young healthy volunteers. Eighteen male volunteers were randomly assigned into three groups of six subjects: low- and high-dose ghrelin groups, who received intravenous injections of 1 and 5 microg/kg ghrelin (acylated form) respectively, and a placebo group who were injected with mannitol instead of ghrelin. Acylated ghrelin disappeared more rapidly from plasma than total ghrelin, with elimination half life (t(1/2)) of 9-13 and 27-31 min respectively. The number of subjects that experienced adverse effects did not significantly differ among the three groups, and all adverse effects were transient and well tolerated. Both the low and high doses of ghrelin strongly stimulated GH release (peak plasma concentration (C(max,0-90 min)): 124.2+/-63.9 and 153.2+/-52.2 ng/ml for 1 and 5 microg/kg ghrelin respectively). Slight alterations of blood glucose and insulin levels after the injection were observed. Although not statistically significant, ghrelin administration tended to increase hunger sensation in a dose-dependent manner. These results suggest that ghrelin is safe, and that clinical trials may be started to assess the usefulness of ghrelin for the treatment of disorders related to GH secretion and appetite.


Effect of freezing bull semen in two non-egg yolk extenders on post-thaw sperm quality.
Traditionally, extenders for bull semen included egg yolk or milk, but recently there has been a move to avoid material of animal origin. The aim of this study was to evaluate the effects of two commercial extenders (based on soya lecithin and liposomes) on bull sperm quality after cryopreservation. Post-thaw sperm quality was evaluated by computer-assisted sperm analysis and flow cytometric assessment of membrane integrity, chromatin integrity, mitochondrial membrane potential, production of reactive oxygen species and tyrosine phosphorylation. Furthermore, an artificial insemination (AI) trial was conducted, and 56-day non-return rates were evaluated. Semen frozen in the liposome-based extender showed similar membrane integrity and higher mitochondrial membrane potential compared to those in the soya lecithin-based extender. Chromatin integrity and production of live H2 O2 + reactive oxygen species were similar in both extenders. Less superoxide was produced in the samples extended with liposome-based extender, with or without menadione stimulation. Chromatin integrity and tyrosine phosphorylation were not affected by either type of extender. No differences in 56-day non-return rate between extenders containing soya lecithin and liposomes were observed in the AI trial (66% ± 0.8 and 65% ± 0.8, respectively). In conclusion, the sperm quality of bull semen frozen in the two extenders that do not contain material of animal origin was similar, although the semen frozen in the liposome-based extender had higher mitochondrial membrane potential. Either extender could be used in situations where extenders containing material of animal origin are to be avoided.


Combined Structural MR and Diffusion Tensor Imaging Classify the Presence of Alzheimer's Disease With the Same Performance as MR Combined With Amyloid Positron Emission Tomography: A Data Integration Approach.
Background: In recent years, classification frameworks using imaging data have shown that multimodal classification methods perform favorably over the use of a single imaging modality for the diagnosis of Alzheimer's Disease. The currently used clinical approach often emphasizes the use of qualitative MRI and/or PET data for clinical diagnosis. Based on the hypothesis that classification of isolated imaging modalities is not predictive of their respective value in combined approaches, we investigate whether the combination of T1 Weighted MRI and diffusion tensor imaging (DTI) can yield an equivalent performance as the combination of quantitative structural MRI (sMRI) with amyloid-PET. Methods: We parcellated the brain into regions of interest (ROI) following different anatomical labeling atlases. For each region of interest different metrics were extracted from the different imaging modalities (sMRI, PiB-PET, and DTI) to be used as features. Thereafter, the feature sets were reduced using an embedded-based feature selection method. The final reduced sets were then used as input in support vector machine (SVM) classifiers. Three different base classifiers were created, one for each imaging modality, and validated using internal (n = 41) and external data from the ADNI initiative (n = 330 for sMRI, n = 148 for DTI and n = 55 for PiB-PET) sources. Finally, the classifiers were ensembled using a weighted method in order to evaluate the performance of different combinations. Results: For the base classifiers the following performance levels were found: sMRI-based classifier (accuracy, 92%; specificity, 97% and sensitivity, 87%), PiB-PET (accuracy, 91%; specificity, 89%; and sensitivity, 92%) and the lowest performance was attained with DTI (accuracy, 80%; specificity, 76%; and sensitivity, 82%). From the multimodal approaches, when integrating two modalities, the following results were observed: sMRI+PiB-PET (accuracy, 98%; specificity, 98%; and sensitivity, 99%), sMRI+DTI (accuracy, 97%; specificity, 99%; and sensitivity, 94%) and PiB-PET+DTI (accuracy, 91%; specificity, 90%; and sensitivity, 93%). Finally, the combination of all imaging modalities yielded an accuracy of 98%, specificity of 97% and sensitivity of 99%. Conclusion: Although DTI in isolation shows relatively poor performance, when combined with structural MR, it showed a surprising classification performance which was comparable to MR combined with amyloid PET. These results are consistent with the notion that white matter changes are also important in Alzheimer's Disease.


Stromelysin-1 (MMP-3)-independent gelatinase expression and activation in mice.
A potential physiological role of stromelysin-1 (MMP-3) in the expression or activation of gelatinase A (MMP-2) or gelatinase B (MMP-9) in the wall of injured arteries was studied with the use of homozygous MMP-3-deficient (MMP-3-/-) mice. One week after perivascular electric injury of the carotid or femoral artery in wild-type (MMP-3+/+) or MMP-3-/- mice, 70 kD and 65 kD proMMP-2 levels were enhanced by twofold to fourfold, with corresponding increases of 20- to 40-fold for active 61 kD and 58 kD MMP-2, and of 10- to 80-fold for 94 kD proMMP-9. Active MMP-2 species represented approximately one third of the total MMP-2 concentration for both MMP-3+/+ and MMP-3-/- mice. Active 83 kD MMP-9 was not detected in noninjured carotid or femoral arteries, whereas one week after injury its contribution to the total MMP-9 level was 11% to 18% for MMP-3+/+ and MMP-3-/- mice. Immunostaining of arterial sections confirmed enhanced expression of both MMP-2 and MMP-9 after vascular injury. Double immunostaining showed colocalization of MMP-9 with macrophages in the adventitia, whereas MMP-2 was also detected mainly in the adventitia but failed to colocalize with smooth muscle cells. Cell culture experiments confirmed comparable ratios of active versus latent MMP-2 in skin fibroblasts and smooth muscle cells derived from MMP-3+/+ and MMP-3-/- mice. Addition of plasmin(ogen) did not significantly affect activation of proMMP-2. In MMP-3+/+ and MMP-3-/- macrophages, comparable levels of 94 kD proMMP-9 were detected, and plasmin(ogen)-mediated conversion to 83 kD MMP-9 was obtained in both genotypes. These data thus indicate that proMMP-2 activation may occur via a plasmin- and MMP-3-independent mechanism, whereas plasmin can directly activate proMMP-9 via a MMP-3-independent mechanism.


[Drug treatment of diabetic retinopathy].
Medical treatment of diabetic retinopathy remains a problem. So far, no drug has produced definitely positive results. Strict metabolic control still has to be considered the sine qua non of a therapeutic approach to diabetic retinopathy. The extent to which multiple daily injections of insulin (insulin pumps) are capable of inhibiting the development of a retinopathy is still under investigation. Prophylactic measures to prevent further complications where retinopathy already exists should not, however, be neglected.


[Contribution of ambulatory digestive surgery].
In France, ambulatory surgery is controlled by specific regulations which outline the organization of the facilities. It is practiced less in France than in other countries, but specific governmental incentive policies have been instituted. The characteristic feature of digestive surgery is the high occurrence of post-operative nausea and vomiting due to the peritoneal incision. New surgical procedures and anesthetic regimens allow ambulatory care in children and adults. But the choice of ambulatory care is based on the patient's characteristics more than on surgical procedure and follows well-known selection criteria. The procedures concerned in general surgery are groin hernia repair, proctologic surgery, and subcutaneous tissue surgery. Laparoscopic cholecystectomy and neck surgery in increasing numbers of patients on an ambulatory basis is the first step before expanding ambulatory surgical procedures toward major surgery. Physicians must have a thorough knowledge of ambulatory surgery as an organizational concept.


HOPE, an open platform for medical data management on the grid.
The paper describes a platform developed for the secure management and analysis of medical data and images in a grid environment. Designed for telemedicine and built upon the EGEE gLite middleware and particularly the metadata catalogue AMGA as well as the GridSphere web portal, the platform provides to healthcare professionals the capacity to upload and query medical information stored over distributed servers. A job submission environment is also available for data analysis. Security features include authentication and authorization by grid certificates, anonymization of medical data and image encryption. The platform is currently deployed on several sites in Europe and Asia and is being customized for applications in the field of telemedicine and medical physics.


Oxidative inactivation of cytochrome P-450 1A (CYP1A) stimulated by 3,3',4,4'-tetrachlorobiphenyl: production of reactive oxygen by vertebrate CYP1As.
Microsomal cytochrome P-450 1A (CYP1A) in a vertebrate model (the teleost fish scup) is inactivated by the aryl hydrocarbon receptor agonist 3,3',4,4'-tetrachlorobiphenyl (TCB). Here, the mechanism of CYP1A inactivation and its relationship to reactive oxygen species (ROS) formation were examined by using liver microsomes from scup and rat and expressed human CYP1As. In vitro inactivation of scup CYP1A activity 7-ethoxyresorufin O-deethylation by TCB was time dependent, NADPH dependent, oxygen dependent, and irreversible. TCB increased microsomal NADPH oxidation rates, and CYP1A inactivation was lessened by adding cytochrome c. CYP1A inactivation was accompanied by loss of spectral P-450, a variable loss of heme and a variable appearance of P-420. Rates of scup liver microsomal metabolism of TCB were < 0.5 pmol/min/mg, 25-fold less than the rate of P-450 loss. Non-heme iron chelators, antioxidant enzymes, and ROS scavengers had no influence on inactivation. Inactivation was accelerated by H(2)O(2) and azide but not by hydroxylamine or aminotriazole. TCB also inactivated rat liver microsomal CYP1A, apparently CYP1A1. Adding TCB to scup or rat liver microsomes containing induced levels of CYP1A, but not control microsomes, stimulated formation of ROS; formation rates correlated with native CYP1A1 content. TCB stimulated ROS formation by baculovirus-expressed human CYP1A1 but not CYP1A2. The results indicate that TCB uncouples the catalytic cycle of CYP1A, ostensibly CYP1A1, resulting in formation of ROS within the active site. These ROS may inactivate CYP1A or escape from the enzyme. ROS formed by CYP1A1 may contribute to the toxicity of planar halogenated aromatic hydrocarbons.


Do heritability estimates of political phenotypes suffer from an equal environment assumption violation? Evidence from an empirical study.
Using data from the Minnesota Twins Political Survey, this paper tests for the presence of unequal environments (EEA) by zygosity in political attitudes. Equal environment measures used include shared bedroom, friends, classes, and dressing alike. Results show an EEA violation at p < .05 in 5% of the models tested. The violations' impact on heritability estimates and their confidence levels appear random in magnitude and direction. No reasonable post hoc explanation emerged for understanding the presence of the violation in some items but not others. This article establishes reasonable priors for the absence of EEA violations in political phenotypes based on the tested environmental components. The findings place the burden on critics to present theoretical work on the specific mechanisms of EEA violations based on which additional empirical assessments could (and should) be conducted.


Spray freeze dried niclosamide nanocrystals embedded dry powder for high dose pulmonary delivery.
Based on the drug repositioning strategy, niclosamide (NCL) has shown potential applications for treating COVID-19. However, the development of new formulations for effective NCL delivery is still challenging. Herein, NCL-embedded dry powder for inhalation (NeDPI) was fabricated by a novel spray freeze drying technology. The addition of Tween-80 together with 1,2-Distearoyl-sn-glycero-3-phosphocholine showed the synergistic effects on improving both the dispersibility of primary NCL nanocrystals suspended in the feed liquid and the spherical structure integrity of the spray freeze dried (SFD) microparticle. The SFD microparticle size, morphology, crystal properties, flowability and aerosol performance were systematically investigated by regulating the feed liquid composition and freezing temperature. The addition of leucine as the aerosol enhancer promoted the microparticle sphericity with greatly improved flowability. The optimal sample (SF- 80D-N20L2D2T1) showed the highest fine particle fraction of ∼47.83%, equivalently over 3.8 mg NCL that could reach the deep lung when inhaling 10 mg dry powders.


Matching for the nonconventional MHC-I MICA gene significantly reduces the incidence of acute and chronic GVHD.
Graft-versus-host disease (GVHD) is among the most challenging complications in unrelated donor hematopoietic cell transplantation (HCT). The highly polymorphic MHC class I chain-related gene A, MICA, encodes a stress-induced glycoprotein expressed primarily on epithelia. MICA interacts with the invariant activating receptor NKG2D, expressed by cytotoxic lymphocytes, and is located in the MHC, next to HLA-B Hence, MICA has the requisite attributes of a bona fide transplantation antigen. Using high-resolution sequence-based genotyping of MICA, we retrospectively analyzed the clinical effect of MICA mismatches in a multicenter cohort of 922 unrelated donor HLA-A, HLA-B, HLA-C, HLA-DRB1, and HLA-DQB1 10/10 allele-matched HCT pairs. Among the 922 pairs, 113 (12.3%) were mismatched in MICA MICA mismatches were significantly associated with an increased incidence of grade III-IV acute GVHD (hazard ratio [HR], 1.83; 95% confidence interval [CI], 1.50-2.23; P < .001), chronic GVHD (HR, 1.50; 95% CI, 1.45-1.55; P < .001), and nonelapse mortality (HR, 1.35; 95% CI, 1.24-1.46; P < .001). The increased risk for GVHD was mirrored by a lower risk for relapse (HR, 0.50; 95% CI, 0.43-0.59; P < .001), indicating a possible graft-versus-leukemia effect. In conclusion, when possible, selecting a MICA-matched donor significantly influences key clinical outcomes of HCT in which a marked reduction of GVHD is paramount. The tight linkage disequilibrium between MICA and HLA-B renders identifying a MICA-matched donor readily feasible in clinical practice.


Suicidality and its association with insight and neurocognition in taiwanese patients with bipolar I disorder in remission.
The aims of this study were to examine the relationships between suicidality, insight, and neurocognition in patients with bipolar I disorder who were in a remitted state. Using the Violence and Suicide Assessment Scale, we evaluated 96 patients with bipolar I disorder in remission to determine their suicidal ideations and attempts over the previous year. We also evaluated their level of insight by using the Schedule of Assessment of Insight (SAI) and its expanded version (SAI-E), as well as their neurocognitive function by a series of neurocognitive function tests. Insight and neurocognitive functions of bipolar subjects who had and who had no suicidal ideations or attempts over the previous year were compared. The results indicated that the remitted bipolar subjects who had suicidal ideations or attempts over the previous year had higher insight scores on all 3 SAI dimensions and on the SAI-E compared with those who had no suicidal ideations or attempts. However, no difference in any neurocognitive function was found between the 2 groups of remitted bipolar subjects. The results of this study suggest clinicians need to be particularly alert to the potential for suicide in bipolar patients with a high level of insight.


Characterization of a new non-Hodgkin's lymphoma cell line (NCEB-1) with a chromosomal (11:14) translocation [t(11:14)(q13;q32)].
A new cell line, NCEB-1, was established by Epstein-Barr virus (EBV) transformation of peripheral blood mononuclear cells from a patient with centroblastic-centrocytic diffuse lymphoma expressing IgM lambda. The transformed cells were lymphoblastoid, with many cells showing a plasmacytoid morphology. The NCEB-1 cells had cytoplasmic Ig (CyIg), with loss of the surface Ig (SIg) expression. Cytogenetic analysis of the cell line demonstrated two clones with variations: a hypodiploid clone, with a complex karyotype including a t(11;14)(q13;q32) similar to the original tumor cells, and a near tetraploid clone with the same markers. Southern blot analysis of DNA from the patient's neoplastic cells and NCEB-1 demonstrated identical Ig heavy chain gene rearrangement, confirming the origin of the cell line. The cell line was not tumorigenic when tested in an in vitro assay using immunosuppressed mice. NCEB-1 has been in continuous culture for 9 months and will be valuable for the in vivo study of non-Hodgkin's lymphoma and EBV transformation.


Quantitation of endogenously occupied and unoccupied binding sites for 1,25-dihydroxyvitamin D3 in rat intestine.
The quantitative reversible dissociation of the 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3]-receptor complex by the mercurial reagent mersalyl was used to develop an assay for endogenously occupied and unoccupied 1,25-(OH)2D3 binding sites. Incubation of intestinal cytosol preparations in buffer containing 50 mM Tris . HCl, 300 mM KCl, and 1.5 mM EDTA, pH 7.4, with 1 mM mersalyl for 60 min was effective in inhibiting 98% of 1,25-(OH)2D3 specific binding activity. Dissociation of bound 1,25-(OH)2[26,27-3H]D3 from the hormone-receptor complex approached completion by 180 min. In cytosol incubated with saturating levels of nonradioactive hormone, 96% of total binding activity was measurable with the hormone binding assay after displacement of bound nonradioactive ligand with 1 mM mersalyl. Endogenously occupied 1,25-(OH)2D3 binding sites contributed 0, 9, and 19% of total binding activity in rats with plasma 1,25-(OH)2D3 levels averaging 2, 121 +/- 36 and 516 +/- 92 pg/ml, respectively. Therefore, the major fraction of cytosolic 1,25-(OH)2D3 specific binding activity is unoccupied in rat intestine. The results suggest that only a small proportion of the measurable receptors are in the bound form to provide maximal 1,25-(OH)2D3-induced calcium transport.


Reliability of maternal recall and reporting of child births and deaths in rural Egypt.
Demographic indicators such as fertility rates and infant mortality rates are often measured in census surveys by interviewing mothers to obtain their pregnancy histories and child deaths. The validity of such surveys depends upon accurate recall of histories, truthful reporting of events and understanding of the questions posed. To measure the reliability of maternal reporting, two census surveys conducted in a rural Egyptian population were compared. Women between 15 and 55 years of age residing in 20 villages were asked their histories of live births, stillbirths and child deaths. An identical set of questions was posed 2 years later. Twice-monthly home visits were conducted in the intervening 2-year interval to identify accurately any new births, stillbirths and deaths occurring in the population. The maternal reports from the first census were combined with the prospectively identified births, stillbirths and deaths and compared with the maternal reports from the second census. For 1502 women, the discrepancies in the total number of births, stillbirths and child deaths reported between the two surveys were 0.6%, 4% and 0.6% respectively. However, when the consistency of responses was analysed, the proportion of women with discordant responses was 10%, 6% and 7% for the same measures. These results suggest that, despite the large number of births and deaths that women may experience in developing countries, maternal interviews provide reliable responses that can be used to estimate mortality and fertility rates in settings where vital records are incomplete or unreliable.


Enhanced Anticancer Effect of a Combination of S-adenosylmethionine (SAM) and Immune Checkpoint Inhibitor (ICPi) in a Syngeneic Mouse Model of Advanced Melanoma.
Immune checkpoint inhibitors (ICPi) targeting the PD-1/PD-L1 pathway have shown marked success in patients with advanced melanoma. However, 60-70% of patients fail to respond, warranting a therapeutic intervention that could increase response rates. We and others have shown that S-adenosylmethionine (SAM), a universal methyl donor, has significant anticancer effects in numerous cancers previously; however, its effect on melanoma progression has not been evaluated. Interestingly, SAM was reported to be essential for T cell activation and proliferation and, thus, could potentially cooperate with ICPi and block melanoma progression. In this study, we examined the antitumor effects of SAM and ICPi alone and in combination in a well-established melanoma mouse model wherein syngeneic C57BL/6 mouse were subcutaneously (orthotopic) injected with B16-F1 cells. Treatment of mice with either SAM or anti-PD-1 antibody alone resulted in significant reduction in tumor volumes and weights; effects that were highest in mice treated with a combination of SAM+anti-PD-1. RNA-sequencing analysis of the primary tumors showed numerous differentially expressed genes (DEGs) following treatment with SAM+anti-PD-1, which was shown to downregulate cancer, MAPK, and tyrosine kinase pathways. Indeed, SAM+anti-PD-1 reversed the aberrant expression of some known melanoma genes. Tumor immunophenotyping revealed the SAM+anti-PD-1 combination was significantly more effective than either SAM or anti-PD-1 as the CD8+ T cells had higher activation, proliferation, and cytokine production compared to all other groups. This study shows that the combination of currently approved agents SAM and ICPi can effectively block melanoma via alteration of key genes/pathways implicated in cancer and immune response pathways, providing the rationale for the initiation of clinical trials with SAM and ICPi.


The effects of chronic nicotine and alcohol use on neurocognitive function.
Despite the extensive literatures on the independent effects of chronic nicotine and alcohol use on neurocognition, little is known about their combined impact. The purpose of this paper was to examine the effects of chronic nicotine and alcohol use on neurocognition in participants who took part in Project MATCH, a study of the efficacies of three behavioral treatments for adults diagnosed with alcohol abuse or dependence. Multiple regression and ANCOVA analyses were conducted to determine the relationship between lifetime weeks of tobacco use and years of alcohol use problems and neuropsychological test performance. Results showed that although years of chronic alcohol use was significantly inversely related to neuropsychological test scores, and chronic nicotine use showed an additive effect, substance use accounted for little variance in neuropsychological functioning. These findings suggest that some protective factors may have helped to attenuate the impact of chronic substance use on neurocognition. The importance of considering individual differences in such research is highlighted. Additional studies on the combined effects of chronic nicotine and alcohol use on neuropsychological function are warranted.


Common variation in the DIO2 gene predicts baseline psychological well-being and response to combination thyroxine plus triiodothyronine therapy in hypothyroid patients.
Animal studies suggest that up to 80% of intracellular T(3) in the brain is derived from circulating T(4) by local deiodination. We hypothesized that in patients on T(4) common variants in the deiodinase genes might influence baseline psychological well-being and any improvement on combined T(4)/T(3) without necessarily affecting serum thyroid hormone levels. We analyzed common variants in the three deiodinase genes vs. baseline psychological morbidity and response to T(4)/T(3) in 552 subjects on T(4) from the Weston Area T(4) T(3) Study (WATTS). Primary outcome was improvement in psychological well-being assessed by the General Health Questionnaire 12 (GHQ-12). The rarer CC genotype of the rs225014 polymorphism in the deiodinase 2 gene (DIO2) was present in 16% of the study population and was associated with worse baseline GHQ scores in patients on T(4) (CC vs. TT genotype: 14.1 vs. 12.8, P = 0.03). In addition, this genotype showed greater improvement on T(4)/T(3) therapy compared with T(4) only by 2.3 GHQ points at 3 months and 1.4 at 12 months (P = 0.03 for repeated measures ANOVA). This polymorphism had no impact on circulating thyroid hormone levels. Our results require replication but suggest that commonly inherited variation in the DIO2 gene is associated both with impaired baseline psychological well-being on T(4) and enhanced response to combination T(4)/T(3) therapy, but did not affect serum thyroid hormone levels.


Studies of the active site of m-calpain and the interaction with calpastatin.
Calpain autolyses in the presence of Ca2+. In the case of m-calpain (80 + 30 kDa) the first product is an 80 + 18 kDa species which has an intact large subunit and the C-terminal Ca(2+)-binding domain of the small subunit. It was possible to bind E64 into the active site of calpain in the presence of Ca2+ before cleavage of either calpain subunit. This suggests that the active site is functional before any autolysis has occurred and that calpain is not a proenzyme. Prolonged autolysis generates several fragments including a 42 kDa active-site domain fragment that showed no proteolytic activity and Ca(2+)-binding domain fragments. Some of the Ca(2+)-binding domain fragments were found to exist as heterodimers (23 + 18 kDa and 22 + 18 kDa), with the Ca(2+)-binding domain of the large subunit interacting with the Ca(2+)-binding domain of the small subunit. These species were true heterodimers, as they showed co-elution of the two Ca(2+)-binding domains on ion-exchange and gel-filtration chromatography, and immunoprecipitation of both polypeptides with an antiserum specific for the small-subunit Ca(2+)-binding domain. The generation of the dimer species after only 15 min autolysis suggests that the interaction between the Ca(2+)-binding domains is present in the native calpain structure. The interaction of calpain with calpastatin was investigated using an assay based on binding to calpastatin-Sepharose and a competitive binding assay. Calpain, active-site-blocked calpain and calpain fragments generated by autolysis were studied. Calpain bound to calpastatin in the presence of Ca2+; however, the isolated active-site-containing 80 kDa large subunit (proteolytically inactive), a 42 kDa active-site-containing fragment (proteolytically inactive) and Ca(2+)-binding domain fragments of calpain did not. Active-site-blocked calpain bound to calpastatin, but with an affinity reduced by approximately two orders of magnitude when compared with native calpain.


Molecular analysis of the third component of canine complement (C3) and identification of the mutation responsible for hereditary canine C3 deficiency.
Genetically determined deficiency of the third component of complement (C3) in the dog is characterized by a predisposition to recurrent bacterial infections and to type 1 membranoproliferative glomerulonephritis. The current studies were undertaken to characterize the cDNA for wild-type canine C3 and identify the molecular basis for hereditary canine C3 deficiency. Amplification, cloning, and sequence analysis indicated that canine C3 is highly conserved in comparison with human, mouse, and guinea pig C3. Southern blot analysis failed to show any gross deletions or rearrangements of DNA from C3-deficient animals. Northern blot analysis indicated that the livers of these animals contain markedly reduced quantities of a normal length C3 mRNA. The full-length 5.1-kb canine C3 cDNA was amplified in overlapping PCR fragments. Sequence analysis of these fragments has shown a deletion of a cytosine at position 2136 (codon 712), leading to a frameshift that generates a stop codon 11 amino acids downstream. The deletion has been confirmed in genomic DNA, and its inheritance has been demonstrated by allele-specific oligonucleotide hybridization.


A Tool for Specifying Data Quality Checks for Clinical Data Management Systems - A Technical Case Report.
Prospective data collection in clinical trials is considered the gold standard of clinical research. Validating data entered in input fields in case report forms is unavoidable to maintain good data quality. Data quality checks include both the conformance of individual inputs to the specification of the data element, the detection of missing values, and the plausibility of the values entered. Besides Libre-/OpenClinica there are many applications for capturing clinical data. While most of them have a commercial approach, free and open-source solutions lack intuitive operation. Our ocRuleTool is made for the specific use case to write validation rules for Open-/LibreClinica, a clinical study management software for designing case report forms and managing medical data in clinical trials. It addresses parts of all three categories of data quality checks mentioned above. The required rules and error messages are entered in the normative Excel specification and then converted to an XML document which can be uploaded to Open-/LibreClinica. The advantage of this intermediate step is a better readability as the complex XML elements are broken down into easy to fill out columns in Excel. The tool then generates the ready to use XML file by itself. This approach saves time, is less error-prone and allows collaboration with clinicians on improving data quality. Our ocRuleTool has proven useful in over a dozen studies. We hope to increase the user base by releasing it to open source on GitHub.


UVA Induced Oxidative Stress Was Inhibited by Paeoniflorin/Nrf2 Signaling or PLIN2.
Photodamages caused by UVA radiation induced oxidative injuries are closely related to photoaging and skin cancer. Paeoniflorin (PF), extracted from the root of Paeonia lactiflora, has been reported to be an effective antioxidant. PLIN2, known as adipose differentiation-related protein, has been previously involved in the regulation of oxidative stress. In this study, we were sought to investigate the photo-protective property of PF and PLIN2 in UVA-radiated human dermal fibroblasts (HDFs). HDFs were pre-treated with PF (800 μM) followed by UVA radiation (22.5 J/cm2). MTS activity, cell apoptosis, ROS, MDA, and SOD were detected, respectively. The expressions of Nrf2, HO-1, NQ-O1, and PLIN2 were determined using RT-qPCR or western blot. Nrf2 was silenced by siRNA, and PLIN2 was overexpressed via lentiviral transduction. Comparing to the UVA radiation, PF pre-treatment could prominently increase the MTS activity, decrease cell apoptosis, reduce the generations of ROS and MDA, increase the activity of SOD and increase the expression of Nrf2 and its target genes HO-1 and NQ-O1. When Nrf2 was knocked down, PF lost above protective properties. In addition, UVA induced oxidative stress led to upregulation of PLIN2 and the latter could be decreased by PF. Overexpression of PLIN2 improved MTS activity and reduced MDA level in HDFs. The combination of PLIN2 overexpression and PF pre-treatment corporately inhibited UVA-induced injury. Besides, we also found that PF and PLIN2 had a compensatory protection against UVA induced oxidative stress. In conclusion, our study demonstrated that UVA induced photodamages could be inhibited by PF via Nrf2/HO-1/NQ-O1 signaling pathway or by PLIN2, and the combination of PLIN2 overexpression and PF played additive effects against UVA-related oxidative stress.


Refractive outcomes and complications after combined copolymer phakic intraocular lens explantation and phacoemulsification with intraocular lens implantation.
To assess safety, efficacy, and predictability of combined Implantable Collamer Lens phakic intraocular lens (pIOL) explantation and phacoemulsification and to report the rate of complications. A secondary aim was to examine whether intraocular lens (IOL) power calculation was affected by the pIOL in situ. Jules-Gonin Eye Hospital, Lausanne, Switzerland. Retrospective case series. The records of patients having pIOL implantation with subsequent phacoemulsification were reviewed. Data analysis was performed between July 1, 2015, and December 31, 2015. The mean corrected distance visual acuity (CDVA), safety, stability of refraction, predictability of IOL power calculation, and postoperative complications were recorded. The study comprised 38 eyes (29 patients). Of the eyes, 32 (82%) were within ±1.00 diopter (D) of the target refraction and 21 (57.9%) were within ±0.50 D. The mean CDVA after IOL implantation was significantly better than before pIOL implantation (P < .001) and similar to after pIOL implantation (P = .87). The safety of this multistepped procedure was good; on average, 1.6 Snellen lines of CDVA were gained over before pIOL implantation. Branch retinal vein occlusion occurred in 1 eye; no other sight-threatening complications were observed. Posterior capsule opacification developed in 15 eyes (39.5%) and was treated successfully with neodymium:YAG capsulotomy. The highest mean CDVA was recorded at the final visit. Good safety, stability, and predictability were also observed. Postoperative complications were few and in all but 1 case, not sight threatening. Overall, results indicate the combination of procedures does not negatively affect the final visual outcome.


Use of extracorporeal membrane oxygenation for eCPR in the emergency room in patients with refractory out-of-hospital cardiac arrest.
Out-of-hospital cardiac arrest (OHCA) refractory to conventional high-quality cardiopulmonary resuscitation (CPR) may be rescued by extracorporeal CPR (eCPR) using veno-arterial extracorporeal membrane oxygenation (V-A ECMO). Even when trying to identify eCPR candidates based on criteria assumed to be associated with a favourable neurological outcome, reported survival rates are frequently below 10%. All patients undergoing implantation of V-A ECMO for eCPR between January 2018 and December 2019 (N = 40) were analysed (age 53±13 years; 75% male). Patients with refractory OHCA and potentially favourable circumstances (initial shockable rhythm, witnessed arrest, bystander CPR, absence of limiting comorbidities, age <75 years) were transported under mechanical chest compression. Candidates for eCPR should have a pH ≥6.9, arterial lactate ≤15 mmol/L and time-to-ECMO should be ≤60 minutes. Overall 30-day survival was 12.5%, with 3 of 5 survivors having a favourable neurological outcome (cerebral performance category (CPC) 1 or 2), representing 7.5% of the total eCPR population. No patient selected for eCPR met all pre-defined criteria (median of unfavourable criteria: 3). Importantly, time-to-ECMO most often (39/40) exceeded 60 minutes (mean 102 ±32 min.), and lactate was >15mmol/L in 30 out of 40 patients. Moreover, 22 out of 40 patients had a non-shockable rhythm on the first ECG. Despite our intention to select patients with potentially advantageous circumstances to achieve acceptable eCPR outcomes, the imminent deadly consequence of withholding eCPR obviously prompted individual physicians to perform the procedure also in presumably more unfavourable settings, resulting in similar mortality rates of eCPR as reported before.


The effects of acute transfer to freshwater on ion transporters of the pharyngeal cavity in European seabass (Dicentrarchus labrax).
Gene expression of key ion transporters (the Na+/K+-ATPase NKA, the Na+, K+-2Cl- cotransporter NKCC1, and CFTR) in the gills, opercular inner epithelium, and pseudobranch of European seabass juveniles (Dicentrarchus labrax) were studied after acute transfer up to 4 days from seawater (SW) to freshwater (FW). The functional remodeling of these organs was also studied. Handling stress (SW to SW transfer) rapidly induced a transcript level decrease for the three ion transporters in the gills and operculum. NKA and CFTR relative expression level were stable, but in the pseudobranch, NKCC1 transcript levels increased (up to 2.4-fold). Transfer to FW induced even more organ-specific responses. In the gills, a 1.8-fold increase for NKA transcript levels occurs within 4 days post transfer with also a general decrease for CFTR and NKCC1. In the operculum, transcript levels are only slightly modified. In the pseudobranch, there is a transient NKCC1 increase followed by 0.6-fold decrease and 0.8-fold CFTR decrease. FW transfer also induced a density decrease for the opercular ionocytes and goblet cells. Therefore, gills and operculum display similar trends in SW-fish but have different responses in FW-transferred fish. Also, the pseudobranch presents contrasting response both in SW and in FW, most probably due to the high density of a cell type that is morphologically and functionally different compared to the typical gill-type ionocyte. This pseudobranch-type ionocyte could be involved in blood acid-base regulation masking a minor osmotic regulatory capacity of this organ compared to the gills.


Adaptive and maladaptive neural compensatory consequences of sensory deprivation-From a phantom percept perspective.
It is suggested that the brain undergoes plastic changes in order to adapt to changing environmental needs. Sensory deprivation results in decreased input to the brain leading to adaptive or maladaptive changes. Although several theories hypothesize the mechanism of these adaptive and maladaptive changes, the course of action taken by the brain heavily depends on the age of incidence of damage. The growing body of literature on the topic proposes that maladaptive changes in the brain are instrumental in creating phantom percepts, defined as the perception of a sensory experience in the absence of a physical stimulus. The current article reviews the mechanisms of adaptive and maladaptive plasticity in the brain in congenital, early, and late-onset sensory deprivation in conjunction with the phantom percepts in the different sensory domains. We propose that the mechanisms of adaptive and maladaptive plasticity fall under a universal construct of updating hierarchical Bayesian prediction errors. This theory of the Bayesian brain hypothesizes that the brain constantly compares its internal milieu with changing environmental cues and either adjusts its predictions or discards the change, depending on the novelty or salience of the external stimulus. We propose that adaptive plasticity reflects both successful bottom-up compensation and top-down updating of the model while maladaptive plasticity reflects failure in one or both mechanisms, resulting in a constant prediction-error. Finally, we hypothesize that phantom percepts are generated by the brain as a solution to this prediction error and are thus a manifestation of unsuccessful adaptation to sensory deprivation.


ADP-ribosyl transferase, rearrangement of DNA, and cell differentiation.
Cell differentiation is the process by which genetic information is selectively expressed to produce cells with various morphologies and functions. The integrated changes necessary for this fundamentally important process have recently been the subject of intense study. This review will summarize data from several laboratories correlating differentiation with the activity of the enzyme ADP-ribosyl transferase and with changes in single-strand DNA breaks in various diverse eukaryotic systems. We will then discuss the implications of these observations for differentiation in general, including the possibility that rearrangement of genetic material is a widespread mechanism for controlling gene expression.


Lecithin:retinol acyltransferase and retinyl ester hydrolase activities are differentially regulated by retinoids and have distinct distributions between hepatocyte and nonparenchymal cell fractions of rat liver.
The cellular distribution of enzymes that esterify retinol and hydrolyze retinyl esters (RE) was studied in liver of vitamin A-sufficient, -deficient, and deficient rats treated with retinoic acid or N-(4-hydroxyphenyl)-retinamide. Livers were perfused and cell fractions enriched in hepatocytes, and nonparenchymal cells were obtained for assays of RE and enzyme activity. The specific activity of lecithin:retinol acyltransferase (LRAT) was approximately 10-fold greater in the nonparenchymal cell than the hepatocyte fraction from both vitamin A-sufficient and retinoid-treated rats. Total RE mass, newly synthesized [3H]RE and LRAT activity were positively correlated in liver and isolated cells of both normal (P < 0.0001) and retinoid-treated rats (P < 0.0002). In nonparenchymal cells, these three constituents were nearly equally enriched as evaluated by their relative specific activity values (RSA, defined as the percentage of recovered activity divided by the percentage of recovered protein), which were each significantly greater than 1.0, with values of 4.3 for total RE mass (P < 0.05), 3.6 for newly synthesized [3H]RE (P < 0.01) and 3.8 for LRAT activity (P < 0.01). In contrast, the specific activities of neutral and acid bile salt-independent retinyl ester hydrolases (REH) did not vary with vitamin A status, and their RSA values were close to 1.0 in both hepatocytes and nonparenchymal cells. These data show that LRAT and REH are differentially regulated by retinoids and that these enzymes also differ in their spacial distribution between liver parenchymal and nonparenchymal cells.


The nasopharynx, face height, and overbite.
In lateral cephalometric films of a group of male subjects, none of whom was a mouth breather, the statistical relationships between overbite and nasopharyngeal dimensions were weak. Moderate relationships were found between face height and nasopharyngeal dimensions. Strongest correlations, with a peak at 10 years, were between--Face height and nasopharyngeal height (+) Face height and nasopharyngeal area (+) Face height and roof angle (-) Dynamic correlations between overbite, face height and nasopharyngeal dimensions were weak. Class II malocclusion subjects on average showed--Smaller Nasopharyngeal and Adenoid Areas. Larger Airways in both real and proportional terms, compared with Class I and normal occlusion subjects.


Maternal and child nutritional supplementation are inversely associated with fasting plasma glucose concentration in young Guatemalan adults.
Cardiovascular disease and diabetes may be programmed early in life by abnormal development associated with undernutrition. We investigated whether maternal nutritional status (MNS; height, pregnancy weight gain, nonpregnant BMI, and prenatal supplementation) or childhood nutritional status (CNS; birth weight, length, ponderal index, height-for-age Z-score at 24 mo, and supplementation from 0 to 24 mo) were related to fasting plasma glucose levels in rural-born Guatemalan adults. We studied 209 men and 220 women (mean age 24.4 y) who were involved in a randomized trial of nutritional supplementation of their mothers during pregnancy and during their early childhoods, conducted from 1969 to 1977. In 2 villages, residents were offered Atole (3.8 MJ and 64 g protein/L); 2 other villages were offered Fresco (1.4 MJ/L, no protein). No associations were observed between anthropometric measures of MNS or CNS and fasting plasma glucose levels. In subgroup analyses, inverse associations (all P < 0.15) with birth size were found among women born to fatter mothers, women with low supplement intake, men born to short mothers, and men more severely stunted at 24 mo. Prenatal supplementation was inversely associated with fasting plasma glucose among women [-0.40 +/- 0.17 mmol/(L. MJ. d), P = 0.02]. Among men, postnatal intake of supplementation of 0.10 to 0.20 MJ/d was associated with up to a 0.56 mmol/L reduction in fasting plasma glucose (P = 0.03), but intake in excess of 0.20 MJ/d provided no added benefit. Among women, the benefit of postnatal supplementation was restricted to those born thin (test for interaction P = 0.10). Improving the nutritional status of undernourished women and children may have positive long-term consequences.


Anemia, Blood Transfusion Requirements and Mortality Risk in Human Immunodeficiency Virus-Infected Adults Requiring Acute Medical Admission to Hospital in South Africa.
Background. Morbidity and mortality remain high among hospitalized patients infected with human immunodeficiency virus (HIV) in sub-Saharan Africa despite widespread availability of antiretroviral therapy. Severe anemia is likely one important driver, and some evidence suggests that blood transfusions may accelerate HIV progression and paradoxically increase short-term mortality. We investigated the relationship between anemia, blood transfusions, and mortality in a South African district hospital. Methods. Unselected consecutive HIV-infected adults requiring acute medical admission to a Cape Town township district hospital were recruited. Admission hemoglobin concentrations were used to classify anemia severity according to World Health Organization/AIDS Clinical Trials Group criteria. Vital status was determined at 90 days, and Cox regression analyses were used to determine independent predictors of mortality. Results. Of 585 HIV-infected patients enrolled, 578 (98.8%) were included in the analysis. Anemia was detected in 84.8% of patients and was severe (hemoglobin, 6.5-7.9 g/dL) or life-threatening (hemoglobin, <6.5 g/dL) in 17.3% and 13.3%, respectively. Within 90 days of the date of admission, 13.5% (n = 78) patients received at least 1 blood transfusion with red cell concentrate and 77 (13.3%) patients died. In univariable analysis, baseline hemoglobin and receipt of blood transfusion were associated with increased mortality risk. However, in multivariable analysis, neither hemoglobin nor receipt of a blood transfusion were independently associated with greater mortality risk. Acquired immune deficiency syndrome-defining illnesses other than tuberculosis and impaired renal function independently predicted mortality. Conclusions. Newly admitted HIV-infected adults had a high prevalence of severe or life-threatening anemia and blood transfusions were frequently required. However, after adjustment for confounders, blood transfusions did not confer an increased mortality risk.


Platelets, arterial thrombosis and cerebral ischemia.
Atherosclerosis is a systemic disease that starts early in life, asymptomatically progressing though adulthood, until clinically manifested. In the last few years, experimental, clinical and pathological investigation has led us to a better knowledge of the pathophysiology of atherothrombotic disease. Atherothrombosis is the result of atherosclerosis progression, and its potentially life-threatening clinical consequences include coronary artery disease, cerebrovascular disease and peripheral artery disease. These events are mostly secondary to atherosclerotic plaque disruption and subsequent in situ thrombus formation which may be embolized and dragged by the arterial flow until occluding distal vessels. The demonstrated beneficial role of antiplatelet drugs in reducing the incidence of nonfatal events in many large clinical trials has demonstrated the major role of platelets in the atherothrombotic process in the arterial tree.


Biologically Plausible, Human-Scale Knowledge Representation.
Several approaches to implementing symbol-like representations in neurally plausible models have been proposed. These approaches include binding through synchrony (Shastri & Ajjanagadde, ), "mesh" binding (van der Velde & de Kamps, ), and conjunctive binding (Smolensky, ). Recent theoretical work has suggested that most of these methods will not scale well, that is, that they cannot encode structured representations using any of the tens of thousands of terms in the adult lexicon without making implausible resource assumptions. Here, we empirically demonstrate that the biologically plausible structured representations employed in the Semantic Pointer Architecture (SPA) approach to modeling cognition (Eliasmith, ) do scale appropriately. Specifically, we construct a spiking neural network of about 2.5 million neurons that employs semantic pointers to successfully encode and decode the main lexical relations in WordNet, which has over 100,000 terms. In addition, we show that the same representations can be employed to construct recursively structured sentences consisting of arbitrary WordNet concepts, while preserving the original lexical structure. We argue that these results suggest that semantic pointers are uniquely well-suited to providing a biologically plausible account of the structured representations that underwrite human cognition.


[Proposal of new trace elements classification to be used in nutrition, oligotherapy and other therapeutics strategies].
1) to propose a new classification of the trace elements based on a study of the recently reported research; 2) to offer detailed and actualized information about trace elements. the analysis of the research results recently reported reveals that the advances of the molecular analysis techniques point out the importance of certain trace elements in human health. A detailed analysis of the catalytic function related to several elements not considered essential o probably essentials up to now is also offered. To perform the integral analysis of the enzymes containing trace elements informatics tools have been used. Actualized information about physiological role, kinetics, metabolism, dietetic sources and factors promoting trace elements scarcity or toxicity is also presented. Oligotherapy uses catalytic active trace elements with therapeutic proposals. The new trace element classification here presented will be of high interest for different professional sectors: doctors and other professions related to medicine; nutritionist, pharmaceutics, etc. Using this new classification and approaches, new therapeutic strategies could be designed to mitigate symptomatology related to several pathologies, particularly carential and metabolic diseases.


Confinement as a tool to probe amorphous order.
We study the effect of confinement on glassy liquids using random first order transition theory as a framework. We show that the characteristic length scale above which confinement effects become negligible is related to the point-to-set length scale introduced to measure the spatial extent of amorphous order in supercooled liquids. By confining below this characteristic size, the system becomes a glass. Eventually, for very small sizes, the effect of the boundary is so strong that any collective glassy behavior is wiped out. We clarify similarities and differences between the physical behaviors induced by confinement and by pinning particles outside a spherical cavity (the protocol introduced to measure the point-to-set length). Finally, we discuss possible numerical and experimental tests of our predictions.


Enhanced susceptibility to chemical induction of ovarian tumors in mice with a germ line p53 mutation.
Mice with a germ line p53 mutation (p53(Ala135Val/wt)) display increased susceptibility to lung, skin, and colon carcinogenesis. Here, we show that p53(Ala135Val/wt) mice developed ovarian tumors significantly more rapidly than their wild-type littermates after 7,12-dimethylbenz(a)anthracene (DMBA) treatment. Approximately 50% of the ovarian tumors in p53(wt/wt) mice and 23% in p53(Ala135Val/wt) mice are adenocarcinomas and the remaining tumors were adenocarcinoma mixed with sarcoma or ovarian sarcomas. All of the p53(Ala135Val/wt) mice had died of ovarian tumors 25 weeks after the initial DMBA treatment, whereas >50% of p53(wt/wt) mice were still alive. These mice not only have a shortened tumor latency but also closely resemble a subset of human ovarian tumors containing the p53 mutation. Microarray and GenMAPP analyses revealed that the mutant p53 (Ala135Val) affected several cellular processes, including the cell cycle, apoptosis, and Wnt pathways. These findings indicate that a germ line p53 mutation significantly enhanced DMBA-induced ovarian tumor development and progression.


Metabolization of Resolvin E4 by ω-Oxidation in Human Neutrophils: Synthesis and Biological Evaluation of 20-Hydroxy-Resolvin E4 (20-OH-RvE4).
Resolvin E4 (RvE4) belongs to the resolvin family of specialized pro-resolving mediators (SPMs). The resolvins are endogenously formed mediators with both potent pro-resolving and anti-inflammatory biological activities and have attracted considerable attention in both inflammation research and drug discovery. Hence, further metabolism of the resolvins is of interest. Gaining knowledge about the structure-function of further metabolites of the resolvins is important due to their interest in drug-discovery efforts. For the first time, the total synthesis and biological evaluations of the ω-20 hydroxylated metabolite of RvE4, named herein 20-OH-RvE4, are presented. RvE4 was converted to 20-OH-RvE4 by human polymorphonuclear leukocytes. LC-MS/MS analysis and UV spectrophotometry reveal that the synthetic 20-OH-RvE4 matched RvE4-converted product 20-OH-RvE4 by human neutrophils. Cellular studies have revealed that RvE4 is formed from eicosapentaenoic acid in physiologic hypoxia by human neutrophils and macrophages, and we herein established that 20-OH-RvE4 is a secondary metabolite formed by the ω-oxidation of RvE4 in human neutrophils. A direct comparison of the biological actions between RvE4 and its metabolic product suggested that 20-OH-RvE4 displayed reduced bioactions in stimulating the efferocytosis of human senescent erythrocytes by human M2-like macrophages. At concentrations down to 0.1 nM, RvE4 increased macrophage erythrophagocytosis, an important pro-resolving function that was diminished due to metabolic transformation. The results provided herein contribute to a novel molecular insight on the further local metabolization of RvE4, the newest member among the SPM superfamily.


Cortisol profiles in sows submitted to an intermittent suckling regime compared with that of abruptly weaned sows.
The aim of this study was to monitor changes in cortisol levels in sows around the time of separation from their piglets, in two different intermittent suckling regimes, compared with that in conventionally weaned sows. Sows were either weaned at 21 days of lactation (CONT) or subjected to an intermittent suckling regime (IS) from 14 days of lactation onwards. Sows in the IS regimes were separated from their piglets for 12 h every day, either from 08:00 to 20:00 hours (IS12) or from 08:00 to 14:00 hours and 20:00 to 02:00 hours (IS6). Separation caused a transient increase in cortisol levels on the first (CONT and IS12) and second (IS12) day of separation, compared with a gradual decline from early morning when the sows were still continuously suckling. In IS6 sows, in contrast, the transient rise in cortisol levels after separation at 08:00 hours was observed on the first 3 days and also on day 7 of the IS regime. Cortisol parameters were correlated with peri-ovulatory characteristics like onset of oestrus, onset of the LH surge and time of ovulation. Onset of the LH surge was delayed in IS6 sows. In conclusion, increase in cortisol levels as a consequence of separation of sows and piglets, is an acute, incidental phenomenon in IS12 and CONT sows, but shows a repeated acute elevation in IS6 sows, possibly placing IS6 sows at a higher risk of influencing peri-ovulatory processes and developing cystic follicles.


Pharmaceutical-grade albumin: impaired drug-binding capacity in vitro.
Albumin is the most abundant protein in blood plasma, and due to its ligand binding properties, serves as a circulating depot for endogenous and exogenous (e.g. drugs) compounds. Hence, the unbound drug is the pharmacologically active drug. Commercial human albumin preparations are frequently used during surgery and in critically ill patients. Recent studies have indicated that the use of pharmaceutical-grade albumin is controversial in critically ill patients. In this in vitro study we investigated the drug binding properties of pharmaceutical-grade albumins (Baxter/Immuno, Octapharma, and Pharmacia & Upjohn), native human serum, and commercially available human serum albumin from Sigma Chemical Company. The binding properties of the various albumin solutions were tested in vitro by means of ultrafiltration. Naproxen, warfarin, and digitoxin were used as ligands. HPLC was used to quantitate the total and free drug concentrations. The data were fitted to a model of two classes of binding sites for naproxen and warfarin and one class for digitoxin, using Microsoft Excel and Graphpad Prism. The drugs were highly bound to albumin (95-99.5%). The highest affinity (lowest K1) was found with naproxen. Pharmaceutical-grade albumin solutions displayed significantly lower drug-binding capacity compared to native human serum and Sigma albumin. Thus, the free fraction was considerably higher, approximately 40 times for naproxen and 5 and 2 times for warfarin and digitoxin, respectively. The stabilizers caprylic acid and N-acetyl-DL-tryptophan used in the manufacturing procedure seem to be of importance. Adding the stabilizers to human serum and Sigma albumin reduced the binding affinity whereas charcoal treatment of the pharmaceutical-grade albumin from Octapharma almost restored the specific binding capacity. This in vitro study demonstrates that the specific binding for warfarin and digitoxin is significantly reduced and for naproxen no longer detectable in pharmaceutical-grade albumin. It further shows that the addition of stabilizers may be of major importance for this effect.


Autopsy study of unruptured incidental intracranial aneurysms.
The autopsy files and preparations of unruptured incidental intracranial aneurysms seen at the Montefiore Medical Center between 1951 and 1987 were reviewed. There were 84 patients with 102 unruptured aneurysms in a total of 10,259 autopsies, giving a prevalence of 0.8%. Sixteen of the 84 (19%) had multiple aneurysms. The thickness of walls of aneurysms could be estimated in 78 of 102 aneurysms, and was determined to be either thin or thin and thick in 71 aneurysms. In this study, four noteworthy factors were found: (1) the incidence of unruptured aneurysms was higher in elderly patients aged 60 years or older, and the peak percentage was 1.2% in the seventh decade; (2) aneurysms occurred more frequently in females than males, with a ratio of 53:31; (3) the most common site of aneurysms was the middle cerebral artery; 37 of 102 aneurysms (36%) occurred on it; and (4) the rate of small aneurysms was very high; 50 of 93 aneurysms (54%) were 4 mm or less in diameter, and 33 aneurysms (35%) were 5-9 mm in diameter. However, relationships could not be found between age distribution and location, size, or thickness of walls; between gender and size or thickness of walls; between location and size or thickness of walls; or between size and thickness of walls. Based on published statistics on subarachnoid hemorrhage and this study, the rupture rate of unruptured aneurysms seems to be very low. Although the risk of rupture may be relatively low in small aneurysms, its low risk probably cannot be explained adequately by morphological examination only.


Olives and olive oil are sources of electrophilic fatty acid nitroalkenes.
Extra virgin olive oil (EVOO) and olives, key sources of unsaturated fatty acids in the Mediterranean diet, provide health benefits to humans. Nitric oxide (•NO) and nitrite (NO2 (-))-dependent reactions of unsaturated fatty acids yield electrophilic nitroalkene derivatives (NO2-FA) that manifest salutary pleiotropic cell signaling responses in mammals. Herein, the endogenous presence of NO2-FA in both EVOO and fresh olives was demonstrated by mass spectrometry. The electrophilic nature of these species was affirmed by the detection of significant levels of protein cysteine adducts of nitro-oleic acid (NO2-OA-cysteine) in fresh olives, especially in the peel. Further nitration of EVOO by NO2 (-) under acidic gastric digestive conditions revealed that human consumption of olive lipids will produce additional nitro-conjugated linoleic acid (NO2-cLA) and nitro-oleic acid (NO2-OA). The presence of free and protein-adducted NO2-FA in both mammalian and plant lipids further affirm a role for these species as signaling mediators. Since NO2-FA instigate adaptive anti-inflammatory gene expression and metabolic responses, these redox-derived metabolites may contribute to the cardiovascular benefits associated with the Mediterranean diet.


Use of the Atlantic nut clam (Nucula proxima) and catworm (Nephtys incisa) in a sentinel species approach for monitoring the health of Bay of Fundy estuaries.
Designing an effective environmental monitoring system for population responses requires knowledge of the biology of appropriate sentinel species and baseline information on the area's physical and chemical characteristics. This study collected information in Saint John Harbor, NB, Canada, for two abundant marine benthic invertebrates, the Atlantic nut clam (Nucula proxima) and the catworm (Nephtys incisa) to characterize their seasonal and spatial variability, determine the ideal sampling time and methods, and develop baseline data for future studies. We also evaluated whether contamination is impacting invertebrates by comparing sediment metal concentrations to responses of benthic infauna. Metals were generally below sediment quality guidelines except for nickel and arsenic. Clam densities were variable between sites but not seasons, whereas catworm densities were not significantly different between sites or seasons. Overall, these species show potential for environmental monitoring, although investigation at more contaminated sites is warranted to assess their sensitivity.


Thermodynamic Bounds on the Ultra- and Infra-affinity of Hsp70 for Its Substrates.
The 70 kDa heat shock protein Hsp70 has several essential functions in living systems, such as protecting cells against protein aggregation, assisting protein folding, remodeling protein complexes, and driving translocation into organelles. These functions require high affinity for nonspecific amino acid sequences that are ubiquitous in proteins. It has been recently shown that this high affinity, called ultra-affinity, depends on a process driven out of equilibrium by ATP hydrolysis. Here, we establish the thermodynamic bounds for ultra-affinity, and further show that the same reaction scheme can in principle be used both to strengthen and to weaken affinities (leading in this case to infra-affinity). We show that cofactors are essential to achieve affinity beyond the equilibrium range. Finally, biological implications are discussed.


Differentiation-related changes in lipid classes with long-chain and very long-chain polyenoic fatty acids in rat spermatogenic cells.
In rat seminiferous tubules (ST), cells that contain polar and neutral lipids with long-chain polyenoic fatty acids (PUFA) and sphingomyelins (SM) and ceramides (Cer) with very long chain (VLC) PUFA of the n-6 series coexist. In this study, pachytene spermatocytes and round spermatids were isolated to determine how these lipids change during spermatogenesis. As the amount per cell of PUFA-rich glycerophospholipids (GPL) decreased with cell size, the 22:5/20:4 ratio increased with cell differentiation. The elovl2 and elovl5 genes, required for 22:5 formation, were expressed (mRNA) in both cell types. Residual bodies- particles with compacted organelles and materials discarded from late spermatids-concentrated cholesterol, 22:5-rich triacylglycerols, and GPL, including plasmalogens and phosphatidylserine. Species of SM and Cer with nonhydroxylated (n-) VLCPUFA (28:4, 30:5, and 32:5) predominated in pachytene spermatocytes, whereas species with the corresponding 2-hydroxy (2-OH) VLCPUFA prevailed in round spermatids. Thus, a dramatic increase in the 2-OH/n-VLCPUFA ratio in SM and Cer was a hallmark of differentiation. A substantial decrease of 2-OH SM occurred between spermatids and mature spermatozoa and 2-OH SM species were collected in residual bodies "en route" to Sertoli cells. Notably, spermatids and spermatozoa gained a significant amount of ceramides devoid of n-VLCPUFA but having 2-OH VLCPUFA as their main fatty acids.


Metabolic regulation of the HBV-specific T cell function.
Chronically HBV infected subjects are more than 260 million worldwide; cirrhosis and liver cancer represent possible outcomes which affect around 700,000 patients per year. Both innate and adaptive immune responses are necessary for viral control and both have been shown to be defective in chronic patients. Metabolic remodeling is an essential process in T cell biology, particularly for T cell activation, differentiation and survival. Cellular metabolism relies on the conversion of nutrients into energy to support intracellular processes, and to generate fundamental intermediate components for cell proliferation and growth. Adaptive immune responses are the central mechanisms for the resolution of primary human infections leading to the activation of pathogen-specific B and T cell functions. In chronic HBV infection the anti-viral immune response fails to contain the virus and leads to persistent hepatic tissue damage which may finally result in liver cirrhosis and cancer. This T cell failure is associated with metabolic alterations suggesting that control of nutrient uptake and intracellular utilization as well as correct regulation of intracellular metabolic pathways are strategic for T cell differentiation during persistent chronic infections. This review will discuss some of the main features of the T cell metabolic processes which are relevant to the generation of an efficient antiviral response, with specific focus on their clinical relevance in chronic HBV infection in the perspective of possible strategies to correct deregulated metabolic pathways underlying T cell dysfunction of chronic HBV patients.


Prevalence of menopausal symptoms in Australian women at midlife: a systematic review.
To systematically review the published data for the prevalence and severity of menopausal symptoms in Australian women. A comprehensive and systematic literature search was done using six databases to extract all English-language, peer-reviewed studies that contained information on the prevalence of menopausal symptoms among women living in Australia. Risk of bias of included studies was assessed using a risk-of-bias tool specifically designed for the systematic review of prevalence studies. Eight independent studies met our inclusion criteria. There was no consistent pattern of vasomotor, psychological, physical or sexual symptom prevalence for the studies that reported symptoms across the menopausal stages. The ranges of the prevalences for the various outcomes were wide. A high level of bias was observed related to both external and internal validities for the included studies. The available data for the prevalence of menopausal symptoms in Australian women are not sufficient to allow conclusive findings. A large, appropriately sampled study using a validated questionnaire is needed to establish the prevalence and severity of menopausal symptoms in Australian women.


Isolation of a novel bone glycosylated phosphoprotein with disulphide cross-links to osteonectin.
An 80 kDa protein was purified from calf bone by HCl-demineralization followed by 0.5 M EDTA/1.0 M NaCl extraction and sequential chromatography on DE-52, hydroxyapatite, and TSK-gel G3000SW HPLC columns. From the DE-52 column the protein was eluted at three different fractions, of which one further separated into two fractions on the hydroxyapatite column, indicating that the protein is present in four different molecular forms designated as 80 k-I-1, k-I-2, k-II, k-III. The N-terminal sequence analysis of all four forms gave the same sequence, SEQYNQEPNNV. Several tryptic internal peptides were also generated, purified and sequenced, leading to the identification of several repeat sequences, IFLGXXEI. Homology searching of the N-terminal and internal sequences indicates that this is a novel protein. Both 80 k-I-2 and k-III had similar amino acid composition with high contents of Asx, Glx and Leu and contained 7 and 16 phosphoserines per 1000 total amino acids, respectively. The 80 k-I-1 and 80 k-II forms were stained with Rhodamine B specific for phosphoproteins. The four forms contained different contents of neutral sugars ranging from 5.5 to 26% (w/w protein) and approximately 1.7% sialic acid. These data indicated that the 80 kDa protein exists in four isomeric forms, at least based on the different post-translational modifications. The evaluation of the 80 kDa glycosylated phosphoprotein under alkylating, reducing and non-reducing conditions indicated that this protein undergoes polymerization through intermolecular disulphide bonds. Furthermore, the 80 kDa protein and osteonectin (ON), both of which are cysteine-rich proteins, can cross-link with each other via disulphide bonds, and this process can be induced to take place in vitro under experimental conditions. The occurrence of such a phenomenon in vivo was confirmed from the presence of similar high Mr components containing both 80 kDa and ON in the same SDS/PAGE bands, detected by the respective antibody reactions in crude bone extracts which were extracted in the presence of alkylating agent.


Cholinergic potentiation of isoproterenol-induced cAMP level in sweat gland.
Although methacholine (MCh) and a Ca2+ ionophore A23187 do not enhance the tissue adenosine 3',5'-cyclic monophosphate (cAMP) level by themselves, they markedly potentiate isoproterenol (ISO)-induced tissue cAMP accumulation in isolated simian eccrine sweat glands in a dose-dependent manner. The agonist concentration producing 50% of the maximal response of such a potentiated cAMP accumulation was 2.1 X 10(-7) M for MCh, 2.5 X 10(-7) M for ISO, and 2.9 X 10(-6) M for A23187. Unlike cAMP accumulation induced by ISO alone, MCh-stimulated ISO-induced cAMP accumulation is dependent on extracellular Ca2+. MCh- plus ISO-induced cAMP level is tripled by 10(-2) M theophylline (TH), and while the ISO-induced cAMP level was also elevated by TH, it was not enhanced to the level of ISO-plus MCh-induced cAMP accumulation, indicating that phosphodiesterase inhibition is not the major mechanism for the augmentative effect of MCh. The augmentative effect of MCh was seen only in the secretory portion, whereas that of A23187 was seen in both the secretory portion and the duct. The data suggest that MCh-induced augmentation of ISO-stimulated cAMP accumulation is due to increased cAMP formation, not decreased cAMP degradation, and that it may be mediated by an elevated intracellular Ca2+.


Influence of beta-naphthoflavone and methoxychlor pretreatment on the biotransformation and estrogenic activity of methoxychlor in channel catfish (Ictalurus punctatus).
The organochlorine pesticide methoxychlor [1,1,1-trichloro-2,2-bis(4-methoxyphenyl) ethane] (MXC) has been classified as a proestrogen in mammals and fish, requiring demethylation prior to eliciting estrogenic activity or binding to the estrogen receptor. While microsomal demethylation occurs readily in the liver of fish, little is known about the enzyme(s) responsible or the effect of cytochrome P450 (CYP) inducers, other than those of CYP1A and CYP2K, on biotransformation. Consequently, male channel catfish were pretreated with MXC or beta-naphthoflavone (BNF), alone and in combination, to determine their effects on CYP protein expression, MXC biotransformation by hepatic microsomes, microsomal protein binding, and MXC estrogenic activity as determined by serum vitellogenin and 17beta-estradiol. Liver microsomes of both treated and untreated mature male catfish catalyzed formation of monodemethylated MXC, bisdemethylated MXC, as well as ring-hydroxylated metabolites. Pretreatment with BNF did not affect MXC metabolite profiles, overall rates of MXC biotransformation, or microsomal proteins recognized by anti-trout CYP2K1, but had the expected effect of inducing CYP1A and associated ethoxyresorufin O-deethylase activity. By contrast, pretreatment with MXC, alone or in combination with BNF, significantly reduced rates of MXC biotransformation and binding to liver microsomal protein. MXC/BNF cotreatment followed by MXC significantly induced serum vitellogenin, whereas MXC treatment alone led to a nonsignificant increase in vitellogenin and a significant increase in serum 17beta-estradiol. Thus, estrogenic activity elicited by cotreatment with MXC and BNF can occur despite diminished capacity of liver microsomes to catalyze formation of estrogenic demethylated metabolites or metabolites that bind microsomal protein. Possible mechanisms of MXC-induced attenuation of CYP-dependent metabolism are discussed.


Ultrasound imaging and guidance in peripheral nerve entrapment: hydrodissection highlighted.
Ultrasound (US) imaging has become one of the most useful modalities to assess peripheral nerve disorders. Nowadays, it is as important as nerve conduction studies and electromyography for peripheral nerve entrapment. Additionally, US is also helpful in guiding a possible intervention. As peripheral nerves are tiny, palpation-guided injections are really challenging for precisely targeting the nerves. By using US, perineural injections have significantly become safe and effective. Recently, US-guided hydrodissection has emerged as the mainstream for nerve entrapment. Accordingly, this review aims to summarize and update the mechanism and evidence regarding this imperative procedure for neuropathic pain management. Furthermore, the pathogenesis, anatomic features, US findings and histological correlations of nerve entrapment syndromes will also be discussed in this article.


Analysis of microarray data for identification of key microRNA signatures in glioblastoma multiforme.
Glioblastoma multiforme (GBM) is one of the most malignant types of glioma known for its reduced survival rate and rapid relapse. Previous studies have shown that the expression patterns of different microRNAs (miRNA/miR) play a crucial role in the development and progression of GBM. In order to identify potential miRNA signatures of GBM for prognostic and therapeutic purposes, we downloaded and analyzed two expression data sets from Gene Expression Omnibus profiling miRNA patterns of GBM compared with normal brain tissues. Validated targets of the deregulated miRNAs were identified using MirTarBase, and were mapped to Search Tool for the Retrieval of Interacting Genes/Proteins, Database for Annotation, Visualization and Integrated Discovery and Kyoto Encyclopedia of Genes and Genomes databases in order to construct interaction networks and identify enriched pathways of target genes. A total of 6 miRNAs were found to be deregulated in both expression datasets studied. Pathway analysis demonstrated that most of the target genes were enriched in signaling cascades connected to cancer development, such as 'Pathways in cancer', 'Focal adhesion' and 'PI3K-Akt signaling pathway'. Of the five target genes that were enriched in the glioblastoma pathway, in the WikiPathway database, both HRas proto-oncogene, GTPase and MET proto-oncogene, receptor tyrosine kinase target genes of hsa-miR-139-5p, were found to be significantly associated with patient survival. The present study may thus form the basis for further exploration of hsa-miR-139-5p, not only as a therapeutic agent, but also as a diagnostic biomarker for GBM as well as a predictive marker for patient survival.