Nitric oxide storage levels modulate vasodilation and the hypotensive effect induced by photobiomodulation using an aluminum gallium arsenide (AlGaAs) diode laser (660 nm).
The aim of this study was to evaluate the participation of nitric oxide (NO) in the hypotensive and vasorelaxation effect induced by PBM using an aluminum gallium arsenide (AlGaAs) diode laser (660 nm). Male Wistar rats were treated with the inhibitor of nitric oxide synthase (L-NAME). A red laser (660 nm; 63 J/cm2; 56 s/point) was applied to the abdominal region at six different points. Thoracic aorta was dissected for vascular reactivity study, and a laser (660 nm; 96 J/cm2; 56 s) was applied after incubation with the NO donor DETA-NO, PBS, or hydroxicobalamin. Endothelial cells (HUVEC) were treated with DETA-NO or CuSO4, and then, PBM (63 J/cm2) was applied, and the nitric oxide was detected. Hypertensive L-NAME rats did not exhibit a decrease in blood pressure after PBM. PBM promoted vasodilation in the aorta isolated from normotensive rats, and less effect in the aorta of L-NAME rats and the addition of the NO donor, DETA-NO, promoted greater vasodilation by PBM in the aorta of L-NAME rats. In endothelial cells, an increase in NO, after PBM, was detected; however, with the addition of CuSO4, which catalyzes the decomposition of NO storage, there was no detection of NO after PBM. The results of this study demonstrate that the hypotensive and vasodilatory effect of PBM with a red laser at 660 nm is modulated by the release of nitric oxide from the storage.


Implication of the neurotrophin receptor p75NTR in vascular diseases: beyond the eye.
The p75 neurotrophin receptor (p75NTR) is a member of TNF-α receptor superfamily that bind all neurotrophins, mainly regulating their pro-apoptotic actions. Ischemia is a common pathology in different cardiovascular diseases affecting multiple organs, however the contribution of p75NTR remains not fully addressed. The aim of this work is to review the current evidence through published literature studying the impact of p75NTR receptor in ischemic vascular diseases. In the eye, several ischemic ocular diseases are associated with enhanced p75NTR expression. Ischemic retinopathy including diabetic retinopathy, retinopathy of prematurity and retinal vein occlusion are characterized initially by ischemia followed by excessive neovascularization. Beyond the eye, cerebral ischemia, myocardial infarction and critical limb ischemia are ischemic cardiovascular diseases that are characterized by altered expression of neurotrophins and p75NTR expression. We surveyed both clinical and experimental studies that examined the impact of p75NTR receptor in ischemic diseases of eye, heart, brain and peripheral limbs. p75NTR receptor is a major player in multiple ischemic vascular diseases affecting the eye, brain, heart and peripheral limbs with significant increases in its expression accompanying neuro-vascular injury. This has been addressed in the current review along with the beneficial vascular outcomes of p75NTR inhibition.


A novel self-evaluation tool to assess the team function of a child protection team.
The aim of this study was to develop a reliable and valid self-evaluation tool for use by child protection team (CPT) members. An online survey was administered to members of 10 CPTs. The survey included the following 3 sections: 1) initial conditions (eg, team composition, resources), 2) enabling conditions (eg, team effort, strategy), and 3) team effectiveness (eg, team cohesion, meeting performance standards). Each section contained multiple subscales. Internal consistency was calculated using Cronbach α. To evaluate construct validity, the subscale scores of the most advanced teams who qualified as centers of excellence (n = 3) were compared with the subscale scores of the other teams (n = 7) to determine whether the tool could distinguish between the two. Of 116 team members, 83 (72%) completed the survey. The subscales exhibited good internal consistency (α = .71-.97). The 3 centers of excellence had significantly higher mean scores than the other 7 CPTs on the following subscales: incentives (in the initial conditions section, 61.46 vs 38.89; P = .003), effort (in the enabling conditions section, 79.31 vs 67.70; P = .003), and professional growth (in the team effectiveness section, 83.89 vs 80.40; P = .004). This novel survey demonstrates satisfactory test characteristics and can be used to assess CPT performance and identify areas for improvement.


Bone changes in children on long-term treatment with etretinate.
Ten children with disorders of keratinization (ichthyosis, 4, pityriasis rubra pilaris, 2, psoriasis, 2, and palmoplantar keratoderma, 2) were treated with etretinate for 0.5 to 9.6 years. The children ranged from 1.5 to 15.7 years of age. The total dose of etretinate ranged from 700 to 52,596 mg and the maintenance dose, from 0.21 to 0.86 mg/kg/day. Radiographic skeletal surveys were performed at various stages during treatment or following treatment. Bone abnormalities were found in eight children. Six children had periosteal thickening, two periosteal bone resorption, two osteoporosis, three disc narrowing, and one slender long bones. Compared with age- and sex-matched normal controls, the children had decreased cortical bone thickness of the second left metacarpal bone. The children's physical growth and laboratory investigations were normal. We recommend that children on long-term etretinate therapy be investigated regularly for bone changes.


Assessment of heavy metal contamination in surface sediments along the Mediterranean coast of Morocco.
In this paper, heavy metal contamination in surface sediments along the Mediterranean coast of Morocco was investigated. Determining pollution degree as well as heavy metal origins were the main objectives of this investigation. For this reason, concentrations of nine heavy metals (Cd, Cu, Cr, Ni, Pb, Zn, Hg, Fe, and Mn) were analyzed at ten stations sampled during three different periods. The obtained concentrations showed significant variation between sampling periods, which was controlled by several environmental and chemical processes. According to contamination indices results using pollution load index (PLI), modified contamination degree (mCd), geoaccumulation index (Igeo), enrichment factor (EF), and potential ecological risk index (RI), sampling stations were classified between uncontaminated and strongly contaminated without detecting any intense heavy metal pollution in surface sediment. Likewise, the EF values were comprised between no enrichment and moderate to severe enrichment. According to sediment quality guidelines, the calculated M-ERM-Q indicated that heavy metal mixtures have between 9 and 49% probability for being toxic. This result revealed lowest to medium-low potential of adverse effects to biota populations. Regarding heavy metal origins, multivariate statistical investigation showed that Cd, Cu, Hg, Pb, and Zn are derived mainly from anthropogenic activities, while Fe, Mn, Cr, and Ni were derived from natural sources. Despite Cd is considered as a typical anthropogenic metal, the very low concentrations obtained in this study support the involvement of natural factor in the enrichment with this metal. Therefore, the surface sediments contamination along the Mediterranean coasts of Morocco is particularly caused by a combination of anthropogenic and natural factors. As a result, the study area can be considered as not significantly enriched by human activities.


Papain-induced asthma--physiological and immunological features.
Increasing reports of respiratory disease associated with exposure to papain prompted clinical, physiological, and immunological studies of the supervisor of a meat tenderizer factory who developed asthma after long-term contact with papain dust. His symptoms were worse at work and better on weekends and vacations. Bronchial inhalation challenges produced both immediate and late asthma to papain but not to the other ingredients in the food product. Immunological studies revealed the presence of specific IgE antibodies by direct and passive transfer skin tests and the radioallergosorbent test, and specific precipitating antibodies by immunodiffusion tests. These findings are indicative of a dual type I and III hypersensitivity. Papain acting as an allergen in an occupational setting is a risk factor for eliciting asthma even in a nontropic individual.


Five new phenolic glycosides from Hedyotis scandens.
Five new phenolic glycosides, hedyotosides A-E (1-5), including a new cyanogenic glycoside (1), along with 10 known compounds (6-15) were isolated from the whole plants of Hedyotis scandens. The structures of compounds 1-5 were established by extensive spectroscopic analyses and acid hydrolysis. All the isolated compounds were evaluated for their in vitro antiviral activity against respiratory syncytial virus (RSV) with cytopathic effect (CPE) reduction assay. Compounds 6 and 15 showed anti-RSV effects with IC(50) values of 20 and 25 μg/mL, respectively.


The Drosophila IMD pathway in the activation of the humoral immune response.
The IMD pathway signaling plays a pivotal role in the Drosophila defense against bacteria. During the last two decades, significant progress has been made in identifying the components and deciphering the molecular mechanisms underlying this pathway, including the means of bacterial sensing and signal transduction. While these findings have contributed to the understanding of the immune signaling in insects, they have also provided new insights in studying the mammalian NF-κB signaling pathways. Here, we summarize the current view of the IMD pathway focusing on how it regulates the humoral immune response of Drosophila.


Ciliary and flagellar structure and function--their regulations by posttranslational modifications of axonemal tubulin.
Eukaryotic cilia and flagella are evolutionarily conserved microtubule-based organelles protruding from the cell surface. They perform dynein-driven beating which contributes to cell locomotion or flow generation. They also play important roles in sensing as cellular antennae, which allows cells to respond to various external stimuli. The main components of cilia and flagella, α- and β-tubulins, are known to undergo various posttranslational modifications (PTMs), including phosphorylation, palmitoylation, tyrosination/detyrosination, Δ2 modification, acetylation, glutamylation, and glycylation. Recent identification of tubulin-modifying enzymes, especially tubulin tyrosine ligase-like proteins which perform tubulin glutamylation and glycylation, has demonstrated the importance of tubulin modifications for the assembly and functions of cilia and flagella. In this chapter, we review recent work on PTMs of ciliary and flagellar tubulins in conjunction with discussing the basic knowledge.


The clinical process in psychiatry: a clinimetric approach.
The aim of this review was to examine the clinical process in psychiatry, with special reference to clinimetrics, a domain concerned with the measurement of clinical phenomena that do not find room in customary taxonomy. A MEDLINE search from inception to August 2010 was performed for English-language articles using the keywords clinical judgment, clinimetric, staging, comorbidity, sequential treatment, and subclinical symptoms in relation to psychiatric illness. It was supplemented by a manual search of the literature. Choice of assessment strategies was based on their established or potential incremental increase in clinical information compared to use of diagnostic criteria. Contributions were evaluated according to the principles of clinimetrics. Several innovative assessment strategies were identified: the use of diagnostic transfer stations with repeated assessments instead of diagnostic endpoints, subtyping versus integration of different diagnostic categories, staging methods, and broadening of clinical information through macroanalysis and microanalysis. The most representative examples were selected. Current assessment strategies in psychiatric research do not reflect the sophisticated thinking that underlies clinical decisions in practice. The clinimetric perspective provides an intellectual home for the reproduction and standardization of these clinical intuitions.


GPR120: A bi-potential mediator to modulate the osteogenic and adipogenic differentiation of BMMSCs.
Free fatty acids display diverse effects as signalling molecules through GPCRs in addition to their involvement in cellular metabolism. GPR120, a G protein-coupled receptor for long-chain unsaturated fatty acids, has been reported to mediate adipogenesis in lipid metabolism. However, whether GPR120 also mediates osteogenesis and regulates BMMSCs remain unclear. In this study, we showed that GPR120 targeted the bi-potential differentiation of BMMSCs in a ligand dose-dependent manner. High concentrations of TUG-891 (a highly selective agonist of GPR120) promoted osteogenesis via the Ras-ERK1/2 cascade, while low concentrations elevated P38 and increased adipogenesis. The fine molecular regulation of GPR120 was implemented by up-regulating different integrin subunits (α1, α2 and β1; α5 and β3). The administration of high doses of TUG-891 rescued oestrogen-deficient bone loss in vivo, further supporting an essential role of GPR120 in bone metabolism. Our findings, for the first time, showed that GPR120-mediated cellular signalling determines the bi-potential differentiation of BMMSCs in a dose-dependent manner. Additionally, the induction of different integrin subunits was involved in the cytoplasmic regulation of a seesaw-like balance between ERK and p38 phosphorylation. These findings provide new hope for developing novel remedies to treat osteoporosis by adjusting the GPR120-mediated differentiation balance of BMMSCs.


Enhanced protein degradation by intracellular delivery of pre-fused PROTACs using lipid-like nanoparticles.
Proteolysis-targeting chimaera (PROTAC) technology is an emerging approach for achieving targeted degradation of a protein of interest (POI) intracellularly. However, the cell permeability of PROTACs is limited by their high molecular weight and total polar surface area. Moreover, the activation of the proteasome-mediated degradation by PROTAC requires the formation of a ternary (three-component) complex, composed of the PROTAC, the POIs, and E3-ligases related proteins (E3Ps). Simplifying the three-component system to two-component system could theoretically increase the efficiency of the formation of ternary complex and enhance the protein degradation efficiency. Herein, we demonstrate that pre-fusion of PROTACs with E3Ps (called "pre-fused PROTACs") before administration could transform the original PROTAC system to two-component system. After delivery by lipid nanoparticles, the degradation of POI by pre-fused PROTACs was dramatically increased and accelerated compared with standard PROTACs. Moreover, we demonstrated that this approach could be generalized to another hydrophobic tag (HyT) degrader by demonstrating the improved targeted protein degradation after pre-fusion the HyT degrader with heat shock protein 70 (HSP70).


B1 integrin/Fak/Src signaling in intestinal epithelial crypt cell survival: integration of complex regulatory mechanisms.
The molecular determinants which dictate survival and apoptosis/anoikis in human intestinal crypt cells remain to be fully understood. To this effect, the roles of beta1 integrin/Fak/Src signaling to the PI3-K/Akt-1, MEK/Erk, and p38 pathways, were investigated. The regulation of six Bcl-2 homologs (Bcl-2, Mcl-1, Bcl-X(L), Bax, Bak, Bad) was likewise analyzed. We report that: (1) Anoikis causes a down-activation of Fak, Src, Akt-1 and Erk1/2, a loss of Fak-Src association, and a sustained/enhanced activation of p38beta, which is required as apoptosis/anoikis driver; (2) PI3-K/Akt-1 up-regulates the expression of Bcl-X(L) and Mcl-1, down-regulates Bax and Bak, drives Bad phosphorylation (both serine112/136 residues) and antagonizes p38beta activation; (3) MEK/Erk up-regulates Bcl-2, drives Bad phosphorylation (serine112 residue), but does not antagonize p38bactivation; (4) PI3-K/Akt-1 is required for survival, whereas MEK/Erk is not; (5) Src acts as a cornerstone in the engagement of both pathways by beta1 integrins/Fak, and is crucial for survival; and (6) beta1 integrins/Fak and/or Src regulate Bcl-2 homologs as both PI3-K/Atk-1 and MEK/Erk combined. Hence, beta1 integrin/Fak/Src signaling translates into integrated mediating functions of p38beta activation and regulation of Bcl-2 homologs by PI3-K/Akt-1 and MEK/Erk, consequently determining their requirement (or not) for survival.


Chronic obstructive pulmonary disease and exacerbations: patient insights from the global Hidden Depths of COPD survey.
Although chronic obstructive pulmonary disease (COPD) is a major global health burden there is a lack of patient awareness of disease severity, particularly in relation to exacerbations. We conducted a global patient survey using an innovative, internet-based methodology to gain insight into patient perceptions of COPD and exacerbations in a real-world sample typical of today's working-age COPD population. Two thousand patients with COPD (53%), chronic bronchitis (52%) and/or emphysema (22%) from 14 countries completed an online questionnaire developed by the authors. The Medical Research Council (MRC) breathlessness scale was used to delineate symptom severity. Over three quarters of patients (77%) had experienced an exacerbation, with 27% of MRC 1 and 2 patients and 52% of MRC 3, 4 and 5 patients requiring hospitalization as a result of an exacerbation. While a majority of MRC 1 and 2 patients (51%) reported being back to normal within a few days of an exacerbation, 23% of MRC 3, 4 and 5 patients took several weeks to return to normal and 6% never fully recovered. A high proportion of patients (39%) took a 'wait and see' approach to exacerbations.Despite the high prevalence of exacerbations and their negative impact on quality of life, 73% of MRC 1 and 2 patients and 64% of MRC 3, 4 and 5 patients felt that they had control of their COPD. However, 77% of all patients were worried about their long-term health, and 38% of MRC 1 and 2 patients and 59% of MRC 3, 4 and 5 patients feared premature death due to COPD. To reduce the adverse effects of COPD on patients' quality of life and address their fears for the future, we need better patient education and improved prevention and treatment of exacerbations.


Enantioselective radical C-H amination for the synthesis of β-amino alcohols.
Asymmetric, radical C-H functionalizations are rare but powerful tools for solving modern synthetic challenges. Specifically, the enantio- and regioselective C-H amination of alcohols to access medicinally valuable chiral β-amino alcohols remains elusive. To solve this challenge, a radical relay chaperone strategy was designed, wherein an alcohol was transiently converted to an imidate radical that underwent intramolecular H-atom transfer (HAT). This regioselective HAT was also rendered enantioselective by harnessing energy transfer catalysis to mediate selective radical generation and interception by a chiral copper catalyst. The successful development of this multi-catalytic, asymmetric, radical C-H amination enabled broad access to chiral β-amino alcohols from a variety of alcohols containing alkyl, allyl, benzyl and propargyl C-H bonds. Mechanistic experiments revealed that triplet energy sensitization of a Cu-bound radical precursor facilitates catalyst-mediated HAT stereoselectivity, enabling the synthesis of several important classes of chiral β-amines by enantioselective, radical C-H amination.


Lack of association between the eNOS rs1800779 (A/G) polymorphism and the myocardial infarction incidence among the Iraqi Kurdish population.
The genetic polymorphisms of the endothelial nitric oxide synthase (eNOS) gene are strongly associated with several cardiovascular diseases (CVDs) in various populations. The current study aimed to investigate the association of the eNOS rs1800779 (A/G) polymorphism with the progress of myocardial infarction (MI). Eighty-five healthy subjects and 80 patients with MI admitted to the Erbil Cardiac Centre in the Kurdistan Region of Iraq were enrolled in the study. All participants were Kurdish from the same ethnic group. The amplification refractory mutation system polymerase chain reaction (ARMS-PCR) was used to determine the rs1800779 (A/G) polymorphism of eNOS, and the nitric oxide (NO) serum level was detected by spectrophotometer. The genotypic frequencies of the eNOS rs1800779 AA (wild type), AG, and GG were 58.75%, 33.75%, and 7.50%, respectively, in the MI patients, and 49.41%, 43.53%, and 7.06%, respectively, for the control group. The frequencies of the A and the G alleles were 75.6% and 24.4%, respectively, in the MI group, and 71.2% and 28.8%, respectively, in the control subjects. The results revealed a lack of association of the rs1800779 genotype distribution with the level of NO serum and increased risk of MI. The study concluded that there is a lack of association between the genotypes and alleles of the rs1800779 eNOS and susceptibility to MI in the studied population.


Sex differences in renal damage induced in the mouse by Amanita virosa.
The sensitivity of male and female mice to Amanita virosa was compared. Dried, homogenized mushroom was given orally by stomach tubing at doses of 100, 200, 400 and 800 mg dried mushroom/kg body weight. Both in males and in females, the kidneys were the only organs showing macroscopical changes. The dose of 100 mg/kg caused renal damage in females, whereas in males the first signs of kidney damage were seen at the dose of 400 mg/kg. The renal lesions observed in the males were located in the cortex, while in the females they were limited to the outer stripe of the outer medullary zone. Testectomy diminished the nephrotoxicity of A. virosa in male mice and caused changes in the localization of renal lesions.


Assessment of free radical-induced damage in brain proteins after ischemia and reperfusion.
Brain damage initiated during global ischemia has been shown to be exacerbated by iron-dependent lipid peroxidation during early reperfusion. We hypothesized that other cellular components might be involved in similar free radical reactions. In this study we examined three brain protein fractions and ribosomal RNA for evidence of free radical damage during post-ischemic reperfusion. Global brain ischemia was induced by 20-min cardiac arrest. Dogs were divided into four groups: (1) non-ischemic controls; (2) 20-min cardiac arrest without reperfusion; (3) 20-min cardiac arrest and 2 h reperfusion; (4) 20-min cardiac arrest and 8 h reperfusion. Soluble proteins and proteins from ribosomes and synaptosomes were assayed by a dinitrophenylhydrazine method for carbonyl groups, which are characteristic products of protein peroxidation. The ribosomal RNA was also examined by electrophoresis. When proteins from each fraction were peroxidized in vitro by Fenton reagents, carbonyl content increased as [Fe2+] was increased from 0 to 100 microM. However, following reperfusion there was no significant accumulation of carbonyl content in either the soluble (ANOVA P = 0.92) or ribosome (P = 0.10) protein fractions. There was a significant decrease in the carbonyl content of the synaptosome protein fraction after 8 h of reperfusion (P = 0.03). Similarly, although ribosomal RNA fragmentation was observed in ethidium stained agarose gels following in vitro reaction with Fenton reagents, there was no evidence of ribosomal RNA fragmentation or cross-linking following reperfusion. These results suggest that reperfusion free radical reactions do not involve these cellular proteins or ribosomal RNA.


A two-photon turn-on probe for glucose uptake.
We report a two-photon turn-on probe (AS1) that can be excited by 780 nm femto-second pulses and visualize glucose uptake and the changes in the intracellular glucose concentration in live cells and tissue by two-photon microscopy.


Glutathione decreases in dopaminergic PC12 cells interfere with the ubiquitin protein degradation pathway: relevance for Parkinson's disease?
Parkinson's disease (PD) is characterized by the presence of proteinaceous neuronal inclusions called Lewy bodies in susceptible dopaminergic midbrain neurons. Inhibition of the ubiquitin-proteasome protein degradation pathway may contribute to protein build-up and subsequent cell death. Ubiquitin is normally activated for transfer to substrate proteins by interaction with the E1 ubiquitin ligase enzyme via a thiol ester bond. Parkinson's disease is also characterized by decreases in midbrain levels of total glutathione which could impact on E1 enzyme activity via oxidation of the active site sulfhydryl. We have demonstrated that increasing reductions in total glutathione in dopaminergic PC12 cells results in corresponding decreases in ubiquitin-protein conjugate levels suggesting that ubiquitination of proteins is inhibited in a glutathione-dependent fashion. Decreased ubiquitinated protein levels appears to be due to inhibition of E1 activity as demonstrated by reductions in endogenous E1-ubiquitin conjugate levels as well as decreases in the production of de novo E1-ubiquitin conjugates when glutathione is depleted. This is a reversible process as E1 activity increases upon glutathione restoration. Our data suggests that decreases in cellular glutathione in dopaminergic cells results in decreased E1 activity and subsequent disruption of the ubiquitin pathway. This may have implications for neuronal degeneration in PD.


Repertoire selection by pre-B-cell receptors and B-cell receptors, and genetic control of B-cell development from immature to mature B cells.
During B-cell development the surrogate light (SL) chain is selectively expressed in progenitor and precursor B cells during the developmental stages of D(H) to J(H) and V(H) to D(H)J(H) rearrangements. Approximately half of all muH chains produced by these rearrangements cannot pair with SL chains and cannot form a pre-B-cell receptor (pre-BCR). A spectrum of affinities between VpreB and individual V(H) domains generates preB cells with pre-BCR of different fitness which, in turn, determines the extent of the pre-B II-cell proliferation and the fidelity of allelic exclusion of the H chain locus. Once pre-BCR is expressed, SL chain expression is turned off. As pre-B II cells proliferate, SL is diluted out, thus limiting pre-BCR formation. As a consequence, pre-B II cells stop proliferating, become small and resting and begin to rearrange the L chain loci. Multiple rearrangements of the kappaL chain alleles are often detected in wild-type small pre-B II cells. Around 20% of the muH chain-expressing small pre-B II cells also express L chains but do not display the Ig on the surface. Hence, it is likely that not all L chains originally generated in resting pre-B II cells can pair with the muH chain previously present in that cell. The best fitting ones are selected preferentially to generate sIg+ B cells. Furthermore, the transition of immature B cells from the bone marrow to spleen and their development to mature cells appear as two separate steps controlled by different genes.


Mechanical thrombectomy versus intravenous alteplase alone in acute isolated posterior cerebral artery occlusion: a systematic review.
Acute isolated posterior cerebral artery occlusions (aPCAOs) were excluded or under-represented in major randomized trials of mechanical thrombectomy (MT). The benefit of MT in comparison to intravenous tissue plasminogen activator (alteplase; IV-tPA) alone in these patients remains controversial and uncertain. We performed a systematic search of PubMed, MEDLINE, and EMBASE databases for articles comparing MT with or without bridging IV-tPA and IV-tPA alone for aPCAO using keywords ('posterior cerebral artery', 'thrombolysis' and 'thrombectomy') with Boolean operators. Extracted data from patients reported in the studies were pooled into groups (MT vs IV-tPA alone) for comparison. Estimated rates for favorable outcome (modified Rankin scale score 0-2), symptomatic intracranial hemorrhage (sICH), and mortality were extracted. Seven articles (201 MT patients, 64 IV-tPA) were included, all retrospective. There was no statistically significant difference between pooled groups in median age, median presentation National Institutes of Health Stroke Scale (NIHSS) score, PCAO segment, and median time from symptom onset to puncture or needle. The recanalization rate was significantly higher in the MT group than the IV-tPA group (85.6% vs 53.1%, p<0.00001). Odds ratios for favorable outcome (OR 1.5, 95% CI 0.8 to 2.5), sICH (OR 1.1, 95% CI 0.2 to 5.5), and mortality (OR 1.4, 95% CI 0.5 to 3.6) did not significantly favor any modality. We found no significant differences in odds of favorable outcome, sICH, and mortality in MT and IV-tPA in comparable aPCAO patients, despite superior MT recanalization rates. Equipoise remains regarding the optimal treatment modality for these patients.


Alkali borosilicate glass by fly ash from a coal-fired power plant.
The possibility of using coal fly ash as a silica source for alkali borosilicate glass was investigated. Alkali borosilicate glasses were prepared from the coal fly ash mixed with 30 wt.% reagents composed of Na(2)O and B(2)O(3) by susceptor-induction heating. Their densities ranged from 2.24 to 2.55 g cm(-3) and decreased as the amount of B(2)O(3) addition increased. However, the Vickers microhardness showed a different tendency with the density since the glass network connectivity improved by boron anomaly, which was identified by a nuclear magnetic resonance analysis. The Vickers microhardness of the glass product, with the addition of 15 wt.% B(2)O(3) and 15 wt.% Na(2)O, was about 4030 MPa. Furthermore, the changes in microstructure were consistent with those in the chemical stability by the toxicity characteristic leaching procedure (TCLP).


Noninvasive measurement of intracranial pressure via the pulsatility index on transcranial Doppler sonography: Is improvement possible?
We hypothesized that using hemodynamic variables could improve the prediction of intracranial pressure (ICP) from the middle cerebral artery pulsatility index (PI) measured with transcranial Doppler sonography. In this prospective study, 39 patients with traumatic brain injury were routinely examined with transcranial Doppler sonography, and the middle cerebral artery PI was calculated. A multivariate model including hematocrit, mean arterial blood pressure, heart rate, and arterial CO2 pressure (PaCO2 ) was evaluated. Thirty-nine comatose patients (16 women and 23 men; age range 18-73 years; median 44 years) were included, and 234 data pairs (consisting of ICP and corresponding PI values) were analyzed. ICP ranged from -3 mmHg to +52 mmHg, and PI from 0.6 to 2.85. We found a significant but weak correlation between PI and the square root of ICP (R(2) between 0.29 and 0.34, p < 0.0001). A slightly stronger correlation was detected when hemodynamic variables were incorporated (R(2) between 0.37 and 0.43). Of these variables, mean arterial blood pressure had the most significant influence. In this study, PI was not a sufficiently strong predictor of ICP to be used in clinical practice. Its reliability did not improve even when hemodynamic variables were considered. Therefore, we recommend abandoning the use of PI for the noninvasive measurement of ICP in clinical practice.


Affinity purification and partial characterization of a yeast multiprotein complex for nucleotide excision repair using histidine-tagged Rad14 protein.
The nucleotide excision repair (NER) pathway of eukaryotes involves approximately 30 polypeptides. Reconstitution of this pathway with purified components is consistent with the sequential assembly of NER proteins at the DNA lesion. However, recent studies have suggested that NER proteins may be pre-assembled in a high molecular weight complex in the absence of DNA damage. To examine this model further, we have constructed a histidine-tagged version of the yeast DNA damage recognition protein Rad14. Affinity purification of this protein from yeast nuclear extracts resulted in the co-purification of Rad1, Rad7, Rad10, Rad16, Rad23, RPA, RPB1, and TFIIH proteins, whereas none of these proteins bound to the affinity resin in the absence of recombinant Rad14. Furthermore, many of the co-purifying proteins were present in approximately equimolar amounts. Co-elution of these proteins was also observed when the nuclear extract was fractionated by gel filtration, indicating that the NER proteins were associated in a complex with a molecular mass of >1000 kDa prior to affinity chromatography. The affinity purified NER complex catalyzed the incision of UV-irradiated DNA in an ATP-dependent reaction. We conclude that active high molecular weight complexes of NER proteins exist in undamaged yeast cells.


Aggressive systemic mastocytosis complicated by protein-losing enteropathy.
A 46-year-old woman was admitted to the hospital with complaints of chronic diarrhoea, vomiting and severe muscle weakness. Clinical examination showed a lethargic, malnourished, dehydrated patient with ascites and bilateral leg oedema. Laboratory evaluation revealed mild normochromic normocytic anaemia and severe hypoproteinaemia with hypoalbuminaemia. Upper gastrointestinal endoscopy showed a thickened, friable duodenal mucosa with multiple erosions. Colonoscopy revealed nodular, pseudopolypoid lesions with patchy erosions in the left hemicolon. Haematoxylin-eosin stained sections from biopsies of endoscopically abnormal bowel segments showed multi-focal aggregates of large, histiocyte-like cells with abundant pale cytoplasm in the lamina propria. These cells were negative on PAS, Ziehl-Neelsen, Giemsa and toluidine blue stains. Their immunophenotype was CD68 (+), c-kit/CD117 (+) and mast cell tryptase (+), which is consistent with mast cells. A trephine biopsy showed diffuse replacement of the bone marrow by atypical, monomorphic, frequently spindle-shaped mast cells. No associated haematopoietic malignancy was detected. The final diagnosis was aggressive systemic mastocytosis with involvement of the gastrointestinal tract complicated by protein-losing enteropathy. This association has not been reported previously. The patient has been treated with prednisolone and interferon-alpha and has since recovered.


Cyclic AMP-elevating agents down-regulate the oxidative burst induced by granulocyte-macrophage colony-stimulating factor (GM-CSF) in adherent neutrophils.
Human neutrophils, plated on fibronectin-precoated wells, were found to release large quantities of superoxide anion (O2-) in response to GM-CSF. O2- production was reduced by prostaglandin E2 (PGE2) and the phosphodiesterase type IV (PDE IV) inhibitor RO 20-1724. Both agents are known to increase intracellular cyclic AMP (cAMP) levels by inducing its production (PGE2) or blocking its catabolism (RO 20-1724). When added in combination, PGE2 and RO 20-1724 had a marked synergistic inhibitory effect, which was reproduced by replacing PGE2 with a direct activator of adenylate cyclase, i.e. forskolin (FK). Moreover, the neutrophil response to GM-CSF was inhibited by a membrane-permeable analogue of cAMP in a dose-dependent manner. As GM-CSF and PGE2 are known to be generated at tissue sites of inflammation, the results suggest the existence of a PGE2-dependent regulatory pathway potentially capable of controlling the neutrophil response to GM-CSF, in turn limiting the risk of local oxidative tissue injury. Moreover, owing to its susceptibility to amplification by RO 20-1724, the PGE2-dependent pathway and in particular PDE-IV may represent a pharmacological target to reduce the generation of histotoxic oxidants by GM-CSF-responding neutrophils.


Endoplasmic Reticulum Stress Instigates the Rotenone Induced Oxidative Apoptotic Neuronal Death: a Study in Rat Brain.
The present study was conducted to evaluate the involvement of endoplasmic reticulum stress in rotenone-induced oxidative neuronal death in rat brain. Rotenone (6 μg/3 μl) was administered intranigrally, unilaterally (right side) in SD rat brain. Neuronal morphology, expression level of tyrosine hydroxylase (TH) and endoplasmic reticulum (ER) stress markers like glucose-regulated protein 78 (GRP78), growth arrest and DNA damage-inducible gene 153 (GADD153), eukaryotic translation initiation factor 2α (p-eIF2α/eIF2α) and cleaved caspase-12 were estimated in the rat brain. Levels of reactive oxygen species (ROS), reduced glutathione (GSH) and enzymatic activities of glutathione peroxidase (GPx) and glutathione reductase (GRd) were estimated to assess the rotenone induced oxidative stress. Apoptotic death of neurons was assessed by estimating the mRNA level of caspase-3. Rotenone administration caused altered neuronal morphology, decreased expression of TH, augmented ROS level, decreased level of GSH and decreased activities of GPx and GRd enzymes which were significantly attenuated with the pretreatment of ER stress inhibitor, salubrinal (1 mg/kg, intraperitoneal). Significantly increased levels of GRP78, GADD, dephosphorylated eIF2α and cleaved caspase-12 was also observed after rotenone administration, which was inhibited with the pretreatment of salubrinal. Rotenone-induced increased mRNA level of caspase-3 was also attenuated by pretreatment of salubrinal. Findings suggested that salubrinal treatment significantly inhibited the rotenone-induced neurotoxicity implicating that ER stress initiates the rotenone-induced oxidative stress and neuronal death.


DLK Activation Synergizes with Mitochondrial Dysfunction to Downregulate Axon Survival Factors and Promote SARM1-Dependent Axon Degeneration.
Axon degeneration is a prominent component of many neurological disorders. Identifying cellular pathways that contribute to axon vulnerability may identify new therapeutic strategies for maintenance of neural circuits. Dual leucine zipper kinase (DLK) is an axonal stress response MAP3K that is chronically activated in several neurodegenerative diseases. Activated DLK transmits an axon injury signal to the neuronal cell body to provoke transcriptional adaptations. However, the consequence of enhanced DLK signaling to axon vulnerability is unknown. We find that stimulating DLK activity predisposes axons to SARM1-dependent degeneration. Activating DLK reduces levels of the axon survival factors NMNAT2 and SCG10, accelerating their loss from severed axons. Moreover, mitochondrial dysfunction independently decreases the levels of NMNAT2 and SCG10 in axons, and in conjunction with DLK activation, leads to a dramatic loss of axonal NMNAT2 and SCG10 and evokes spontaneous axon degeneration. Hence, enhanced DLK activity reduces axon survival factor abundance and renders axons more susceptible to trauma and metabolic insult.


Quinolines as extremely potent and selective PDE5 inhibitors as potential agents for treatment of erectile dysfunction.
In a continuing effort to discover novel chemotypes as potent and selective PDE5 inhibitors for the treatment of male erectile dysfunction (ED), we have found that 4-benzylaminoquinoline derivatives are very potent and selective PDE5 inhibitors. Some compounds in this series had PDE5 IC(50)'s as low as 50 pM. While an electron withdrawing group at the C6-position of the quinoline substantially improved PDE5 potency, an ethyl group at the C8-position not only improved the PDE5 potency but also the isozyme selectivity. Substitutents at the C3-position can incorporate a variety of different groups. The synthesis and primary structure-activity relationship of this new series of potent PDE5 inhibitors are described.


Parallel engineering and activity profiling of a base editor system.
Selecting the most suitable existing base editors and engineering new variants for installing specific base conversions with maximal efficiency and minimal undesired edits are pivotal for precise genome editing applications. Here, we present a platform for creating and analyzing a library of engineered base editor variants to enable head-to-head evaluation of their editing performance at scale. Our comprehensive comparison provides quantitative measures on each variant's editing efficiency, purity, motif preference, and bias in generating single and multiple base conversions, while uncovering undesired higher indel generation rate and noncanonical base conversion for some of the existing base editors. In addition to engineering the base editor protein, we further applied this platform to investigate a hitherto underexplored engineering route and created guide RNA scaffold variants that augment the editor's base-editing activity. With the unknown performance and compatibility of the growing number of engineered parts including deaminase, CRISPR-Cas enzyme, and guide RNA scaffold variants for assembling the expanding collection of base editor systems, our platform addresses the unmet need for an unbiased, scalable method to benchmark their editing outcomes and accelerate the engineering of next-generation precise genome editors.


Studies of Lassa Virus Cell Entry.
Host cell entry is the first and most fundamental step of every virus infection and represents a major barrier for zoonotic transmission and viral emergence. Targeting viral entry appears further as a promising strategy for therapeutic intervention. Several cellular receptors have been identified for Lassa virus, including dystroglycan, TAM receptor tyrosine kinases, and C-type lectins. Upon receptor binding, LASV enters the host cell via a largely unknown clathrin- and dynamin-independent endocytotic pathway that delivers the virus to late endosomes, where fusion occurs after engagement of a second, intracellular receptor, the late endosomal/lysosomal resident protein LAMP1. Here, we describe a series of experimental approaches to investigate LASV cell entry and to test candidate inhibitors for their action at this early and decisive step of infection.


A prospective and randomized comparison of the safety and effects of therapeutic levels of enoxaparin versus unfractionated heparin in chronically anticoagulated patients undergoing elective cardiac catheterization.
Consecutive patients with atrial fibrillation and/or prosthetic heart valves, receiving chronic anticoagulation with phenprocoumon and scheduled to undergo cardiac catheterization, were randomized to subcutaneous enoxaparin twice daily (n = 32) or intravenous UFH (n = 36). Cardiac catheterization was performed at an international normalized ratio <1.5. Activated partial thromboplastin times and levels of anti-Factor Xa activity were measured daily. The time until effective anticoagulation (primary endpoint) was significantly shorter for enoxaparin than for UFH (1.1 +/- 0.4 days versus 3.7 +/- 2.5 days, p<0.0001). The percentage of days of effective anticoagulation was significantly higher in the enoxaparin group than in the UFH group (93.3 +/- 9.5% versus 53.7 +/- 26.6%, p <0.0001). In conclusion, enoxaparin achieves therapeutic levels of anticoagulation more rapidly and consistently than UFH in chronically anticoagulated patients with prosthetic heart valves and/or atrial fibrillation undergoing cardiac catheterization.


Role of melatonin in the treatment of COVID-19; as an adjuvant through cluster differentiation 147 (CD147).
COVID-19 caused by the SARS-CoV-2 outbreak quickly has turned into a pandemic. However, no specific antiviral agent is yet available. In this communication, we aimed to evaluate the significance of CD147 protein and the potential protective effect of melatonin that is mediated by this protein in COVID-19. CD147 is a glycoprotein that is responsible for the cytokine storm in the lungs through the mediation of viral invasion. Melatonin use previously was shown to reduce cardiac damage by blocking the CD147 activity. Hence, melatonin, a safe drug, may prevent severe symptoms, reduce symptom severity and the adverse effects of the other antiviral drugs in COVID-19 patients. In conclusion, the use of melatonin, which is reduced in the elderly and immune-compromised patients, should be considered as an adjuvant through its CD147 suppressor and immunomodulatory effect.


Spatial and temporal coordination of insulin granule exocytosis in intact human pancreatic islets.
Insulin secretion is widely studied because it plays a central role in glucose homeostasis and diabetes. Processes from insulin granule fusion in beta cells to in vivo insulin secretion have been elucidated, but data at the cellular level do not fully account for several aspects of the macroscopic secretory pattern. Here we investigated how individual secretory events are coordinated spatially and temporally within intact human islets. We used the fluorescent probe neuropeptide Y (NPY)-pHluorin to visualise insulin granule secretion in isolated intact human islets. We found that individual beta cells respond to increases in glucose concentration by releasing insulin granules in very discrete bursts with periods consistent with in vivo pulsatile insulin secretion. In successive secretory bursts during prolonged exposure to high glucose levels, secretory events progressively localised to preferential release sites, coinciding with the transition to second phase insulin secretion. Granule secretion was very synchronised in neighbouring beta cells, forming discrete regional clusters of activity. These results reveal how individual secretory events are coordinated to produce pulsatile insulin secretion from human islets.


Insulin lispro low mixture twice daily versus basal insulin glargine once daily and prandial insulin lispro once daily in patients with type 2 diabetes requiring insulin intensification: a randomized phase IV trial.
To compare the efficacy and safety of two insulin intensification strategies in patients with type 2 diabetes inadequately controlled on basal insulin glargine with metformin and/or pioglitazone. A multinational, randomized, open-label trial that compared insulin lispro low mixture (LM25; n = 236) twice daily with a basal-prandial regimen of insulin glargine once daily and insulin lispro once daily (IGL; n = 240) over 24 weeks in patients with HbA1c 7.5-10.5% and fasting plasma glucose ≤ 6.7 mmol/l. The primary objective was to assess non-inferiority [per-protocol (PP) population], and then superiority [intention-to-treat (ITT) population], of LM25 versus IGL according to change in HbA1c after 24 weeks (non-inferiority margin 0.4%, two-sided significance level 0.05). Estimated change [least squares (LS) mean (95% CI)] in HbA1c after 24 weeks: -1.30 (-1.44, -1.16)% with LM25 and -1.08 (-1.22, -0.94)% with IGL. Non-inferiority was shown [LS mean (95% CI) HbA1c treatment difference -0.21 (-0.38, -0.04) (PP population)]; gated superiority assessment showed a statistically significant advantage for LM25 (p = 0.010; ITT population). Mean blood glucose, glycaemic variability, overall tolerability and hypoglycaemic episodes per patient-year did not show significant differences between treatments during the study. In patients with type 2 diabetes inadequately controlled on once-daily basal insulin glargine and metformin and/or pioglitazone, intensification with LM25 was superior to a basal-prandial approach in terms of reduction in HbA1c after 24 weeks and did not increase hypoglycaemia episodes.


Multisystem Involvement in Post-Acute Sequelae of Coronavirus Disease 19.
The purpose of this study was to describe cerebrovascular, neuropathic, and autonomic features of post-acute sequelae of coronavirus disease 2019 ((COVID-19) PASC). This retrospective study evaluated consecutive patients with chronic fatigue, brain fog, and orthostatic intolerance consistent with PASC. Controls included patients with postural tachycardia syndrome (POTS) and healthy participants. Analyzed data included surveys and autonomic (Valsalva maneuver, deep breathing, sudomotor, and tilt tests), cerebrovascular (cerebral blood flow velocity [CBFv] monitoring in middle cerebral artery), respiratory (capnography monitoring), and neuropathic (skin biopsies for assessment of small fiber neuropathy) testing and inflammatory/autoimmune markers. Nine patients with PASC were evaluated 0.8 ± 0.3 years after a mild COVID-19 infection, and were treated as home observations. Autonomic, pain, brain fog, fatigue, and dyspnea surveys were abnormal in PASC and POTS (n = 10), compared with controls (n = 15). Tilt table test reproduced the majority of PASC symptoms. Orthostatic CBFv declined in PASC (-20.0 ± 13.4%) and POTS (-20.3 ± 15.1%), compared with controls (-3.0 ± 7.5%, p = 0.001) and was independent of end-tidal carbon dioxide in PASC, but caused by hyperventilation in POTS. Reduced orthostatic CBFv in PASC included both subjects without (n = 6) and with (n = 3) orthostatic tachycardia. Dysautonomia was frequent (100% in both PASC and POTS) but was milder in PASC (p = 0.002). PASC and POTS cohorts diverged in frequency of small fiber neuropathy (89% vs 60%) but not in inflammatory markers (67% vs 70%). Supine and orthostatic hypocapnia was observed in PASC. PASC following mild COVID-19 infection is associated with multisystem involvement including: (1) cerebrovascular dysregulation with persistent cerebral arteriolar vasoconstriction; (2) small fiber neuropathy and related dysautonomia; (3) respiratory dysregulation; and (4) chronic inflammation. ANN NEUROL 2022;91:367-379.


Authorship Disparities in International Neurosurgical Research Collaborations: A Bibliometric Analysis.
Research is pivotal to neurosurgical training and practice. The objectives of this study were to quantify neurosurgical research output by authors from low-income countries (LICs), using author affiliation as a proxy, and to understand the patterns of collaboration between LIC authors and their international partners. PubMed, CINAHL, and EMBASE were searched for neurosurgical literature published by authors from the 27 World Bank LICs from 2010 to 2020. These articles were screened for relevance. Information about publication type, study design, and author demographics was then extracted from included articles. Scopus was subsequently used to determine the H-indices of the authors. The rate at which LIC authors have been publishing neurosurgical research has increased significantly from 2010 to 2020. Overall, 19 of the 27 LICs were represented. The LICs with the most research output includes Uganda, Ethiopia, and Rwanda. When LIC authors collaborated with authors from middle-or-high-income countries, these LIC researchers were solely listed in a middle authorship position more than 70% of the time. On average, the H-index of LIC authors was 4.9, compared with average H-indices of 8.7 and 16.8 for their MIC and HIC collaborators, respectively. The positioning of LIC researchers as middle author contributors revealed significant authorship disparities in international neurosurgical research collaborations. The average H-indices of authors from middle-or-high-income countries were more than 3 times greater than those of LIC authors. Quantifying this issue allows neurosurgical organizations to understand the current landscape and to set concrete goals for research capacity building in LICs.


Microvascular complications and prevalence of urgency incontinence in Japanese patients with type 2 diabetes mellitus: The dogo study.
Diabetes was significantly positively associated with urgency incontinence in several epidemiological studies. We examine the association between diabetic neuropathy, which we defined based on neuropathic symptoms, the absence of the Achilles reflex, and/or abnormal vibration perception, and urgency incontinence among Japanese patients with type 2 diabetes mellitus. Study subjects were 742 Japanese patients with type 2 diabetes mellitus, aged 19-70 years, who had undergone blood tests at our institutions. A self-administered questionnaire was used to collect information on the variables under study. Urgency incontinence was defined as present when a subject answered "once a week or more" to the question: "Within one week, how often do you leak urine because you cannot defer the sudden desire to urinate ?". Diabetic neuropathy was diagnosed if the patients showed two or more of the following three characteristics: neuropathic symptoms, the absence of the Achilles reflex, and/or abnormal vibration perception. Adjustment was made for sex, age, body mass index, duration of type 2 diabetes mellitus, current smoking, hypertension, dyslipidemia, glycated hemoglobin, stroke, coronary artery disease, insulin therapy, diabetic retinopathy, diabetic nephropathy, and diabetic neuropathy. The prevalence of urgency incontinence was 8.6%. Diabetic neuropathy was independently positively associated with urgency incontinence: the adjusted OR was 2.20 (95%CI: 1.16-4.36). Associations between diabetic retinopathy or nephropathy and the prevalence of urgency incontinence were not significant. In Japanese patients with type 2 diabetes mellitus, only diabetic neuropathy was independently positively associated with urgency incontinence. Neurourol. Urodynam. 35:1024-1027, 2016. © 2015 Wiley Periodicals, Inc.


Hyperbaric oxygen therapy ameliorates TNBS-induced acute distal colitis in rats.
This study investigated the therapeutic effects of hyperbaric oxygen in experimental acute distal colitis focusing on its effect on the production of pro-inflammatory cytokines, nitric oxide and hypoxia-inducible factor 1alpha. Colitis was induced with a rectal infusion of 150 mg/kg of TNBS under anesthesia with Ketamine (50 mg/kg) and Xylazine (10 mg/kg). Control animals received only rectal saline. After colitis induction, animals were subjected to two sessions of hyperbaric oxygen and were then euthanized. The distal intestine was resected for macroscopic analysis, determination of myeloperoxidase activity, western-blotting analyses of inducible nitric oxide synthase and cyclooxygenase-2 expression and immunohistochemical analysis of hypoxia-inducible factor 1alpha and cyclooxygenase-2. Cytokines levels in the distal intestine were measured using an enzyme-linked immunosorbent assay. Hyperbaric oxygen therapy attenuated the severity of acute distal colitis, with reduced macroscopic damage score. This effect was associated with prevention in the increase of pro-inflammatory cytokine production; myeloperoxidase activity, in the expression of inducible nitric oxide synthase and cyclooxygenase-2. Finally, hyperbaric oxygen inhibited the acute distal colitis-induced up-regulation of hypoxia-inducible factor 1alpha. The results indicate that hyperbaric oxygen attenuates the severity of acute distal colitis through the down-regulation of pro-inflammatory events.


Global epidemiological study of variation of premenstrual symptoms with age and sociodemographic factors.
To determine patterns of premenstrual symptom experiences across the reproductive age range, and the effects of other factors on premenstrual symptoms. Global cross-sectional survey. Sample A total of 7226 women aged 15-49 recruited by random sampling with approximately 400 each from France, Germany, Hungary, Italy, Spain, UK, Brazil, Mexico, Hong Kong, Pakistan and Thailand. Approximately 1000 women in Japan and Korea and 500 Australian women were found using Internet panels. Questionnaire of 23 premenstrual symptoms, sociodemographic and lifestyle variables. Four different patterns of premenstrual symptoms were found with ageing: symptoms showing no change with age, symptoms decreasing with age, symptoms reaching a maximum just before or around age 35, and symptoms reaching a maximum around age 40-44. Smoking was associated with an increase in most of the premenstrual symptoms. Increasing parity was associated with a decrease in some of the symptoms. These results suggest that the premenstrual symptoms most associated with the years of regular ovulatory cycles are skin disorders, lack of energy, appetite problems, joint/muscle/back pain, anger, abdominal bloating, depressed mood and irritability. Other symptoms which peaked later (swelling of extremities, breast tenderness/pain, weight gain) may reflect exacerbation by hormonal changes of the late reproductive/early menopausal transition. Smokers report more problematic premenstrual symptoms.


Visualizing the Dynamics of Inactive X Chromosomes in Living Cells Using Antibody-Based Fluorescent Probes.
The inactive X chromosome (Xi) harbors characteristic epigenetic features, including the enrichment of histone H3 lysine 27 trimethylation (H3K27me3) and H4 lysine 20 monomethylation (H4K20me1) as well as a lack of histone acetylation. Recently, these modifications have been visualized not only in fixed specimen, but also in living cells via probes derived from modification-specific antibodies. The probes include fluorescently labeled antigen binding fragments (Fabs), which can be loaded into cells, as well as genetically encoded single-chain variable fragments tagged with the green fluorescent protein. We refer to the latter as modification specific intracellular antibodies, or "mintbodies" for short. By using Fabs or mintbodies to target Xi-specific modifications, the dynamics of Xi in living cells can be visualized.


Increased airway reactivity and hyperinsulinemia in obese mice are linked by ERK signaling in brain stem cholinergic neurons.
Obesity is a major risk factor for asthma, which is characterized by airway hyperreactivity (AHR). In obesity-associated asthma, AHR may be regulated by non-TH2 mechanisms. We hypothesized that airway reactivity is regulated by insulin in the CNS, and that the high levels of insulin associated with obesity contribute to AHR. We found that intracerebroventricular (ICV)-injected insulin increases airway reactivity in wild-type, but not in vesicle acetylcholine transporter knockdown (VAChT KD(HOM-/-)), mice. Either neutralization of central insulin or inhibition of extracellular signal-regulated kinases (ERK) normalized airway reactivity in hyperinsulinemic obese mice. These effects were mediated by insulin in cholinergic nerves located at the dorsal motor nucleus of the vagus (DMV) and nucleus ambiguus (NA), which convey parasympathetic outflow to the lungs. We propose that increased insulin-induced activation of ERK in parasympathetic pre-ganglionic nerves contributes to AHR in obese mice, suggesting a drug-treatable link between obesity and asthma.


CYSTEINE-RICH RECEPTOR-LIKE PROTEIN KINASES: their evolution, structure, and roles in stress response and development.
Plant-specific receptor-like protein kinases (RLKs) are central components for sensing the extracellular microenvironment. CYSTEINE-RICH RLKs (CRKs) are members of one of the biggest RLK subgroups. Their physiological and molecular roles have only begun to be elucidated, but recent studies highlight the diverse types of proteins interacting with CRKs, as well as the localization of CRKs and their lateral organization within the plasma membrane. Originally the DOMAIN OF UNKNOWN FUNCTION 26 (DUF26)-containing extracellular region of the CRKs was proposed to act as a redox sensor, but the potential activating post-translational modification or ligands perceived remain elusive. Here, we summarize recent progress in the analysis of CRK evolution, molecular function, and role in plant development, abiotic stress responses, plant immunity, and symbiosis. The currently available information on CRKs and related proteins suggests that the CRKs are central regulators of plant signaling pathways. However, more research using classical methods and interdisciplinary approaches in various plant model species, as well as structural analyses, will not only enhance our understanding of the molecular function of CRKs, but also elucidate the contribution of other cellular components in CRK-mediated signaling pathways.


Renal cell carcinoma: the Ochsner Medical Institution experience (1945-1978).
We reviewed 161 patients operated upon for renal cell carcinoma between 1945 and 1978. Life table and survival analyses were computed to compare the effects of stage, tumor differentiation, cell type, surgical technique, renal vein involvement and sex on the years of survival. Patients with stage I and well differentiated tumors had the best prognosis. All patients surviving 10 years or more had well differentiated tumors. The type of nephrectomy did not affect survival and lymphadenectomy was only of value in staging the disease. The stage and differentiation of the tumor were more important to outcome than choice of therapy.


[On the number and morphometrical parameters of the nucleus ambiguous neurons after the injury and regeneration of the recurrent laryngeal nerve in the rat].
In laryngeal nerves injuries it is essential to know the morpho-functional reorganization of the neurons which supply the larynx in order to be successful with the clinical techniques of functional reinnervation and/or orthotopic transplant. Due to the lack of this type of studies in the literature, we investigated the organization of laryngeal motoneurons in the nucleus ambiguous (NA) after the axotomy and regeneration of the recurrent laryngeal nerve (RLN) in adult rats. We used biotinylated dextran amines (BDA, 3 kDa), this fact is an innovation in the field, because this is a novel methodological approach to this model. We studied a control group of 14 animals and four experimental groups of between 10 and 16 animals each one. In the experimental groups we studied the regeneration of the axotomized nerve in four different intervals of time after the injury: 21-28 days, 42-60 days, 90-120 days, and 150-180 days. In the control group we traced the RLN without injury while in the experimental groups we traced the axotomized RLN after each regeneration interval. The number of traced neurons in the control group was 143 +/- 38; in the experimental groups the number was always lower than in the control (21-28 days: 14 +/- 23 neurons; 42-60 days: 46 +/- 49; 90-120 days: 55 +/- 57; 150-180 days: 61 +/- 60). The morphologic parameters studied within the neuronal bodies in the experimental groups were no statistically different when compared with those in the control group. Results show that the tracing of the RLN after its axotomy and regeneration, in the adult rat, involves a decrease in the number of traced neurons within the NA but no changes in their size or shape during the analysed periods.


ECGI in atrial fibrillation: A clinician's wish list.
Have we a challenge of credibility in the invasive treatment of atrial fibrillation (AFIB)? The incidence of AFIB in the European Union (EU) is about 600,000 cases per year, while only 100,000 get an invasive treatment with a failure rate of close to 40%. Those that remain in AFIB need two times more hospitalizations and three times more re-hospitalizations. AFIB accounts for over 1% of the EU health care costs. Too many patients are not referred and the indication rates of cardiac ablation are inexplicably variable throughout the EU. A reflection is made on how electrocardiographic imaging (ECGI) can contribute to bringing the therapy to an advanced level by achieving a higher success rate, efficiently increasing access to ablation therapies, a better patient selection and therapy planning, personalization and follow up.


New developments in the treatment of esophageal cancer.
Esophageal cancer is a rare but highly virulent malignancy occurring in the United States and other Western countries. Adenocarcinoma of the esophagus has had the most rapid rate of increase of any solid tumor malignancy in the United States. Controversy continues about the optimal surgical approach for the treatment of locally advanced esophageal cancer. Combined chemoradiotherapy is the standard of care in the nonsurgical management of esophageal cancer. Trials of preoperative chemotherapy followed by surgery have not shown a consistent benefit for the addition of chemotherapy to surgery. Preoperative chemoradiotherapy followed by surgery continues to be actively studied in the surgical management of locally advanced esophageal cancer. Pathologic complete responses are seen in 20-40% of patients, with 5-year survival achieved in 30-35% of patients. Recent randomized trials evaluating the role of surgery after completion of chemoradiotherapy have not clearly shown a benefit for the routine use of surgery after chemoradiotherapy. The toxicity of conventional fluorouracil, cisplatin, and radiation have led researchers to evaluate newer agents, such as the taxanes and irinotecan, in combined chemoradiotherapy trials. These trials have indicated promising antitumor activity and therapy tolerance, depending on the dose and schedule of therapy administered. Increasing the dose of radiotherapy or adding a brachytherapy boost to chemoradiotherapy has not improved the outcome of treatment in clinical trials. The advent of newer, targeted therapies, including agents directed against growth factor receptor pathways, tumor angiogenesis, and tumor invasion and metastasis, is leading to a new generation of clinical trials combining these agents with conventional cytotoxic chemotherapy and radiation.


Living retinal nematode (filarial-like) destroyed with photocoagulation.
A motile worm creating tracks upon the pigment epithelium of the retina was observed in each of two patients. The fundus findings resembled pseudoretinitis pigmentosa. Unilateral macular degeneration resulted in one patient. The worm was a nematode, probably a filaria. This report represents the third and fourth intraocular filariae found in the United States. To our knowledge, it is the first report of living, intraretinal filarial-like worms destroyed by photocoagulation.


Triacylglyceride, antioxidant and antimicrobial features of virgin Camellia oleifera, C. reticulata and C. sasanqua Oils.
Virgin oils obtained from seeds of Camellia oleifera (CO), Camellia reticulata (CR) and Camellia sasanqua (CS) were studied for their triacylglyceride composition, antioxidant and antimicrobial activities. Levels of fatty acids determined by ¹H-nuclear magnetic resonance analysis were similar to those reported for olive oils (82.30%-84.47%; 5.69%-7.78%; 0.26%-0.41% and 8.04%-11.2%, for oleic, linoleic, linolenic and saturated acids, respectively). The CR oil showed the best antioxidant potential in the three in vitro models tested. With regard to EC₅₀ values (µg/mL), the order in DPPH radical-scavenging was CR (33.48) < CO (35.20) < CS (54.87). Effectiveness in reducing power was CR (2.81) < CO (3.09) < CS (5.32). IC₅₀ for LPO inhibition were 0.37, 0.52 and 0.75 µg/mL for CR, CO and CS, respectively. All the oils showed antimicrobial activity, and exhibited different selectivity and MICs for each microorganism tested (E. coli, B. cereus and C. albicans). B. cereus was the less sensitive species (MIC: 52.083 ± 18.042 for CO; 41.667 ± 18.042 for CR; 104.167 ± 36.084 for CS mg/mL) and the E. coli was the most sensitive to camellia oil's effect. The standard gentamicin presented higher MIC for E. coli (4.2) than the CR (MIC= 2.6) and CO (MIC = 3.9) oils.


Isothermal Titration Calorimetry for Quantification of Protein-Carbohydrate Interactions.
Isothermal titration calorimetry allows the determination of thermodynamic parameters for the interaction between a protein and mono- or oligosaccharides in solution. For the study of protein-carbohydrate interactions, it is a robust way to determine the stoichiometry and affinity, as well as the enthalpic and entropic contributions to this interaction, without the use of labeled proteins or substrates. Here we describe a standard multiple-injection titration experiment for measuring the binding energetics between a carbohydrate-binding protein and an oligosaccharide.


Safety and efficacy of repeated injections of botulinum toxin A in peripheral neuropathic pain (BOTNEP): a randomised, double-blind, placebo-controlled trial.
Data from previous studies suggest that botulinum toxin A has analgesic effects against peripheral neuropathic pain, but the quality of the evidence is low. We aimed to assess the safety and efficacy of repeated administrations of botulinum toxin A in patients with neuropathic pain. We did a randomised, double-blind, placebo-controlled trial at two outpatient clinics in France (Clinical Pain Centre, Ambroise Paré Hospital, APHP, Boulogne-Billancourt, and Neurological Centre, Hôpital Dupuytren, Limoges) and one in Brazil (Neurological Department, Hospital das Clínicas da FMUSP, São Paulo). Patients aged 18-85 years with peripheral neuropathic pain were randomly assigned (1:1) by block randomisation, according to a centralised schedule, to receive two subcutaneous administrations of botulinum toxin A (up to 300 units) or placebo, 12 weeks apart. All patients and investigators were masked to treatment assignment. The primary outcome was the efficacy of botulinum toxin A versus placebo, measured as the change from baseline in self-reported mean weekly pain intensity over the course of 24 weeks from the first administration. The primary efficacy analysis was a mixed-model repeated-measures analysis in the intention-to-treat population. This trial is registered with ClinicalTrials.gov, NCT01251211. Between Oct 2, 2010, and Aug 2, 2013, 152 patients were enrolled, of whom 68 were randomly assigned (34 per group), and 66 (37 [56%] men) were included in the primary analysis (34 in the botulinum toxin A group and 32 in the placebo group). Botulinum toxin A reduced pain intensity over 24 weeks compared with placebo (adjusted effect estimate -0·77, 95% CI -0·95 to -0·59; p<0·0001). Pain on injection was the only adverse effect reported, and occurred in 19 (56%) participants in the botulinum toxin A group and 17 (53%) of those in the placebo group (p=1·0). Severe pain was experienced by ten (29%) participants in the botulinum toxin A group and 11 (34%) in the placebo group (p=0·8). Two administrations of botulinum toxin A, each of which comprised several injections, have a sustained analgesic effect against peripheral neuropathic pain. Several factors, such as the presence of allodynia and a limited thermal deficit, may be useful in predicting treatment response and should be investigated further. Institut National de la Santé et de la Recherche Médicale (INSERM) and Fondation CNP (France).


Possible alpha-adrenoceptor blocking activity of arotinolol (S-596), a new beta-adrenoceptor blocking agent in isolated dog coronary artery.
Effects of arotinolol on dog coronary arteries were investigated in vitro. In distal portions of left circumflex coronary arteries contracted with 3 X 10(-2)M KCl, norephinephrine relaxed the strips in a concentration-dependent fashion. Propranolol (10(-6)M) converted the norepinephrine-induced relaxations to contractions, and arotinolol (10(-6)-10(-5)M) inhibited the relaxations induced by norepinephrine in a concentration-dependent manner. In proximal portions of the strips after potassium-contracture, norephinephrine produced concentration-dependent contractions which were augmented by propranolol (10(-6)M) and inhibited by arotinolol (10(-6)-10(-5)M). These results suggest that arotinolol has an alpha-adrenoceptor blocking activity in addition to a beta-adrenoceptor blocking action in dog coronary arteries.


First Report of Wohlfahrtiimonas chitiniclastica Isolation from a Patient with Cellulitis in the United States.
We report the first documented isolation of Wohlfahrtiimonas chitiniclastica from a human in the United States. Initially misidentified as Acinetobacter lwoffii by Vitek-2, the isolate was subsequently identified as W. chitiniclastica by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) and 16S rRNA gene sequencing. While the clinical significance of the isolate in this case is unclear, it highlights the superior performance of MALDI-TOF MS for bacterial identification.


Development of a digital platform to improve community response to overdose and prevention among harm reduction organizations.
The overdose crisis in the USA remains a growing and urgent public health concern. Over 108,000 people died due to overdose during 2021. Fatal and non-fatal overdoses are under-reported in the USA due to current surveillance methods. Systemic gaps in overdose data limit the opportunity for data-driven prevention efforts and resource allocation. This study aims to improve overdose surveillance and community response through developing a digital platform for overdose reporting and response among harm reduction organizations. We used a community-engaged, user-center design research approach. We conducted qualitative interviews with N = 44 overdose stakeholders including people who use drugs and harm reductionists. Results highlighted the need for a unified, multilingual reporting system uniquely tailored for harm reduction organizations. Anonymity, data transparency, protection from legal repercussions, data accuracy, and community-branded marketing emerged as key themes for the overdose platform. Emergent themes included the need for real-time data in a dashboard designed for community response and tailored to first responders and harm reduction organizations. This formative study provides the groundwork for improving overdose surveillance and data-driven response through the development of an innovative overdose digital platform.


The Rectangle Target Plot: A New Approach to the Graphical Presentation of Accuracy of Systems for Self-Monitoring of Blood Glucose.
The measurement accuracy of systems for self-monitoring of blood glucose (SMBG) is usually analyzed by a method comparison in which the analysis results are displayed using difference plots or similar graphs. However, such plots become difficult to comprehend as the number of data points displayed increases. This article introduces a new approach, the rectangle target plot (RTP), which aims to provide a simplified and comprehensible visualization of accuracy data. The RTP is based on ISO 15197 accuracy evaluations of SMBG systems. Two-sided tolerance intervals for normally distributed data are calculated for absolute and relative differences at glucose concentrations <100 mg/dL and ≥100 mg/dL. These tolerance intervals provide an estimator of where a 90% proportion of results is found with a confidence level of 95%. Plotting these tolerance intervals generates a rectangle whose center indicates the systematic measurement difference of the investigated system relative to the comparison method. The size of the rectangle depends on the measurement variability. The RTP provides a means of displaying measurement accuracy data in a simple and comprehensible manner. The visualization is simplified by reducing the displayed information from typically 200 data points to just 1 rectangle. Furthermore, this allows data for several systems or several lots from 1 system to be displayed clearly and concisely in a single graph.


Could HPV Testing on Self-collected Samples Be Routinely Used in an Organized Cervical Screening Program? A Modeled Analysis.
Cervical screening on self-collected samples has mainly been considered for targeted use in underscreened women. Updated evidence supports equivalent sensitivity of PCR-based human papillomavirus (HPV) testing on self-collected and clinician-collected samples. Using a well-established model, we compared the lifetime impact on cancer diagnoses and deaths resulting from cervical screening using self-collected samples only, with and without the existing restriction in Australia to women aged 30+ years and ≥2 years overdue, compared with the mainstream program of 5-yearly HPV screening on clinician-collected samples starting at 25 years of age. We conservatively assumed sensitivity of HPV testing on self-collected relative to clinician-collected samples was 0.98. Outcomes were estimated either in the context of HPV vaccination ("routinely vaccinated cohorts;" uptake as in Australia) or in the absence of HPV vaccination ("unvaccinated cohorts"). In unvaccinated cohorts, the health benefits of increased participation from self-collection outweighed the worst case (2%) loss of relative test sensitivity even if only 15% of women, who would not otherwise attend, used it ("additional uptake"). In routinely vaccinated cohorts, population-wide self-collection could be marginally (0.2%-1.0%) less effective at 15% additional uptake but 6.2% to 12.4% more effective at 50% additional uptake. Most (56.6%-65.0%) of the loss in effectiveness in the restricted self-collection pathway in Australia results from the requirement to be 2 or more years overdue. Even under pessimistic assumptions, any potential loss in test sensitivity from self-collection is likely outweighed by improved program effectiveness resulting from feasible levels of increased uptake. Consideration could be given to offering self-collection more widely, potentially as an equal choice for women.See related commentary by Lim, p. 245.


The benefits of being self-determined in promoting physical activity and affective well-being among women recently treated for breast cancer.
In this study, changes in motivational regulations in women following treatment for breast cancer were described. Changes in motivational regulations as predictors of subsequent change in light and moderate-to-vigorous physical activity (PA) and affect were also examined. Women [n = 150; M(age) = 54.41 (SD = 10.87) years] completed self-report questionnaires and wore an accelerometer for 7 days at Time 1 [M = 3.94 (SD = 3.08) months following primary treatment], as well as 3 (Time 2) and 6 (Time 3) months later. Data were analyzed using repeated-measures analysis of variance and path analysis using residual change scores. Identified regulation and self-determined motivation (i.e., combined intrinsic motivation and identified regulation) scores decreased over time (p < 0.05). In the path model [χ(2)(4) = 5.66, p = 0.22, root mean square error of approximation = 0.05 (90% CI: 0.0; 0.15), comparative fit index = 0.99, standardized root mean square of the residuals = 0.03], ΔTime(1-2) in external regulation was associated with ΔTime(2-3) in positive affect (β = -0.16), ΔTime(1-2) in introjected (β = 0.25) and amotivation (β = 0.19) were related to ΔTime(2-3) in negative affect, and ΔTime(1-2) in self-determined motivation was related to ΔTime(2-3) in positive affect (β = 0.40) and moderate-to-vigorous PA (β = 0.21). Changes in motivational regulations were related to changes in PA and affect in the aftermath of breast cancer. Given the benefits of self-determined motivation, additional research is needed to develop and test interventions aimed at enhancing this type of motivation.


New insights into the influences of baking and storage on the nonvolatile compounds in oolong tea: A nontargeted and targeted metabolomics study.
A nontargeted and targeted metabolomics method was applied to comprehensively investigate the influences of baking and storage on chemical constituents in fresh-, strong-, and aged-scent types of Foshou oolong teas. The contents of N-ethyl-2-pyrrolidone-substituted flavanols (EPSFs), flavone C-glycosides, gallic acid, and most lipids increased after baking and storage, while the contents of cis-flavanols, alkaloids, flavonol O-glycosides, and most amino acids decreased. Degradation, epimerization, and interaction with theanine were main pathways for the decrease in cis-flavanols. Approximately 20.7%, 12.8%, and 11.6% of epigallocatechin gallate were degraded, epimerized, and interacted with theanine after baking, respectively; 22.5% and 8.71% of epigallocatechin gallate were degraded and interacted with theanine after 10-year storage, respectively. Simulated reactions confirmed that the increases in EPSFs and apigenin C-glycosides were caused by interactions between theanine and flavanols and between apigenin aglycone and glucose, respectively. This study offers novel insights into chemical changes during baking and storage of oolong tea.


Predicting return to work from health related welfare following low intensity cognitive behaviour therapy.
The aim of this study was to identify predictors of return to work in the short and long term following condition management cognitive-behavioural therapy (CM-CBT). All participants (N=3794) were disability welfare claimants, unemployed due to the presence of a physical or mental health condition. CM-CBT consisted of a seven session group cognitive-behavioural psychoeducational programme, with participants followed-up at 3 and 12-30 months. The primary employment outcome measure was a categorical measure of either returned to work, made progress towards work or remained on welfare. Results index an incremental progress and return to work rate, increasing from 34.41% at short-term follow-up to 53.07% at long-term follow-up. Clinically, 17.40% were classed as recovered following CM-CBT. Reliable psychological change during CM-CBT predicted successful return to work and remaining on welfare was associated with psychological regression over time. The results are discussed in terms of identified methodological weaknesses and the potential of CBT in enabling return to work for the health related unemployed.


Prevalence of rhinitis and related-symptoms in schoolchildren from different cities in Brazil.
By using the standard written questionnaire (WQ), designed for the International Study of Asthma and Allergies in Childhood (ISAAC) we determined the prevalence of rhinitis and its related-symptoms, in Brazilian children and adolescents, living in different cities of the country. The WQ was answered by the parents of 11,403 children aged 6-7 years from five Brazilian cities: Porto Alegre (South, N = 2,846), São Paulo (Southeast, N = 3,005) Uberlândia (Southeast, N = 2,991), Itabira (Southeast, N = 1,151) and Recife (Northeast, N = 1,410). The WQ was also applied to 20,587 adolescents (13-14 years old) living in: Porto Alegre (South, N = 3,195), Curitiba (South, N = 3,004), São Paulo (Southeast, N = 3,008), Uberlândia (Southeast, N = 2,998), Itabira (Southeast, N = 2,134), Salvador (Northeast, N = 3,162) and Recife (Northeast, N = 3,086). The mean response rates were 75% and 95%, for the 6-7 year-old children and for the adolescents, respectively. The data was transcribed to a database (Epi-Info) and analyzed regarding the answers to questions of rhinitis module. The mean prevalence of rhinitis (affirmative response to question 2) was 26.6% and 34.2% in the groups of 6-7 and 13-14 year-old, respectively. Applying the criteria that evaluate the association between nasal and ocular symptoms (affirmative response to question 3) the mean prevalence of allergic rhinitis were 12.8% for the 6-7 year-old children and 18.0% for the adolescents. In conclusion, the prevalence of rhinitis and its related symptoms among children and adolescents living in different Brazilian cities was as high as the prevalence observed in other areas of the world.


Functional interferon system is required for clearance of lassa virus.
Lassa virus (LASV) is the causative agent of Lassa hemorrhagic fever (LF) in humans, a deadly disease endemic to West Africa that results in 5,000 to 10,000 deaths annually. Here we present results demonstrating that functional type I and type II interferon (IFN) signaling is required for efficient control of LASV dissemination and clearance.


Reconstruction of a 4-cm human median nerve gap by including an autogenous nerve slice in a bioabsorbable nerve conduit: case report.
Nerve gaps up to 3 cm in length can be reconstructed in the human with a bioabsorbable nerve conduit; however, nerve gaps greater than 3 cm have not been reconstructible in animal models unless a neurotrophic factor, like cultured Schwann cells, was added to the nerve conduit. It is the purpose of this article to present the result of human nerve regeneration across a 4-cm nerve gap in the median nerve at the wrist when a slice of autogenous normal nerve was placed into the middle of a bioabsorbable nerve conduit.


Copper nanoparticles induced, trimesic acid grafted cellulose-an effective, non-hazardous processing approach for multifunctional textile with low chemical induction.
Cross-linkers have great importance in textile due to the widespread utilization of cellulosic fibers for clothing. Unfortunately, the acute toxicity of formaldehyde-based resins and the poor performance of non-formaldehyde resins still keep the research door open for scientists in this area. Herein, we demonstrated copper nanoparticles induced trimesic acid grafted cellulose as a sustainable solution for multifunctional easy-care clothing. Our treated fabric presents crease recovery angle value of 248° comparable to that of most promising citric acid-based cross-linkers at the chemical concentration of trimesic acid as low as 2% with a sweeping improvement of around 30% in strength retention, not reported earlier. The relatively low fabric stiffness, without any yellowing, is contributing to the comfort and aesthetic demand while nanoparticles induction promoting utmost antimicrobial need. For the first time, the superiority of the development was validated by interlacing the fabric/finish traits with sustainability building blocks that provide the step forward for rapid industrialization. Furthermore, environmental, health, and safety mapping comparison provides a better understanding of the intensity of hazards that different finishing crosslinkers pose on the environment and public health. With improved performance and superior sustainability, such fabric can act as a preferable alternative to the multifunctional easy-care fabric market. The online version contains supplementary material available at 10.1007/s10570-021-04251-5.


Challenges in the testing of non-heart-beating cadavers for viral markers: implications for the safety of tissue donors.
Natural changes that occur in blood and tissue after death may result in false positive results in antigen and antibody detection tests performed to identify markers of viral infection in potential tissue donors. Such tissue, which might otherwise be acceptable for therapeutic purposes, would not meet current standards for safe tissue banking. This is especially important in the context of insufficiency in the tissue supply. In this study, a series of blood samples collected during routine post-mortem examination was assayed using a range of commercially available kits for the detection of HBsAg, anti-HCV and anti-HIV 1 + 2 antibody/antigen. Results of tests on 104 samples collected from 97 individuals indicate that some kits result in a higher number of initial reactive samples than others. Approximately 40% of samples were reactive in one or more HBsAg assay, less than 10% in at least one anti-HIV kit and only 1 sample at low level on an anti-HCV kit. Liver or lymph node samples from individuals whose serum sample gave reactive results in antigen/antibody assays were tested for viral nucleic acid in the corresponding nucleic acid amplification test. Only one individual's sample was confirmed to test positive for HBsAg in a confirmatory neutralisation test and by nucleic acid amplification technology, and a second individual whose serum was scored reactive for anti-HCV, but negative for HBsAg, had a liver sample which was HBV DNA positive and HCV RNA negative. The results of the study indicate that antibody/antigen assays are not as specific as NAT using state of the art DNA extraction techniques. Both types of assay complement each other and used together will help assure the safety of tissues for transplantation.


Training the Millennial learner through experiential evolutionary scaffolding: implications for clinical supervision in graduate education programs.
They are the Millennials--Generation Y. Over the next few decades, they will be entering genetic counseling graduate training programs and the workforce. As a group, they are unlike previous youth generations in many ways, including the way they learn. Therefore, genetic counselors who teach and supervise need to understand the Millennials and explore new ways of teaching to ensure that the next cohort of genetic counselors has both skills and knowledge to represent our profession well. This paper will summarize the distinguishing traits of the Millennial generation as well as authentic learning and evolutionary scaffolding theories of learning that can enhance teaching and supervision. We will then use specific aspects of case preparation during clinical rotations to demonstrate how incorporating authentic learning theory into evolutionary scaffolding results in experiential evolutionary scaffolding, a method that potentially offers a more effective approach when teaching Millennials. We conclude with suggestions for future research.


Up to date on primary ciliary dyskinesia in children.
Primary ciliary dyskinesia (PCD) is a congenital, clinically and ultrastructurally heterogeneous disease due to abnormal structure and/or function of cilia, with impaired mucociliary transport leading to several respiratory disorders. PCD can be diagnosed by the combination of thorough clinical examination with functional and ultrastructural analysis of the cilia. This paper shows progresses in PCD diagnosis obtained by ciliogenesis in culture evaluation of ciliated respiratory cells and by genetic analysis of mutations in candidate genes. Moreover, since to date no specific treatments are available to correct the ciliary dysfunction, the paper shows the proper therapeutical approach by the use of respiratory physiotherapy and regular exercise to favour airways clearance, by antibiotics administration to control acute airway infections. Macrolides administration as antiinflammatory option is suggested.


[New autoanti-bodies in rheumatoid arthritis: anti-citrullinated protein or peptide autoanti-bodies and the others].
New treatment strategies require that rheumatoid arthritis (RA) be diagnosed as early as possible. New diagnostic markers were required, because rheumatoid factors (RF), until now criteria for classification of RA, are not sufficiently specific and sometimes appear late, thereby limiting their diagnostic usefulness. The objective of this review is to describe the current state of knowledge and more particularly to analyze the interest of new RA autoanti-bodies, called anti-peptide or anti-citrullinated protein anti-bodies (ACPA). Other autoanti-bodies have been described, including anti-Sa, anti-alpha enolase, and anti-calpastatin autoanti-bodies. Nonetheless, their diagnostic value remains limited compared to ACPA. Accordingly, in daily practice today, the only autoanti-bodies that must be tested for to diagnose RA are the ACPAs and RFs. The discovery of ACPA (initially called anti-keratin and anti-perinuclear anti-bodies) was a major step forward for the laboratory diagnosis of RA. The tests most often used routinely areenzyme-linked immunosorbent assays(ELISA) with cyclic citrullinated peptides, whence the name anti-CCP autoanti-bodies. Accordingly, the two terms ACPA and anti-CCP can both be used. The diagnostic value, in particular their specificity, is on the order of 95%, regardless of the method of identification. These markers are very useful and are often present earlier than RF. These ACPA also have prognostic value because they are associated with more aggressive forms of RA. On the other hand, their value over time, in particular, their fluctuation as a function of treatment, is more controversial. In practice, it is recommended to test for both RF and ACPA in a diagnostic work-up for early RA. During follow-up, the value of testing for these autoanti-bodies has not been demonstrated, but additional studies are still necessary with the anti-CCP autoanti-bodies and the new anti-citrullinated protein autoanti-bodies.


Microscopic investigation of the resonant mechanism for the implementation of nc-MRI at ultra-low field MRI.
In this paper the possible use of the resonant mechanism between some spectral components of the neuronal activity and the spin dynamics in ultra-low field MRI experiments--for the implementation of the nc-MRI techniques and proposed by Kraus et al., 2008--is investigated by means of "realistic" simulations of the neuronal activity of a modelled neuronal network. Previously characterized digital neurons are used to reproduce neuronal currents based on biophysical details and the distribution of the local magnetic field inside a MRI cubic voxel (having a dimension of 1.2 mm) is evaluated. The properties of the water proton spin dynamics as a consequence of the neuronal field and of external applied fields are extrapolated integrating the Bloch equations. The characteristics of the expected MR signals are discussed in relation to the specifics of the NMR sequence used and to the properties of the neuronal activity. The great potentialities of the technique are provided by: a) the possible easy implementation of the technique, b) the possible cheap instrumentation required; c) the flexibility of the ultra-low field systems.


Coagulation and inflammation in cancer: Limitations and prospects for treatment.
The development of so-called immune checkpoint inhibitors (ICIs), which target specific molecular processes of tumour growth, has had a transformative effect on cancer treatment. Widespread use of antibody-based medicines to inhibit tumour cell immune evasion by modulating T cell responses is becoming more common. Despite this, response rates are still low, and secondary resistance is an issue that arises often. In addition, a wide range of serious adverse effects is triggered by enhancing the immunological response. As a result of an increased mortality rate, a higher prevalence of thrombotic complications is connected with an increased incidence of immunological reactions, complement activation, and skin toxicity. This suggests that the tumour microenvironment's interaction between coagulation and inflammation is important at every stage of the tumour's life cycle. The coagulation system's function in tumour formation is the topic of this review. By better understanding the molecular mechanisms in which tumour cells circulate, plasmatic coagulation and immune system cells are engaged, new therapy options for cancer sufferers may be discovered.


The age-related decrease in material properties of BALB/c mouse long bones involves alterations to the extracellular matrix.
One possibility for the disproportionate increase in fracture risk with aging relative to the decrease in bone mass is an accumulation of changes to the bone matrix which deleteriously affect fracture resistance. In order to effectively develop new targets for osteoporosis, a preclinical model of the age-related loss in fracture resistance needs to be established beyond known age-related decreases in bone mineral density and bone volume fraction. To that end, we examined long bones of male and female BALB/c mice at 6-mo. and 20-mo. of age and assessed whether material and matrix properties of cortical bone significantly differed between the age groups. The second moment of area of the diaphysis (minimum and maximum principals for femur and radius, respectively) as measured by ex vivo micro-computed tomography (μCT) was higher at 20-mo. than at 6-mo. for both males and females, but ultimate moment as measured by three-point bending tests did not decrease with age. Cortical thickness was lower with age for males, but higher for old females. Partially accounting for differences in structure, material estimates of yield, ultimate stress, and toughness (left femur) were 12.6%, 11.1%, and 40.9% lower, respectively, with age for both sexes. The ability of the cortical bone to resist crack growth (right femur) was also 18.1% less for the old than for the young adult mice. These decreases in material properties were not due to changes in intracortical porosity as pore number decreased with age. Rather, age-related alterations in the matrix were observed for both sexes: enzymatic and non-enzymatic crosslinks by high performance liquid chromatography increased (femur), volume fraction of bound water by 1H-nuclear magnetic resonance relaxometry decreased (femur), cortical tissue mineral density by μCT increased (femur and radius), and an Amide I sub-peak ratio I1670/I1640 by Raman spectroscopy increased (tibia). Overall, there are multiple matrix changes to potentially target that could prevent the age-related decrease in fracture resistance observed in BALB/c mouse.


AKT inhibitor Hu7691 induces differentiation of neuroblastoma cells.
While neuroblastoma accounts for 15% of childhood tumor-related deaths, treatments against neuroblastoma remain scarce and mainly consist of cytotoxic chemotherapeutic drugs. Currently, maintenance therapy of differentiation induction is the standard of care for neuroblastoma patients in clinical, especially high-risk patients. However, differentiation therapy is not used as a first-line treatment for neuroblastoma due to low efficacy, unclear mechanism, and few drug options. Through compound library screening, we accidently found the potential differentiation-inducing effect of AKT inhibitor Hu7691. The protein kinase B (AKT) pathway is an important signaling pathway for regulating tumorigenesis and neural differentiation, yet the relation between the AKT pathway and neuroblastoma differentiation remains unclear. Here, we reveal the anti-proliferation and neurogenesis effect of Hu7691 on multiple neuroblastoma cell lines. Further evidence including neurites outgrowth, cell cycle arrest, and differentiation mRNA marker clarified the differentiation-inducing effect of Hu7691. Meanwhile, with the introduction of other AKT inhibitors, it is now clear that multiple AKT inhibitors can induce neuroblastoma differentiation. Furthermore, silencing AKT was found to have the effect of inducing neuroblastoma differentiation. Finally, confirmation of the therapeutic effects of Hu7691 is dependent on inducing differentiation in vivo, suggesting that Hu7691 is a potential molecule against neuroblastoma. Through this study, we not only define the key role of AKT in the progression of neuroblastoma differentiation but also provide potential drugs and key targets for the application of differentiation therapies for neuroblastoma clinically.