Optimization of the simultaneous determination of imatinib and its major metabolite, CGP74588, in human plasma by a rapid HPLC method using D-optimal experimental design.
A simple, rapid and specific HPLC method has been developed and validated for the simultaneous determination of imatinib, a tyrosine kinase inhibitor, and its major metabolite, CGP74588, in human plasma. The optimization of the HPLC procedure involved several variables, of which the influences of each was studied. After a series of preliminary-screening experiments, the composition of the mobile phase and the pH of the added buffer solution were set as the investigated variables, while the resolution between imatinib and CGP74588 peaks, the retention time and the imatinib peak width were chosen as the dependent variables. Applying D-optimal design, the optimal chromatographic conditions for the separation were defined. The method proved to show good agreement between the experimental data and predictive values throughout the studied parameter range. The optimum assay conditions were achieved with a Chromolith™ Performance RP-8e 100 mm × 4.6 mm column and a mixture of methanol/acetonitrile/triethylamine/diammonium hydrogen phosphate (pH 6.25, 0.048 mol L(-1)) (20:20:0.1:59.9, v/v/v/v) as the mobile phase at a flow rate of 2 mL min(-1) and detection wavelength of 261 nm. The run time was less than 5 min, which is much shorter than the previously optimized methods. The optimized method was validated according to FDA guidelines to confirm specificity, linearity, accuracy and precision.


In vitro function and in vivo viability of stored platelet concentrates. Effect of a secondary plasticizer component of PVC storage bags.
Previous studies have shown that CL-2399 (Cutter) and PL-130 (Fenwal) polyvinyl chloride (PVC) plastic bags are unsatisfactory for storage of platelet concentrates (PC) at 22 degrees C. In an effort to explain the effects of plastic bags, the chemical make-up of CL-2399 and PL-130 PVC films was determined and compared with that of P1-146 (Fenwal) PVC, which is satisfactory for PC storage at 22 degrees C. The only significant difference between the three materials was the incorporation of tetrahydrofurfuryl oleate (THFO) as a secondary plasticizer in CL-2399 and PL-130. The response of platelets to aggregating agents, uptake of serotonin, recovery from hyptonic stress, and serotonin release during storage following storage in a modified CL-2399 plastic prepared without THFO and designated CL-3000 (Cutter) was equivalent to PL-146 and far superior to CL-2399. In vivo studies in two laboratories of platelets stored in CL-3000 bags showed satisfactory recovery (56 +/- 4.2% and 46.7 +/- 2.7%) and survival (6.4 +/- 0.4 days and 7.4 +/- 0.6 days). From these studies we conclude that the THFO secondary plasticizer component of PL-130 and CL-2399 is the cause of the poor platelet viability of platelets stored in these plastic bags. The mechanism of impairment is not known. The causative agent(s) may be degradation products of THFO (formed during manufacture of the PVC film) that are leached from the plastic into PC during storage.


Cytochrome P450 3A4 activity in premenopausal and postmenopausal women, based on 6-beta-hydroxycortisol:cortisol ratios.
To characterize cytochrome P450 (CYP) 3A4 activity in premenopausal and postmenopausal women by evaluating the urinary 6-beta-hydroxycortisol:cortisol ratio. Prospective study Thirteen premenopausal and 13 postmenopausal women who were healthy and not receiving drugs known to affect CYP3A4 activity Beginning on day 2 of menses, premenopausal women collected first morning urine samples every other day for a complete menstrual cycle. Postmenopausal women collected first morning urine every other day for 28 days. Mean weekly 6-beta-hydroxycortisol:cortisol ratios did not differ during the phase (week) of the menstrual cycle. Daily ratios did not differ in postmenopausal women. No difference between premenopausal and postmenopausal women was found on comparing overall median ratios. Cytochrome P450 3A4 activity as measured by 6-beta-hydroxy cortisol:cortisol ratio did not differ by week of menstrual cycle, suggesting no menstrual cycle-related changes. Menopause does not appear to be associated with differences in CYP3A4 activity, compared with premenopause.


Anticancer mechanisms on pyroptosis induced by Oridonin: New potential targeted therapeutic strategies.
Pyroptosis is a type of inflammatory cell death that is triggered by the formation of pores on the cell membrane by gasdermin (GSDM) family proteins. This process activates inflammasomes and leads to the maturation and release of proinflammatory cytokines such as interleukin-1β (IL-1β) and interleukin-18 (IL-18). Pyroptosis, a form of programmed cell death, has been found to be associated with various biomolecules such as caspases, granzymes, non-coding RNA (lncRNA), reactive oxygen species (ROS), and NOD-like receptor protein 3 (NLRP3). These biomolecules have been shown to play a dual role in cancer by affecting cell proliferation, metastasis, and the tumor microenvironment (TME), resulting in both tumor promotion and anti-tumor effects. Recent studies have found that Oridonin (Ori) has anti-tumor effects by regulating pyroptosis through various pathways. Ori can inhibit pyroptosis by inhibiting caspase-1, which is responsible for activating pyroptosis of the canonical pathway. Additionally, Ori can inhibit pyroptosis by inhibiting NLRP3, which is responsible for activating pyroptosis of the noncanonical pathway. Interestingly, Ori can also activate pyroptosis by activating caspase-3 and caspase-8, which are responsible for activating pyroptosis of the emerging pathway; Ori has been found to be effective in inhibiting pyroptosis by blocking the action of perforin, which is responsible for facilitating the entry of granzyme into cells and activating pyroptosis. Additionally, Ori plays a crucial role in regulating pyroptosis by promoting the accumulation of ROS while inhibiting the ncRNA and NLRP3 pathways. It is worth noting that all of these pathways ultimately regulate pyroptosis by influencing the cleavage of GSDM, which is a key factor in the process. These studies concludes that Ori has extensive anti-cancer effects that are related to its potential regulatory function on pyroptosis. The paper summarizes several potential ways in which Ori participates in the regulation of pyroptosis, providing a reference for further study on the relationship between Ori, pyroptosis, and cancer.


Safety of Use of a Sheet-Type Adhesion Barrier (Interceed®) During Liver Surgery.
Adhesion barriers are increasingly used in hepatobiliary surgery. However, there has been no solid evidence yet in support of their safety. Incidences of global postoperative morbidities and major abdominal morbidities were compared between 101 consecutive patients who received a sheet-type adhesion barrier (Interceed®) and 134 patients who did not receive any adhesion barriers during hepatectomy. Propensity score (PS) adjustment was used to account for potential bias to receive Interceed. In the PS-adjusted population, the incidences of both global postoperative morbidities and major abdominal morbidities showed no significant difference between the Interceed group and the control group (17.9% vs. 17.6%; P = 0.948 and 7.8% vs. 9.1%; P = 0.813, respectively). Multivariate analysis showed that age + 10 years (odds ratio [OR], 1.70; 95% CI, 1.15-2.50; P = 0.007), estimated blood loss + 100 mL (OR, 1.05; 95% CI, 1.01-1.09, P = 0.009), and laparoscopic approach (OR, 0.10; 95% CI, 0.01-0.75; P = 0.026) were independent predictors for global postoperative morbidities and operation time + 1 h (OR, 1.56; 95% CI, 1.23-1.96; P < 0.001) was a risk factor for major abdominal morbidity, while no specific association between the use of Interceed and the risk of postoperative morbidity was observed. Use of Interceed does not increase the risk of postoperative morbidities after hepatectomy.


Randomized clinical trial of topical betaxolol for persistent macular edema after vitrectomy and epiretinal membrane removal.
To report the efficacy and safety of topical betaxolol for treatment of persistent macular edema. Randomized clinical trial. Thirty-seven eyes (37 patients) with best-corrected visual acuity between 20/200 and 20/50 and macular edema that remained for 3 months after vitrectomy and removal of epiretinal membrane were prospectively, randomly assigned to receive betaxolol or placebo. Nineteen eyes of 19 patients received betaxolol twice daily, and 18 eyes of 18 patients received placebo as a randomized comparison group. The patients were followed up for 6 months. This study evaluated the effect of betaxolol on best-corrected visual acuity and area of macular edema, which was digitally measured on serial fluorescein angiogram. Calculations of mean best-corrected visual acuity were based on logarithm of the minimal angle of resolution (logMAR). To assess changes in area of edema, the initial (pretreatment) size of the edema was set to 100%, and all posttreatment measurements were normalized relative to the initial size. Mean best-corrected visual acuity at baseline was 0.216 (20 of 92.6) and 0.244 (20 of 82.0) in the treatment and control group, respectively. Mean area of macular edema was 2.271 +/- 1.629 mm(2) and 2.273 +/- 1.209 mm(2) in the treatment and control group; there was no significant difference. The visual acuity at 6 months after the start of the follow-up was 0.471 (20 of 42.5) in the treatment group and 0.236 (20 of 84.7) in the control group. Mean changes in logMAR of visual acuity for 3- and 6-month follow-up were -0.282 +/- 0.191 and -0.337 +/- 0.197 in the treatment group, and -0.016 +/- 0.186 and +0.015 +/- 0.267 in the control group; a significant difference was found (P <.0001; P <.0001). Areas of macular edema at 6 months after the start of the follow-up were 1.492 +/- 1.357 mm(2) in the treatment group and 2.125 +/- 1.434 mm(2)in the control group. Mean change in area of the edema for 6 months were 76.5% +/- 24.1% and 63.4% +/- 28.3% in the treatment group and 92.9% +/- 15.4% and 87.4% +/- 25.6% in the control group; treated patients showed a significantly larger reduction than untreated patients at each examination (P =.0193; P =.0102). No complication associated with treatment or placebo was found. Topical betaxolol appeared to have a favorable treatment effect in eyes with macular edema that remained after vitrectomy and removal of epiretinal membrane. Further investigation of more cases and longer follow-up are needed.


Fractures of the shafts of the phalanges of the hand.
Fractures of the shafts of the phalanges are common and often disabling. Little detailed information is available as to the results of treatment, against which new methods of treatment may be compared. Thirty-nine comminuted fractures and 109 which were not comminuted were followed-up and their results are presented in detail. In both groups only about 57% obtained satisfactory results. Possible methods of improving these are discussed and the importance of accurate reduction of non comminuted fractures is emphasised.


Halomonas socia sp. nov., isolated from high salt culture of Dunaliella salina.
A moderately halophilic bacteria designed strain NY-011(T) was isolated from the high salt culture of Dunaliella salina in Chengdu of Sichuan Province, China. The isolate was Gram-negative, nonmotile, rod-shaped and 12.5-21.6 μm in length. Colonies on solid media are circular, wet, smooth and cream. The strain grew optimally at 37 °C, pH 7.0 and in the presence of 8 % NaCl. Acid was produced from glycerol, D-arabinose, glucose, trehalose, inositol, mannose, mannitol, sucrose, maltose and sorbitol. Catalase is produced but not oxidase. The major fatty acids are C18: 1ω7c (37.59 %), C19: 0 cyclo ω8c (18.29 %), C16: 0 (16.05 %) and C6: 0 (12.43 %). The predominant respiratory lipoquinone found in strain NY-011(T) is ubiquinone with nine isoprene units (Q-9). The genomic DNA G + C content of strain NY-011(T) was 62.7 mol%. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain NY-011(T) belonged to the genus Halomonas. The highest levels of 16S rRNA gene sequence similarity were found between the strain NY-011(T) and H. pantelleriensis (sequence similarity 98.43 %). However, the levels of DNA-DNA relatedness between them were only 23.1 %. In addition, the strain NY-011(T) had a phenotypic profile that readily distinguished it from H. pantelleriensis. The strain NY-011(T) therefore represents a new species of the genus Halomonas, for which the name Halomonas socia sp. nov. is proposed, with NY-011(T) (=CCTCC AB 2011033(T) = KCTC 23671(T)) as the type strain.


Treatment of hepatocellular carcinoma with tamoxifen: a double-blind placebo-controlled trial in 120 patients.
The progression of hepatocellular carcinoma may be influenced by estrogens. This has offered the rationale for evaluating the therapeutic usefulness of estrogen-receptor blockers; it is being debated whether long-term tamoxifen administration improves survival in patients with this neoplasm. The aim of this study was to assess the efficacy of tamoxifen administration in the treatment of hepatocellular carcinoma. One hundred twenty patients with this neoplasm who were not suitable for surgery, ethanol injection, or transarterial embolization were included in a placebo-controlled trial and randomized to tamoxifen, 20 mg/day per os, (group A, n = 58) or placebo (group B, n = 62). Patients with terminal diseases were excluded. Both groups were similar with regard to sex, age, liver function (Child-Pugh's score, 6.5 +/- 1.4 vs. 6.4 +/- 1.4), baseline performance status, and tumor stage. Tamoxifen had no antitumoral effect with no differences in the survival between groups (1- and 2-year actuarial rate: group A, 51% and 27%; and group B, 43% and 29%; P = 0.75), even when stratifying patients according to baseline status. Furthermore, there were no differences in the probability of disease progression (P = 0.46) and baseline performance status maintenance (P = 0.93) during follow-up. Tamoxifen has no efficacy in the treatment of patients with advanced hepatocellular carcinoma.


Generalized data model for real-world capacitated lot-sizing problems with linked lot sizes and backorders.
This data article establishes the first data model for the Multi-Level Capacitated Lot-Sizing Problem with Linked Lot Sizes and Backorders (MLCLSP-L-B) for tablets manufacturing processes. The dataset consists out of nine tables and collects all required meta-information for capacitated lot-sizing problems with linked lot sizes and backorders. Furthermore, it is compatible to store information for single- and multi-level lot-sizing problem instance information. An exact field description and field type is provided for each table. Anonymized real-world data represents four single-level packaging and five multi-level tablets manufacturing processes that are shared with lot-sizing research community by the standardized data template.


A phenol-chloroform protocol for extracting DNA from ancient samples.
The myriad downstream applications of ancient DNA (aDNA) analysis all ultimately require that sequence data are generated from extracts of ancient material. DNA extraction from tissues known to contain preserved biomolecules (e.g. teeth, hair, tissue, bone) relies on subtle modifications of a basic technique that has been in use for nearly two decades. Multiple DNA extraction protocols have been introduced, with varying levels of success depending on tissue type and the long-term preservation environment to which the ancient tissue was exposed. Here, we describe the phenol-chloroform method for extracting aDNA from any tissue type. This commonly employed method allows for the recovery of total nucleic acid content with minimal loss of low molecular weight double-stranded DNA.


Repair of dural defects with electrospun bacterial cellulose membranes in a rabbit experimental model.
To evaluate the advantages and mechanisms involved in repairing rabbit dural defect with a novel electrospun bacterial cellulose (EBC) membrane, a series of experiments were carried out in vitro and in vivo. Compared with common bacterial cellulose (BC) membrane, a more dispersed and regular fiber structure and a better porosity and water holding capacity were found in the EBC membrane, which also had superior degradability. However, the biomechanical properties were slightly decreased. The results demonstrated that BC and EBC membranes had little effect on proliferation and apoptosis of mouse fibroblast cells. There were no complications such as infection, cerebrospinal fluid leakage, epilepsy and brain swelling after BC and EBC membrane repairs in rabbit models. Using real-time quantitative polymerase chain reaction (RT-qPCR) and western blot, the early inflammatory reactions in the EBC group were shown to be lower than in the BC group, and were close to the autologous dura mater group. Histological observations and western blot revealed more collagen fibers evenly distributed on the outer side of EBC membranes than in the BC and unpatched groups, and fewer brain tissue adhesions and epidural scars were found in the EBC group. Compared with common BC membrane, the EBC membrane had better biophysical properties and biocompatibility. It is expected to be a suitable alternative material for the repair of damaged dura mater.


Recruitment of efferent lymph during infusion of 20 % albumin.
Intravenous infusion of hyper-oncotic 20 % albumin expands the plasma volume by approximately twice the infused volume. We investigated whether the recruited fluid stems from accelerated flow of efferent lymph, which would add protein to the plasma, or from reversed transcapillary solvent filtration, where the solvent is expected to be low in protein. We analyzed data from 27 intravenous infusions of 20 % albumin (3 mL/kg; approximately 200 mL) over 30 min given to 27 volunteers and patients. Twelve of the volunteers were also given a 5 % solution and served as controls. The pattern of blood hemoglobin, colloid osmotic pressure, and the plasma concentrations of two immunoglobulins (IgG and IgM) were studied over 5 h. A decrease of the difference between the plasma colloid osmotic pressure and plasma albumin occurred during the infusions and was almost four times greater for 5 % albumin than for 20 % albumin at 40 min (P < 0.0036), which indicates that non-albumin protein enriched the plasma when 20 % was infused. Moreover, the difference between the infusion-derived dilution of the blood plasma based on hemoglobin and the two immunoglobulins amounted to -1.9 % (-6 to +0.2) for 20 % albumin and to -4.4 % (25th-75th percentile range - 8.5 to +0.2) during experiments with 5 % albumin (P < 0.001). This supports that the plasma was enriched by immunoglobulins, probably via the lymph, when 20 % was infused. Between half and two-thirds of the extravascular fluid that was recruited during infusion of 20 % albumin in humans consisted of protein-containing fluid consistent with efferent lymph.


All-Inside PCL Reconstruction, Double Bundle, With Internal Brace Augmentation.
PCL reconstructive techniques are constantly evolving, and further clinical studies are needed to definitively understand the potential benefits of internal brace augmentation and anatomic double-bundle PCL reconstruction. This Technical Note reports an arthroscopic all-inside anatomic double-bundle PCL reconstruction with internal brace augmentation that is effective and reproducible.


Peptide loaded polymeric nanoparticles by non-aqueous nanoprecipitation.
It is always a challenge to encapsulate water-soluble peptides in polymer nanoparticle (NP) systems. We establish and validate our newly developed non-aqueous nanoprecipitation method to encapsulate neuro-peptides drugs such as oxytocin and Luteinizing hormone-releasing hormone (LHRH) in poly(sebacic anhydride) (PSA) NPs. NPs were prepared by a solvent-antisolvent process under a strict anhydrous environment to obtain high drug loading and to avoid premature PSA degradation and drug release. Dynamic light scattering (DLS) and Scanning Electron Microscopy (SEM) reveal the size for both drug loaded PSA NPs to ∼ 300 nm. The drug loaded NPs were dispersible and spherical in shape with uniform morphology. The in vitro release profile of oxytocin from PSA NPs occurs with the burst release of ∼ 50% within the first hour in the aqueous release medium, whereas LHRH release is comparatively slow. Thus, looking into the fast degrading properties of PSA and drug release behavior, the developed NPs can be used for direct delivery of the neuropeptides to the olfactory epithelium using a refillable nasal atomizer that deposits mist onto the olfactory neuro-epithelium. We also applied our developed method to prepare NPs of poly(lactic-co-glycolic acid) (PLGA), polylactic acid (PLA), and poly(ε-caprolactone) (PCL). A Thyrotropin releasing hormone (TRH) was used as the sample neuropeptide drug to validate our non-aqueous method. The results reveal the formation of TRH loaded PLGA, PLA and PCL NPs with 100% drug loading. TEM analysis shows the formation of spherical NPs, having similar release properties as those of PSA NPs. Overall, we report that our developed method is suitable for co-encapsulating hydrophilic drugs in polymer NPs with high drug loading and release properties.


Frontal lobe ischemic stroke presenting with peripheral type facial palsy: A crucial diagnostic challenge in emergency practice.
Here, we illustrate a 69-year old female admitting with weakness on left side of the face who firstly considered peripheral facial palsy in the forefront. However, detailed neurological examination and cranial MRI findings finally yielded the proper diagnosis of right hemisphere ischemic stroke. Via this remarkable presentation, we point out the clinical challenges in evaluation processes of patients with facial palsy in emergency practice and emphasize the importance of detailed examination for the proper diagnosis as well as initiation of appropriate treatment agents without delay.


Effects of high iron and glucose concentrations over the relative expression of Bcl2, Bax, and Mfn2 in MIN6 cells.
Type 2 diabetes is characterized by hyperglycemia and oxidative stress. Hyperglycemia is linked to mitochondrial dysfunction and reduced β-cell mass due to the reduced expression of genes such as Mfn2 as well as the participation of the Bcl2 gene family, responsible for increased apoptosis. The purpose of this study was to describe the effect of different iron and/or glucose concentrations over Mfn2, Bax, and Bcl2 expressions in a β-pancreatic cell line (MIN6 cells). MIN6 cells were pre-incubated with different iron and/or glucose concentrations, and the relative mRNA abundance of the Bcl2/Bax ratio and of Mfn2 genes was measured by qRT-PCR. Heme oxygenase (HO) activity, iron uptake, superoxide dismutase activity, and glutathione content were also determined. The Bcl2/Bax ratio increased and Mfn2 expression decreased in MIN6 cells after glucose stimulation. These effects were higher when glucose and iron were incubated together. Additionally, treatment with glucose/iron showed a higher HO activity. Our study revealed that high glucose/Fe concentrations in MIN6 cells induced an increase of the Bcl2/Bax ratio, an indicator of increased cell apoptosis.


Revealing the substitution preference of zinc in ordinary Portland cement clinker phases: A study from experiments and DFT calculations.
Ordinary Portland cement (OPC) clinker mainly consist four minerals, tricalcium silicate (C3S), dicalcium silicate (C2S,), tricalcium aluminate (C3A), and tetracalcium aluminoferrite (C4AF). To learn the doping behaviors of Zn in OPC clinker, a series of samples were prepared by calcinating the mixtures of CaCO3, SiO2, Al2O3, Fe2O3, and ZnO. Our results from energy-dispersive spectroscopy, X-ray diffraction and density functional theoretical simulations show that a small amount of ZnO enter C3S and C2S by replacing Ca ions while most incorporate into C4AF by substituting Fe atoms, resulting in a decrease of C3A in OPC as dosage increases. Further analyses from partial density of states and distributions of bond order-bond length indicate that the doping preference can be ascribed to the similar electron contributions and small structure distortions between host and guest ions. Unlike the strong Fe‒O bond, the newly formed Zn‒O is much weaker. The weak Zn‒O may be responsible for the limited solubility of Zn in C4AF. These results provide a possibility of increasing solubility of Zn in OPC clinker by increasing the contents of C3A and C4AF, thus will be very meaningful in the synthesis of OPC clinker by utilizing Zn-bearing alternative raw materials.


Re-examining the origins of spectral blinking in single-molecule and single-nanoparticle SERS.
Single metal nanoparticles and nanoaggregates are known to emit intense bursts of surface-enhanced Raman scattering (SERS) in an intermittent on and off fashion. The characteristic "blinking" timescales range from milliseconds to seconds. Here we report detailed temperature dependence (both heating and cooling) and light-intensity studies to further examine the origins of this intriguing phenomenon. The results indicate that blinking SERS contains both a thermo-activated component and a light-induced component. Several lines of evidence suggest that the observed fluctuations are caused by thermally activated diffusion of individual molecules on the particle surface, coupled with photo-induced electron transfer and structural relaxation of surface active sites or atomic-scale roughness features.


Reduction of hypoglycaemia, lifestyle modifications and psychological distress during lockdown following SARS-CoV-2 outbreak in type 1 diabetes.
To assess changes in glucose metrics and their association with psychological distress and lifestyle changes in patients with type 1 diabetes (T1D) using flash glucose monitoring (FGM) during lockdown following severe acute respiratory syndrome coronavirus 2 outbreak. Single-centre, observational, retrospective study enrolling T1D patients who attended a remote visit on April 2020 at the Endocrinology division of the University Hospital Policlinico Consorziale, Bari, Italy. Lockdown-related changes in physical activity level and dietary habits were assessed on a semi-quantitative basis. Changes in general well-being were assessed by the General Health Questionnaire-12 items with a binary scoring system. Glucose metrics were obtained from the Libreview platform for the first 2 weeks of February 2020 (T0) and the last 2 weeks before the phone visit (T1). Out of 84 patients assessed for eligibility, 48 had sufficient FGM data to be included in the analysis. FGM data analysis revealed significant reductions in coefficient of variation, number of hypoglycaemic events, and time below range, while no changes were found in time in range, time above range, mean sensor glucose, and glucose management indicator. Moreover, the frequency of sweets consumption was inversely related to the occurrence of hypoglycaemic events during lockdown. Lockdown-related lifestyle changes, albeit unhealthy, may lead to reduction in FGM-derived measures of hypoglycaemia and glycaemic variability in patients with T1D.


FOXP1 and PAX5 are rare but recurrent translocations partners in acute lymphoblastic leukemia.
Here, we report the case of a 57-year-old man, who was diagnosed with B-cell acute lymphoblastic leukemia (B-ALL). His diagnostic workup identified a translocation t(3;9)(p13;p13). This is the fifth case reported to date that involved the forkhead box P1 gene (FOXP1) and paired box gene 5 (PAX5). The PAX5-FOXP1 translocation is a nonrandom aberration, which is recurrent in both childhood and in adult B-ALL, and may contribute to leukemogenesis by blocking differentiation of hematopoietic cells into mature B-cells.


The rational design of an AIDS vaccine.
The extraordinary genetic diversity and immune evasion of human immunodeficiency virus (HIV) pose significant challenges for vaccine development. AIDS vaccine design requires a scientifically driven, rational approach that encompasses the latest advances in viral molecular genetics, structural biology, and immunology.


Sodium sulfide prevents water diffusion abnormality in the brain and improves long term outcome after cardiac arrest in mice.
Sudden cardiac arrest (CA) is one of the leading causes of death worldwide. Previously we demonstrated that administration of sodium sulfide (Na(2)S), a hydrogen sulfide (H(2)S) donor, markedly improved the neurological outcome and survival rate at 24 h after CA and cardiopulmonary resuscitation (CPR) in mice. In this study, we sought to elucidate the mechanism responsible for the neuroprotective effects of Na(2)S and its impact on the long-term survival after CA/CPR in mice. Adult male mice were subjected to potassium-induced CA for 7.5 min at 37°C whereupon CPR was performed with chest compression and mechanical ventilation. Mice received Na(2)S (0.55 mgkg(-1) i.v.) or vehicle 1 min before CPR. Mice that were subjected to CA/CPR and received vehicle exhibited a poor 10-day survival rate (4/12) and depressed neurological function. Cardiac arrest and CPR induced abnormal water diffusion in the vulnerable regions of the brain, as demonstrated by hyperintense diffusion-weighted imaging (DWI) 24 h after CA/CPR. Extent of hyperintense DWI was associated with matrix metalloproteinase 9 (MMP-9) activation, worse neurological outcomes, and poor survival rate at 10 days after CA/CPR. Administration of Na(2)S prevented the development of abnormal water diffusion and MMP-9 activation and markedly improved neurological function and long-term survival (9/12, P<0.05 vs. Vehicle) after CA/CPR. These results suggest that administration of Na(2)S 1 min before CPR improves neurological function and survival rate at 10 days after CA/CPR by preventing water diffusion abnormality in the brain potentially via inhibiting MMP-9 activation early after resuscitation.


Rubella virus: mechanism of attenuation in the vaccine strain (HPV77).
The vaccine type (HPV77 strain) of rubella virus replicates slower and manifests a delayed appearance of cytopathic effect in Vero-76 cells as compared to wild-type virus (M33). The change in cytopathic effect coincides with the delayed appearance of both genomic and subgenomic RNA as well as viral structural proteins in the cell. The delay in the appearance of the viral proteins in the cells was also evident when the cells infected with the vaccine-type virus were treated with the lysosomotropic agent such as chloroquine. Binding studies using [35S]methionine-labeled virus showed that the vaccine-type virus bound to the cells poorly and the binding was not completely competed out with the cold virus.


Sex and Prediction Error, Part 3: Provoking Prediction Error.
In parts 1 and 2 of this Lacanian neuropsychoanalytic series, surplus prediction error was presented as a neural correlate of the Lacanian concept of jouissance. Affective consciousness (a key source of prediction error in the brain) impels the work of cognition, the predictive work of explaining what is foreign and surprising. Yet this arousal is the necessary bedrock of all consciousness. Although the brain's predictive model strives for homeostatic explanation of prediction error, jouissance "drives a hole" in the work of homeostasis. Some residual prediction error always remains. Lacanian clinical technique attends to this surplus and the failed predictions to which this jouissance "sticks." Rather than striving to eliminate prediction error, clinical practice seeks its metabolization. Analysis targets one's mode of jouissance to create a space for the subject to enjoy in some other way. This entails working with prediction error, not removing or tolerating it. Analysis aims to shake the very core of the subject by provoking prediction error-this drives clinical change. Brief clinical examples illustrate this view.


Participation of endogenous neuropeptide Y in the suppression of baroreceptor reflex response by locus coeruleus in the rat.
We evaluated the potential participation of endogenous brain neuropeptide Y (NPY) in the suppression of baroreceptor reflex (BRR) response by locus coeruleus (LC), using adult male Sprague-Dawley rats anesthetized with pentobarbital sodium (40 mg/kg, i.p.). Bilateral microinjection of an antiserum against NPY (1:20, 20 nl) into the caudal one-third level of the nucleus tractus solitarii (NTS), the terminal site for baroreceptor afferent fibers, significantly reversed the suppressive effect of electrical or chemical activation of the LC on the BRR response. Treatments with NPY (4.65 pmol, 20 nl), normal rabbit serum, aCSF and heat-inactivated NPY or NPY antiserum, on the other hand, were ineffective. The LC-promoted inhibition of the BRR response was also attenuated by the alpha 1-adrenoceptor antagonist prazosin (50 pmol, 20 nl), either microinjected alone or in combination with NPY antiserum into the bilateral NTS. Mathematical treatment of our data revealed that the depressive effect on the BRR response of NPY or NE released at the NTS following LC activation manifested different time-course and magnitude. The one by endogenous NPY maximized at 40 min and amounted to no more than 20% of, whereas that by NE peaked at 10 min and contributed no less than 30% to, the suppression. These results suggest that both endogenous NPY and NE may participate in the suppression of BRR response by the LC at the NTS.


Scanning-integrating cytophotometric analyses of brain neuronal RNA and acetylcholinesterase in acute soman toxicated rats.
Cytophotometric analyses of RNA and acetylcholinesterase responses of caudate and cerebrocortical neurons of soman toxicated rats were conducted to characterize impairments in regulatory aspects of neuronal metabolism occurring in the acute phase of cholinesterase impairment. There was a severe and dose-dependent suppression (20-60%) in neuronal acetylcholinesterase activity in both a.m. and p.m.-treated rats; no diurnal differences were apparent in control acetylcholinesterase levels or neuronal acetylcholinesterase responsiveness to soman toxication. RNA levels, however, were markedly higher in p.m. than in a.m. saline-treated controls. Soman depressed caudate neuron RNA contents in the afternoon, but not in the morning. Cerebrocortical neuron RNA levels were suppressed in both a.m. and p.m.-toxicated rats, although this RNA depletion was more severe in the afternoon. These results indicate that soman can elicit marked alterations in neuronal transcriptional-translational capabilities and that there are diurnal variations in cellular metabolic responsiveness to soman toxication. Although functional relationships between soman-induced cholinesterase inhibition and RNA depletion remain to be elucidated, depressed RNA metabolism appears to be a maladaptive response preventing rapid regeneration of cholinesterase following poisoning.


Synergistic oligodeoxynucleotide strongly promotes CpG-induced interleukin-6 production.
Bacterial genomes span a significant portion of diversity, reflecting their adaptation strategies; these strategies include nucleotide usage biases that affect chromosome configuration. Here, we explore an immuno-synergistic oligodeoxynucleotide (iSN-ODN, named iSN34), derived from Lactobacillus rhamnosus GG (LGG) genomic sequences, that exhibits a synergistic effect on immune response to CpG-induced immune activation. The sequence of iSN34 was designed based on the genomic sequences of LGG. Pathogen-free mice were purchased from Japan SLC and maintained under temperature- and light-controlled conditions. We tested the effects of iSN34 exposure in vitro and in vivo by assessing effects on mRNA expression, protein levels, and cell type in murine splenocytes. We demonstrate that iSN34 has a significant stimulatory effect when administered in combination with CpG ODN, yielding enhanced interleukin (IL)-6 expression and production. IL-6 is a pleotropic cytokine that has been shown to prevent epithelial apoptosis during prolonged inflammation. Our results are the first report of a bacterial-DNA-derived ODN that exhibits immune synergistic activity. The potent over-expression of IL-6 in response to treatment with the combination of CpG ODN and iSN34 suggests a new approach to immune therapy. This finding may lead to novel clinical strategies for the prevention or treatment of dysfunctions of the innate and adaptive immune systems.


Mediators of asthma outcomes.
Patient adherence, the level of asthma self-management skills, exposure to stress, and depression can have considerable influence on a wide range of asthma outcomes and thus are considered asthma outcome mediators. National Institutes of Health institutes and other federal agencies convened an expert group to recommend standardized measures for 7 domains of asthma clinical research outcomes measures. Although the review of mediators of these outcomes was not within the scope of any specific outcome topic, a brief summary is presented so that researchers might consider potential mediators. We prepared a summary of key mediators of asthma outcomes based on expertise and knowledge of the literature. The rationale for including measures of adherence, self-management skills, and exposures to stress in asthma clinical research is presented, along with a brief review of instruments for collecting this information from clinical research participants. Appropriate measurement of adherence, self-management skills, and exposures to stress will enhance characterization of study participants and provide information about the potential impact these factors can have on mediating the effects of treatment interventions.


Tandem and triple high-dose chemotherapy with autologous stem cell rescue in metastatic breast cancer.
The purpose of this phase II study was to evaluate the therapeutic efficacy and toxicity of a tandem or triple high-dose chemotherapy (HDC) with autologous peripheral blood stem cell transplantation (PBSCT) in patients with metastatic breast cancer (MBC) as first line chemotherapy. Conventional chemotherapy consisted of two cycles of epirubicin 120 mg/m2 and ifosfamide 7500 mg/m2 in the case of tandem HDC and one cycle of paclitaxel 135 mg/m2, epirubicin 90 mg/m2 and ifosfamide 6000 mg/m2 in the case of triple HDC. Tandem HDC was composed of two cycles of epirubicin 180 mg/m2, ifosfamide 12000 mg/m2 and carboplatin 900 mg/m2. In the case of triple HDC, paclitaxel 180 mg/m2, etoposide 1500 mg/m2 and thiotepa 600 mg/m2 was added as the third cycle. Patients with tandem HDC (n = 20) were evaluable for both survival and toxicity, and patients with triple HDC (n = 21) only for toxicity because of short-term follow-up. Both tandem and triple HDC were well tolerated and could be safely administered. Non-hematological WHO grade 3 or 4 toxicities were mucositis (8), temporary renal insufficiency (1), myocardial infarction (1), and neuropathy (1). No toxic death occurred. The Kaplan-Meier estimates for 44-months without progression and the overall survival were 12% and 38% respectively. The median survival was 22 months (95% CI: 7.4-51.7 months) and the median progression-free interval 14 months (95% CI: 5.1-43.7 months). In a population with an unfavorable prognosis, tandem HDC showed similar efficacy as to that described in other phase II studies. Triple HDC seems not to improve patient outcome compared to tandem HDC, but a long-term follow up is required.


A review on cationic starch and nanocellulose as paper coating components.
Starch and derivatives thereof have proven their usefulness in paper coating processes. Among these derivatives, cationic starch has been widely used in the paper industry as a flocculation, dispersion and ink fixing agent. In another context, nanoscale cellulosic materials have been shown to improve the strength, retention of fillers, the barrier properties of packaging paper products, and printing qualities. This review summarizes the recent studies on the general components used in paper coating, describes the conventional and alternative synthetic processes of cationic starches and nanocellulose, and deals with their current and potential applications in papermaking, focusing primarily on surface treatments. Moreover, environmental applications have been considered to expand the understanding and usefulness of these materials. Further research on modified polysaccharides is encouraged to replace, in a feasible way, petro-based components of coating formulations, and to provide paper surfaces with new properties.


Exposure to exogenous DNA can modify the sensitivity of the Fas apoptotic pathway.
Gene-transfer techniques are commonly employed for both in vitro and in vivo studies. However, modifications of the target cell following the introduction of the gene of interest are not often examined. These modifications can alter the immunogenicity and/or the susceptibility of the target cell to apoptosis and may produce unwanted consequences in vivo. Gene transfer into the murine fibroblastic Psi-CRIP packaging cell line was performed using calcium phosphate precipitation, cationic liposome-DNA complexes or a retroviral RNA-mediated method. After gene transfer, Fas expression, cytokine production, and sensitivity to Fas ligand (FasL)-mediated death were assessed. Following transfection of a FasL expression vector by calcium phosphate precipitation, an unexpected increase was observed in apoptotic cell death in previously Fas-resistant Psi-CRIP cells. This apoptosis was due to Fas upregulation and an increase of sensitivity to FasL-mediated death. Other plasmids coding non-cytotoxic factors also modulated this apoptotic pathway. The co-stimulatory molecule CD80 was also upregulated. Exposure to naked DNA alone elicited the same response. The effect was not dependent on the methylation status of exogenous DNA, but was found to be dependent on the target cell type and might be avoided by the use of an RNA-mediated retroviral system. Plasmid transfection or simple exposure to naked DNA can increase sensitivity to apoptosis. The generation of FasL packaging cell lines is therefore limited by an increase in FasL/Fas-mediated apoptosis. These findings should be considered when using genetically modified transplantable cells in order to prevent elimination by host cytotoxic cells and in particular when cells are engineered using FasL.


Effect of day 3 embryo morphometrics and morphokinetics on survival and implantation after slow freezing-thawing and after vitrification-warming: a retrospective cohort study.
Morphometric and morphokinetic evaluation of in vitro cultured human embryos allows evaluation without time restriction and reduces intra- and inter-observer variability. Even though these technologies have been reported to improve the quality of cleavage stage embryo evaluation during fresh culture, possible advantages in the evaluation of cryopreserved embryos have been scarcely explored. This study aims to compare morphometric and morphokinetic parameters between slow frozen and vitrified embryos and to determine their relationship to embryo survival and implantation rate (IR) after thawing/warming. During fresh culture, morphometric characteristics (Total Cell Volume (TCV), symmetry, fragmentation and number of blastomeres) were measured in 286 thawed/warmed embryos. Likewise, after thawing/warming, similar morphometric characteristics were measured in 135 survived embryos. Moreover, morphokinetic parameters (time to mitosis resumption and time to compaction) were measured in 90 embryos after thawing/warming. Then, using linear regression, we investigated the differences between vitrified and slow frozen embryos and the relation of the measured characteristics to embryo survival and IR. Statistical corrections were applied to account for data clustering and for multiple testing. Vitrified embryos resume mitosis and start compaction significantly earlier than slow frozen embryos. Mitosis resumption rate was 82% for vitrified and 63% for slow frozen embryos and median time to mitosis resumption was 7.6 h and 13.1 h (p = 0.02), respectively. Compaction rate was 62% in vitrified and only 23% in slow frozen embryos. Median time to compaction was 18.1 h for vitrified embryos but, for slow frozen could not be computed since less than half of the slow frozen embryos reached compaction (p = 0.0001). Moreover, intact embryos resume mitosis significantly earlier than not intact ones regardless of the freezing method (rate: 79% vs. 66%, median time: 7.6 h vs 14.6 h, respectively, p = 0.03). Regarding morphometrics, slow frozen embryos showed lower TCV and higher blastomere symmetry after thawing than vitrified embryos despite having similar blastomere number. IR was related to blastomere number at cryopreservation in slow frozen embryos, but not in vitrified ones. Interestingly, vitrified/warmed embryos undergo mitosis resumption and compaction significantly earlier than slow frozen/thawed embryos. However, the clinical use of this morphokinetic parameters still remains to be investigated in larger studies. Retrospectively registered on December 15, 2015 NCT02639715 .


Sustained delivery of rhMG53 promotes diabetic wound healing and hair follicle development.
MG53 is an essential component of the cell membrane repair machinery, participating in the healing of dermal wounds. Here we develop a novel delivery system using recombinant human MG53 (rhMG53) protein and a reactive oxygen species (ROS)-scavenging gel to treat diabetic wounds. Mice with ablation of MG53 display defective hair follicle structure, and topical application of rhMG53 can promote hair growth in the mg53 -/- mice. Cell lineage tracing studies reveal a physiological function of MG53 in modulating the proliferation of hair follicle stem cells (HFSCs). We find that rhMG53 protects HFSCs from oxidative stress-induced apoptosis and stimulates differentiation of HSFCs into keratinocytes. The cytoprotective function of MG53 is mediated by STATs and MAPK signaling in HFSCs. The thermosensitive ROS-scavenging gel encapsulated with rhMG53 allows for sustained release of rhMG53 and promotes healing of chronic cutaneous wounds and hair follicle development in the db/db mice. These findings support the potential therapeutic value of using rhMG53 in combination with ROS-scavenging gel to treat diabetic wounds.


Effect of trehalose on the phase properties of hydrated and lyophilized dipalmitoylphosphatidylcholine multilayers.
The structure and thermal behavior of hydrated and lyophilized dipalmitoylphosphatidylcholine (DPPC) multilayers in the presence of trehalose were investigated by differential scanning calorimetry and X-ray diffraction methods. Trehalose enters the aqueous space between hydrated bilayers and increases the interbilayer separation (from 0.36 to 1.37 nm in the different DPPC phases at 1 M trehalose). It does not affect the lipid chain packing and also the slow isothermal conversion at 4 degrees C of the metastable L beta' phase into the equilibrium crystalline Lc phase. Addition of trehalose leads to a slight upward shift (about 1 degrees C at 1 M trehalose) of the three phase transitions (sub-, pre-, and main transition) in fully hydrated DPPC while their other properties (enthalpy, excess specific heat, and transition width) remain unchanged. The effect of trehalose on the thermal behavior of DPPC multilayers freeze-dried from an initially completely hydrated state is qualitatively similar to that of water. These data support the "water replacement" hypothesis about trehalose action. It is suggested that trehalose prevents the formation of direct interbilayer hydrogen bonds in states of low hydration.


Characterization of benign and malignant solid breast masses in harmonic 3D power Doppler imaging.
The authors assessed the characteristics of benign and malignant solid breast tumors in harmonic three-dimensional (3D) power Doppler imaging and proposed decision models to classify benign and malignant breast tumors. A total of 86 malignant and 97 benign harmonic 3D power Doppler US images were analyzed. All the harmonic 3D power Doppler images were obtained using a Voluson730 US system (GE, Zipf, Austria) equipped with a RSP 6-12 transducer and tissue harmonic imaging modalities. Imaging analysis was performed using the Virtual Organ Computer-aided Analysis (VOCAL)-imaging program. Histogram indices, the vascularization index (VI), flow index (FI) and vascularization-flow index (VFI), were calculated for the intra-tumor and for shells with an outside thickness of 3mm surrounding the breast tumors. The receiver operating characteristic (ROC) curves were calculated to estimate the diagnostic performances. The results revealed that the choice of decision model comprised the parameters of patient age, intra-tumor VI, and tumor volume to classify benign and malignant breast tumors. The area under the ROC curve (Az) was 0.910, accuracy was 81.4%, and sensitivity and specificity were 81.4% and 81.4%, respectively. The parameter intra-tumor VI was the choice for all of the histogram indices in differentiating between malignant and benign lesions. The decision model, which was composed of patient age, tumor volume and intra-tumor VI, and a cut-off value for intra-tumor VI at the upper end of patient age and tumor volume, was recommended in clinical application.


The relationship between discharge medications and falls in post-hospitalised older adults: A 6-month follow-up.
To identify discharge medications, especially psychotropic medications that are associated with falls, amongst older adults within 6 months following hospitalisation. Negative binomial regression was used to examine relationships between discharge medications and falls in older post-hospitalised adults. Multiple regression that considered falls risk factors at discharge was performed. Data for 267 participants showed that discharge medications were not independently associated with falls postdischarge after adjustment for other falls risk factors. Male gender (adjusted incidence rate ratio [95% confidence interval, CI]) 2.15 [1.36-3.40]), higher depression scores (1.14 [1.05-1.25]) and co-morbidity of neurological disease other than stroke (5.98 [3.08-11.60]) were independently associated with an increased rate of falls. Higher depression scores (1.20 [1.11-1.31]) and co-morbidity of cancer (1.97 [1.20-3.25]) were independently associated with an increased rate of injurious falls in the 6 months postdischarge. Falls prevention strategies, other than hospital discharge medication management in the postdischarge older adults, warrant investigation.


Meta-analysis of randomized controlled trials on effect of cilostazol on restenosis rates and outcomes after percutaneous coronary intervention.
Cilostazol is a generic drug with antiplatelet and antiproliferative effects. It is unclear whether adding cilostazol to standard dual antiplatelet therapy (aspirin and clopidogrel) after percutaneous coronary intervention reduces restenosis and improves the outcomes. We, therefore, conducted a systematic review and meta-analysis. We systematically searched the Cochrane Library, EMBASE, and MEDLINE databases for randomized controlled trials comparing dual antiplatelet therapy with and without cilostazol after percutaneous coronary intervention. The data were pooled using random-effects models and stratified into short-term (1-month), midterm (1- to 12-month), and long-term (≥12-month) follow-up durations. Twelve randomized controlled trials involving 5,655 patients met our inclusion criteria. The addition of cilostazol to dual antiplatelet therapy was not associated with a significant change in target lesion revascularization (TLR) and target vessel revascularization (TVR) at short-term follow-up. However, TLR and TVR were significantly reduced at midterm follow-up (relative risk 0.57, 95% confidence interval 0.39 to 0.84, and relative risk 0.62, 95% confidence interval 0.47 to 0.83, respectively). Data regarding TLR and TVR at long-term follow-up were limited and inconclusive. We did not find a difference in myocardial infarction, mortality, or major bleeding at any follow-up duration. In conclusion, the addition of cilostazol to dual antiplatelet therapy after percutaneous coronary intervention has favorable effects on TLR and TVR at 1 to 12 months, with no differences in adverse outcomes at any follow-up duration.


Is the incidence of congenital toxoplasmosis declining?
Prenatal infection with the protozoan parasite Toxoplasma gondii can cause congenital toxoplasmosis (CT), an often fatal or lifelong-disabling condition. Several studies of human populations have reported temporal decreases in seroprevalence, suggesting declining CT incidence. However, the consistency of this trend among diverse populations remains unclear, as does its implication for prenatal screening programmes, the major intervention against CT. Using temporally resolved data on the seroprevalence of T. gondii in various countries, we discuss how the parasite's changing epidemiology may affect trends in CT incidence in varying and counterintuitive ways. We argue that parasite stage-specific serology could be helpful for understanding underlying causes of secular changes in seroprevalence. Furthermore, we highlight the importance of updating cost-effectiveness estimates of screening programmes, accounting for neuropsychiatric sequelae.


Structural studies of the O-antigen of the lipopolysaccharide from an avirulent strain (M4S) of Pseudomonas solanacearum.
The structure of the O-antigen of the lipopolysaccharide from an avirulent strain (M4S) of Pseudomonas solanacearum has been investigated by methylation analysis, n.m.r. spectroscopy, and N-deacetylation-deamination, followed by analysis and controlled Smith-degradation of the product. These studies demonstrate that the O-antigen is composed of a tetrasaccharide repeating-unit having the following structure: ----3)-alpha-D-GlcpNAc-(1----2)-alpha-L-Rhap-(1----2)-alpha- L-Rhap-(1----3)- alpha-L-Rhap-(1----.


Visual deprivation effects on human motor cortex excitability.
Single and paired-pulse transcranial magnetic stimulation (TMS) were applied to the motor cortex of 12 healthy volunteers, who were instructed to relax under eyes-open and eyes-closed conditions with room lights on and after 30 min of blindfolding. Compared to the eyes-open condition, significantly larger motor-evoked potentials and less intracortical inhibition were observed during blindfolding. Visual deafferentation changes resting human motor cortex excitability and might be a novel way to promote brain plasticity. These results raise the issue of how widespread the effects of temporary deafferentation may be and whether they are mediated by discrete or diffuse systems. These findings also illustrate an important potential confound in TMS studies of the motor cortex.


Pioneers in migration, pioneering in dementia: first generation immigrants in a European metropolitan memory clinic.
By reviewing the clinical files of 1,058 consecutive newly admitted outpatients of a Brussels-based memory clinic between 2005 and 2012, this study aims to document the demographic and clinical characteristics of European and non-European first generation immigrants. They accounted for 18.6% of the patients, of which 8.6% came from outside Europe (mostly from Morocco, Turkey and the Democratic Republic of Congo). Immigrants with AD tended to be younger, and there was a higher proportion of males among non-European ethnic minorities. There was a higher proportion of Parkinson-related cognitive disorders and Lewy Body disease among European immigrants, whereas non-Europeans had more often a psychiatric diagnosis. Even after correction for education, non-European immigrants had lower MMSE scores, and they did report longer delays between first symptoms and diagnosis, although this last difference was not statistically significant. These results suggest that non-European immigrants with cognitive problems consult later.


Introducing EMR children to arithmetical operations: a program involving pictorial problems and distinctive-feature prompts.
This study assessed a program involving distinctive-feature prompts for teaching four EMR children to use the appropriate arithmetical operations (addition, subtraction, multiplication, and division) in the presence of multicomponent pictorial problems. The prompts highlighted critical features of the problems and cued the operations implied. The operations were computed by means of pocket calculators adapted to the response requirements. Training was carried out according to a modified multiple-probe design. Data indicate that the program was effective in establishing the use of operations with all subjects. This behavior was also displayed on problems portraying actions not included in training. The findings are discussed in terms of distinctive-feature prompting, orienting responses, and self-cueing. Attention is given to the generalization of responding across problems, and to educational implications of the program.


Auditory brainstem evoked responses in brainstem compression due to posterior fossa tumors.
Five patients with unilateral posterior fossa tumors were investigated with Auditory Brainstem Evoked Responses (B.E.R.). The B.E.R., recorded from the ear ipsilateral to the tumor, were consistent with a lesion of the acoustic nerve or the cochlear nuclei in all patients. The contralaterally evoked responses in four patients with large tumors and radiological signs of brainstem compression showed normal latencies and amplitudes of the first three waves, while waves IV and V were definitely abnormal with respect to amplitudes and latencies. (The 4th patient had a normal wave IV and a delayed, small wave V). In one patient with a relatively small acoustic neurinoma the contralateral B.E.R. were normal. It is concluded that the upper pontine and lower midbrain level, presumed sites of generation of wave IV and V of the B.E.R., are more susceptible to brainstem compression due to unilateral posterior fossa tumors, than the lower parts of the brainstem.


Analysis of carbohydrates in glycoproteins by high-performance liquid chromatography and high-performance capillary electrophoresis.
We describe two methods for the analysis of oligosaccharide chains in glycoproteins by high-performance liquid chromatography (HPLC) and high-performance capillary electrophoresis (HPCE). O- and N-glycosidically linked oligosaccharides released from glycoproteins can be identified as their borohydride-reduced forms by anion-exchange HPLC with pulsed amperometric detection. N-Glycosidically linked oligosaccharides can also be analyzed as 2-aminopyridine derivatives by HPCE in direct zone electrophoresis mode in an acidic phosphate buffer and zone electrophoresis mode as borate complexes in an alkaline buffer. We also present a convenient procedure for the analysis of the constituent monosaccharides of these oligosaccharides chains by HPLC based on reversed-phase partition mode as 1-phenyl-3-methyl-5-pyrazolone derivatives.


Association of Dynamic Changes in the CD4 T-Cell Transcriptome With Disease Severity During Primary Respiratory Syncytial Virus Infection in Young Infants.
Nearly all children are infected with respiratory syncytial virus (RSV) within the first 2 years of life, with a minority developing severe disease (1%-3% hospitalized). We hypothesized that an assessment of the adaptive immune system, using CD4+ T-lymphocyte transcriptomics, would identify gene expression correlates of disease severity. Infants infected with RSV representing extremes of clinical severity were studied. Mild illness (n = 23) was defined as a respiratory rate (RR) < 55 and room air oxygen saturation (SaO2) ≥ 97%, and severe illness (n = 23) was defined as RR ≥ 65 and SaO2 ≤ 92%. RNA from fresh, sort-purified CD4+ T cells was assessed by RNA sequencing. Gestational age, age at illness onset, exposure to environmental tobacco smoke, bacterial colonization, and breastfeeding were associated (adjusted P < .05) with disease severity. RNA sequencing analysis reliably measured approximately 60% of the genome. Severity of RSV illness had the greatest effect size upon CD4 T-cell gene expression. Pathway analysis identified correlates of severity, including JAK/STAT, prolactin, and interleukin 9 signaling. We also identified genes and pathways associated with timing of symptoms and RSV group (A/B). These data suggest fundamental changes in adaptive immune cell phenotypes may be associated with RSV clinical severity.


NFIB-MLL1 complex is required for the stemness and Dlx5-dependent osteogenic differentiation of C3H10T1/2 mesenchymal stem cells.
Despite significant progress in our understanding of the molecular mechanism of mesenchymal stem cell (MSC) differentiation, less is known about the factors maintaining the stemness and plasticity of MSCs. Here, we show that the NFIB-MLL1 complex plays key roles in osteogenic differentiation and stemness of C3H10T1/2 MSCs. We find that depletion of either NFIB or MLL1 results in a severely hampered osteogenic potential and failed activation of key osteogenic transcription factors, such as Dlx5, Runx2, and Osx, following osteogenic stimuli. In addition, the NFIB-MLL1 complex binds directly to the promoter of Dlx5, and exogenous expression of Myc-Dlx5, but not the activation of either the BMP- or the Wnt-signaling pathway, is sufficient to restore the osteogenic potential of cells depleted of NFIB or MLL1. Moreover, chromatin immunoprecipitation (ChIP) and ChIP-sequencing analysis showed that the NFIB-MLL1 complex mediates the deposition of trimethylated histone H3K4 at both Dlx5 and Cebpa, key regulator genes that function at the early stages of osteogenic and adipogenic differentiation, respectively, in uncommitted C3H10T1/2 MSCs. Surprisingly, the depletion of either NFIB or MLL1 leads to decreased trimethylated histone H3K4 and results in elevated trimethylated histone H3K9 at those developmental genes. Furthermore, gene expression profiling and ChIP-sequencing analysis revealed lineage-specific changes in chromatin landscape and gene expression in response to osteogenic stimuli. Taken together, these data provide evidence for the hitherto unknown role of the NFIB-MLL1 complex in the maintenance and lineage-specific differentiation of C3H10T1/2 MSCs and support the epigenetic regulatory mechanism underlying the stemness and plasticity of MSCs.


Induction of ER Stress-Mediated Apoptosis by α-Lipoic Acid in A549 Cell Lines.
α-Lipoic acid (α-LA) has been studied as an anticancer agent as well as a therapeutic agent for diabetes and obesity. We performed this study to evaluate the anticancer effects and mechanisms of α-LA in a lung cancer cell line, A549. α-LA-induced apoptosis of A549 cells was detected by fluorescence-activated cell sorting analysis and a DNA fragmentation assay. Expression of apoptosis-related genes was analyzed by western blot and reverse transcription-polymerase chain reaction analyses. α-LA induced apoptosis and DNA fragmentation in A549 cells in a dose- and time-dependent manner. α-LA increased caspase activity and the degradation of poly (ADP-ribose) polymerase. It induced expression of endoplasmic reticulum (ER) stress-related genes, such as glucose-regulated protein 78, C/EBP-homologous protein, and the short form of X-box binding protein-1, and decreased expression of the anti-apoptotic protein, X-linked inhibitor of apoptosis protein. Reactive oxygen species (ROS) production was induced by α-LA, and the antioxidant N-acetyl-L-cysteine decreased the α-LA-induced increase in expression of apoptosis and ER stress-related proteins. α-LA induced ER stress-mediated apoptosis in A549 cells via ROS. α-LA may therefore be clinically useful for treating lung cancer.


Cux/CDP homeoprotein is a component of NF-muNR and represses the immunoglobulin heavy chain intronic enhancer by antagonizing the bright transcription activator.
Nuclear matrix attachment regions (MARs) flanking the immunoglobulin heavy chain intronic enhancer (Emu) are the targets of the negative regulator, NF-muNR, found in non-B and early pre-B cells. Expression library screening with NF-muNR binding sites yielded a cDNA clone encoding an alternatively spliced form of the Cux/CDP homeodomain protein. Cux/CDP fulfills criteria required for NF-muNR identity. It is expressed in non-B and early pre-B cells but not mature B cells. It binds to NF-muNR binding sites within Emu with appropriate differential affinities. Antiserum specific for Cux/CDP recognizes a polypeptide of the predicted size in affinity-purified NF-muNR preparations and binds NF-muNR complexed with DNA. Cotransfection with Cux/CDP represses the activity of Emu via the MAR sequences in both B and non-B cells. Cux/CDP antagonizes the effects of the Bright transcription activator at both the DNA binding and functional levels. We propose that Cux/CDP regulates cell-type-restricted, differentiation stage-specific Emu enhancer activity by interfering with the function of nuclear matrix-bound transcription activators.


Production of monoclonal antibodies to thyroglobulin by in vitro immunization with a free synthetic peptide.
A synthetic peptide corresponding to amino acids 1-19 of thyroglobulin was used to test the possibility of generating protein-reactive monoclonal antibodies by immunization in vitro with a synthetic peptide as antigen. Splenocytes from non-immunized Balb/c mice were cultured in serum-free medium for 3 days in the presence of thymocyte-conditioned medium and the synthetic peptide prior to fusion with SP2/0 murine myeloma cells. The synthetic peptide was used in its free form, i.e. not coupled to a protein carrier. Hybridomas secreting monoclonal antibodies reactive with the synthetic peptide were obtained after immunization in vitro with as little as 10 ng/ml of the synthetic peptide. Between 50 and 70% of the primary clones obtained in different experiments produced monoclonal antibodies also reactive with the intact protein. Six stable hybridomas were isolated; all produced antibodies of the IgM class. We conclude that immunization in vitro with a free synthetic peptide is an efficient method for the generation of monoclonal antibodies reactive with the intact protein.


Histology of Pregnancy-Associated Breast Cancer.
Breasts are one of the most common sites of neoplastic lesions in women during pregnancy and lactation. This chapter reviews carcinomas of the breast during pregnancy and lactation while focusing on histologic features, biomarker profiles and some involved molecular pathways. Also, a brief review of previous studies on this field is performed.


Prevalence of occult hepatitis B virus infection among patients receiving haemodialysis in Sana'a city.
Occult hepatitis B virus infection (OBI) is a challenging entity. Due to the increase in invasive procedures, blood transfusions, and difficulties in diagnosing OBI, patients are more likely to acquire OBI. This cross-sectional study was conducted to assess the prevalence of OBI by hepatitis B virus (HBV)-DNA detection, the prevalence of HBV infection by total hepatitis B core antibody (HBcAb) detection, and the potential risk factors for HBV infection in patients receiving haemodialysis regularly. This study included 80 patients receiving haemodialysis regularly, without acute or chronic HBV infection. They were selected from the dialysis units in Sana'a city, Yemen from June 2016 to June 2017. Patients who were positive for hepatitis B surface antigen and hepatitis B surface antibody were excluded from this study. Blood samples were taken prior to each haemodialysis session, and serological markers of HBV were included. HBcAb was measured by enzyme-linked immunosorbent assay, and HBV-DNA was measured by polymerase chain reaction. HBV-DNA was detected in four patients (5%) and HBcAb was detected in 38 patients (47.5%). There was a significant association between HBV-DNA and HBcAb in patients receiving haemodialysis regularly. Patients who test positive for HBcAb should undergo additional HBV-DNA testing to allow accurate HBV screening and prevent infection of other patients.


[Rheological properties of the blood and erythrocyte suspension after exposure to zootoxins].
The rheological properties of blood (viscosity, suspension stability and deformation of red blood cells) have been studied under the influence of zootoxins. It has been shown that the toxins Apis mellifera, Mesobuthus eupeus, Agkistrodon blomhoffii in vitro exert essential effect on haemorheological indexes.


Drug membrane transporters and CYP3A4 are affected by hypericin, hyperforin or aristoforin in colon adenocarcinoma cells.
Our previous results have shown that the combination of hypericin-mediated photodynamic therapy (HY-PDT) at sub-optimal dose with hyperforin (HP) (compounds of Hypericum sp.), or its stable derivative aristoforin (AR) stimulates generation of reactive oxygen species (ROS) leading to antitumour activity. This enhanced oxidative stress evoked the need for an explanation for HY accumulation in colon cancer cells pretreated with HP or AR. Generally, the therapeutic efficacy of chemotherapeutics is limited by drug resistance related to the overexpression of drug efflux transporters in tumour cells. Therefore, the impact of non-activated hypericin (HY), HY-PDT, HP and AR on cell membrane transporter systems (Multidrug resistance-associated protein 1-MRP1/ABCC1, Multidrug resistance-associated protein 2-MRP2/ABCC2, Breast cancer resistance protein - BCRP/ABCG2, P-glycoprotein-P-gp/ABCC1) and cytochrome P450 3A4 (CYP3A4) was evaluated. The different effects of the three compounds on their expression, protein level and activity was determined under specific PDT light (T0+, T6+) or dark conditions (T0- T6-). We found that HP or AR treatment affected the protein levels of MRP2 and P-gp, whereas HP decreased MRP2 and P-gp expression mostly in the T0+ and T6+ conditions, while AR decreased MRP2 in T0- and T6+. Moreover, HY-PDT treatment induced the expression of MRP1. Our data demonstrate that HP or AR treatment in light or dark PDT conditions had an inhibitory effect on the activity of individual membrane transport proteins and significantly decreased CYP3A4 activity in HT-29 cells. We found that HP or AR significantly affected intracellular accumulation of HY in HT-29 colon adenocarcinoma cells. These results suggest that HY, HP and AR might affect the efficiency of anti-cancer drugs, through interaction with membrane transporters and CYP3A4.


ERBB2-induced inflammation in lung carcinogenesis.
ERBB2/HER2/NEU, a member of the epidermal growth factor receptor family, is overexpressed in more than 25 % of non-small cell lung cancer and is considered to be a significant and independent prognostic factor in lung cancer. Here we generated a lung specific HER2 overexpressing transgenic mouse model. In this model, HER2 was driven by the human surfactant protein-C promoter to investigate the role of the HER2 oncogene in pulmonary carcinogenesis and progression. Notably, significant pathological changes, including lymphocyte infiltration and mesenchymal cells hyperplasia, were found in the lung tissue of transgenic mice aged from 4 to 12 months. The occurrence and severity of those lesions increased as the mice aged. Some inflammatory factors, such as tumor necrosis factor, interleukin 1 and interleukin 6, were upregulated in lung tissue of transgenic mice compared with that of wild-type mice, implying that long-term HER2 overexpression could induce serious lung inflammation and some precancerous lesions. This model would be useful for studying the mechanism of HER2 involvement in lung carcinogenesis and for understanding the relationship between carcinogenesis and inflammation.


Synthesis, structure, and biological activity of novel (oxdi/tri)azoles derivatives containing 1,2,3-thiadiazole or methyl moiety.
To develop novel inhibitors of ketol-acid reductoisomerase, a series of (oxdi/tri)azoles derivatives was synthesized and characterized by (1)H NMR, MS, elemental analyses, and crystallography. According to the biological activities of these compounds obtained both in vivo and in vitro, compound 4-cyclopropyl-3-((4-fluorobenzyl)thio)-5-methyl-4H-1,2,4-triazole showed excellent KARI inhibitory activity (100% at 100 μg mL (-1) in vitro). In addition, most of the title compounds exhibited good herbicidal activity against Brassica campestris in vivo.


Differential effects of a school-based obesity prevention program: A cluster randomized trial.
School-based healthy living interventions are widely promoted as strategies for preventing obesity. The peer-led Healthy Buddies™ curriculum has been shown to improve obesity-related outcomes in school-aged children. We examined whether these improvements existed among subgroups of children stratified by sex, income level and urban/rural geography. In a cluster-randomized controlled trial, elementary schools in Manitoba, Canada, were randomly allocated to Healthy Buddies™ (10 schools, 340 students) or standard curriculum (10 schools, 347 students). Healthy Buddies™ participants had 21weekly lessons on healthy eating, physical activity and self-efficacy, delivered by children age 9-12 to children age 6-8. We assessed pre- and post-intervention body mass index (BMI) z-scores, waist circumference, healthy living knowledge, dietary intake and self-efficacy among the younger children. Compared to standard curriculum (n = 154), Healthy Buddies™ participants (n = 157) experienced a greater reduction in waist circumference (-1.7 cm; 95% confidence interval [CI][-2.8, -0.5 cm]) and improved dietary intake (4.6; 95% CI [0.9, 8.3]), healthy living knowledge (5.9; 95% CI [2.3, 9.5]) and self-efficacy (5.3; 95% CI [1.0, 9.5]) scores. In subgroup analyses, effects for waist circumference (-2.0 cm; 95% CI [-3.6, -0.5]), healthy living knowledge (9.1; 95% CI [4.4, 13.8]) and self-efficacy (8.3; 95% CI [3.3, 13.3]) were significant among boys. Dietary intake (10.5; 95% CI [5.5, 15.4]), healthy living knowledge (9.8; 95% CI [4.5, 15.0]) and self-efficacy (6.7; 95% CI [0.7, 12.7]) improved among urban-dwelling but not rural-dwelling children. Healthy Buddies™ was effective for boys and children living in urban settings. Enhanced curricula may be needed to improve program effectiveness for select subgroups of school-aged children.


Excretion of hydroflumethiazide in bile and urine of man.
Biliary and urinary excretion of hydroflumethiazide (HFT) and its metabolite, 2,4-disulfamyl-5-trifluoromethylaniline (DTA), was investigated in 5 otherwise healthy patients with a T-drain in the common bile duct after cholecystectomy and choledochotomy. After a single oral dose of HFT 302--453 micromoles, a mean of 0.051% (range 0.028--0.075) of the dose was excreted in bile and 34.9% (range 23.7--45.4) in urine during the first 6 hours after administration. Biliary excretion appeared to be of minor importance in the elimination of HFT by man. DTA could not be detected in bile.


Effect of calving to timed artificial insemination interval on fertility of beef cows.
The objectives of this study were to determine the effect of calving to timed artificial insemination (C-TAI) interval on fertility of Bos taurus and Bos indicus cows and to determine the best C-TAI interval to include postpartum cows in TAI programs in each genotype (B. taurus and B. indicus). In experiment 1, a retrospective study was performed, with data from 2,709 TAI in Bos taurus and Bos indicus cows with different C-TAI intervals. There was a positive linear effect of C-TAI interval on the pregnancy probability (p < .0001). However, there was no effect or interaction with the genotype. When C-TAI intervals were grouped in classes, pregnancy rate differed in groups (p < .05). Receiver operating characteristic (ROC) analysis was performed, and it was observed that 52 days would be the better C-TAI interval to maximize fertility in beef cows. According to the previous result, we further stratified into two C-TAI interval groups (≤ 52 days or > 52 days). Cows with C-TAI longer than 52 days (C-TAI > 52) had greater pregnancy rate (56%) than cows with short C-TAI (46%; p < .01). In experiment 2, 18 postpartum beef cows were subjected to TAI protocol at different C-TAI intervals (n = 6 cows per group): 40, 60 or 80 days postpartum. Cows with 60 days of C-TAI had a greater preovulatory follicle diameter than cows with 80 days postpartum (p < .05). The C-TAI interval did not affect (p > .05) the ovulation rate nor the proportion of endometrial polymorphonuclear leucocytes. We conclude that C-TAI interval affects pregnancy rate in both Bos taurus and Bos indicus. We recommend a C-TAI interval longer than 52 days to increase fertility of beef cows included in TAI programs.


Discriminating between rate heterogeneity and interspecific recombination in DNA sequence alignments with phylogenetic factorial hidden Markov models.
A recently proposed method for detecting recombination in DNA sequence alignments is based on the combination of hidden Markov models (HMMs) with phylogenetic trees. Although this method was found to detect breakpoints of recombinant regions more accurately than most existing techniques, it inherently fails to distinguish between recombination and rate variation. In the present paper, we propose to marry the phylogenetic tree to a factorial HMM (FHMM). The states of the first hidden chain represent tree topologies, whereas the states of the second independent hidden chain represent different global scaling factors of the branch lengths. Inference is done in terms of a hierarchical Bayesian model, where parameters and hidden states are sampled from the posterior distribution with Gibbs sampling. We have tested the proposed model on various synthetic and real-world DNA sequence alignments. The simulation results suggest that as opposed to the standard phylogenetic HMM, the phylogenetic FHMM clearly distinguishes between recombination and rate heterogeneity and thereby avoids the prediction of spurious recombinant regions. The proposed method has been implemented in a MATLAB package that extends Kevin Murphy's HMM toolbox. Software and data used in our study are available from http://www.bioss.sari.ac.uk/~dirk/Supplements


Mammary Organoids and 3D Cell Cultures: Old Dogs with New Tricks.
3D cell culture methods have been an integral part of and an essential tool for mammary gland and breast cancer research for half a century. In fact, mammary gland researchers, who discovered and deciphered the instructive role of extracellular matrix (ECM) in mammary epithelial cell functional differentiation and morphogenesis, were the pioneers of the 3D cell culture techniques, including organoid cultures. The last decade has brought a tremendous increase in the 3D cell culture techniques, including modifications and innovations of the existing techniques, novel biomaterials and matrices, new technological approaches, and increase in 3D culture complexity, accompanied by several redefinitions of the terms "3D cell culture" and "organoid". In this review, we provide an overview of the 3D cell culture and organoid techniques used in mammary gland biology and breast cancer research. We discuss their advantages, shortcomings and current challenges, highlight the recent progress in reconstructing the complex mammary gland microenvironment in vitro and ex vivo, and identify the missing 3D cell cultures, urgently needed to aid our understanding of mammary gland development, function, physiology, and disease, including breast cancer.


Junction-free Flat Copper Nanofiber Network-based Transparent Heater with High Transparency, High Conductivity, and High Temperature.
Transparent conducting electrodes (TCE) are widely used in a variety of applications including displays, light-emitting diodes (LEDS), and solar cells. An important factor in TCE design is active control of the sheet resistance and transparency; as these are inversely proportional, it is essential to develop a technology that can maintain high transparency, while actively controlling sheet resistance, for a range of applications. Here, a nanofiber network was fabricated based on direct electrospinning onto a three-dimensional (3-D) complex substrate; flat metal electrodes without junction resistance were produced using heat treatment and electroless deposition. The fabricated transparent electrode exhibited a transparency of over 90% over the entire visible light range and a sheet resistance of 4.9 ohms/sq. Adhesion between the electrode and substrate was superior to other electrospinning-based transparent electrodes. The performance of the transparent electrode was verified by measurements taken while using the electrode as a heater; a maximum temperature of 210 °C was achieved. The proposed copper nanofiber-based heater electrode offers the advantages of transparency as well as application to complex 3-D surfaces.


Spontaneous formation of gold nanoparticles on MoS2 nanosheets and its impact on solution-processed optoelectronic devices.
Understanding size-dependent properties of 2D materials is crucial for their optimized performance when incorporated through solution routes. In this work, the chemical nature of MoS2 as a function of nanosheet size is investigated through the spontaneous reduction of chloroauric acid. Microscopy studies suggest higher gold nanoparticle decoration density in smaller nanosheet sizes, resulting from higher extent of reduction. Further corroboration through surface-enhanced Raman scattering using the gold-decorated MoS2 nanosheets as substrates exhibited an enhancement factor of 1.55 × 106 for smaller nanosheets which is 7-fold higher as compared to larger nanosheets. These plasmonic-semiconductor hybrids are utilized for photodetection, where decoration is found to impact the photoresponse of smaller nanosheets the most, and is optimized to achieve responsivity of 367.5 mAW-1 and response times of ∼17 ms. The simplistic modification via solution routes and its impact on optoelectronic properties provides an enabling platform for 2D materials-based applications.


Cyclic parenteral nutrition in hospitalized adult patients: a 9-year experience.
We are reporting our 9-year experience of cyclic parenteral nutrition with 200 in-patients (mean age 52 years) treated 64 +/- (SEM)3 days (range: 15-230) for Gl diseases. During the first period (A, n = 38), all-in-one nutritive bags with intralipid 10% were used; in a second period (B, n = 135), intralipid 10% was infused separately over the first 6 h of nutrition; in the final period (C, n = 27), Intralipid 20% was used in all-in-one nutritive bags. Indications for parenteral nutrition were non-malignant digestive diseases in 89.5% of the cases. The frequency of complications appearing during cyclic parenteral nutrition was the following: catheter-related sepsis 8%, catheter obstruction 8%, hypertriglyceridemia 33%, hypercholesterolemia 14.7%, liver function test abnormalities 28% and biliary sludge and/or lithiasis 31.6%. 5 patients, with chronic obstructive small bowel disease, developed jaundice with no identifiable cause other than parenteral nutrition. The prevalence of catheter obstruction significantly decreased (P < 0.01) during period B, as compared with periods A and C. The prevalence of liver function test abnormalities decreased significantly (P < 0.01) during periods B and C, as compared with period A. Mortality rate related to cyclic parenteral nutrition was 3%. These results suggest that a) the separate infusion of lipid emulsion reduces the prevalence of catheter obstruction; b) as compared with 10% intralipid in all-in-one nutritive bags, the separate infusion of 10% Intralipid or the 20% Intralipid given in all-in-one nutritive bags is associated with a decreased prevalence of liver function test abnormalities; c) chronic small intestine obstruction seems to play a key role in parenteral-associated jaundice.


Pulmonary alveolar proteinosis: new insights from a single-center cohort of 70 patients.
Pulmonary alveolar proteinosis (PAP) is a rare syndrome characterized by the intra-alveolar accumulation of surfactant lipids and proteins. The aim of the study is to describe the epidemiologic, clinical, physiologic, and laboratory features of PAP in a large single-center cohort of patients with PAP. Over 30 years, 70 patients with PAP were managed at our institution, 64 with primary and 6 with secondary PAP. The mean age at diagnosis was 43 years with a male to female ratio of 1.3. BAL was the most commonly applied diagnostic method, performed in 83% of cases. A history of smoking was seen in 79%, and of dust exposure in 54%, most commonly to aluminum, silica and sawdust. GM-CSF autoantibody correlated with clinical outcome and KL-6 with diffusing capacity. The number of whole lung lavages (WLL) necessary to reach remission was higher in current smokers. This study shows that the use of BAL for the diagnosis of PAP can reduce the need of histological confirmation. A history of dust or fume inhalation is strongly associated with PAP, also with the autoimmune form. Smoking seems to influence the response to treatment, increasing the number of WLL necessary to reach remission.


Prospectively estimating the recoverability of renal function after relief of unilateral urinary obstruction by measurement of renal parenchymal volume.
Renal parenchymal volume (RPV) has been suggested as an indicator of the potential functional residual capacity for a given kidney. The goal of this study was to determine whether the recoverability of renal function could be predicted by RPV as estimated by computed tomography (CT) before an operation. Eighty-two adult patients diagnosed with long-term chronic unilateral ureteral obstruction and a normal contralateral kidney were recruited for evaluation. RPV was measured by nonenhanced CT. Glomerular filtration rate (GFR) was measured by radioisotope renal scan. Animal models were used to validate use of the CT method to measure RPV. RPV and GFR values for all patients were obtained before surgical relief of the urinary obstruction and compared with those values obtained at 12 months postsurgery. There was no statistically significant difference found between RPV measured by CT or by the water displacement method. Among patient age, sex, and pre-RPV and pre-GFR of obstructed and contralateral kidney, pre-RPV and pre-GFR of obstructed kidney were the independent factors that best indicated recoverability of renal function. Pre-RPV correlated well with post-GFR (r = 0.68, P < .01). The cut-off point of pre-RPV to predict recoverability of renal function after the relief operation was 58.2 mL, as determined by receiver operating characteristic curve analysis. Pre-RPV was the independent factor that determines recoverability of renal function. Renal function may stabilize or improve after relief of urinary obstruction when the pre-RPV value is ≥58.2 mL.


Comprehensive genetic screening of early-onset dementia patients in an Austrian cohort-suggesting new disease-contributing genes.
Early-onset dementia (EOD), with symptom onset before age 65, has a strong genetic burden. Due to genetic and clinical overlaps between different types of dementia, whole-exome sequencing (WES) has emerged as an appropriate screening method for diagnostic testing and novel gene-finding approaches. We performed WES and C9orf72 repeat testing in 60 well-defined Austrian EOD patients. Seven patients (12%) carried likely disease-causing variants in monogenic genes, PSEN1, MAPT, APP, and GRN. Five patients (8%) were APOE4 homozygote carriers. Definite and possible risk variants were detected in the genes TREM2, SORL1, ABCA7 and TBK1. In an explorative approach, we cross-checked rare gene variants in our cohort with a curated neurodegeneration candidate gene list and identified DCTN1, MAPK8IP3, LRRK2, VPS13C and BACE1 as promising candidate genes. Conclusively, 12 cases (20%) carried variants relevant to patient counseling, comparable to previously reported studies, and can thus be considered genetically resolved. Reduced penetrance, oligogenic inheritance and not yet identified high-risk genes might explain the high number of unresolved cases. To address this issue, we provide complete genetic and phenotypic information (uploaded to the European Genome-phenome Archive), enabling other researchers to cross-check variants. Thereby, we hope to increase the chance of independently finding the same gene/variant-hit in other well-defined EOD patient cohorts, thus confirming new genetic risk variants or variant combinations.


68 Ga-FAPI PET/CT Detection of Non-FDG-Avid Poorly Differentiated Adenocarcinoma of the Trachea.
Primary tracheal adenocarcinoma is relatively rare in clinical practice. We describe the 68 Ga-FAPI-04 and 18 F-FDG PET/CT findings of primary poorly differentiated tracheal adenocarcinoma in a 48-year-old woman. In the present case, the tumor showed increased 68 Ga-FAPI uptake but not significant 18 F-FDG activity.


Ovarian masses in the child and adolescent: An American Pediatric Surgical Association Outcomes and Evidence-Based Practice Committee systematic review.
The treatment of ovarian masses in pediatric patients should balance appropriate surgical management with the preservation of future reproductive capability. Preoperative estimation of malignant potential is essential to planning an optimal surgical strategy. The American Pediatric Surgical Association Outcomes and Evidence-Based Practice Committee drafted three consensus-based questions regarding the evaluation and treatment of ovarian masses in pediatric patients. A search of PubMed, the Cochrane Library, and Web of Science was performed and Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines were followed to identify articles for review. Preoperative tumor markers, ultrasound malignancy indices, and the presence or absence of the ovarian crescent sign on imaging can help estimate malignant potential prior to surgical resection. Frozen section also plays a role in operative strategy. Surgical staging is useful for directing chemotherapy and for prognostication. Both unilateral oophorectomy and cystectomy have been used successfully for germ cell and borderline ovarian tumors, although cystectomy may be associated with higher rates of local recurrence. Malignant potential of ovarian masses can be estimated preoperatively, and fertility-sparing techniques may be appropriate depending on the type of tumor. This review provides recommendations based on a critical evaluation of recent literature. Systematic review of level 1-4 studies. Level 1-4 (mainly 3-4).


Avoiding a feared stimulus: Modelling costly avoidance of learnt fear in a sensory preconditioning paradigm.
Avoidance of learnt fear prevents the onset of a feared stimulus and the threat that follows. In anxiety-related disorders, it turns pathological given its cost and persistence in the absence of realistic threat. The current study examined the acquisition of costly avoidance of learnt fear in healthy individuals (n = 45), via a sensory preconditioning paradigm. Two neutral preconditioning stimuli (PSs) were paired with two neutral conditioned stimuli (CSs). One CS then came to predict an aversive outcome whereas the other CS came to predict safety. In test, participants engaged in stronger avoidance to the PS associated with the fear-related CS than the PS associated with the safety-related CS. Of note, executing behavioral avoidance led to missing out a competing reward, thus rendering avoidance costly. The results also provide preliminary evidence that threat anticipation and a negative change in valence play a role in driving costly avoidance of learnt fear. Future studies should examine how avoidance of learnt fear maintains pathological anxiety.


Isolation of microcellulose from timoho fiber using the process of delinigfication and maceration: Evaluation of physical, chemical, structural, and thermal properties.
Timoho fiber (TF) is a biodegradable bio-fiber rich in cellulose and a renewable agro textile fiber rich in cellulose polysaccharides. Isolation of micro cellulose from TF is an alternative reinforcement approach in a bio-composite application. This study characterized TF obtained from isolation employing FTIR, XRD, SEM-EDS, and TGA using hydrolysis. Pretreatment was done using alkaline, bleaching, and hydrolysis. FTIR confirmed that the maximum amount of noncellulosic materials had been removed from TF. The XRD result showed that hydrolysis treatment increased the crystal size and crystallinity index of the TF (88.68 %). TGA also revealed that the highest thermal stability had been found in the hydrolyzed TF. The micro cellulose TF represented an interesting source of cellulose reinforcing materials, and it might be continued in the form of nano cellulose.


Integrity in Guardianship Decision Making: Applying the Will and Preferences Paradigm.
Guardianship laws have changed significantly over the 20th and 21st centuries to shift from the "best interests" model to the "supported decision-making" model. Such changes reflect the importance of supporting the dignity of risk of persons who lack decision-making capacity for some decisions. Recent changes to the Victorian Guardianship and Administration Act 2019 (Vic) in Australia require guardians to give effect to the "will and preferences" of the represented person, as far as practicable. The changes reveal a compromise between the competing rights to autonomy and to safety. Here, we explore the meaning of "will and preferences" before considering the personal factors that may contribute to the understanding of an individual's will and preferences. The practical challenge of ascertaining, interpreting, and giving effect to a person's will and preferences are also discussed. Finally, we consider the practical challenge of identifying when a person needs decision-making support.


Endogenous opioids contribute to insensitivity to pain in humans and mice lacking sodium channel Nav1.7.
Loss-of-function mutations in the SCN9A gene encoding voltage-gated sodium channel Nav1.7 cause congenital insensitivity to pain in humans and mice. Surprisingly, many potent selective antagonists of Nav1.7 are weak analgesics. We investigated whether Nav1.7, as well as contributing to electrical signalling, may have additional functions. Here we report that Nav1.7 deletion has profound effects on gene expression, leading to an upregulation of enkephalin precursor Penk mRNA and met-enkephalin protein in sensory neurons. In contrast, Nav1.8-null mutant sensory neurons show no upregulated Penk mRNA expression. Application of the opioid antagonist naloxone potentiates noxious peripheral input into the spinal cord and dramatically reduces analgesia in both female and male Nav1.7-null mutant mice, as well as in a human Nav1.7-null mutant. These data suggest that Nav1.7 channel blockers alone may not replicate the analgesic phenotype of null mutant humans and mice, but may be potentiated with exogenous opioids.


Evidence against a hydrodynamic function for fish schools.
THERE are numerous explanations for the formation of fish schools(1,2), one of the most popular being that the members gain hydrodynamic advantage over solitary individuals(3-6). The model proposed by Weihs(5,6) for the first time makes precise predictions about school structure which can be verified. We report here the first empirical test of this model, and demonstrate that three species of schooling fish do not swim in appropriate positions to gain hydrodynamic advantage.


Locking classical correlations in quantum States.
We show that there exist bipartite quantum states which contain a large locked classical correlation that is unlocked by a disproportionately small amount of classical communication. In particular, there are (2n+1)-qubit states for which a one-bit message doubles the optimal classical mutual information between measurement results on the subsystems, from n/2 bits to n bits. This phenomenon is impossible classically. However, states exhibiting this behavior need not be entangled. We study the range of states exhibiting this phenomenon and bound its magnitude.


Distribution of primary afferent fibres from the forelimb of the rat to the upper cervical spinal cord in relation to the location of spinothalamic neuron populations.
Half the monosynaptic spinothalamic neurons in the rat spinal cord lie in the first 4 cervical segments, predominantly in the internal basilar column and ventral horn. The present study shows that both these regions receive primary afferents from the forelimb. The superficial dorsal horn receives few, and the region of the lateral cervical nucleus no primary afferents from the brachial plexus. We suggest that internal basilar column cells may represent localised relays concerned with the limbs, and that the gracile and cuneate nuclei are not unique, but are paralleled by other neuron populations in the upper cord.


Gastrins and cholecystokinins release acetylcholine but not substance P from neurons in the guinea-pig taenia coli.
Gastrins and cholecystokinins contract the isolated taenia coli of the guinea-pig. Porcine CCK-39 produced the greatest contractile response and human gastrin-17 I and -34 the weakest. Pentagastrin had the highest affinity to the receptors and non-sulphated CCK-8 the lowest. The contractions produced by the CCK peptides were reduced by the neuronal blocker tetrodotoxin and by the muscarinic blocker atropine but not by the substance P antagonist [D-Pro2,D-Trp7,9]SP. It is concluded that gastrin/CCK peptides act directly on smooth muscle cells and that in addition these peptides, notably sulphated forms of CCK, are capable of exciting cholinergic neurons (but not SP neurons) to cause smooth muscle contraction.


Structure and function of lipid droplet assembly complexes.
Cells store lipids as a reservoir of metabolic energy and membrane component precursors in organelles called lipid droplets (LDs). LD formation occurs in the endoplasmic reticulum (ER) at LD assembly complexes (LDAC), consisting of an oligomeric core of seipin and accessory proteins. LDACs determine the sites of LD formation and are required for this process to occur normally. Seipin oligomers form a cage-like structure in the membrane that may serve to facilitate the phase transition of neutral lipids in the membrane to form an oil droplet within the LDAC. Modeling suggests that, as the LD grows, seipin anchors it to the ER bilayer and conformational shifts of seipin transmembrane segments open the LDAC dome toward the cytoplasm, enabling the emerging LD to egress from the ER.