Processing of nonstructural proteins NS4A and NS4B of dengue 2 virus in vitro and in vivo.
The production, from polyprotein precursors, of two hydrophobic nonstructural proteins of dengue 2 (DEN2) virus, NS4A and NS4B, was analyzed both in cell-free systems and in infected cells. In DEN2-infected cells, NS4B is first produced as a peptide of apparent size 30 kDa; NS4B is then post-translationally modified, in an unknown way, to produce a polypeptide of apparent size 28 kDa. The rate and extent of NS4B modification was found to be cell-dependent; in BHK cells the half-time for the conversion of the 30-kDa form to the 28-kDa form was 90 min. N-terminal sequence analysis of NS4B suggests that the N-terminus is produced by an enzyme with a specificity similar to that of signalase. Low levels of a putative polyprotein, NS4AB, were also found in mammalian cells, but not mosquito cells, infected with DEN2, suggesting that a small proportion of DEN2 4A/4B cleavage can occur post-translationally or that some nonstructural polyproteins escape normal processing. Cleavage of the 4A/4B bond in infected cells required expression of DEN2 sequences in addition to those in NS4A and NS4B, as NS4AB produced in cells by a vaccinia expression system was not cleaved. NS4AB produced in cells by a vaccinia expression system was modified post-translationally, presumably in the same way as NS4B. We show that upon translation of DEN2 polyproteins in a cell-free system, the N-terminus of NS4A is generated by cleavage by the viral nonstructural proteinase NS3 and that processing of DEN2 polyproteins occurs with a preferred, but nonobligatory order.


Improvements in total hip prosthesis implantation technique. A cement-proof seal for the lower medullary cavity and a dihedral self-stabilizing trochanteric osteotomy.
The anchorage of the femoral component of a total hip prosthesis can be improved by the following two refinements to the surgical technique: 1. The femoral medullary cavity is plugged with a bone core taken from the excised femoral head or with a polyethylene bung. The plug prevents the bone cement from being displaced distally into the lower part of the medullary cavity. It contains a small hole through which blood can escape, thus preventing the latter from accumulating in the upper part of the medullary cavity and impairing the bond between the cement and the bone wall of the cavity. 2. The greater trochanter is no longer sliced off with a single cut in one plane but is now removed by dihedral osteotomy, i.e., by making cuts in two planes which intersect each other so as to form a ridge. On compressing this osteotomy by tension band fixation all movement between the fragments is eliminated, including that in the parasagittal plane which results from flexion and extension of the hip. In this manner loosening and pseudarthrosis of the osteotomy are prevented.


Identification of disulfiram as a potential antifungal drug by screening small molecular libraries.
Candida albicans and Candida auris strains are common causative species of Candidiasis. The limited number of antifungal drugs and the current situation of resistance to existing antifungals force us to search for new antifungal alternatives. In this work, primary screening of small molecule libraries (Metabolism Compound Library and Epigenetics Compound Library) consisting of 584 compounds against Candida albicans SC5314 was performed. The dose-response assays, XTT assays, scanning electron microscopy and confocal laser scanning microscopy were used to confirm the antifungal activities of the selected compounds against Candida strains. Through the primary screening, we identified five compounds (U73122, disulfiram, BSK805, BIX01294, and GSKJ4) that inhibited strains growth ≥ 80% for dose-response assays. Disulfiram was identified as the most potent repositionable antifungal drug with 50% growth inhibition detected at a concentration as low as 1 mg/L. The further results showed the antifungal activity of disulfiram against biofilm formation of Candida strains with a 50% minimum inhibitory concentration ranging from 32 to 128 mg/L. Further observations by scanning electron microscopy and confocal laser scanning microscopy confirmed the destruction of biofilm architecture and the change of biofilm morphology after being exposed to disulfiram. The study indicated the potential clinical application of disulfiram as a promising antifungal drug against candidiasis.


Method development and validation for quantification of six bioactive compounds (andrographolide, columbin, piperine, gallic, paracoumaric and oleanolic acids) by HPTLC.
During the spread of pandemic diseases, immunity boosting herbal drugs are taken as a preventive medicine. Kapacurak Kuṭinīr Cūraṇam is a Siddha drug used for flu like viral infections, cold and fever. Developing an analytical method to estimate the content of active phytoconstituents in such antiviral immune boosting drug will be useful in the phyto pharmaceutical industry. A precise, reliable and sensitive ordinary phase high performance thin layer chromatography (HPTLC) method has been developed and validated for identification and simultaneous estimation of six bioactive components namely like andrographolide, columbin, gallic acid, ρ-coumaric acid, piperine and oleanolic acid from any Indian traditional medicine, medicinal plant, drugs and food materials etc. The separation was achieved on silica gel 60F254 TLC plates using toluene: ethyl acetate: formic acid (7:3:0.5, v/v) as mobile phase. The gallic acid, ρ-coumaric acid, piperine markers were estimated using the densitometric scanning in absorption mode at 254 nm. The densitometric scanning was done after derivatization (vanillin-sulphuric acid reagent) at λ=520 nm for andrographolide, columbin and oleanolic acid. The linear regression analysis data for the calibration plots showed a correlation coefficient in the concentration range 1-5 μg per band for the bioactive markers with respect to area. The method was validated for accuracy, precision, limit of detection (LOD), and quantitation of limit (LOQ). Developed method was accurate, precise and fast to ensure the quality of Kapacurak Kuṭinīr Cūraṇam.


Downregulation of Pin1 in human atherosclerosis and its association with vascular smooth muscle cell senescence.
Pin1 is prevalently overexpressed in human cancers and implicated to regulate cell growth and apoptosis. Thus far, however, no role for Pin1 has been described in modulating vascular smooth muscle cell (VSMC) senescence. Immunohistochemistry and Western blotting were used to assess Pin1 protein level in human normal and atherosclerotic tissues. β-galactosidase staining, cumulative population doubling level, telomerase activity, and relative telomere length measurement were used to confirm VSMC senescence. The expressions of Pin1 and other genes involved in this research were analyzed by quantitative reverse-transcription polymerase chain reaction and Western blotting in VSMCs. Apolipoprotein E gene-deleted mice (ApoE-/-) fed a high-fat diet were treated with juglone or 10% ethanol, respectively, for 3 weeks. The extent of atherosclerosis was evaluated by Oil Red O, Masson trichrome staining, and immunohistology. Pin1 protein level decreased in human atherosclerotic tissues and VSMCs, synchronously with increased VSMC senescence. Adenoviral-mediated Pin1 overexpression rescued cellular senescence in atherosclerotic VSMCs, with concurrent down-regulation of P53, p21, growth arrest and DNA-damage-inducible protein 45-alpha (Gadd45a), phosphorylated retinoblastoma (p-pRb), p65 and upregulation of cyclin subfamilies (cyclin B, D, and E), and cyclin-dependent kinase subfamilies (2, 4, and 6), whereas Pin1 knockdown resulted in the converse effects, indicating that VSMC senescence mediated by Pin1 is an integrated response to diverse signals. In vivo data from ApoE-/- mice showed that treatment of juglone led to accelerated atherosclerosis development. Altogether this work supports a role for Pin1 as a vital modulator of VSMC senescence, thereby providing a novel target for regulation and control of atherosclerosis.


Design of Hydrogen Storage Alloys/Nanoporous Metals Hybrid Electrodes for Nickel-Metal Hydride Batteries.
Nickel metal hydride (Ni-MH) batteries have demonstrated key technology advantages for applications in new-energy vehicles, which play an important role in reducing greenhouse gas emissions and the world's dependence on fossil fuels. However, the poor high-rate dischargeability of the negative electrode materials-hydrogen storage alloys (HSAs) limits applications of Ni-MH batteries in high-power fields due to large polarization. Here we design a hybrid electrode by integrating HSAs with a current collector of three-dimensional bicontinuous nanoporous Ni. The electrode shows enhanced high-rate dischargeability with the capacity retention rate reaching 44.6% at a discharge current density of 3000 mA g(-1), which is 2.4 times that of bare HSAs (18.8%). Such a unique hybrid architecture not only enhances charge transfer between nanoporous Ni and HSAs, but also facilitates rapid diffusion of hydrogen atoms in HSAs. The developed HSAs/nanoporous metals hybrid structures exhibit great potential to be candidates as electrodes in high-performance Ni-MH batteries towards applications in new-energy vehicles.


Mesial temporal lobe seizures may activate thermoregulatory mechanisms in humans: an infrared study of facial temperature.
Facial temperature T(f), an important link in cerebral thermoregulation and an ignored index of autonomic activity, was studied in three patients with mesial temporal epilepsy. Sets of infrared images were obtained automatically during seizures and also hourly, round-the-clock. For each patient, scan sets were sorted into: (1) ictal; (2) interictal with negligible motion; and (3) interictal with motion. Comparisons of mean, variance, and trend in each scan set among these groups showed considerable variability. However, significant trends in T(f) occurred (P<0.05) in two-thirds of seizures in one patient, three-fourths in another, and two-ninths in the third, consisting of steady decreases in T(f) beginning at peak seizure intensity R(max) and continuing into the postictus. Maximal ictal T(f) was greater than interictal T(f) (P<0.05), but it negatively correlated with R(max) in two subjects. The ictal changes in T(f), which may have a homeostatic role, suggest activation of thermoregulatory mechanisms during mesial temporal seizures.


Preclinical characterisation of NSAIDs in ultradeformable carriers or conventional topical gels.
We compared in vivo transport and biodistribution of ketoprofen applied on the skin in ultradeformable carriers (Diractin) or a conventional topical gel (Gabrilen) with oral drug (Oruvail); for reference we used in vitro study data. The drug from Gabrilen diffuses into body with low bioavailability (<10%) and limited regio-selectivity (AUC(deep muscle/plasma) approximately 45/0.8 (t=0-8h), reaching maximum concentration in subcutaneous tissues and plasma at similar time (t(max) approximately 3-4h). The apparent drug elimination half-life is then similar to oral ketoprofen (t1/2,a) approximately 2 h). In contrast, Diractin containing ultradeformable carriers (Transfersome vesicles) delivers the drug more efficiently (>50%) and more directly into peripheral muscles (AUC(deep muscle/plasma) approximately 447/0.7 (652/1.4) for t=0-8 (0-24)h; tmax approximately 1 h), arguably in non-diffusive fashion. Ketoprofen from Diractin moreover disappears from body periphery slower (t1/2,a) approximately 4-6 h), owing to sustained drug release from the carriers in target tissue. Final clearance always proceeds via plasma (tmax approximately 4 h). Epicutaneous application of ketoprofen in conventional gels or the carrier-based formulation thus leads to different local accumulations and clearances. Ketoprofen from Diractin achieves more desirable biodistribution and clearance, arguably due to spontaneous carrier-mediated drug transport across the skin, which ensures local and relatively long-lasting drug deposition into peripheral target tissues.


Microcin E492 antibacterial activity: evidence for a TonB-dependent inner membrane permeabilization on Escherichia coli.
The mechanism of action of microcin E492 (MccE492) was investigated for the first time in live bacteria. MccE492 was expressed and purified to homogeneity through an optimized large-scale procedure. Highly purified MccE492 showed potent antibacterial activity at minimal inhibitory concentrations in the range of 0.02-1.2 microM. The microcin bactericidal spectrum of activity was found to be restricted to Enterobacteriaceae and specifically directed against Escherichia and Salmonella species. Isogenic bacteria that possessed mutations in membrane proteins, particularly of the TonB-ExbB-ExbD complex, were assayed. The microcin bactericidal activity was shown to be TonB- and energy-dependent, supporting the hypothesis that the mechanism of action is receptor mediated. In addition, MccE492 depolarized and permeabilized the E. coli cytoplasmic membrane. The membrane depolarization was TonB dependent. From this study, we propose that MccE492 is recognized by iron-siderophore receptors, including FepA, which promote its import across the outer membrane via a TonB- and energy-dependent pathway. MccE492 then inserts into the inner membrane, whereupon the potential becomes destabilized by pore formation. Because cytoplasmic membrane permeabilization of MccE492 occurs beneath the threshold of the bactericidal concentration and does not result in cell lysis, the cytoplasmic membrane is not hypothesized to be the sole target of MccE492.


Anatomy of the 'venous foot pump'.
The gross anatomy of the deep plantar veins has been examined in 14 cadaveric feet in order to assess their function as a 'venous foot pump'. The lateral plantar vein was found to be larger than the medial and is double in its proximal segment. Both plantar veins contain valves which face proximally and have a convoluted and intermuscular course. These features suggest that the deep plantar veins act as a pump that empties during contraction of the intrinsic foot muscles during the stance phase of gait.


Phase 1 Trial of Stereotactic Body Radiation Therapy Neoadjuvant to Radical Prostatectomy for Patients With High-Risk Prostate Cancer.
This study aimed to evaluate the feasibility and safety of prostate stereotactic body radiation therapy (SBRT) neoadjuvant to radical prostatectomy (RP) in a phase 1 trial. The primary endpoint was treatment completion rate without severe acute surgical complications. Secondary endpoints included patient-reported quality of life and physician-reported toxicities. Patients with nonmetastatic high-risk or locally advanced prostate cancer received 24 Gy in 3 fractions to the prostate and seminal vesicles over 5 days, completed 2 weeks before RP. Patients with pN1 disease were treated after multidisciplinary discussion and shared decision making. Patient-reported quality of life (International Prostate Symptom Score and Expanded Prostate Cancer Index Composite 26-item version questionnaires) and physician-reported toxicity (Common Terminology Criteria for Adverse Events, version 4.03) were assessed before SBRT, immediately before surgery, and at 3-month intervals for 1 year. Twelve patients were enrolled, and 11 completed treatment (1 patient had advanced disease on prostate-specific membrane antigen positron emission tomography after enrollment but before treatment). There were no significant surgical complications. After RP, 2 patients underwent additional radiation therapy to nodes with androgen suppression for pN1 disease. Median follow-up after completion of treatment was 20.1 months, with 9 of 11 patients having a follow-up period of >12 months. Two patients had biochemical recurrence (prostate-specific antigen ≥0.05) within the first 12 months, with an additional 2 patients found to have biochemical recurrence after the 12-month period. The highest Common Terminology Criteria for Adverse Events genitourinary grades were 0, 1, 2, and 3 (n = 1, 4, 4, and 2, respectively), and the highest gastrointestinal grades were 0, 1, and 2 (n = 9, 1, and 1, respectively). At 12 months, incontinence was the only grade ≥2 toxicity. One and 2 of 9 patients had grade 2 and 3 incontinence, respectively. On the Expanded Prostate Cancer Index Composite (26-item version), the mean/median changes in scores from baseline to 12 months were -32.8/-31.1 for urinary incontinence, -1.6/-6.2 for urinary irritative/obstructive, -2.1/0 for bowel, -34.4/-37.5 for sexual function, and -10.6/-2.5 for hormonal. The mean/median change in International Prostate Symptom Score from baseline to 12 months was 0.5/0.5. RP after neoadjuvant SBRT appears to be feasible and safe at the dose tested. The severity of urinary incontinence may be higher than RP alone.


Retinal vasculitis associated with HLA DR4. Brief definitive report.
Inflammation of retinal blood vessels may be associated with a variety of systemic immune diseases. Despite the fact that a number of immunological abnormalities have been reported in patients with retinal vasculitis (RV), previous studies have failed to demonstrate an immunogenetic predisposition to this disease. HLA A, B, and DR locus typing of 25 patients (14 females) with well-characterized RV revealed an increased incidence of HLA DR4 (corrected p value = 0.04, relative risk = 3.5). The HLA DR4 antigen was increased in patients with both central and peripheral RV as well as in patients with idiopathic disease (14 patients) and in those with Behcet's syndrome (8 patients). The results of this study indicate that immune response genes may be involved in the pathogenesis of RV.


Combined liver-kidney transplantation with delayed kidney implantation: A case report.
In rare cases, patients require a combined liver-kidney transplant. However, the peri- and postoperative care of liver transplant recipients differs from kidney transplant recipients, which can lead to conflicts of interest. In the case of poor coagulation status and/or instable hemodynamics of the patient, liver transplantation, followed by delayed kidney transplantation can lead to better postoperative recovery. In our case report, we present a 48-year old man with Alagille syndrome and IgA nephropathy with bilirubin-associated acute kidney injury, causing him to develop both end-stage liver and kidney disease. He underwent a combined liver-kidney transplant as the first patient in the Netherlands, in which the donor kidney was transplanted one day after the liver transplantation. One-year post-transplant patient is in good clinical condition, with normal liver function and an eGFR of 57 ml/min. Combined liver-kidney transplantation with delayed kidney implantation in a medical center with no previous experience with this technique is feasible and safe. This could be better for both the patient and the kidney graft.


Prevalence of exclusive breastfeeding practice and its association with maternal employment in Ethiopia: a systematic review and meta-analysis.
Exclusive breastfeeding is defined as the practice of providing only breast milk for an infant for the first 6 months of life without the addition of any other food or water, except for vitamins, mineral supplements, and medicines. Findings are inconsistent regarding the prevalence of exclusive breastfeeding in Ethiopia. Full-time maternal employment is an important factor contributing to the low rates of practice of exclusive breastfeeding. Empowering women to exclusively breastfeed, by enacting 6 months' mandatory paid maternity leave can increase the rate of exclusive breastfeeding in the first 6 months of life up to 50%. The purpose of this review was to estimate the pooled prevalence of exclusive breastfeeding and its association with full-time maternal employment in the first 6 months of life for infants in the context of Ethiopia. The Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guideline was used in this systematic review and meta-analysis. All observational studies reporting the prevalence of exclusive breastfeeding and its association with maternal employment in Ethiopia were considered. The search was conducted from 6 November 2020 to 31 December 2020 and all papers published in the English language from 1 January 2015 to 31 December 2020 were included in this review. Forty-five studies were included in the final analysis after reviewing 751 studies in this meta-analysis yielding the pooled prevalence of EBF 60.42% (95% CI 55.81, 65.02) at 6 months in Ethiopia. Those full-time employed mothers in the first 6 months were 57% less likely to practice exclusive breastfeeding in comparison to mothers not in paid employment in Ethiopia (OR 0.43; 95% CI 0.31, 0.61). Full-time maternal employment was negatively associated with the practice of exclusive breastfeeding in comparison to unemployed mothers. The prevalence of exclusive breastfeeding in Ethiopia is low in comparison to the global recommendation. The Ethiopian government should implement policies that empower women. The governmental and non-governmental organizations should create a conducive environment for mothers to practice exclusive breastfeeding in the workplace.


Conformation, orientation, and adsorption kinetics of dermaseptin B2 onto synthetic supports at aqueous/solid interface.
The antimicrobial activity of cationic amphipathic peptides is due mainly to the adsorption of peptides onto target membranes, which can be modulated by such physicochemical parameters as charge and hydrophobicity. We investigated the structure of dermaseptin B2 (Drs B2) at the aqueous/synthetic solid support interface and its adsorption kinetics using attenuated total reflection Fourier transform infrared spectroscopy and surface plasmon resonance. We determined the conformation and affinity of Drs B2 adsorbed onto negatively charged (silica or dextran) and hydrophobic supports. Synthetic supports of differing hydrophobicity were obtained by modifying silica or gold with omega-functionalized alkylsilanes (bromo, vinyl, phenyl, methyl) or alkylthiols. The peptide molecules adsorbed onto negatively charged supports mostly had a beta-type conformation. In contrast, a monolayer of Drs B2, mainly in the alpha-helical conformation, was adsorbed irreversibly onto the hydrophobic synthetic supports. The conformational changes during formation of the adsorbed monolayer were monitored by two-dimensional Fourier transform infrared spectroscopy correlation; they showed the influence of peptide-peptide interactions on alpha-helix folding on the most hydrophobic support. The orientation of the alpha-helical Drs B2 with respect to the hydrophobic support was determined by polarized attenuated total reflection; it was around 15 +/- 5 degrees. This orientation was confirmed and illustrated by a molecular dynamics study. These combined data demonstrate that specific chemical environments influence the structure of Drs B2, which could explain the many functions of antimicrobial peptides.


The melanocortin ACTH 1-39 promotes protection of oligodendrocytes by astroglia.
Damage to myelin and oligodendroglia (OL) in multiple sclerosis (MS) results from a wide array of mechanisms including excitotoxicity, neuroinflammation and oxidative stress. We previously showed that ACTH 1-39, a melanocortin, protects OL in mixed glial cultures and enriched OL cultures, inhibiting OL death induced by staurosporine, ionotropic glutamate receptors, quinolinic acid or reactive oxygen species (ROS), but not nitric oxide (NO) or kynurenic acid. OL express melanocortin receptor 4 (MC4R), suggesting a direct protective effect of ACTH 1-39 on OL. However, these results do not rule out the possibility that astroglia (AS) or microglia (MG) also play roles in protection. To investigate this possibility, we prepared conditioned medium (CM) from AS and MG treated with ACTH, then assessed the protective effects of the CM on OL. CM from AS treated with ACTH protected OL from glutamate, NMDA, AMPA, quinolinic acid and ROS but not from kainate, staurosporine, NO or kynurenic acid. CM from MG treated with ACTH did not protect from any of these molecules, nor did CM from AS or MG not treated with ACTH. While protection of OL by ACTH from several toxic molecules involves direct effects on OL, ACTH can also stimulate AS to produce mediators that protect against some molecules but not others. Thus the cellular mechanisms underlying the protective effects of ACTH for OL are complex, varying with the toxic molecules.


Modeling air pollution by integrating ANFIS and metaheuristic algorithms.
Air pollution is increasing for many reasons, such as the crowding of cities, the failure of planning to consider the benefit of society and nature, and the non-implementation of environmental legislation. In the recent era, the impacts of air pollution on human health and the ecosystem have become a primary global concern. Thus, the prediction of air pollution is a crucial issue. ANFIS is an artificial intelligence technique consisting of artificial neural networks and fuzzy inference systems, and it is widely used in estimating studies. To obtain effective results with ANFIS, the training process, which includes optimizing its premise and consequent parameters, is very important. In this study, ANFIS training has been performed using three popular metaheuristic methods: Genetic Algorithm (GA), Particle Swarm Optimization (PSO), and Differential Evolution (DE) for modeling air pollution. Various air pollution parameters which are particular matters: PM2.5 and PM10, sulfur dioxide (SO2), ozone (O3), nitrogen dioxide (NO2), carbon monoxide (CO), and several meteorological parameters such as wind speed, wind gust, temperature, pressure, and humidity were utilized. Daily air pollution predictions in Istanbul were obtained using these particular matters and parameters via trained ANFIS approaches with metaheuristics. The prediction results from GA, PSO, and DE-trained ANFIS were compared with classical ANFIS results. In conclusion, it can be said that the trained ANFIS approaches are more successful than classical ANFIS for modeling and predicting air pollution.


Improved method for cytogenetic studies of solid tumors.
Solid human tumors were dissociated with collagenase, cultured for 16-48 hours, and harvested for cytogenetic preparation. Of the 19 tumors used, 14 showed sufficient numbers of metaphases to be useful for chromosome analysis.


New evidence for the essential role of arginine residues in anion transport across the red blood cell membrane.
2,3-Butanedione, in the dark and in the presence of borate, reacts rapidly to inactivate the sulfate equilibrium exchange across the human red cell membrane. Reactivation occurs spontaneously after the removal of borate, indicating the reaction of butanedione with essential arginine residues. The inactivation of the transport system depends on the concentration of the reagent, on the incubation time and exhibits pseudo-first-order kinetics. Chloride ions are able to protect the transport system against inactivation with the reagent. This would suggest the participation of the modified residue in the substrate binding site. When the transport system is inhibited to 50-60% by butanedione, the transporter can still bind covalently the anion transport inhibitor 2H2DIDS up to 85 +/- 12% of its total binding capacity. 3H2DIDS concentration was either 3.15, 10 or 20 microM. Modification of resealed ghosts with 50 mM butanedione under conditions where the transport system is to more than 75% inhibited, causes a reduction of only about 30% of the reversibly bound 3H2DIDS.


Transvaginal laparoscopic donor nephrectomy.
Laparoscopic donor nephrectomy (LDN) has numerous advantages over open donor nephrectomy. The cosmetic issues and pain that arise due to the 5 to 6-cm incisions on the abdominal wall in LDN have led to transvaginal laparoscopic donor nephrectomy (TVLDN). Between May and August 2012, we performed seven donor nephrectomies via a transvaginal approach. The mean age of the donors was 53.0 ± 9.52 years. The mean operative time was 97.29 ± 39.47 minutes and mean warm ischemia time, 220.71 ± 55.49 seconds. Donors were mobilized, began oral intake at 8 hours postoperative, and were all discharged within the first 24 hours. Except one dose of analgesic applied immediately after the operation, no additional medication was required. No infectious complications were encountered in any recipient. TVLDN may be a good alternative for female donors. Compared with LDN, TVLDN has benefits of less postoperative pain, faster recovery, shorter hospital stay, and excellent cosmetic results.


Effects of long-term infusion of growth hormone (GH)-releasing factor on pulsatile GH secretion in the male rat.
The effects of long-term infusion of GRF on pulsatile GH secretion were investigated in conscious, freely-moving male rats. GH-releasing factor (GRF) was intravenously administered for 14 days using an osmotic minipump. Six-hour iv infusion of 0.5 microgram/h GRF significantly increased mean GH concentration in the conscious rats. In contrast, 14-day iv infusion of 0.5 microgram/h GRF significantly inhibited pulsatile GH secretion. The pituitary GH content was significantly increased by the 14-day treatment. The GH response to bolus injection of 1 microgram GRF under urethane anesthesia was enhanced by the 14-day infusion of GRF. Pretreatment with somatostatin (SS) antiserum did not restore the inhibited GH secretion in rats treated with 14-day infusion of GRF. These results suggest that neither pituitary GH depletion nor high somatostatin tone is involved in the GRF-induced GH inhibition.


The effect of base of support stability on shoulder muscle activity during closed kinematic chain exercises.
A total of thirty eligible subjects (17 female and 13 male, age = 22.26 ± 0.99 years, height = 170.96 ± 8.42 cm, weight = 61.63 ± 9.92 kg) were tested in six different randomly ordered positions. Surface Electromyography (EMG) was recorded from the upper trapezius (UT), lower trapezius (LT), serratus anterior (SA), long head of the biceps (LB), teres major (TM) and posterior deltoid (PD) muscles in the dominant shoulder in 6 different closed kinetic chain (CKC) positions. To investigate changes in muscular activity of the shoulder muscles at different base of support stability levels. Muscle activity was greater in the most stable position for all muscles except UT (P < 0.01). Shoulder muscle activity did not increase in parallel with a reduction in base of support stability in the present study.


Health-care utilization and respiratory morbidities in preterm infants with pulmonary hypertension.
To assess health-care utilization and risk of respiratory morbidities in preterm infants with bronchopulmonary dysplasia (BPD) and pulmonary hypertension (PH). Retrospective data were obtained from subjects (n=109) attending a BPD clinic. Subjects were stratified by the presence or absence of PH before and after 2 months of age. Analytic methods included t-tests, χ(2) tests and regression. Subjects with BPD and PH present after 2 months of age were hospitalized for 2.2 months longer than those without PH (P=0.02). These subjects were 4.5 times more likely to receive home supplemental oxygen or mechanical ventilation (P=0.03). No difference in the risk of respiratory morbidities after initial hospital discharge was seen with PH. PH in preterm infants is associated with longer initial hospitalizations and a higher likelihood of requiring home respiratory support. This has implications for counseling families and reducing the medical, psychosocial, and economic burden of BPD and PH.


Cervical cytopathology image refocusing via multi-scale attention features and domain normalization.
Cervical cytopathology image refocusing is important for addressing the problem of defocus blur in whole slide images. However, most of current deblurring methods are developed for global motion blur instead of local defocus blur and need a lot of supervised re-training for unseen domains. In this paper, we propose a refocusing method for cervical cytopathology images via multi-scale attention features and domain normalization. Our method consists of a domain normalization net (DNN) and a refocusing net (RFN). In DNN, we adopt registration-free cycle scheme for normalizing the unseen unsupervised domains into the seen supervised domain and introduce gray mask loss and hue-encoding mask loss to ensure the consistency of cell structure and basic hue. In RFN, combining the locality and sparseness characteristics of defocus blur, we design a multi-scale refocusing network to enhance the reconstruction of cell nucleus and cytoplasm, and introduce defocus intensity estimation mask to strengthen the reconstruction of local blur. We integrate hybrid learning strategy on the supervised and unsupervised domains to make RFN achieving well refocusing on the unsupervised domain. We build a cervical cytopathology image refocusing dataset and conduct extensive experiments to demonstrate the superiority of our method compared with current deblurring state-of-the-art models. Furthermore, we prove that the refocused images help improve the performance of subsequent high-level analysis tasks. We release the refocusing dataset and source codes to promote the development of this field.


Taurine Pretreatment Prevents Isoflurane-Induced Cognitive Impairment by Inhibiting ER Stress-Mediated Activation of Apoptosis Pathways in the Hippocampus in Aged Rats.
Isoflurane, a commonly used inhalation anesthetic, may induce neurocognitive deficits, especially in elderly patients after surgery. Recent study demonstrated that isoflurane caused endoplasmic reticulum (ER) stress and subsequent neuronal apoptosis in the brain, contributing to cognitive deficits. Taurine, a major intracellular free amino acid, has been shown to inhibit ER stress and neuronal apoptosis in several neurological disorders. Here, we examined whether taurine can prevent isoflurane-induced ER stress and cognitive impairment in aged rats. Thirty minutes prior to a 4-h 1.3 % isoflurane exposure, aged rats were treated with vehicle or taurine at low, middle and high doses. Aged rats without any treatment served as control. The brains were harvested 6 h after isoflurane exposure for molecular measurements, and behavioral study was performed 2 weeks later. Compared with control, isoflurane increased expression of hippocampal ER stress biomarkers including glucose-regulated protein 78, phosphorylated (P-) inositol-requiring enzyme 1, P-eukaryotic initiation factor 2-α (EIF2α), activating transcription factor 4 (ATF-4), cleaved ATF-6 and C/EBP homologous protein, along with activation of apoptosis pathways as indicated by decreased B cell lymphoma 2 (BCL-2)/BCL2-associated X protein, increased expressions of cytochrome-c and cleaved caspase-3. Taurine pretreatment dose-dependently inhibited isoflurane-induced increase in expression of ER stress biomarkers except for P-EIF2α and ATF-4, and reversed isoflurane-induced changes in apoptosis-related proteins. Moreover, isoflurane caused spatial working memory deficits in aged rats, which were prevented by taurine pretreatment. The results indicate that taurine pretreatment prevents anesthetic isoflurane-induced cognitive impairment by inhibiting ER stress-mediated activation of apoptosis pathways in the hippocampus in aged rats.


[Expression and significance of human leukocyte antigen DR and intercellular adhesion molecule-1 in normal and pathological corneas].
To study the expression of human leukocyte antigen DR (HLA-DR) and intercellular adhesion molecule-1 (ICAM-1) in normal and pathological corneas and their significance in corneal inflammation and rejection after keratoplasty. The expression of HLA-DR and ICAM-1 were detected in 10 normal corneas and 48 pathological corneas by using immunohistochemical techniques. HLA-DR and ICAM-1 were not expressed or only slightly expressed in normal corneas and markedly expressed in pathological corneas, especially in inflammatory corneas. In general, HLA-DR and ICAM-1 were co-expressed in similar regions. The expression rates of HLA-DR and ICAM-1 in the 7 cases of keratoplasty rejection were higher than that in the 21 cases of non-rejection. The overexpression of HLA-DR and ICAM-1 is related to corneal inflammation and can promote the occurrence of keratoplasty rejection.


A critical assessment of visual identification of marine microplastic using Raman spectroscopy for analysis improvement.
Identification and characterisation of microplastic (MP) is a necessary step to evaluate their concentrations, chemical composition and interactions with biota. MP ≥10μm diameter filtered from below the sea surface in the European and subtropical North Atlantic were simultaneously identified by visual microscopy and Raman micro-spectroscopy. Visually identified particles below 100μm had a significantly lower percentage confirmed by Raman than larger ones indicating that visual identification alone is inappropriate for studies on small microplastics. Sixty-eight percent of visually counted MP (n=1279) were spectroscopically confirmed being plastic. The percentage varied with type, colour and size of the MP. Fibres had a higher success rate (75%) than particles (64%). We tested Raman micro-spectroscopy applicability for MP identification with respect to varying chemical composition (additives), degradation state and organic matter coating. Partially UV-degraded post-consumer plastics provided identifiable Raman spectra for polymers most common among marine MP, i.e. polyethylene and polypropylene.


Elucidation of the fragmentation pathways of a complex 3,7-O-glycosyl flavonol by CID, HCD, and PQD on an LTQ-Orbitrap Velos Pro hybrid mass spectrometer.
The present study was designed to systematically investigate the ESI-MS(n) behavior of a complex 3, 7-O-glycosyl flavonol, kaempferol 3-O-α-L-[2,3-di-O-β-D-(6-E-p-coumaroyl)glucopyranosyl]-rhamnopyranosyl-7-O-α-L-rhamnopyranoside (KO) isolated from Epimedium wushanense, and to address the elimination priority among different glycosylation sites and different sugars/substituents. The direct-infusion ESI-MS(n) experiment of KO was performed on a hybrid LTQ-Orbitrap Velos Pro mass spectrometer in both negative and positive ion modes by three different fragmentation mechanisms (CID, HCD, and PQD). The CID, HCD, and PQD analyses of KO exhibited remarkable discrimination in respect of the scan range, richness, and distribution of product ions through the entire spectra. KO experienced different fragmentation pathways between two ionization modes: the negative mode CID of KO eliminated the glycosyl portions (priority: 7-sugar > 3-substituent and terminal substituents > inner sugar) and produced aglycone product ions at m/z 284.03/285.04; however, abundant sodium-adduct B(3)2 together with subsequent (i,j)X(3)0 cleavages were found characteristic for the positive mode CID-MS(n). The fragmentation pathways by CID for KO were proposed by analyzing the high accuracy ESI-MS(n) data. Complementary structural information of KO regarding the aglycone and glycosyl portions was obtained by analyzing the ESI-MS(n) data in both ionization modes. In conclusion, the LTQ-Orbitrap method facilitates highly reliable qualitative analysis of bioactive flavonoids with three alternative fragmentation modes.


Redox-responsive polyurethane-polyurea nanoparticles targeting to aortic endothelium and atherosclerosis.
Aortic endothelial cell dysfunction is an early trigger of atherosclerosis, the major cause of the cardiovascular disease (CVD). Nanomedicines targeting vascular endothelium and lesions hold great promise as therapeutic solutions to vascular disorders. This study investigates the vascular delivery efficacy of polyurethane-polyurea nanocapsules (Puua-NCs) with pH-synchronized shell cationization and redox-triggered release. Fluorescent lipophilic dye DiI was encapsulated into Puua-NCs of variable sizes and concentrations. In vitro cellular uptake studies with human aortic endothelial cells showed that these Puua-NCs were taken up by cells in a dose-dependent manner. In apolipoprotein E-deficient mice fed a Western diet, a model of atherosclerosis, circulating Puua-NCs were stable and accumulated in aortic endothelium and lesions within 24 hours after intravenous administration. Treatment with thiol-reducing and oxidizing reagents disrupted the disulfide bonds on the surface of internalized NCs, triggering disassembly and intracellular cargo release. Ultimately, Puua-NCs are a potential redox-controllable cardiovascular drug delivery system.


Preservation and phallotoxin-staining of the microfilament system in Amoeba proteus.
The spatial organization of the microfilament system as the main component of the cytoskeleton in Amoeba proteus was preserved by a glutaraldehyde-lysine-fixation and visualized with fluorescent phallotoxins (NBD- phallacidin , R-phalloidin). Results obtained by means of this method coincide exactly with observations gained from immunocytochemical, ultrastructural and molecular cytochemical studies, i.e., the microfilament system is mainly displayed beneath the cell membrane, at the hyalo - granuloplasmic border and around the cell nucleus. The preparation procedure employed is suitable for the rapid demonstration of cytoplasmic microfilaments in cells difficult to preserve by chemical fixation.


Correction: Preferring more e-cigarette flavors is associated with e-cigarette use frequency among adolescents but not adults.
[This corrects the article DOI: 10.1371/journal.pone.0189015.].


Neural shifts in alpha rhythm's dual functioning during empathy maturation.
Empathy is a social-cognitive process that operates by relying mainly on the suppression of the cortical alpha rhythm. This phenomenon has been evidenced in dozens of electrophysiological studies targeting adult human subjects. Yet, recent neurodevelopmental studies indicated that at a younger age, empathy involves reversed brain responses (e.g., alpha enhancement patterns). In this multimodal study, we capture neural activity at the alpha range, and hemodynamic response and target subjects at approximately 20 years old as a unique time window in development that allows investigating both low-alpha suppression and high-alpha enhancement. We aim to further investigate the functional role of low-alpha power suppression and high-alpha power enhancement during empathy development. Brain data from 40 healthy individuals were recorded in two consecutive sessions of magnetoencephalography (MEG) and functional magnetic resonance imaging (fMRI) while subjects perceived vicarious physical pain or no pain. MEG revealed that the alpha pattern shift during empathy happens in an all-or-none pattern: power enhancement before 18 and suppression after 18 years of age. Additionally, MEG and fMRI highlight a correspondence between high-alpha power increase and blood-oxygen-level-dependent (BOLD) decrease before 18, but low-alpha power decrease and BOLD increase after 18. Importantly, this neurodevelopmental transition was not revealed by four other measures: self-reported (a) ratings of the task stimuli, (b) ratings of naturalistic vignettes of vicarious pain, (c) trait empathy, or neural data from (d) a control neuroimaging task. Findings suggest that at the critical age of around 18, empathy is underpinned by an all-or-none transition from high-alpha power enhancement and functional inhibition to low-alpha power suppression and functional activation in particular brain regions, possibly indicating a marker of maturation in empathic ability. This work advances a recent neurodevelopmental line of studies and provides insight into the functional maturation of empathy at the coming of age.


Preparation of chain-end clickable recombinant protein and its bio-orthogonal modification.
Introducing unique functional group into protein is an attractive approach for site-selective protein modification applications. In this report, we systemically investigated four site-selective strategies to introduce azide functionality into recombinant thrombomodulin (TM456), via direct recombinant expression with unnatural amino acid, chemical, and enzymatic modification for its bio-orthogonal modification application. First, a straightforward recombinant method to express TM456 with azide functionality near C-terminus by replacing methionine with azidohomoanlanine from methionine auxotroph Escherichia coli cell was investigated. Next, a sortase-mediated ligation (SML) method to incorporate azide functionality into the C-terminus of recombinant TM456 was demonstrated. The third is to add azide functionality to the N-terminal amine of recombinant TM456via amidation chemistry, and the fourth is tyrosine selective three-component Mannich reaction to introduce azide functionality to recombinant TM456. Overall, SML of recombinant protein affords the highest overall yield for incorporating azide functionality into the C-terminus recombinant TM456 since the key protein expression step uses natural amino acids. Also, single site modification facilitates the highest TM456 activity.


Listeriosis infection of an abdominal aortic aneurysm in a diabetic patient.
A rare case of an abdominal aortic aneurysm (AAA) infected by Listeria monocytogenes in a 72-year-old male diabetic farmer, is reported. Our patient had a history of a recent pneumonia that could have been caused by Listeria too. Aneurysm infection was manifested by fever and abdominal and back pain, which prompted investigation with CT scanning that revealed a 4.9 cm AAA with typical signs of infection. He underwent urgent AAA repair with aortobifemoral bypass grafting and had an uneventful course. Aneurysm content microbiology revealed Listeria monocytogenes and following a 9-week course of antibiotics our patient remains asymptomatic 11 months later.


Application of real-time confocal microscopy to intracellular calcium ion dynamics in rat arterioles.
The regulation of cytosolic Ca(2+) homeostasis is essential for cells, including vascular smooth muscle cells. Arterial tone, which underlies the maintenance of peripheral resistance in the circulation, is a major contributor to the control of blood pressure. Confocal microscopy was employed to study the alteration in intracellular calcium ion concentration ([Ca(2+)](i)) in arterioles (external diameters <100 microm) with respect to selected modifying reagents. 5-Hydroxytryptamine (1 microM), ATP (10 microM), and endothelin 1-3 (5 nM) elicited an increase in [Ca(2+)](i) in most arteriole smooth muscle cells. The [Ca(2+)](i) increase sometimes propagated in an intercellular manner. When noradrenaline (10 microM) was used as a stimulant, [Ca(2+)](i) increase was observed only in a portion of the smooth muscle cells. It was also noted that the reaction of these cells with respect to ATP is different between testis and brain arterioles; the [Ca(2+)](i) increase in testicular arterioles is dependent on Ca(2+) influx from extracellular space, whereas in cerebral arterioles it plays a role in both the influx of extracellular Ca(2+) and the release of Ca(2+) from intracellular stores (i.e., sarco/endoplasmic reticulum). These results indicate that arterioles in different tissues may differ greatly in their responses. Real-time confocal microscopy was found to be a useful tool for investigating the structural and functional changes in living tissues.


Developmental defects of enamel in primary teeth: prevalence and associated factors.
Studies on the prevalence of enamel defects in the primary dentition as a whole are scarce, as most investigations examine specific population groups. The aim of this study was to evaluate the prevalence of enamel defects in primary teeth and determine whether prematurity, birthweight, and socio-demographic variables are associated with such defects. A cross-sectional study was carried out with 381 children aged 3-5 years. Data were collected through clinical examinations and interviews with parents during the National Immunisation Day. The diagnosis of enamel defects was performed using the Developmental Defects of Enamel (DDE) Index. Through interviews, information was collected on socio-demographic aspects, pregnancy, birthweight, prematurity, and breastfeeding. Statistical analysis was performed using the SPSS program for Windows and involved descriptive analysis, Fisher's exact test, the chi-square test, and Poisson regression. The prevalence of developmental defects of enamel was 29.9%. Demarcated opacity was the most frequent type of defect. Children with a history of very low birthweight had a greater prevalence of enamels defects (PR, 2.7; 95% CI, 1.66-4.61). Prematurity and socio-demographic variables were not associated with enamel defects. Children with a history of very low birthweight had a greater frequency of enamel defects in primary teeth.


Deoxyglucose demonstration of in-utero hearing in the guinea pig foetus.
2-Deoxyglucose (2-[14C]DG) autoradiography was used to demonstrate central auditory function in the foetal guinea pig in-utero. The major advantage of this approach is that the experiment is carried out with the foetuses within the intact amniotic sac. Using pure tone stimuli at around 100 dB SPL, isofrequency bands of elevated metabolic activity were observed in the inferior colliculus of the majority of foetuses within the last 10 days of gestation.


Pre-operative stenting and complications following pancreatoduodenectomy for pancreatic cancer: an analysis of the ACS-NSQIP registry.
Historically, pre-operative biliary stenting has been associated with higher infectious complication rates following pancreatoduodenectomy. However, alleviation of biliary obstruction is necessary for consideration of pre-operative chemotherapy, which may improve disease-free survival, or for mitigation of symptoms while awaiting surgery. Our aim is to compare contemporary post-operative complication risk among patients with pre-operative endoscopic retrograde cholangiopancreatography (ERCP) stenting compared to those without. Patients who underwent a pancreatoduodenectomy for pancreatic cancer with biliary obstruction within the ACS-NSQIP registry from 2014 to 2017 were identified. The primary outcome was to compare the risk of 30-day complication (composite outcome) between patients with and without pre-operative ERCP stenting. Propensity score matching was used to ensure balanced baseline characteristics and log-binomial regression models were used to estimate risk ratios for overall perioperative complication between groups. From 6073 patients with obstructive jaundice undergoing pancreatoduodenectomy for pancreatic cancer, 92% (5564) were eligible for the study. After performing a propensity score matching on 20 baseline characteristics, 952 patients without stenting were matched to up to four patients who received pre-operative ERCP stenting (n = 3467) for a matched cohort of 4419. A total of 1901 (55%) patients with pre-operative ERCP stenting experienced a post-operative complication compared to 501 (53%) patients without stenting (risk ratio 1.04, 95% CI 0.97-1.11, p = 0.23). Pre-operative ERCP stenting was not associated with an increased risk of post-operative complication in patients undergoing pancreatoduodenectomy with obstructive jaundice. Biliary stenting may be safely considered for symptom relief and to potentially facilitate pre-operative chemotherapy for pancreatic cancer.


Computational study on 2D three-phase lag bioheat model during cryosurgery using RBF meshfree method.
Biological tissue has a multidimensional and non-homogeneous inner structure by nature. The temperature distribution and freezing front locations in biological tissue are crucial to optimizing the damage to tissue during cryosurgery. There is a need for a good mathematical model and effective simulation techniques to predict the effectiveness of the therapy. The present study concerns the numerical study of phase change phenomena during cryosurgery using the three-phase lag (TPL) bioheat model in arbitrary soft tissue domains, i.e., circular (Γ1), ameba-like (Γ2), and multiconnected (Γ3). We employ the effective heat capacity formulation to solve the nonlinear governing equation. The Gaussian radial basis function and Crank-Nicolson finite difference approximation are applied for spatial and time derivatives, respectively. Using the present algorithm, we study the impact of phase lag (τv) due to thermal displacement involved in the TPL model on phase change interface position and thermal distribution in all three domains. The obtained results may be beneficial in the field of oncology.


Tunable photoluminescence and energy transfer of Eu3+,Ho3+-doped Ca0.05Y1.93-xO2 nanophosphors for warm white LEDs applications.
A series of Eu3+ ions doped Ca0.05Y1.93-xO3:0.02Ho3+ (CYO:Ho3+,xEu3+) nanophosphors having multicolour tuneability have been synthesised by following a simplistic solution combustion approach. The synthesised samples have been characterised by employing X-ray diffraction (XRD), Transmission electron microscope (TEM), and Fourier transforms infrared spectroscopy (FTIR). The optical properties have been engrossed by UV-visible and photoluminescent excitation and emission spectra, and decay lifetimes measurements. The characteristic emission, which occurs due to the f-f transition of Ho3+ and Eu3+ has been observed in emission spectra with excitation of 448 nm. By adjusting the doping ratio of Ho3+/Eu3+, the as-synthesized nanophosphor accomplishes multicolour tunability from green-yellow to red. Emission spectra and decay lifetime curve recommend dipole-dipole interaction causes energy transfer from Ho3+ → Eu3+. The energy transfer process from Ho3+ to Eu3+ has been confirmed through electric dipole-dipole interaction with critical distance 15.146 Å. Moreover, temperature dependent emission spectra show the high thermal stability with an activation energy ⁓ 0.21 eV, with the quantum efficiency of 83.6%. CIE coordinate illustrates that the singly doped Ho3+ and Eu3+ lie in the green and red region, respectively, while the as-synthesized CYO:Ho3+,xEu3+shows tunability from green to red with low CCT and high colour purity values. Hence, the CYO:Ho3+,xEu3+nanophosphor may be a near-UV excited multicolour colour-tunable pertinent candidate with potential prospects for multicolour- display and near-ultraviolet lighting applications.


Life-history variation in a hybrid species complex ofDaphnia.
Life-history variation was examined among members of theDaphnia longispina group, which consists ofD. galeata, D. hyalina, D. cucullata, and hybrids. Factorial experiments were conducted at two temperatures (14° and 20° C) and two food concentrations (0.2 and 1.0 mg Cl-1). Differences in life-history features (size at maturity, age at first reproduction, size of first clutch, offspring size in first clutch) under the different environmental conditions were assessed among eightDaphnia clones, which represented members of this species complex. Significant differences between parentals and hybrids for most life-history features were observed under various treatments; generally, hybrid clones showed intermediate life-history traits when compared with parentals. When comparisons were made among clones within a given species (i.e.D. galeata, D. galeata xcucullata, D. cucullata), clonal differences were also noted for certain life-history traits. The data are discussed with reference to the formation and maintenance of hybrid species complexes in nature.


Involved site radiation therapy in stage I-III nasopharyngeal carcinoma with limited lymph node burden (ISRT-NPC) or elective region irradiation: a study protocol for a multicenter non-inferiority randomized controlled phase III clinical trial.
Current radiotherapy guidelines and consensus statements uniformly recommend elective region irradiation (ERI) as the standard strategy for nasopharyngeal carcinoma (NPC). However, given the scarcity of skip-metastasis, the improved assessment accuracy of nodal involvement, and the striking advancements in chemotherapy for NPC, a one-fits-all delineation scheme for clinical target volumes of the nodal region (CTVn) may not be appropriate anymore, and modifications of the CTVn delineation strategy may be warranted. Involved site irradiation (ISI) covering merely the initially involved nodal site and potential extranodal extension has been confirmed to be as effective as ERI with decreased radiation-related toxicities in some malignancies, but has not yet been investigated in NPC. This study aims to compare the regional control, survival outcomes, radiation-related toxicities, and quality of life (QoL) of ISI with conventional ERI in NPC patients with a limited nodal burden. ISRT-NPC is a prospective, multicenter, open-label, noninferiority, phase III randomized controlled trial. A total of 414 patients will be randomly assigned in a 1:1 ratio to receive ISI or ERI. Randomization will be stratified by institution scale and N stage. Generally, in the ISI group, the high-risk CTV1 (dose: 60 Gy) includes a 1-cm expansion of the positive LN as well as the VIIa and the retrostyloid space above the bilateral transverse process of the atlantoaxial spine (C1), regardless of N status. The low-risk CTV2 (dose: 50 Gy) covers the cervical nodal region with a 3-cm caudal expansion below the transverse process of C1 for N0 disease and a 3-cm expansion below the positive LN for positive LNs. The results of this trial are expected to confirm that ISI is a non-inferior strategy to ERI in stage I-III patients with low LN burden, enabling the minimization of treatment-related toxicity and improvement of long-term QoL without compromising regional control. ClinicalTrails.gov, NCT05145660. Registered December 6, 2021.


Post-operative intravenous fluid administration for infant cleft surgery: An observational study.
To investigate post-operative intravenous fluid administration and length of stay in a single site cleft centre. Previous publications have linked increased length of stay following primary cleft surgery to the administration of intravenous fluids post-operatively. One hundred and ten primary cleft operations were conducted from May 2015 to April 2016 on non-syndromic infants. At West Midlands Cleft Centre, there are three cleft surgeons and 20 paediatric anaesthetists. This observational study compares classification of cleft type and surgical procedure with intravenous fluid administration, time taken to tolerate oral feeding, and length of stay. Cleft lip repair infants had the shortest length of stay in hospital, 25 h 8 min (median) and 33% had intravenous fluids. The palate repair only and lip and palate repair children had a median length of stay of 29 h 20 min and 29 h 0 min respectively, A total of 79% and 76% of these groups had intravenous fluids administered. Cleft lip repair infants fed in significantly less time than palate alone or lip and palate operations (p values 0.00 and 0.03, respectively). Cleft lip repair only infants feed well post-operatively and rarely require intravenous fluids. Infants having cleft repair involving the palate are slower to feed orally, and may require fluids due to poor oral intake. Intravenous fluids following lip repair is associated with longer hospital stay. We suggest intravenous fluids may not be needed routinely following cleft lip repair, but should always be considered following repair involving the palate.


Subcellular and subsynaptic localization of group I metabotropic glutamate receptors in the nucleus accumbens of cocaine-treated rats.
There is significant pharmacological and behavioral evidence that group I metabotropic glutamate receptors (mGluR1a and mGluR5) in the nucleus accumbens play an important role in the neurochemical and pathophysiological mechanisms that underlie addiction to psychostimulants. To further address this issue, we undertook a detailed ultrastructural analysis to characterize changes in the subcellular and subsynaptic localization of mGluR1a and mGluR5 in the core and shell of nucleus accumbens following acute or chronic cocaine administration in rats. After a single cocaine injection (30 mg/kg) and 45 min withdrawal, there was a significant decrease in the proportion of plasma membrane-bound mGluR1a in accumbens shell dendrites. Similarly, the proportion of plasma membrane-bound mGluR1a was decreased in large dendrites of accumbens core neurons following chronic cocaine exposure (i.e. 1-week treatment followed by 3-week withdrawal). However, neither acute nor chronic cocaine treatments induced significant change in the localization of mGluR5 in accumbens core and shell, which is in contrast with the significant reduction of plasma membrane-bound mGluR1a and mGluR5 induced by local intra-accumbens administration of the group I mGluR agonist, (RS)-3,5-dihydroxyphenylglycine (DHPG). In conclusion, these findings demonstrate that cocaine-induced glutamate imbalance has modest effects on the trafficking of group I mGluRs in the nucleus accumbens. These results provide valuable information on the neuroadaptive mechanisms of accumbens group I mGluRs in response to cocaine administration.


Extreme makeover: converting one cell into another.
Cells of adult mammals can be converted (reprogrammed) to new cells. In one approach, adult cells are converted to pluripotent stem cells, followed by differentiation to regenerate new cell types. Alternatively, adult cells may be directly converted into other mature cells or progenitors. We discuss and compare these two approaches with particular emphasis on the latter and its relevance for regenerative medicine.


Informal E-waste recycling practices and environmental pollution in Africa: What is the way forward?
Globally, e-waste is the fastest growing and most valuable waste-stream. While countries worldwide are increasingly acknowledging the e-waste problem and introducing policies and regulations that deal with e-waste, large quantities of e-waste still go undocumented. Much of these global e-wastes are accumulating in open-dumpsites and landfills in African-countries where they are recycled informally resulting in significant environmental and public-health concerns. Although, there is a plethora of studies on e-waste management and disposal, only a few-studies have focused on African-countries who are major recipients of e-waste. Moreover, despite the attempts to mitigate the problem of e-waste in African-countries, e-waste has remained a major-concern and there are currently very limited workable solutions. This study examines informal e-waste recycling, environmental pollution and the extent of environmental and health impacts in major countries of concern including Ghana, Nigeria, Egypt, Kenya and South Africa. The global e-waste Waste Atlas Report, 2020 identified these countries as major recipients of e-waste. To achieve the aims of this research, previous studies from 2005 to 2022 are collected from various databases and analyzed. Accordingly, this study focuses on environmental pollution and public-health impacts resulting from e-waste dumping and informal recycling practices, illegal transboundary shipment of e-waste to the selected countries, and the interventions of governments and international organizations in reducing the impact of e-waste pollution and informal recycling practices in Africa. Based on the outcomes of this study, practical approaches on the way-forward are recommended. The findings of this study contribute to a growing-body of research on informal e-waste recycling practices in Africa to document that individuals working within e-waste sites and residents in nearby communities are exposed to a number of toxic-substances, some at potentially concerning levels.


Cost and effectiveness of treatment options for childhood obesity.
What is already known about this subject Approximately one-fifth of children in the UK are obese. There are currently few, effective interventions available in the UK. There are very little data on relative cost-effectiveness of childhood obesity interventions, which hampers the commissioning of future services. What this study adds Simple multi-component obesity interventions can be provided at relatively low cost per 0.1 body mass index standard deviation score (BMI SDS) improvement. More intensive and effective interventions incur greater cost per 0.1 BMI SDS reduction but this may be justified given the improved overall BMI SDS reduction attained. To describe the costs and outcomes of three models of care for childhood obesity previously evaluated in two 2-arm pilot randomized trials in England. The treatments were (i) a hospital clinic (control in both trials), comprising a multidisciplinary team of consultant, dietitian and exercise specialist; (ii) a nurse-led primary care clinic replicating the service provided by the hospital and (iii) an intensive intervention using Mandometer®, a behaviour modification tool aimed at encouraging slower eating and better recognition of satiety. Patient-level data on resources used to deliver each intervention were collected during the trials. Apart from the cost of the Mandometer® the majority of cost was staff time, dependent on discipline and grade. Outcome for both trials was body mass index standard deviation score (BMI SDS) measured at 12 months. Cost and outcome data were available for 143 children in total. Cost per child was £1749 (SD £243) in the Mandometer® group, £301 (£76) in the primary care group, and £263 (£88) and £209 (£81) in the hospital groups. Mean reduction in BMI SDS was 0.40 (0.35), 0.17 (0.26), 0.15 (0.25) and 0.14 (0.32), respectively. Intensive management using Mandometer® was effective but costly (£432 per 0.1 reduction in BMI SDS) compared to conventional care (range £153-£173). A total of 26% children receiving conventional care achieved a clinically meaningful reduction in BMI SDS; however, use of Mandometer® training may be justified in children not responding to conventional lifestyle interventions.


Reversal of floppy eyelid syndrome with treatment of obstructive sleep apnoea.
To describe the beneficial effects of treatment of obstructive sleep apnoea on the symptoms and signs of floppy eyelid syndrome. A case of sleep apnoea with associated floppy eyelid syndrome is reported. A 32-year-old man presented with left floppy eyelid syndrome and a known diagnosis of obstructive sleep apnoea. He underwent treatment for 4 years with continuous positive airways pressure by mask during sleep and the symptoms and signs of his floppy eyelid syndrome disappeared. Treatment of obstructive sleep apnoea may reverse the changes of floppy eyelid syndrome.


How sensitive are cost-effectiveness analyses to choice of parametric distributions?
Cost-effectiveness analyses of clinical trial data are based on assumptions about the distributions of costs and effects. Cost data usually have very skewed distributions and can be difficult to model. The authors investigate whether choice of distribution can make a difference to the conclusions drawn. The authors compare 3 distributions for cost data-normal, gamma, and lognormal-using similar parametric models for the cost-effectiveness analyses. Inferences on the cost-effectiveness plane are derived, together with cost-effectiveness acceptability curves. These methods are applied to data from a trial of rapid magnetic resonance imaging (rMRI) investigation in patients with low back pain. The gamma and lognormal distributions fitted the cost data much better than the normal distribution. However, in terms of inferences about cost-effectiveness, it was the normal and gamma distributions that gave similar results. Using the lognormal distribution led to the conclusion that rMRI was cost-effective for a range of willingness-to-pay values where assuming a gamma or normal distribution did not. Conclusions from cost-effectiveness analyses are sensitive to choice of distribution and, in particular, to how the upper tail of the cost distribution beyond the observed data is modeled. How well a distribution fits the data is an insufficient guide to model choice. A sensitivity analysis is therefore necessary to address uncertainty about choice of distribution.


Mesenchymal stem cell derived microvesicles: trophic shuttles for enhancement of sperm quality parameters.
Diminishing sperm quality during cryopreservation process ends up in a complete or partial loss of sperm's fertilizing potential. Rehabilitation of such affected sperm is crucial to improve their fertilizing potential. A variety of evidence has indicated that secreted microvesicles (MVs) from mesenchymal stem cells (MSCs) are involved in regeneration of injured endogenous cells via shuttling MSC trophic molecules. Sperm obtained from cauda epididymides of adult male Wistar rats were equally assigned to four separate groups. Following suspension in cryoprotectant extender, sperm were untreated or treated with increasing concentrations of MSC-derived MVs (25, 50 and 100μg). After incubation in successive steps, sperm were cryopreserved. The frozen-thawed sperm were assessed for viability, motility and antioxidant capacity parameters. Consequently, expression levels of surface adhesion molecules (CD29, CD44, ICAM-I and VCAM-I) involved in sperm fusogenic and signaling properties, were assessed by flow cytometry. Results showed an enhanced quality parameters and adhesive properties of cryopreserved sperm following treatment with MSC-derived MVs.


Efficacy of intravenous amiodarone as short-term treatment for refractory ventricular tachycardia.
The antiarrhythmic efficacy and safety of intravenous amiodarone were evaluated in 13 patients with recurrent sustained ventricular tachycardia. All patients had been refractory to at least three prior antiarrhythmic agents (mean, 3.3 per patient), and 11 had multiple direct current cardioversions. Intravenous amiodarone suppressed sustained tachycardia in seven patients; the concurrent use of procainamide resulted in suppression in two additional patients. Ventricular tachycardia recurred in three patients, two within 2 hours of initiation of amiodarone. Adverse effects necessitated the discontinuation of amiodarone in one patient because of significant hemodynamic deterioration. The use of intravenous amiodarone was associated with a significant fall in heart rate and cardiac output. Plasma amiodarone levels were high after initial bolus loading but declined to low or less than therapeutic levels within 60 minutes. Thus although we found intravenous amiodarone to be effective in the short-term treatment of refractory ventricular tachycardia, administration of amiodarone can be associated with significant hemodynamic deterioration. The dosing protocol used should be modified to a lower initial bolus and a higher early maintenance infusion rate.


Transcriptomic Signature of the CD24hi CD38hi Transitional B Cells Associated With an Immunoregulatory Phenotype in Renal Transplant Recipients.
The role of B cells after transplant regarding allograft rejection or tolerance has become a topic of major interest. Recently, in renal transplant recipients, a B cell signature characterized by the overexpression of CD19+ CD38hi CD24hi transitional B cells has been observed in operationally tolerant patients and in belatacept-treated patients with significantly lower incidence of donor-specific antibodies. The phenotypic and functional characterization of these transitional B cells is far from exhaustive. We present the first transcriptomic and phenotypic analysis associated with this cell phenotype. Three populations were studied and compared: (i) transitional CD24hi CD38hi , (ii) CD24+ CD38- , and (iii) CD24int CD38int B cells. Transcriptome bioinformatic analysis revealed a particular signature for the CD24hi CD38hi population. Phenotypic analysis showed that CD24hi CD38hi transitional B cells also expressed CD9, CD10, CD1b and inducible T cell costimulator ligand (ICOS-L) markers. In addition, we found enrichment of IL-10+ cells among CD24hi CD38hi cells expressing ICOS-L and CD1b, the latter showing regulatory properties. Renal transplant recipients treated with belatacept exhibited significant expression of CD1b. Our results show that transitional CD24hi CD38hi B cells exhibit a distinct and specific profile, and this could be helpful for understanding of immune-regulatory mechanisms and immune monitoring in the field of organ transplant and autoimmune disease.


Assessment of ochratoxin A and tenuazonic acid in Canadian ice-wines.
Twenty-six samples of commercial ice-wine made from late-harvested grapes were assayed for the mycotoxins ochratoxin A and tenuazonic acid. Canadian wines originated in British Columbia (18) and Ontario (8). For comparison two German wines from Hesse were also studied. Four additional samples of research ice-wine originating in were also studied. In all wine samples, assays using immuno-affinity chromatography and fluorescence liquid chromatography indicated ochratoxin A below 0.15 μg/L, the limit of determination of the method. Tenuazonic acid was determined by solidphase micro-extraction and liquid chromatography and was below the limit of determination (70 μg/L) in all samples. The European Union food tolerance limit for ochratoxin A in wine is 2 μg/L. A tolerance for tenuazonic acid has not yet been established.


Can quantitative diffusion-weighted MR imaging differentiate benign and malignant cold thyroid nodules? Initial results in 25 patients.
The characterization of cold nodules of the thyroid gland is mandatory because approximately 20% of these nodules are of malignant origin. The purpose of this study was to evaluate the distinction of cold thyroid nodules by using quantitative diffusion-weighted MR imaging (DWI). In 25 patients with cold nodules on scintigraphy and suggestive findings at fine-needle aspiration, thyroid carcinoma was suggested. In these patients, cold nodules and the normal parenchyma of the contralateral thyroid lobe were prospectively investigated with quantitative DWI (echo-planar imaging sequence; maximum b-value, 800 s/mm(2)) before surgery. The differences in the mean apparent diffusion coefficient (ADC) values in benign and malignant nodules were tested by using a Mann-Whitney U test. Histologically, there were 20 carcinomas with a minimum size of 8 mm and 5 adenomas. The mean ADC values (in 10(-3) mm(2)/s) differed significantly among carcinoma, adenoma, and normal parenchyma (P < .05). The ranges (95% confidence interval) of the ADC values for carcinoma (2.43-3.037), adenoma (1.626-2.233), and normal parenchyma (1.253-1.602) showed no overlap. When an ADC value of 2.25 or higher was used for predicting malignancy, the highest accuracy of 88%, with 85% sensitivity and 100% specificity, was obtained. Quantitative DWI seems to be a feasible tool with which to differentiate thyroid carcinomas from adenomas; however, further studies are required including larger numbers of patients to confirm our results.


Biosynthesis of chick type VI collagen. II. Processing and secretion in fibroblasts and smooth muscle cells.
The biosynthesis of type VI collagen was studied in "matrix-free" chick embryo smooth muscle cells and fibroblasts. Omission of ascorbate from the culture affected to a great extent the secretion in fibroblasts but had a very minor effect on smooth muscle cells. Quantitative analysis of the secretion process in continuous time course and in pulse-chase experiments confirmed that fibroblasts and smooth muscle cells secreted type VI collagen with the same chain composition but with different kinetics: after 4 h of chase more than 60% of the labeled type VI collagen was present in the culture medium of fibroblasts, whereas at the same time interval less than 25% was secreted by smooth muscle cells. The different kinetics depends on intrinsic properties of the cells, since it was detected also in adherent cells. However, even in fibroblasts, secretion of type VI collagen was much slower than secretion of fibronectin, of which more than 50% was already in the cell medium after 1 h of chase. Treatment of the cells with inhibitors of hydroxylation and glycosylation caused a shift in mobility that revealed a size heterogeneity in the Mr = 260,000 subunit. No evidence of processing was observed in chick cells for any of the subunits that were synthesized and secreted uncleaved. In addition, after several days of chase the Mr of the subunits of type VI collagen isolated from the matrix remained unchanged, thus excluding that in the chick even a partial or incomplete processing takes place.


Physical activity in hospitalised stroke patients.
The aim of this paper was to examine the amount and type of physical activity engaged in by people hospitalised after stroke. Method. We systematically reviewed the literature for observational studies describing the physical activity of stroke patients. Results. Behavioural mapping, video recording and therapist report are used to monitor activity levels in hospitalised stroke patients in the 24 included studies. Most of the patient day is spent inactive (median 48.1%, IQR 39.6%-69.3%), alone (median 53.7%, IQR 44.2%-60.6%) and in their bedroom (median 56.5%, IQR 45.2%-72.5%). Approximately one hour per day is spent in physiotherapy (median 63.2 minutes, IQR 36.0-79.5) and occupational therapy (median 57.0 minutes, IQR 25.1-58.5). Even in formal therapy sessions limited time is spent in moderate to high level physical activity. Low levels of physical activity appear more common in patients within 14 days post-stroke and those admitted to conventional care. Conclusions. Physical activity levels are low in hospitalised stroke patients. Improving the description and classification of post stroke physical activity would enhance our ability to pool data across observational studies. The importance of increasing activity levels and the effectiveness of interventions to increase physical activity after stroke need to be tested further.


Effects of creatine supplementation on memory in healthy individuals: a systematic review and meta-analysis of randomized controlled trials.
From an energy perspective, the brain is very metabolically demanding. It is well documented that creatine plays a key role in brain bioenergetics. There is some evidence that creatine supplementation can augment brain creatine stores, which could increase memory. A systematic review and meta-analysis of randomized controlled trials (RCTs) was conducted to determine the effects of creatine supplementation on memory performance in healthy humans. The literature was searched through the PubMed, Web of Science, Cochrane Library, and Scopus databases from inception until September 2021. Twenty-three eligible RCTs were initially identified. Ten RCTs examining the effect of creatine supplementation compared with placebo on measures of memory in healthy individuals met the inclusion criteria for systematic review, 8 of which were included in the meta-analysis. Overall, creatine supplementation improved measures of memory compared with placebo (standard mean difference [SMD] = 0.29, 95%CI, 0.04-0.53; I2 = 66%; P = 0.02). Subgroup analyses revealed a significant improvement in memory in older adults (66-76 years) (SMD = 0.88; 95%CI, 0.22-1.55; I2 = 83%; P = 0.009) compared with their younger counterparts (11-31 years) (SMD = 0.03; 95%CI, -0.14 to 0.20; I2 = 0%; P = 0.72). Creatine dose (≈ 2.2-20 g/d), duration of intervention (5 days to 24 weeks), sex, or geographical origin did not influence the findings. Creatine supplementation enhanced measures of memory performance in healthy individuals, especially in older adults (66-76 years). PROSPERO registration no. 42021281027.


Leber congenital amaurosis - a model for efficient genetic testing of heterogeneous disorders: LXIV Edward Jackson Memorial Lecture.
To critically evaluate our experience in molecular testing of Leber congenital amaurosis (LCA) and to use this information to devise a general approach to heterogeneous recessive disorders. Careful clinical and molecular characterization of large cohorts of patients affected with inherited eye diseases will be an essential step in the development of effective therapy for these diseases, especially when the therapy involves gene replacement. A molecular genetic case-control study. Six hundred forty-two unrelated individuals with the clinical diagnosis of LCA and 200 unrelated control individuals were screened for disease-causing sequence variations in eight genes using various combinations of single-strand conformational polymorphism analysis (SSCP), automated DNA sequencing, multiplex allele-specific ligation analysis (SNPlex), and high-density solid-phase single nucleotide polymorphism genotyping. Four hundred forty instances of 189 different disease-causing sequence variations were observed in this study, 98 of which have not been previously reported. One hundred forty-six of the 189 variations (77%) were observed in only a single individual. The observed variations were not evenly distributed among the LCA patients or among the eight genes. Empirical analysis of this uneven distribution was used to devise a multi-platform mutation detection strategy that is four times more efficient than a more conventional strategy of completely sequencing all of the coding regions of all LCA genes in all subjects. Hardy-Weinberg analysis of the observed mutations suggests that these eight genes are collectively responsible for about 70% of the cases of LCA in North America. The carrier frequency of the most common LCA allele (an intron 26 variation in CEP290) was found to be 2/3,248, which suggests that the overall prevalence of LCA in this population is about 1/81,000. An allele-specific ligation assay (SNPlex) was designed to detect 68 of the most common LCA-causing alleles, and semi-quantitative analysis of the data from this assay also revealed examples of gene deletion and isodisomy in the cohort. The data demonstrates that a tiered screening strategy combining allele-specific detection with automated DNA sequencing can increase the efficiency of autosomal recessive mutation detection four-fold when compared with DNA sequencing alone. However, the very high rate of unique mutations observed in this study (77%) suggests that DNA sequencing will remain an important part of the overall strategy if high sensitivity is to be achieved.


Serological studies of an acid-labile O-polysaccharide of Proteus vulgaris OX19 lipopolysaccharide using human and rabbit antibodies.
In a Weil-Felix test, sera from patients infected with Rickettsia sp. agglutinate Proteus OX types of bacteria and Proteus lipopolysaccharide (LPS) are responsible for the cross-reaction. Data on the character of LPS of one of the OX group strains, Proteus vulgaris OX19, are contradictory, and it remained unclear whether it has an O-polysaccharide (OPS) and is thus LPS of the smooth type (S) or not (rough-type LPS). Our studies showed that P. vulgaris OX19 (strain PZH-24) produces a smooth-type LPS that contains a long-chain OPS, but it undergoes depolymerization during mild acid hydrolysis conventionally used for LPS delipidation and loses the serological activity. An elucidation of the complete structure of OPS demonstrated the presence of a glycosyl phosphate linkage responsible for the acid-lability of the polysaccharide chain. In ELISA, both IgM type antibodies in a Weil-Felix test with human anti-Rickettsia typhi sera and rabbit anti-P. vulgaris OX19 antibodies reacted with OPS. Rabbit antibodies did not inhibit the cross-reaction with human antibodies and thus bind to different epitopes.


Conservative management of early-onset severe preeclampsia: comparison between randomized and observational studies a systematic review.
To compare maternal and perinatal outcomes between randomized trials and observational studies in which conservative management was performed for more than 48 h in patients with early-onset severe preeclampsia. We searched PubMed, LILACS, Cochrane and Google Scholar. The studies were divided in two groups: randomized and observational studies, from 1990 to 2018 that included patients with severe preeclampsia before 34 weeks of gestation with pregnancy prolongation ≥48 h but that did not include fetal growth restriction or HELLP syndrome at the beginning. The main variables recorded were maternal and perinatal complications. Forty-four studies met the inclusion criteria, and 5 of these were randomized. The average pregnancy prolongation was 9 days, with no difference between groups. Maternal complications were significantly more common in observational studies, RR = 0.71, 95% CI (0.54-0.93), p = .009. Perinatal complications were also significantly more common in observational studies (RR = 0.89, 95% CI (0.80-0.98), p = .01) at the expense of stillbirth and neonatal deaths. The percentages of cesarean sections were significantly higher in randomized studies, RR = 1.54, 95% CI (1.46-1.64). There were 2 maternal deaths, both in observational studies. Observational studies in which conservative management of early-onset preeclampsia is performed and do not include patients with fetal growth restriction or patients with HELLP syndrome and where at least 2 days of pregnancy prolongation is achieved are associated with significantly more maternal and perinatal complications.


Decreased monocyte class II MHC expression following major abdominal surgery in children is related to operative stress.
Monocyte class II major histocompatibility complex (MHC) expression is necessary for antigen presentation and stimulation of T-cells. The aim of this study was to correlate monocyte class II MHC response to operative stress in children and the possible influence of cytokines in the postoperative period. We studied 21 children undergoing elective abdominal surgery. Operative stress score (OSS) was calculated. Monocyte class II MHC expression was measured preoperatively, immediately after surgery, 24 and 48 h postoperatively, using flow cytometry. Class II MHC is expressed as mean fluorescence intensity (MFI) of monocytes expressing MHC (mean +/- SD). Cytokine levels (interleukins 1ra, 6, and 10, and tumor necrosis factor-alpha) were also measured. Data between time points were compared using repeated measures ANOVA. There was an immediate postoperative decrease in class II MHC expression, with lowest levels 24 h postoperatively (preoperative 50 +/- 23.6, 24 h 18.2 +/- 9.4, P < 0.0001 vs. preoperative). At 48 h there was partial recovery in class II MHC, but levels were still significantly lower than preoperative (23.9 +/- 11.1, P < 0.001). The degree of monocyte depression was related to the magnitude of operative stress. Patients who had OSS <10 displayed some recovery in expression at 48 h 25.5 +/- 11.1), whereas in patients with OSS > or = 10 (severe surgical stress), expression further decreased at 48 h (MFI 14.0 +/- 0.1). There was an elevation of interleukin-1ra in the immediate postoperative period in both groups. There was no elevation in the other cytokines. Abdominal surgery in children decreases monocyte MHC expression. Class II MHC depression was related to magnitude of surgical trauma, implying that more severe immuneparesis follows surgery of greater magnitude. This may predispose to postoperative infection.


Changes in Laminin Expression Pattern during Early Differentiation of Human Embryonic Stem Cells.
Laminin isoforms laminin-511 and -521 are expressed by human embryonic stem cells (hESC) and can be used as a growth matrix to culture these cells under pluripotent conditions. However, the expression of these laminins during the induction of hESC differentiation has not been studied in detail. Furthermore, the data regarding the expression pattern of laminin chains in differentiating hESC is scarce. In the current study we aimed to fill this gap and investigated the potential changes in laminin expression during early hESC differentiation induced by retinoic acid (RA). We found that laminin-511 but not -521 accumulates in the committed cells during early steps of hESC differentiation. We also performed a comprehensive analysis of the laminin chain repertoire and found that pluripotent hESC express a more diverse range of laminin chains than shown previously. In particular, we provide the evidence that in addition to α1, α5, β1, β2 and γ1 chains, hESC express α2, α3, β3, γ2 and γ3 chain proteins and mRNA. Additionally, we found that a variant of laminin α3 chain-145 kDa-accumulated in RA-treated hESC showing that these cells produce prevalently specifically modified version of α3 chain in early phase of differentiation.


The Use of Probiotic-Loaded Single- and Double-Layered Microcapsules in Cake Production.
To date, the probiotic product development studies have mostly focused on dairy-based foods. However, endowing bakery products with probiotic properties not only provides a variety in food selection but would also potentially improve public health when the consumption rates are taken into consideration. This study aimed to incorporate single- and double-layered microcapsules containing Saccharomyces boulardii, Lactobacillus acidophilus, and Bifidobacterium bifidum, produced by spray drying and chilling, in cake production. Microcapsules were added after baking to the three different types of cakes (cream-filled, marmalade-filled, and chocolate-coated). Additionally, the microcapsules were injected into the center of the cake mix and baked at 200 °C for 20 min, for plain cake only. After baking of plain cakes, the count of S. boulardii and L. acidophilus as determined in the double-layered microcapsules produced by spray chilling was 2.9 log cfu/g. The survivability rates of S. boulardii and L. acidophilus were also determined as 67.4 and 70.7% in this microcapsule, respectively. However, there were no viable B. bifidum detected after baking. The free forms of these probiotics did not survive in any plain cake experiments. Single-layered microcapsules produced by spray chilling provided a better protective effect on the probiotics in cream-filled and marmalade-filled cake samples during storage, particularly the cream-filled cakes. This study showed that combined spray chilling and spray drying microencapsulation techniques (double-layered microcapsules) could increase the survivability of probiotic microorganisms during the cake baking process. During storage, the cake samples had a near neutral pH value, and the textural properties deteriorated due to staling. However, cake staling had a limited effect on the sensorial attributes of the cakes and the samples could be readily consumed after storage for 90 days.


Branch vein occlusion. Retinal neovascularization outside the involved segment.
Three hundred sixty-six cases of branch vein occlusion entered within the multicenter Branch Vein Occlusion Randomized Clinical Trial have been reviewed. Four cases that developed retinal neovascularization outside of the retinal area involved with the occlusion have been documented by photography and fluorescein angiography.


Sexual differences in the excretion of organic and inorganic mercury by methyl mercury-treated rats.
Adult male and female Long Evans rats received 1 mumole of methyl (203Hg) mercuric chloride per kilogram sc. Whole-body retention of mercury and excretion of organic and inorganic mercury in urine and feces were monitored for 98 days after dosing. Females cleared mercury from the body more rapidly than did males. The major route of mercury excretion was feces. By 98 days after dosing, cumulative mercury excretion in feces accounted for about 51% of the dose in males and about 54% of the dose in females. For both sexes, about 33% of the dose was excreted in feces as inorganic mercury. Cumulative excretion of organic mercury in feces accounted for about 18 and 21% of the dose in males and females, respectively. Urinary excretion of mercury was quantitatively a smaller route for mercury clearance but important sexual differences in loss by this route were found. Over the 98-day experimental period, males excreted in urine about 3.2% of the dose and females excreted 7.5%. Cumulative organic Hg excretion in urine accounted for 1.8% of the dose in males and 5.3% of the dose in females. These sexual differences in urinary and fecal excretion of organic and inorganic mercury following methyl mercury treatment were consistent with previous reports of sexual differences in mercury distribution and retention in methyl mercury-treated rats, particularly sexual differences in organic mercury uptake and retention in the kidney. Relationships between body burdens of organic or inorganic Hg and output of these forms of Hg in urine and feces were also found to be influenced by the interval after MeHg treatment and by sex. Relationship between concentration of Hg in liver and feces and in kidney and urine differed for organic and inorganic Hg and depended upon sexual status and interval after MeHg treatment. These findings emphasize that sexual differences in distribution, retention, and metabolism of methyl mercury are factors to be considered in estimations of hazards associated with exposure to this agent.


Does renewable energy improve environmental quality? Evidence from RECAI countries.
Since 1990, the ecological footprints have been increasing significantly with a continuous increase rate, which led to challenges to environmental quality. The basis for economic growth was said to be the shift of energy and environmental strategies toward a sustainable future. Indeed, it became a matter of proclaimed acceptance that environmental challenges nurtured expansion, innovation, and competitiveness. Climate change is the most pressing issue being faced by the world due to an increase in ecological footprint from 7.0 billion to 20.6 billion GHA. It indicates the seriousness of environmental degradation; therefore, nations need to ensure environmental sustainability. Keeping this in mind, the present research mainly aims to examine the impact of renewable energy utilization on the ecological footprints of RECAI economies, spanning the period from 1990 to 2020. To significantly achieve the research objective, we utilized panel econometric methods for empirical analysis. The results of long-run elasticities indicate that both renewable energy utilization as well as trade openness significantly controls the ecological footprints, while higher conventional energy utilization and economic growth significantly impede environmental sustainability. The empirical findings provide new insights for policymakers on renewable energy for the betterment of environmental quality in RECAI countries.


Intestinal Phospholipid Disequilibrium Initiates an ER Stress Response That Drives Goblet Cell Necroptosis and Spontaneous Colitis in Mice.
Patients with ulcerative colitis have low concentrations of the major membrane lipid phosphatidylcholine (PC) in gastrointestinal mucus, suggesting that defects in colonic PC metabolism might be involved in the development of colitis. To determine the precise role that PC plays in colonic barrier function, we examined mice with intestinal epithelial cell (IEC)-specific deletion of the rate-limiting enzyme in the major pathway for PC synthesis: cytidine triphosphate:phosphocholine cytidylyltransferase-α (CTαIKO mice). Colonic tissue of CTαIKO mice and control mice was analyzed by histology, immunofluorescence, electron microscopy, quantitative polymerase chain reaction, Western blot, and thin-layer chromatography. Histopathologic colitis scores were assigned by a pathologist blinded to the experimental groupings. Intestinal permeability was assessed by fluorescein isothiocyanate-dextran gavage and fecal microbial composition was analyzed by sequencing 16s ribosomal RNA amplicons. Subsets of CTαIKO mice and control mice were treated with dietary PC supplementation, antibiotics, or 4-phenylbutyrate. Inducible loss of CTα in the intestinal epithelium reduced colonic PC concentrations and resulted in rapid and spontaneous colitis with 100% penetrance in adult mice. Colitis development in CTαIKO mice was traced to a severe and unresolving endoplasmic reticulum stress response in IECs with altered membrane phospholipid composition. This endoplasmic reticulum stress response was linked to the necroptotic death of IECs, leading to excessive loss of goblet cells, formation of a thin mucus barrier, increased intestinal permeability, and infiltration of the epithelium by microbes. Maintaining the PC content of IEC membranes protects against colitis development in mice, showing a crucial role for IEC phospholipid equilibrium in colonic homeostasis. SRA accession number: PRJNA562603.


C5a delays apoptosis of human neutrophils via an extracellular signal-regulated kinase and Bad-mediated signalling pathway.
We recently demonstrated that complement fragment C5a delays apoptosis of human neutrophils via induction of the phosphatidylinositol-3 kinase (PI 3-K) pathway. In the present study, we examined whether C5a modulates neutrophil survival through the extracellular signal-regulated kinase (ERK) and Bad-mediated signalling pathway. Human neutrophils were isolated by percoll gradient and preincubated for 1 h with or without PD98059 (20 microM), a specific ERK inhibitor, followed by incubation with C5a (1 microg mL(-1)) for 24 h. Apoptosis was quantified by flow cytometry, using propidium iodide nuclear staining. Extracellular signal-regulated kinase downstream signalling events were evaluated by measuring the expression of cytosolic total and phosphorylated p44/p42 proteins, and Bad phosphorylation using immunoblot analyses. These time-dependent analyses were performed over a brief exposure to C5a (0-30 min). Modulation of cytosolic caspase-9 and caspase-3 activity was measured by Western blot analyses. C5a inhibited neutrophil apoptosis (P=0.04), which was abrogated in the presence of PD98059 (P=0.04). Time-dependent effect of C5a on p44/p42 phosphorylation was rapid, peaked at 5 min, and was abrogated by the ERK inhibitor (P=0.04). In addition, brief stimulation of neutrophils with C5a induced phosphorylation of Bad, which was inhibited by the ERK inhibitor (P=0.03). Further, C5a suppressed the proteolytic cleavage of caspase-9 and caspase-3, which was reversed by ERK inhibition. Finally, blockade of both the ERK (with PD98059) and PI 3-K (with wortmannin) pathways did not induce additive inhibition of neutrophil apoptosis by C5a. This study demonstrates that in addition to the PI 3-K pathway, C5a also inhibits neutrophil apoptosis via an ERK-signalling pathway, resulting in phosphorylation of Bad and blockade of proteolytic cleavage of caspases. The activation of this additional survival-signalling pathway may be another important cellular mechanism that enhances neutrophil survival in inflammatory states.