Overexpression of VEGF in dermal fibroblast cells accelerates the angiogenesis and wound healing function: in vitro and in vivo studies.
Fibroblasts are the main cells of connective tissue and have pivotal roles in the proliferative and maturation phases of wound healing. These cells can secrete various cytokines, growth factors, and collagen. Vascular endothelial growth factor (VEGF) is a unique factor in the migration process of fibroblast cells through induces wound healing cascade components such as angiogenesis, collagen deposition, and epithelialization. This study aimed to create VEGF165 overexpressing fibroblast cells to evaluate angiogenesis function in wound healing. In vitro, a novel recombinant expression vector, pcDNA3.1(-)-VEGF, was produced and transfected into the fibroblast cells. Following selecting fibroblast cells with hygromycin, recombinant cells were investigated in terms of VEGF expression by quantifying and qualifying methods. Mechanical, physical, and survival properties of polyurethane-cellulose acetate (PU-CA) scaffold were investigated. Finally, in vivo, the angiogenic potential was evaluated in four groups containing control, PU-CA, PU-CA with fibroblast cells, and VEGF-expressing cells on days 0, 2, 5, 12 and 15. Wound biopsies were harvested and the healing process was histopathologically evaluated on different days. qRT-PCR showed VEGF overexpression (sevenfold) in genetically-manipulated cells compared to fibroblast cells. Recombinant VEGF expression was also confirmed by western blotting. Manipulated fibroblast cells represented more angiogenesis than other groups on the second day after surgery, which was also confirmed by the antiCD31 antibody. The percentage of wound closure area on day 5 in genetically-manipulated Hu02 and Hu02 groups showed a significant reduction of wound area compared to other groups. These findings indicate that overexpression of VEGF165 in fibroblast cells results in enhanced angiogenesis and formation of granulated tissue in the early stage of the healing process, which can show its therapeutic potential in patients with impaired wound healing and also provide functional support for gene therapy.


The efficacy and safety of venetoclax therapy in elderly patients with relapsed, refractory chronic lymphocytic leukaemia.
Elderly chronic lymphocytic leukaemia (CLL) patients treated outside of trials have notably greater toxicity with the Bruton's tyrosine kinase inhibitor ibrutinib compared to younger patients. It is not known whether the same holds true for the B-cell lymphoma 2 inhibitor venetoclax. We provide a comprehensive analysis of key safety measures and efficacy in 342 patients comparing age categories ≥75 and <75 years treated in the relapsed, refractory non-trial setting. We demonstrate that venetoclax has equivalent efficacy and safety in relapsed/refractory CLL patients who are elderly, the majority of whom are previous ibrutinib-exposed and therefore may otherwise have few clear therapeutic options.


Neuroendocrine aspects of the relief of melancholia by induced seizures (electroconvulsive treatment).
Some important features of the antidepressant efficacy of ECT are described. Two principal hypotheses of this action, a neuroendocrine and a neurohumoral, are discussed in detail. The merits of the neuroendocrine hypothesis of convulsive therapy for studies of melancholia, catatonia, and new antidepressant treatments are discussed.


Surgical decision-making for managing complex intracranial aneurysms.
The treatment of complex intracranial aneurysms remains a therapeutic challenge. These lesions are frequently not amenable to selective clipping or coiling or other endovascular procedures and surgery still has a predominant role.We illustrate our "surgical decision making" for managing complex intracranial aneurysmal lesions. The best strategy is decided on the basis of pre-operative neuroradiological and intra-operative main determinants such as anatomical location, peri-aneurysmal angioanatomy (branch vessels, critical perforators), broad neck, intraluminal thrombosis, aneurysmal wall atherosclerotic plaques and calcifications, absence of collateral circulation, and previous treatment. The surgical strategy encompasses one of the following treatment possibilities: (1) Direct clip reconstruction; (2) Complete trapping ("classic" or "variant"); (3) Partial trapping (proximal "inflow" or distal "outflow" occlusion). Because the goal of any aneurysm treatment is both (1) aneurysm exclusion and (2) blood flow replacement, cerebral revascularization represents a major management option whenever definitive or temporary vessel occlusion is needed.Cerebral revascularization can therefore be used temporarily as a "protective" bypass, or definitively as a "flow replacement" bypass.Complete and partial trapping strategies are associated with flow "replacement" bypass surgery, to preserve blood flow into the territory supplied by the permanently trapped vessel. The construction of the "ideal" bypass depends on several factors, the most important of which are amount of flow needed, recipient vessel, donor vessel, and microanastomosis technique.The choice between "complete" or "partial" trapping depends on angioanatomical criteria as well. A complete trapping is always favored, as it has the advantage of immediate aneurysm exclusion. When perforating vessels arise from the aneurysmal segment or when the inspection of all the angioanatomy of the aneurysm is considered inadvisable and risky, "partial trapping" strategies are of interest. Partial trapping may consist either of proximal or distal occlusion. We discuss the rationale behind these treatment modalities and illustrate it with a case series of seven patients successfully treated for complex intracranial aneurysmal lesions (location: 1 ICA, 1 ACom, 3 MCA, 2 PICA).


The management of bereavement on intensive care units.
To investigate the management of the bereaved on Intensive Care Units (ICU) throughout the United Kingdom, and to identify inadequacies that may exist either in the provision of staff training in dealing with bereavement or in the facilities or support available for the bereaved. Questionnaires were sent to the senior nurse and senior doctor in all general ICUs with more than four beds nationwide. The questions asked about nursing and medical practice around the time of a patient's death, as well as about staff attitudes towards, and training in, dealing with bereavement and the support they received for this role. We obtained a 68% (293/430) response rate. Most ICUs had facilities for relatives, but little for the specific needs of the bereaved. Only 6% of doctors and 21% of nurses had training in dealing with bereavement and grieving. A staff support group was available in 23% of ICUs, and 75% of the remainder thought it would be useful to have one. Lack of staff training and poor facilities for relatives were identified as the major concerns of ICU staff. Many doctors and nurses working in Intensive Care Units feel inadequately trained to deal confidently with the bereaved. A minority of ICUs have support mechanisms available for their staff, inspite of the perceived need for them. Furthermore, many ICU staff feel the facilities they are able to offer the bereaved are inadequate. We have identified the major inadequacies and the needs of ICU staff for improved training. Meeting these needs would play a significant role not only in reducing staff stress but also minimising the morbidity in surviving relatives.


A search for novel thermoluminescent radiation dosimeter media.
We describe two example pilot efforts to help define new thermoluminescent dosimeter media. The first concerns ZnS:Mn nanophosphors, prepared by chemical precipitation using zinc and sodium sulfate, doped with manganese sulfate at concentrations varying from 1 to 3mol. The second concerns chemical vapor deposited diamond, produced as a thin film or as amorphous carbon on a single-crystal silicon substrate, each deposited under the same conditions, use being made of the hot filament-chemical vapor deposition (HFCVD) technique. The gas concentrations used were 1% CH4 in 99% H2 and 25% CH4 in 75% H2. Characterization of formations used FESEM, XRD and EDX. The nanophosphors consisted of particles of sizes in the range 85-150nm, the thermoluminescence (TL)-based radiation detection medium giving rise to a single peaked glow curve of maximum yield at a temperature of 250°C at a heating rate of 5°C/s. The TL response increased linearly with radiation dose, ZnS doped to 2mol of Mn being found the most sensitive. Regarding chemical vapor deposited (CVD) carbon, inappreciable TL was found for the resultant ball-like amorphous carbon films, graphite, and the silicon substrate, whereas CVD diamond films showed a promising degree of linearity with dose. For both the ZnS and diamond samples, TL signal fading was appreciable, being some 40% per day for ZnS and>50% per day for CVD films even under storage in the dark at room temperature, making it apparent that there is need to adjust parameters such as the size of nanoparticles.


Prophylaxis against thromboembolism in spinal surgery.
Thromboprophylaxis in spinal surgery is a controversial topic. In the current environment of risk management, it seems reasonable to review and critically appraise the risks and benefits of different prophylaxis methods in spinal surgery.


Perfluorinated compounds in soils from Liaodong Bay with concentrated fluorine industry parks in China.
With the shift of manufacturing plants from more industrialized countries to China, occurrences of Perfluorinated compounds (PFCs) in the environment has attracted more attention. Concentrations of PFCs in 14 soils collected from the coast of Liaodong Bay were determined and spatial distributions used to identify potential sources of PFCs. A mass balance approach was introduced to quantify concentrations of Total Fluorine (TF) and Extractable Organic Fluorine (EOF). Concentrations of ∑PFCs ranged from <MDL to 3.14 ng g(-1). Perfluorooctanoic acid (PFOA) and Perfluoroundecanoic acid (PFUdA) were predominant with maximum concentrations of 0.47 ng g(-1) and 0.42 ng g(-1). Statistical analysis indicated that ∑PFCs and EOF account for a very small portion in TF and there is limited correlation among ∑PFCs, EOF and TF in soils. Two fluorine industry parks have recently been established in the vicinity of the study site, where electrochemical fluorination (ECF) is used to synthesize ammonium perfluorooctanoate (APFO) and PFOA. The majority of the PFCs detected had chain lengths of 8 carbons or more, which suggests that direct emission from plants in the two parks was a major source of PFCs in soils of the coastal area of Liaodong Bay.


Influence of multi-walled carbon nanotubes on the effects of roxithromycin in crucian carp (Carassius auratus) in the presence of natural organic matter.
Carbon nanotubes are increasingly entering the aquatic environment and may interact with other co-existing contaminants, such as antibiotics. However, whether these interactions may affect their bioavailability in aquatic organisms is the subject of considerable debate. The primary objective of this study was to assess the risks arising from the coexistence of roxithromycin (ROX) and multi-walled carbon nanotubes (MWCNTs) in waters containing natural organic matter (NOM), focusing on the distribution and bioaccumulation of ROX in crucian carp (Carassius auratus), and the related biochemical status. There were no significant differences in ROX bioaccumulation in fish following exposure to ROX with and without NOM. However, the further addition of MWCNTs significantly facilitated the bioaccumulation of ROX in the liver (32-80%), gill (15-74%), intestine (51-113%), and bile (15-67%) in different exposure periods. Meanwhile, a 0.3-fold increase in the metabolic enzyme activity and oxidative stress in the liver were markedly accelerated by the co-exposed MWCNTs compared to ROX alone. The findings imply that the ROX adsorbed on MWCNTs may be a higher threat to fish than ROX alone. The high and fast release of ROX from MWCNTs in bile salts and serum albumin may contribute to the enhancement in bioaccumulation and bioactivity of ROX in fish with MWCNTs.


Relative strength of photodynamic biologic tissue glue in penetrating keratoplasty in cadaver eyes.
To compare the relative strength of photodynamic biologic tissue glue (PBTG) with that of 10-0 nylon sutures in closing penetrating keratoplasty (PKP) wounds in cadaver eyes. Corneal Research Laboratory, University of Chicago, Chicago, Illinois, USA. A 7 mm central corneal trephination was performed on 8 recently enucleated human eyes. Each corneal button was removed and then resutured in its original position, using 4 interrupted 10-0 nylon cardinal sutures and PBTG (n = 7) or 16 interrupted 10-0 nylon sutures (n = 1). Wound leak and incisional bursting pressures were determined. The average pressure at which wound leak and iris prolapse occurred was 124 mm Hg (range 70 to 180) and 185 mm Hg (range 90 to 300), respectively. The pressure required to create wound leak increased as the time between glue application and pressurization of the globe lengthened (mean 22 minutes, range 5 to 60 minutes), indicating that the bonding and stability of the adhesive increased with time. In contrast, the control eye developed wound leak at 70 mm Hg and iris prolapse at 300 mm Hg. Photodynamic biologic tissue glue was comparable to sutures in providing adequate corneal wound strength in PKP in a cadaver eye model. Wound closure with PBTG may reduce the number of sutures required in corneal transplantation and decrease the incidence of suture-related complications and allograft rejection.


The effects of ergonomics training on the knowledge, attitudes, and practices of teleworkers.
The rapid growth of teleworking has raised several social and legal issues regarding an employer's responsibility for the safety of an employee's home office. In this paper, researchers discuss the need for safety training for teleworkers and the effectiveness of a home office ergonomics training program. Study participants (N = 50) were randomly assigned into a treatment or control group. The treatment group completed the ergonomics training and a pre- and posttest. The control group completed the pre- and posttests without training. The study demonstrated the need for teleworker ergonomics training. More than 85% of participants had not received teleworker training before, and 44% had experienced pain or discomfort while teleworking. Participants who completed the training significantly improved their scores on knowledge, attitude, and practices subtests. In a follow-up survey, participants indicated that they had made ergonomic changes to their offices based on the training. Several participants indicated that the pain or discomfort that they had been experiencing was eliminated or reduced as a result of the training.


Prophylaxis of supraventricular tachyarrhythmia after coronary bypass surgery with oral verapamil: a randomized, double-blind trial.
This study investigated the efficacy of oral administration of verapamil, started 24 hours after coronary artery bypass grafting (CABG), in reducing the incidence of postoperative supraventricular tachyarrhythmia (SVT). Two hundred patients were randomly assigned in a double-blind fashion to receive a one-week course of either a placebo or 80 mg of verapamil every 6 hours. Overall, SVT developed in 23 control and 14 verapamil-treated patients, a 39% reduction in incidence (p less than 0.10). Of the patients who received at least four doses and continued to receive the study drug, 17 in the control and 7 in the verapamil group experienced SVT, a 53% decrease in incidence (p less than 0.06). Atrial fibrillation constituted 34 of the 37 SVT episodes and was associated with a slower ventricular response in the group given verapamil (115 +/- 8 versus 156 +/- 4 beats per minute; p less than 0.001). No evidence was found linking postoperative SVT with the withdrawal of beta-blocking drugs. Adverse effects required that 20 patients in the verapamil and 6 in the placebo group be removed from the study. Hypotension or pulmonary edema or both developed in 13 of the patients receiving verapamil, but in only 1 of the control patients (p less than 0.001). We conclude that although verapamil has potential merit for the prophylaxis of SVT after CABG, its use in this setting is associated with a high incidence of unacceptable hemodynamic side effects.


Sensory Modulation Abilities in Healthy Preterm-Born Children: An Observational Study Using the Sensory Processing and Self-Regulation Checklist (SPSRC).
This study aimed to investigate prematurity as a risk factor for sensory processing disorders, using the Italian Version of Sensory Processing and Self-Regulation Checklist (SPSRC-IT), based on a sample of healthy Italian children born preterm in comparison with a sample of typical full-term children. Two groups of caregivers of Italian healthy preschooler children were recruited. The first group comprised 37 caregivers of full-term children (FT), while the second group consisted of 37 caregivers of preterm children (PT) (gestational age < 37 weeks). Significant differences between the groups in several subsections and factors of the SPSRC-IT were found, specifically in the Physiological Conditions section, in the Gustatory and Olfactory Sense section, in the Vestibular Sense section, and in the Proprioceptive Sense section, with lower scores in the PT group. Moreover, children born at a lower gestational age or with lower weights had a higher risk of dysfunctions in processing gustatory and olfactory, vestibular, and proprioceptive stimuli. In conclusion, the SPSRC-IT suggested a potential link between prematurity and challenges in the development of sensory processing and self-regulation skills, especially in children with a very low birth weight and very low gestational age.


Massive lipid exudation and retinal detachment after combined brachytherapy and transpupillary thermotherapy in choroidal melanoma.
A 59 year-old male with choroidal melanoma in the left eye who underwent plaque brachytherapy (iodine 125). One week after surgery, massive exudation with retinal detachment and lipid exudation was observed. Evolution was assessed with funduscopy and ultrasound every month. Nine months after surgery transpupillary thermotherapy (TTT) was performed over the fluid-free irradiated residual tumour. Three months after this procedure, new retinal breaks appeared in the treated area with vitreous seeding that required enucleation. Combined treatment with plaque brachytherapy and TTT may associate severe complications that may require enucleation of the involved eye.


Spinal versus epidural anesthesia for vesicovaginal fistula repair surgery in a rural sub-Saharan African setting.
To compare spinal anesthesia with epidural anesthesia in patients undergoing surgery for vesicovaginal fistula (VVF) repair. Nonrandomized, prospective, clinical, pilot study. Rural African hospital with 165 beds. 60 ASA physical status I and II patients undergoing VVF repair surgery. 30 patients were included in each study group. Spinal (1.5-2 mL hyperbaric bupivacaine 0.75%) or lumbar epidural (20-24 mL bupivacaine 0.5%) anesthesia was administered to the patients. Demographics, quality of anesthesia, duration of postoperative analgesia, as well as preoperative, intraoperative, and postoperative data, were all recorded. Data were compared between study groups using unpaired Student's t test for continuous variables and chi(2) and Fisher's exact tests for categorical data. Quality of anesthesia was different between groups (P = 0.009). Good anesthesia quality was significantly more frequent in the spinal (86.7%) than the epidural group (50%, P = 0.005). Postoperative analgesia quality was comparable (P = 0.347). There were no differences between groups in hemodynamic parameters before, during, or after surgery. Spinal anesthesia proved to be the better anesthetic technique for VVF repair surgery when compared with epidural anesthesia in a rural, sub-Saharan African setting.


Retraction Note to: Effect of whole-body vibration exercise on lumbar bone mineral density, bone turnover, and chronic back pain in post-menopausal osteoporotic women treated with alendronate.
The Editor-in-Chief has retracted this article [1] because an investigation by Keio University has concluded that there are inaccuracies in the data reported.


Hypervolemia does not cause degradation of the endothelial glycocalyx layer during open hysterectomy performed under sevoflurane or propofol anesthesia.
Fluid-induced hypervolemia may stimulate the release of natriuretic peptides and cause degradation (shedding) of the endothelial glycocalyx layer. Sevoflurane is believed to protect the glycocalyx, but the importance of using sevoflurane to prevent shedding during routine surgery is unclear. The plasma concentrations of brain natriuretic peptide and two biomarkers of glycocalyx shedding, syndecan-1, and heparan sulfate, were measured in 26 patients randomized to receive general anesthesia with sevoflurane or propofol during open abdominal hysterectomy. The fluid therapy consisted of 25 mL/kg (approximately 2 L) of Ringer´s lactate over 30 minutes. Blood hemoglobin and plasma albumin were used to indicate plasma volume expansion and capillary leakage. The plasma concentrations of brain natriuretic peptide and shedding products showed low levels throughout the surgery (median brain natriuretic peptide, 21 ng/L; syndecan-1, 12.9 ng/mL; and heparan sulfate, 6.5 µg/mL), but the heparan sulfate concentration increased 2 hours post-operatively (to 17.3 µg/mL, P < .005). No differences were noted between the propofol and sevoflurane groups in any of the measured parameters. Albumin was apparently recruited to the bloodstream during the first 20 minutes, when the intravascular retention of infused fluid was almost 100%. The urine flow was <1 mL/min, despite the vigorous volume loading. No relevant elevations of brain natriuretic peptide or degradation products of the glycocalyx layer were observed when hypervolemia was induced during open abdominal hysterectomy performed with sevoflurane or propofol anesthesia. Plasma volume expansion from Ringer´s lactate was pronounced.


Updates from the Evidence Base Examining Association between Periodontal Disease and Type 2 Diabetes Mellitus: Current Status and Clinical Relevance.
Epidemiological surveillance documents an escalating epidemic prevalence of both type 2 diabetes (T2DM) and periodontal disease (PD). The principal goals of this review are to: 1) re-examine the clinical significance of associations between PD and T2DM, based on strength of collective evidence as determined by systematic review and meta-analysis, and 2) review findings of the systematic reviews and meta-analyses in light of the current understanding of PD-associated pathophysiology and intersection with T2DM pathophysiology. Tooth loss predicts risk for chronic disease and mortality. PD is significantly associated with complications of diabetes, including retinopathy. Based on systematic reviews and meta-analyses, the adjunctive use of certain antibiotics enhances non-surgical periodontal treatment (NSPT) in patients with T2DM. Systematic reviews and meta-analyses support NSPT efficacy in achieving metabolic control. Systematic reviews and meta-analyses support the association between PD and T2DM, albeit the effect size may be modest. PD-T2DM interactions have important clinical implications.


Quantification and distribution of big conductance Ca2+-activated K+ channels in kidney epithelia.
Big conductance Ca2+ activated K+ channels (BK channels) is an abundant channel present in almost all kind of tissue. The accurate quantity and especially the precise distribution of this channel in kidney epithelia are, however, still debated. The aim of the present study has therefore been to examine the presence of BK channels in kidney epithelia and determine the actual number and distribution of these channels. For this purpose, a selective peptidyl ligand for BK channels called iberiotoxin or the radiolabeled double mutant analog 125I-IbTX-D19Y/Y36F has been employed. The presence of BK channels were determined by a isotope flux assay where up to 44% of the total K+ channel activity could be inhibited by iberiotoxin indicating that BK channels are widely present in kidney epithelia. Consistent with these functional studies, 125I-IbTX-D19Y/Y36F binds to membrane vesicles from outer cortex, outer medulla and inner medulla with Bmax values (in fmol/mg protein) of 6.8, 2.6 and 21.4, respectively. These studies were performed applying rabbit kidney epithelia tissue. The distinct distribution of BK channels in both rabbit and rat kidney epithelia was confirmed by autoradiography and immunohistochemical studies. In cortical collecting ducts, BK channels were exclusively located in principal cells while no channels could be found in intercalated cells. The abundant and distinct distribution in kidney epithelia talks in favor for BK channels being important contributors in maintaining salt and water homeostasis.


Amino acid transport in the retina.
The uptake, exit, homoexchange, inhibitory pattern, and kinetic analysis of transport of three amino acids were studied in the isolated retina of adult rat under different metabolic conditions. Only in the case of glycine, uptake and exit were shown to duplicate the processes observed in brain slices. In the case of lysine, glucose and oxygen showed an inhibitory effect, but with glutamate spontaneous exit could not be measured. It was also found that the rate of homoexchange for glycine and glutamate, but not for lysine, increases in the presence of oxygen and glucose.


S-D-lactoylglutathione in resting and activated human neutrophils.
Zymosan particles opsonised with human serum factors functionally activate human neutrophils and induce a substantial modification of the human neutrophil cytosolic glyoxalase system. The activity of glyoxalase I increases and the activity of glyoxalase II decreases by 20-40% of their resting cell activities during the initial 10 min of activation. The cellular concentration of the glyoxalase intermediate S-D-lactoylglutathione increases by ca. 100% of resting cell levels during this period. This modification may be related to the ability of S-D-lactoylglutathione to stimulate the assembly of microtubules.


A new method of attachment of isolated mammalian ventricular myocytes for tension recording: length dependence of passive and active tension.
The study of the Frank-Starling's law in mammalian single cells has been hindered by a lack of an easily performed method of stretching cells. Some authors have succeeded in this but their methods required a great deal of technical expertise and in most cases they have not had much success. We have developed an easy method of stretching mammalian ventricular cells from slack sarcomere length (S.L.) (Lo, 1.77 +/- 0.05 microns) to about 117% of this length. Thin carbon fibers (12 microns in diameter) which can be bound electrochemically to the cell membrane surface have been used. A flexible long fiber of known compliance (80 microns/microN) was attached to one end of the cell and a stiff double fiber (4 microns/microN) to the other end. The cell attachment was relatively easy to perform and successful results were obtained in 80% of the attempts. The displacement of the flexible fiber allows the quantitative measurements of the resting tension in a group of non-stimulated cells and of auxotonic contractions developed upon stimulation in another group of cells. Increasing S.L. from Lo to 105-106% of Lo, an increase in active tension from 0.21 +/- 0.03 mN/mm to 0.26 +/- 0.01 mN/mm (n = 4) could be noticed with a stimulation frequency of 0.5 Hz. An increase in active tension was also observed at 1 Hz. Staircase kinetics were accelerated with stretching; this confirms at the single cell level the hypothesis of an effect of length-dependent activation on the staircase. Eulerian differential stiffness constant was calculated and found to be 13.5 +/- 1.2, a value which is comparable to that described in intact heart. Thus the important stiffness found in the whole heart may be due to intracellular component(s) such as myofilament and/or connectin.


A C. elegans screening platform for the rapid assessment of chemical disruption of germline function.
Despite the developmental impact of chromosome segregation errors, we lack the tools to assess environmental effects on the integrity of the germline in animals. We developed an assay in Caenorhabditis elegans that fluorescently marks aneuploid embryos after chemical exposure. We qualified the predictive value of the assay against chemotherapeutic agents as well as environmental compounds from the ToxCast Phase I library by comparing results from the C. elegans assay with the comprehensive mammalian in vivo end point data from the ToxRef database. The assay was highly predictive of mammalian reproductive toxicities, with a 69% maximum balanced accuracy. We confirmed the effect of select compounds on germline integrity by monitoring germline apoptosis and meiotic progression. This C. elegans assay provides a comprehensive strategy for assessing environmental effects on germline function.


Macroalgal-derived alginate/wastepaper hydrogel to alleviate sunflower drought stress.
Alginate/wastepaper hydrogel mitigated sunflower drought stress by increasing the water holding capacity of the soil and decreasing phosphate leaching. The hydrogel promoted sunflower growth and decreased stress related biomarkers. There is a growing interest in the development of biodegradable hydrogels for the alleviation of drought stress on plants. A novel hydrogel based on brown algal-derived alginate and wastepaper was developed and tested as a soil supplement for sunflower growth under moderate (75% field capacity (FC)) and extreme (50% FC) water-deficit stress. The hydrogel showed fast swelling in water, which obeyed the pseudo-first order kinetics. Besides, it increased the water holding capacity of the soil and exhibited a good phosphate adsorption (37 mg PO4- g-1 hydrogel after 6 days) in the soil, and more than 67% of the adsorbed phosphate was desorbed after 20 days. Thus, the phosphate leaching from the hydrogel-amended soil in a column experiment was only 2.77 mg after 4 times of over-irrigation, compared to 11.91 mg without the hydrogel. The hydrogel application promoted various root traits such as fresh and dry biomass, area, and length by > 2-, > 1.6-, > 1.35-, and > 1.3-folds under both water regimes in relation to the no-hydrogel treatments at the same conditions. Furthermore, the sunflower shoots exhibited similar proline contents to the well-watered control (100% FC), with > 50% reduction in relation to the drought-stressed plants under the same conditions. Similarly, the malondialdehyde contents were lowered by > 15%. The analysis of the antioxidant enzymes also indicated a marked reduction in the specific activities of catalase and ascorbate peroxidase under both 75 and 50% FC compared to the respective controls. Additionally, the hydrogel promoted the uptake of phosphate by sunflower roots. These results implied that the developed biodegradable hydrogel could be effectively applied as a soil additive to alleviate drought stress on crops.


Transference effects of prior non-contingent reinforcement on the acquisition of temporal control on fixed-interval schedules.
In two experiments we examined the influence of response and time factors on the speed of acquisition of temporal control on FI schedules. In Experiment 1, prior exposure to FT accelerated the development of temporal control on FI schedules of the same temporal value. It was also found that the slower acquisition on FI with prior RT was similar to that of rats with prior standard training. In Experiment 2, prior exposure to FT accelerated the development of temporal control on a FI schedule with a threefold increase in temporal value. Additionally, it was found that with prior FI 30s training, acquisition of temporal control on FI 90s was even faster than with prior FT 30s. Measures of head-entries into the feeder along the experiments indicated that temporal control was already developed during the periodic but not during the non-periodic histories and that this control transferred to lever press during FI testing phase.


When birds and sias fly: A neural indicator of inferring a word meaning in context.
Inference generation is a crucial skill in language comprehension. Recent research suggests that readers use both the contents from prior written text and their background knowledge, stored in long-term memory, to generate predictive inferences about what will come up next in a sentence. We recorded Event-Related Potentials (ERPs) to examine the reader's ability to make online inferences even in the presence of pseudowords (orthographically legal, but meaningless letter strings), that is, in the presence of referents with no a priori match to vocabulary stored knowledge. As expected, a large and sustained negativity (250-900ms) was elicited by the target word 'fly' when preceded by the pseudoword 'Sias' in the sentence 'Sias fly.' relative to when preceded by 'Birds' in the sentence 'Birds fly'. However, when readers were provided with an initial statement inviting to make an inference: 'Sias have wings', the word 'fly' in 'Sias fly' only elicited a negative voltage deflection over 100ms period (250-350), rapidly falling down to baseline. This result indicates that participants rapidly generated online inferences even with a hindered access to a referent's meaning (i.e. not knowing what 'Sias' are). Remarkably, brainwave traces to the access to a word's meaning in long-term memory (access to a well-known fact such as 'Birds fly') only diverged from ERPs for an inferred-from-reading knowledge ('Sias fly') for 100ms. We conclude that a fundamental search for across sentence coherence drives fast inference making processes in reading tasks. This pattern of brain response is critical to understand the rapid acquisition of new vocabulary when learning first and second languages.


In vitro megakaryocyte expansion in patients with delayed platelet engraftment after autologous stem cell transplantation.
Increasing the number of megakaryocytic cells in stem cell transplants by ex vivo expansion culture may provide an approach to accelerate platelet engraftment after high-dose chemotherapy. However, it is unknown if a relationship exists between the expansion potential of progenitor cells and the time to platelet engraftment in vivo. Therefore, we questioned if those patients who potentially would benefit most from expanded cell supplements are able to generate megakaryocytic cells efficiently in vitro. The in vitro megakaryocyte proliferation was analyzed from 19 leukapheresis samples from a group of multiple myeloma patients who all showed rapid neutrophil engraftment, but varied from 7 to 115 days post-transplant to achieve platelet levels >20x10(9)/l. CD34+ cells were isolated and analyzed for their potential to form megakaryocytic colonies (CFU-Mk) in colony assays and megakaryocytic (CD61+) cells in suspension cultures. The frequency and size of CFU-Mk and the expansion potential of CD61+ cells varied eightfold between individual patients. A similar range was found with CD34+ cells isolated from normal bone marrow (n=9). Rapid platelet engraftment occurred in patients receiving both high or low CFU-Mk doses and with high and low expansion of CD61+ cells. Four patients who experienced prolonged (>3 weeks) thrombocytopenia received low CFU-Mk doses, but the expansion potential was around median values or higher. Therefore, we conclude that the megakaryocyte proliferation is not impaired and that in vitro expansion could increase the number of megakaryocytic cells, although other factors could be more relevant in platelet engraftment in this group of patients.


Bioimpedance Measurements in Adolescents with Polycystic Ovary Syndrome: A Pilot Study.
Limited data are available on the body composition of adolescent women with polycystic ovary syndrome (PCOS). The aim of this study was to examine differences in body composition indices of metabolism, homeostasis and inflammation, between Greek adolescent females suffering from PCOS and age- and body mass index (BMI)-matched non-PCOS controls. Thirteen PCOS patients and nine non-PCOS controls, aged 13-24 years participated in this cross-sectional study. Study participants underwent assessment by a novel dual frequency bioimpedance device (BIA-ACC). The following body composition indices were measured in each adolescent: extra cellular water (ECW) as inflammation marker, total body water (TBW) as homeostasis marker, extracellular mass to body cell mass ratio (ECM/BCM), fat mass (FM), fat-free mass (FFM) and intracellular water (ICW) as markers of body mass composition and metabolism. Non-linear analysis showed no statistically significant differences in the body composition characteristics between PCOS patients and controls. Further studies with larger sample sizes are needed to confirm whether adolescents with PCOS actually have similar body composition profile with their non-PCOS peers.


Sentinel node biopsy versus elective neck dissection. Which is more cost-effective? A prospective observational study.
The aim of this prospective not randomized observational study was to determine the costs and outcomes of sentinel lymph node biopsy (SNB) vs elective neck dissection (END) among patients with early oral squamous cell carcinoma (OSCC). Seventy-three consecutive patients were divided according to neck staging method. Patients took the decision themselves after receiving detailed information of both suggested treatment tools. False negative (FN) and negative predictive value (NPV) were assessed. Log-rank test was used to compare disease-free survival (DFS) and overall survival (OS). Only direct costs were analysed. Cost information derived from volumes for hospital stay, surgery and neck outcome were obtained from an internal database of tertiary health care center. Thirty-two patients underwent SNB and 41 underwent an END (levels I-III). Average follow-up time was 48.2 months (range 7-80). Five neck recurrences were recorded in the SNB group (range 11-21). Seven neck recurrences occurred in the END group (range 9-16). No significant differences were found in DFS or OS. True negative patients in SNB group incurred in 42% less costs than END group. FN regardless of radiotherapy, was also lower in the SNB group. However, pN+ patients generated 23% more costs in the SNB group. In this not randomized observational study with an average follow-up period of 48.2 months, SNB appear to confer less cost than END, with similar prognosis.


Diversity and metal tolerance of nematode-trapping fungi in Pb-polluted soils.
The diversity of nematode-trapping fungi (NTF) in two lead (Pb) mines in Yunnan Province, China was investigated in 2004. In total, 20 species belonging to five genera were identified from 500 samples collected at the Lanping and the Huize mines. Pb concentrations ranged from 216 approximately 7,150 mg/kg for the former and 132 approximately 13,380 mg/kg for the latter, respectively. The fungi were divided into five groups based on different trapping mechanisms. The trapping-net producer group contained the largest number of species, with nine. Two predators, Dactylellina ellipsosporum and Arthrobotrys oligospora, were found at frequencies of 32.85% and 15.41%, respectively. The diversity indexes of NTF were positively correlated with Pb pollution levels in both the Lanping Mine (r=0.66) and the Huize Mine (r=0.72), suggesting that the distribution of NTF was not negatively affected by Pb contamination. For most strains of a given species, there was no significant difference (P>0.01) in the Pb tolerance between the strains isolated from habitats with low or high Pb concentrations. However, Pb toxicity exerted adverse effects on trap formation and predacious capability of fungi. We discuss the possible metal tolerance mechanisms and their relationships to the survival strategy of NTF in Pb-polluted environments.


CCK receptor subtype in insulin-producing cells: a combined functional and in situ hybridization study in rat islets and a rat insulinoma cell line.
Cholecystokinin (CCK) stimulates insulin secretion. It is, however, not established whether CCK receptors are expressed in insulin-producing cells. We therefore investigated, by in situ hybridization, whether CCK-A or CCK-B receptor mRNA could be detected in normal rat pancreatic islets and in the rat insulinoma cell line, RINm5F. The CCK-A, but not the CCK-B, receptor transcript was detected in both islets and RINm5F cells. Islet CCK-A receptors were mostly confined to the center of the islets corresponding to the distribution of the B cells. In RINm5F cells, insulin release was not significantly affected by cholecystokinin (CCK)-8-S (10(-13) to 10(-7) M), which is in contrast to the insulinotropic effect of CCK-8-S in normal rat islets. Similarly, in FURA-2AM-loaded cells, CCK-8-S (10(-11) to 10(-7) M) was without effect on the intracellular Ca2+ concentration ([Ca2+]ic) in RINm5F cells, whereas CCK-8-S (10(-7) M) markedly increased [Ca2+]ic (by 366+/-2 nM (P < 0.001) in normal rat islet cells. We conclude that the CCK-A, but not the CCK-B, receptor subtype is expressed in both normal rat islets and in the rat insulinoma-derived cell line RINmS5F. There is, however, a functional difference between normal islets and the RINm5F cells with respect to effects of CCK-8-S on insulin release and [Ca2+]ic.


Association Between Hepatic Steatosis, Measured by Controlled Attenuation Parameter, and Fibrosis Burden in Chronic Hepatitis B.
The interaction between chronic hepatitis B (CHB) and hepatic steatosis is poorly understood. We investigated whether measurement of controlled attenuation parameter (CAP), a non-invasive method to quantify steatosis, can assist in monitoring patients with CHB. We performed transient elastography, to measure liver stiffness, and made CAP measurements in 1606 patients with CHB (898 treated with nucleoside analogues, for a median 75.4 months) in Hong Kong, from January 2015 through September 2016. We also collected information on patients' medical history, current treatment, and smoking and alcohol habits, anthropometric measurements. We obtained and analyzed fasting blood samples. Severe liver fibrosis was defined, according to guidelines, as a liver stiffness measurement greater than 9.0 kPa in patients with normal level of alanine aminotransferase (ALT) or greater than 12.0 kPa in patients with a level of ALT 1-5-fold the upper limit of normal. Steatosis was defined as a CAP measurement of 248 dB/m or more, and severe steatosis as a CAP measurement or 280 dB/m more. We performed multivariate analysis to identify factors associated with severe fibrosis. The prevalence of steatosis, severe steatosis, and severe fibrosis in our cohort were 40.8%, 22.6%, and 14.1%, respectively. A higher proportion of patients with severe steatosis had severe fibrosis (21.4% vs 11.9% in the overall cohort; P < .001). In multivariate analysis, severe steatosis was associated with severe fibrosis in treatment-naïve patients (odds ratio, 3.60, 95% CI, 1.21-10.75) and in patients receiving treatment (odds ratios: 1.95 [1.06-3.61] for 3 or more years of treatment, 2.28 [1.13-4.61] for 5 or more years of treatment, and 2.79 [1.17-6.62] for 7 or more years of treatment). With every increase in CAP value of 10 dB/m, the risk of severe fibrosis increased by 15% in treatment-naïve patients and by 7%-8% in patients receiving treatment. Severe steatosis, determined by CAP measurement, is associated with severe fibrosis in treatment-naïve patients with CHB and in patients receiving treatment. Longitudinal studies are required to investigate if steatosis control, in addition to antiviral treatment, can reduce the burden fibrosis in patients with CHB.


Cell-to-cell movement of plant viruses. Insights from amino acid sequence comparisons of movement proteins and from analogies with cellular transport systems.
Cell-to-cell movement is a crucial step in plant virus infection. In many viruses, the movement function is secured by specific virus-encoded proteins. Amino acid sequence comparisons of these proteins revealed a vast superfamily containing a conserved sequence motif that may comprise a hydrophobic interaction domain. This superfamily combines proteins of viruses belonging to all principal groups of positive-strand RNA viruses, as well as single-stranded DNA containing geminiviruses, double-stranded DNA-containing pararetroviruses (caulimoviruses and badnaviruses), and tospoviruses that have negative-strand RNA genomes with two ambisense segments. In several groups of positive-strand RNA viruses, the movement function is provided by the proteins encoded by the so-called triple gene block including two putative small membrane-associated proteins and a putative RNA helicase. A distinct type of movement proteins with very high content of proline is found in tymoviruses. It is concluded that classification of movement proteins based on comparison of their amino acid sequences does not correlate with the type of genome nucleic acid or with grouping of viruses based on phylogenetic analysis of replicative proteins or with the virus host range. Recombination between unrelated or distantly related viruses could have played a major role in the evolution of the movement function. Limited sequence similarities were observed between i) movement proteins of dianthoviruses and the MIP family of cellular integral membrane proteins, and ii) between movement proteins of bromoviruses and cucumoviruses and M1 protein of influenza viruses which is involved in nuclear export of viral ribonucleoproteins. It is hypothesized that all movement proteins of plant viruses may mediate hydrophobic interactions between viral and cellular macromolecules.


Intermittent-access operant alcohol self-administration promotes binge-like drinking and drinking despite negative consequences in male and female heterogeneous stock rats.
The study of Alcohol Use Disorders (AUD) in preclinical models is hampered by difficulty in training rodents to voluntarily consume high levels of alcohol. The intermittency of alcohol access/exposure is well known to modulate alcohol consumption (e.g., alcohol deprivation effect, intermittent-access two-bottle-choice) and recently, intermittent access operant self-administration procedures have been used to produce more intense and binge-like self-administration of intravenous psychostimulant and opioid drugs. In the present study, we sought to systematically manipulate the intermittency of operant self-administered alcohol access to determine the feasibility of promoting more intensified, binge-like alcohol consumption. To this end, 24 male and 23 female NIH Heterogeneous Stock rats were trained to self-administer 10% w/v ethanol, before being split into three different-access groups. Short Access (ShA) rats continued receiving 30-min training sessions, Long Access (LgA) rats received 16-h sessions, and Intermittent Access (IntA) rats received 16-h sessions, wherein the hourly alcohol-access periods were shortened over sessions, down to 2 min. IntA rats demonstrated an increasingly binge-like pattern of alcohol drinking in response to restriction of alcohol access, while ShA and LgA rats maintained stable intake. All groups were tested on orthogonal measures of alcohol-seeking and quinine-punished alcohol drinking. The IntA rats displayed the most punishment-resistant drinking. In a separate experiment, we replicated our main finding, that intermittent access promotes a more binge-like pattern of alcohol self-administration using 8 male and 8 female Wistar rats. In conclusion, intermittent access to self-administered alcohol promotes more intensified self-administration. This approach may be useful in developing preclinical models of binge-like alcohol consumption in AUD.


CD4+ Cytotoxic T cells - Phenotype, Function and Transcriptional Networks Controlling Their Differentiation Pathways.
The two major subsets of peripheral T cells are classically divided into the CD4+ T helper cells and the cytotoxic CD8+ T cell lineage. However, the appearance of some effector CD4+ T cell populations displaying cytotoxic activity, in particular during viral infections, has been observed, thus breaking the functional dichotomy of CD4+ and CD8+ T lymphocytes. The strong association of the appearance of CD4+ cytotoxic T lymphocytes (CD4 CTLs) with viral infections suggests an important role of this subset in antiviral immunity by controlling viral replication and infection. Moreover, CD4 CTLs have been linked with anti-tumor activity and might also cause immunopathology in autoimmune diseases. This raises interest into the molecular mechanisms regulating CD4 CTL differentiation, which are poorly understood in comparison to differentiation pathways of other Th subsets. In this review, we provide a brief overview about key features of CD4 CTLs, including their role in viral infections and cancer immunity, and about the link between CD4 CTLs and immune-mediated diseases. Subsequently, we will discuss the current knowledge about transcriptional and epigenetic networks controlling CD4 CTL differentiation and highlight recent data suggesting a role for histone deacetylases in the generation of CD4 CTLs.


Hydrolysates of citrus plants stimulate melanogenesis protecting against UV-induced dermal damage.
The sun-tanning process occurs as a spontaneous response to ultraviolet (UV) irradiation. UV will induce tanning and DNA damage, processes that can lead to photoaging and skin disorders such as hyperpigmentation and cancer. The pigment melanin protects skin from UV damage; therefore, an efficient melanin-promoting suntan lotion could be highly beneficial. In this study, a process was developed to increase the content of naringenin in citrus extracts and to determine whether a higher naringenin content of citrus would induce melanogenesis. Melanin content and tyrosinase expression in mouse B16 melanoma cells were assayed after treatment with citrus plant extracts and their hydrolysates. The results indicate that hydrolysis increased the naringenin content in citrus extracts and that citrus preparations stimulated cellular melanogenesis and tyrosinase expression. It is suggested that this method is applicable to the industrial production of melanin-promoting suntan lotions with antiphotocarcinogenic properties derived from citrus rind and citrus products.


Guidelines for measuring and reporting environmental parameters for experiments in greenhouses.
The importance of appropriate, accurate measurement and reporting of environmental parameters in plant sciences is a significant aspect of quality assurance for all researchers and their research. There is a clear need for ensuring research across the world can be compared, understood and where necessary replicated by fellow researchers. A common set of guidelines to educate, assist and encourage comparativeness is of great importance. On the other hand, the level of effort and attention to detail by an individual researcher should be commensurate with the particular research being conducted. For example, a researcher focusing on interactions of light and temperature should measure all relevant parameters and report a measurement summary that includes sufficient detail allowing for replication. Such detail may be less relevant when the impact of environmental parameters on plant growth and development is not the main research focus. However, it should be noted that the environmental experience of a plant during production can have significant impact when subsequent experiments investigate plants at a molecular, biochemical or genetic level or where species interactions are considered. Thus, researchers are encouraged to make a critical assessment of what parameters are of primary importance in their research and these parameters should be measured and reported. This paper brings together a collection of parameters that the authors, as members of International Committee on Controlled Environment Guidelines (ICCEG) in consultation with members of our three parent organizations, believe constitute those which should be recorded and reported when publishing scientific data from experiments in greenhouses. It provides recommendations to end users on when, how and where these parameters should be measured along with the appropriate internationally standardized units that should be used.


C-reactive protein-induced activated partial thromboplastin time prolongation in heparinized samples is attenuated by elevated factor VIII.
Activated partial thromboplastin time (aPTT) and antifactor Xa (anti-Xa) activity are used to monitor unfractionated heparin therapy in children on extracorporeal membrane oxygenation (ECMO). Elevated C-reactive protein (CRP) can prolong aPTT and cause discrepancy between these two assays. We aimed to evaluate CRP effect on aPTT and anti-Xa assays in the presence of heparin and to determine whether elevated CRP affects laboratory monitoring in pediatric ECMO patients. Citrated normal specimens were spiked with CRP, heparin, and recombinant factor VIII (FVIII) and followed by measurement of aPTT and anti-Xa activity. Additionally, aPTT, anti-Xa activity, FVIII, fibrinogen, and CRP were measured in 18 ECMO specimens. Elevated CRP prolonged aPTT in normal specimens with or without heparin, but did not affect anti-Xa assay. In contrast, ECMO specimens showed similar aPTT and anti-Xa values regardless of CRP level. Elevated CRP in specimens was accompanied by increased fibrinogen and FVIII activity. Additional in vitro experiments confirmed that FVIII spiked simultaneously with CRP attenuated CRP-induced aPTT prolongation in heparinized specimens. In vitro CRP-induced aPTT prolongation is not observed in pediatric ECMO samples due to concomitant FVIII increase. Discordant changes of CRP and FVIII in plasma could contribute to aPTT/anti-Xa discrepancies observed during heparin therapy in the pediatric population. The anti-Xa assay is preferable for heparin monitoring in pediatric ECMO settings.


Preferential sexual transmission of pseudorabies virus in feral swine populations may not account for observed seroprevalence in the USA.
This paper compares the behavior of two competing models for the transmission of pseudorabies virus in feral swine in the USA. In first model, horizontal (non-sexual) density dependent transmission is the only transmission modality. In the second model, the only transmission modality is sexual transmission between mature males and females. The comparison of model behavior was carried out to test the hypothesis that preferential sexual transmission of PRV in feral swine can account for the seroprevalence observed in the field. The observed range of seroprevalence of PRV in mature feral swine in the USA is consistent with a preferential sexual transmission only if the feral swine mating system is a random mating system or a polygynous system in which there is a relatively large rate of acquisition of new mates. The observed range of seroprevalence of PRV in mature feral swine in the USA is not consistent with a preferential sexual transmission if there is mate guarding. This is important because the National Pseudorabies Surveillance Plan deems monitoring the risk of PRV introduction from feral swine to be a "minor objective" both in terms of the scope of the plan and with respect to the resources allocated. The rationale for this statement was derived from experimental studies, which suggested that the PRV indigenous to feral swine in the USA is preferentially sexually transmitted.


Correction to: Prevention by Regular Exercise of Acute Sleep Deprivation-Induced Impairment of Late Phase LTP and Related Signaling Molecules in the Dentate Gyrus.
The original version of this article unfortunately does not include the second affiliating institution of Dr. Munder A. Zagaar. "Department of Pharmacy Pracce and Clinical Health Sciences, Texas Southern University, Houston, TX 77004" should have been included on the paper.


Liposome-bound Zn (II)-phthalocyanine. Mechanisms for cellular uptake and photosensitization.
In the present study, cellular uptake of a liposomal formulation of ZnPc (CGP 55847) has been studied in human cervix carcinoma cells of the line NHIK 3025. The cellular uptake of ZnPc is found to be completed after 4-8 h of incubation. The maximum level of ZnPc in the cells after incubation with 1 microgram/ml ZnPc in E2a medium containing 3% serum is 60 ng/mg protein. The cellular uptake is attenuated by the presence of serum and at low temperature of the incubation medium, but the activation energy (30 kJ/mol) and fluorescence microscopic analysis of cells incubated with ZnPc at 0 degree C indicate that ZnPc is taken up into cells by a diffusion-mediated pathway. Measurements of subcellular marker enzymes have been performed immediately after light exposure of ZnPc-treated cells. The mitochondrial marker enzyme (cytochrome c oxidase) and the marker enzyme for the Golgi apparatus (UDP galactosyl transferase), but not those for lysosomes (beta-N-acetyl-D-glucosaminidase) and endoplasmic reticulum (NADPH cytochrome c reductase), are inactivated upon photodynamic treatment. These results indicate that ZnPc is mainly located in the Golgi apparatus and the mitochondria of NHIK 3025 cells. In contrast, photoactivated Photofrin is found to reduce the activity of UDP galactosyl transferase, but not that of NADPH cytochrome c reductase. The tetraphenylporphine TPPS2a and light reduce the activity of NADPH cytochrome c reductase, without influencing the activity of UDP galactosyl transferase. TPPS4 and light do not attenuate the activities of UDP galactosyl transferase and NADPH cytochrome c reductase.


Short-term morbidity of the upper limb after sentinel lymph node biopsy or axillary lymph node dissection for Stage I or II breast carcinoma.
The goals of sentinel lymph node biopsy (SLNB) are to improve axillary staging and reduce unnecessary axillary lymph node dissections (ALND), thereby reducing treatment-related upper-limb morbidity. In the current prospective study, short-term upper-limb morbidity was assessed after SLNB and/or ALND. The study comprised 204 patients with Stage I/II breast carcinoma. Mean patient age was 55.6 years (standard deviation, 11.6). Sixty-six patients (32%) underwent SLNB only, and 138 (68%) underwent a Level I-II ALND. Assessment (preoperative [t0] and 6 weeks postoperative [t1]) included evaluation of shoulder range of motion, muscle strength, grip strength, pain, upper/forearm circumference, shoulder disability, and activities of daily life (ADL). Considerable treatment-related upper-limb morbidity was observed. Significant (P < 0.001) changes were found for pain, range of motion in forward flexion, abduction and abduction/external rotation, strength of shoulder abductors and elbow flexors, and in perceived disability in ADL. However, no significant difference in change of upper-limb function and ADL was found between the SLNB and ALND groups. Significant short-term treatment-related upper-limb morbidity exists after SLNB or ALND. There is no significant difference in short-term treatment-related morbidity between SLNB and ALND.


Overproduction of anti-Tn antibody MLS128 single-chain Fv fragment in Escherichia coli cytoplasm using a novel pCold-PDI vector.
Overproduction of recombinant proteins in Escherichia coli is often hampered by their failure to fold correctly, leading to their accumulation within inclusion bodies. To overcome the problem, a variety of techniques aimed at soluble expression have been developed including low temperature expression and/or fusion of soluble tags and chaperones. However, a general protocol for bacterial expression of disulfide bond-containing proteins has hitherto not been established. Single chain Fv fragments (scFvs) are disulfide bond-containing proteins often difficult to express in soluble forms in E. coli. We here examine in detail the E. coli expression of a scFv originating from an anti-carbohydrate MLS128 antibody as a model system. We combine three techniques: (1) tagging scFv with thioredoxin, DsbC and protein disulfide isomerase (PDI), (2) expressing the proteins at low temperature using the pCold vector system, and (3) using Origami E. coli strains with mutations in the thioredoxin reductase and glutathione reductase genes. We observed a high expression level of soluble MLS128-scFv in the Origami strain only when PDI is used as a tag. The recombinant protein retains full binding activity towards synthetic carbohydrate antigens. The developed "pCold-PDI" vector has potential for overproduction of other scFvs and disulfide-containing proteins in the Origami strains.


[The necessity for treatment paths for lower limb prosthetics : All that glitters is not gold!]
Frequently occurring problems in the fitting of leg prostheses are either, for a prosthetic fitting, insufficiently resilient leg stumps or insufficiently adapted prostheses. For this reason, there has been a demand at the political level to establish treatment pathways and a quality standard for the care of people after leg amputations. How can a joint, interdisciplinary approach to the provision of aids be developed? Does interdisciplinary care of amputees lead to an improvement in the quality of care? A compendium "Quality standard in the field of lower extremity prostheses" was compiled and presented at the OT World in Leipzig in 2018. The basic principle is treatment pathways for the interdisciplinary care of all amputation levels of the lower extremities. The result is a compendium that represents the current "gold standard" in orthopaedic care after lower limb amputations. The compendium not only describes the treatment paths in detail but also makes demands on the quality of the result of the surgical amputation, as well as the goals of residual limb conditioning and the demands on the quality of the fitting of the prosthesis. A traffic light system is used to make recommendations, restrictions and rejections, thus defining the gold standard. If all the professions involved (doctors, CPOs, physiotherapists) do not work separately but rather together with the necessary expertise and routine, the quality of care for amputees will improve, and at the same time, the health care system will become more cost efficient.


Can humanization theory contribute to the philosophical debate in public health?
This paper will explore the humanization value framework for research, policy and practice with regard to its relevance for public health, specifically the reduction of inequities in health. This proposed framework introduces humanizing values to influence research, policy and practice. The framework is articulated through eight specific constituents of what it is to be human. These dimensions are articulated as humanizing and dehumanizing dimensions that have the potential to guide both research and practice. The paper will then go on to consider these dimensions in relation to the emergent qualities of the potential 'fifth-wave' of public health intervention. The humanization dimensions outlined in this paper were presented as emerging from Husserl's notion of lifeworld, Heidegger's contemplations about human freedom and being with others, and Merleau-Ponty`s ideas about body subject and body object. Husserl's ideas about the dimensions that make up 'lifeworld', such as embodiment, temporality and spatiality, underpin the suggested dimensions of what it is to be human. They are proposed in the paper as together informing a value base for considering the potentially humanizing and dehumanizing elements in systems and interactions. It is then proposed that such a framework is useful when considering methods in public health, particularly in relation to developing new knowledge of what is both humanizing and dehumanizing within research and practice.


[Comparative effectiveness of chrysanol and auranofin in rheumatoid arthritis].
A randomized study of 46 patients with classic and definite rheumatoid arthritis on chrysotherapy (25 patients--auranofin orally and 21--allochrysine intramuscularly) showed a therapeutic effect of both drugs. Allochrysine turned out more effective, it was not abandoned in any patients as a result of its inefficacy. However as compared to allochrysine auranofin was slightly better tolerated.


Acaricidal and anthelmintic action of ethanolic extract and essential oil of Achyrocline satureioides.
Although Brazilian livestock is considered one of the most productive in the world, the losses caused by parasites still greatly limit these activities and its incorrect control through traditional synthetic chemicals has caused problems such as parasitic resistance. The present study evaluated the anthelmintic and acaricidal action of ethanolic extract (EE) and essential oil (OE) obtained from Achyrocline satureioides inflorescences against an endo and an ectoparasite of great importance in sheep and cattle. The action against Haemonchus contortus was evaluated by the Larval Development Test (LDT) and the Egg Hatching Test (EHT), while the acaricidal potential against Rhipicephalus (Boophilus) microplus was evaluated by the Adult Immersion Test (AIT) and Larval Package Test (LPT). Chemical analyzes were performed by liquid chromatography coupled to mass of spectrometry (LC-MS) and gas chromatography coupled to mass spectrometry (GC-MS) in order to evaluate the chemical composition of EE and EO, respectively. Luteolin (m/z 285), quercetin (m/z 301) and methyl quercetin (m/z 315) were the major compounds detected in the A. satureioides EE, while 41 compounds were identified in EO. Among them, the major compounds were caryophyllene oxide (15.4%), limonene (10.9%), trans-calamenene + δ-cadinene (8.0%), (E)-caryophyllene (5.5%), spathulenol (5.1%) and α-pinene (4.5%). Against H. contortus, by EHT, the EE and EO showed LC50 of 83.14 mg mL-1 and 10.42 mg mL-1, respectively. In LDT, LC50 were 0.16 mg mL-1 and 0.42 mg mL-1, respectively by EE and EO. In the AIT assay for R. (B.) microplus tick LC50 obtained for EE and EO were 76.43 mg mL-1 and 114.35 mg mL-1, respectively. EE at 100 mg mL-1 completely inhibited egg laying, while the EO at 50 mg mL-1 completely inhibited larval hatching. No significant larvicidal effect was observed in LPT for both extracts. The EE and EO presented a high efficacy at low concentrations in the inhibition of egg hatching and larval development of H. contortus. Although both extracts did not show consistent larvicidal action, the results were significant in inhibiting the oviposition of R. (B.) microplus engorged females.


Decoupling trend and emission reduction potential of CO2 emissions from China's petrochemical industry.
This paper aims to study the decoupling status and emission reduction potential of China's petrochemical industry from 1996 to 2019. Firstly, the IPCC method is used to calculate the CO2 emissions of the petrochemical industry in China, then the logarithmic mean Divisia index (LMDI) method is used to identify the influencing factors of CO2 emissions, then the decoupling index is constructed to analyze the dependence of economic development on CO2 emissions, and finally the emission reduction potential model is established by using the influencing factors to reflect the CO2 emission reduction potential of the petrochemical industry. The results reveal that (1) the CO2 emissions can be divided into two stages of slow decline (1996-2000), (2015-2019), and one stage of rapid growth (2000-2015). (2) The energy intensity effect is the most effective factor to restrain CO2 emission, the economic growth effect is the key factor to promote CO2 emission. (3) From 1996 to 2019, there was a weak decoupling relationship between CO2 emission of petrochemical industry and economic development. Strong decoupling only occurred in 1996-2000 and 2015-2019. The CO2 emissions show only three decoupling score: I, II, and III. (4) CO2 mitigation occurred in four sub periods: 1996-2000, 2005-2010, 2010-2015, and 2015-2019. Therefore, the government should establish an energy-saving and environment-friendly industrial production system, intensify the use of clean energy, and optimize the labor force structure. It not only effectively strengthens the decoupling between the petrochemical industry and economic development, but also provides an empirical example for the carbon emission reduction and economic sustainable development of the petrochemical industry in other countries in the world.


Intravoxel incoherent motion diffusion-weighted imaging of the pancreas: Characterization of benign and malignant pancreatic pathologies.
To evaluate the diagnostic value of apparent diffusion coefficient (ADC) and intravoxel incoherent motion (IVIM) parameters in differentiating patients with either a normal pancreas (NP), pancreatic ductal adenocarcinoma (PDAC), neuroendocrine tumor (NET), solid pseudopapillary tumor (SPT), acute pancreatitis (AcP), vs. autoimmune pancreatitis (AIP). In all, 84 pathologically confirmed pancreatic tumors (60 PDACs, 15 NETs, 9 SPTs), 20 pancreatitis (13 AcPs, 7 AIPs), and 30 NP subjects underwent IVIM diffusion-weighted imaging using 10 b-values (0-900 sec/mm2 ) at 1.5T. The ADC, pure molecular diffusion coefficient (Dslow ), perfusion fraction (f), and perfusion-related diffusion coefficient (Dfast ) were calculated and compared using a Kruskal-Wallis test and post-hoc Dunn procedure. Receiver operating characteristic (ROC) analysis was performed to assess diagnostic performance. The f and Dfast of the PDAC were significantly lower than that of the NP (f = 0.10 vs. 0.24; Dfast = 42.21 vs. 71.74 × 10-3 mm2 /sec; P < 0.05). In ROC analysis, f showed the best diagnostic performance (area-under-the-curve, 0.919) among all parameters in differentiating PDAC from NP (P ≤ 0.001). The f values of AcP (0.11) and AIP (0.13) and the Dfast values of SPT (20.48 × 10-3 mm2 /sec) and AcP (24.49 × 10-3 mm2 /sec) were significantly lower compared with NP (f = 0.24; Dfast = 71.74 × 10-3 mm2 /sec; P < 0.05). For NET, the f (0.21) was significantly higher than that of PDAC (0.10, P < 0.01). Perfusion-related parameters f and Dfast are more helpful in characterizing pancreatic diseases than ADC or Dslow . The PDCA, SPT, AcP, and AIP were characterized by reduced f and Dfast values compared with normal pancreas. The f value might help in differentiating between PDAC and NET. 3 J. Magn. Reson. Imaging 2017;45:260-269.


Harnessing carbon monoxide-releasing platforms for cancer therapy.
Carbon monoxide (CO) is a gaseous molecule that has drawn growing concerns for cancer treatment, due to its multifaceted regulation of cellular function and the tumor microenvironment. The past decades have witnessed extensive efforts into this emerging field, centered in the development of intelligent CO donors and CO delivery based on nanomaterials. However, CO delivery is beset with biosafety problems arising from burst release and the demand for high therapeutic concentrations in tumor tissues. In this review, we provide an overview of physiological and therapeutic effects of CO, and discuss the recent progress in CO application for cancer treatment. The main emphasis is placed on CO-releasing platforms to overcome the challenges in delivery and underlying synergistic mechanism against cancer. Eventually, the obstacles confronted and future directions of CO therapy are also highlighted for future clinical translation.


When a Common Language Is Not Enough: Transcreating Cancer 101 for Communities in Puerto Rico.
In Puerto Rico (PR), cancer is the leading cause of death. Previous research has identified the need for cancer education in PR. Using culturally adapted cancer curricula to train local health educators may effectively increase cancer education and reduce health disparities. This article describes the three-phase process used to transcreate the Cancer 101 curriculum to train Master of Public Health (MPH) students to educate PR communities. First, an expert panel collaboratively reviewed the curriculum for content, legibility, utility, and colloquialisms. Recommendations included incorporating local references and resources, replacing words and examples with culturally relevant topics, and updating objectives and evaluation items. Subsequent focus groups with 10 MPH students assessed the adaptation's strengths, weaknesses, and utility for future trainees. Participants were satisfied with the curriculum's overall adaptation, ease of use, and listed resources; further improvements were suggested for all modules. Final expert panel revisions highlighted minor feedback, with the final curriculum containing nine transcreated modules. The transcreation process identified the need for changes to content and cultural translation. Changes were culturally and literacy-level appropriate, represented PR's social context, and were tailored for future trainees to successfully deliver cancer education. Findings highlight the importance of adapting Spanish educational materials across Hispanic sub-groups.


The good, the bad, and the ugly in diabetic diets.
In this article we have focused on the evolving pattern of nutritional management of the person with diabetes. Before the advent of insulin in 1922, it was sufficient to identify a meal plan that would keep people alive until they could be rescued from mortality due to diabetic ketoacidosis (the major killer of the era) by pharmacologic means. Now, the life expectancy of people with diabetes is close to that of the general population and focus has turned to combating the new threats of macrovascular disease and kidney failure. Over recent years the susceptibility of NIDDM patients to macrovascular events has been established and the twofold increase in risk of a heart attack in diabetic men is outshadowed by the four- to fivefold risk in diabetic women and the 13- to 17-fold greater risk in diabetics under the age of 30 years compared with their nondiabetic counterparts. The mechanism behind the susceptibility to macrovascular disease has generated a veritable plethora of investigations focusing on the atherogenic profile of diabetic dyslipidemia. Hyperinsulinemia, insulin resistance, and overtreatment of the diabetic with insulin have been claimed as contributors to the development of premature atherosclerosis. The hallmark of the diabetic dyslipidemia is the tendency to elevated VLDL triglyceride levels and the closely linked reduction in HDL cholesterol. Although there is some controversy on the relationship between triglyceride levels and the incidence of CAD, there is no doubt that HDL is an independent risk factor. It can now be safely said that elevated triglycerides are a risk factor in women and that in men elevated triglycerides constitute a risk factor if accompanied by a reduced HDL level. For these reasons, any approach to nutritional management of the diabetic must attempt not only to normalize glycemia but to make every effort to reduce the atherogenic profile. In the accompanying algorithm (Fig. 4), we consider the risk factors conducive to a reduction in life expectancy and offer a meal plan that is appropriate for the individual with diabetes. For the 80% of NIDDM patients who are obese, a diet with a reduction of 500 to 1000 kcal is in order and this may be achieved by a periodic VLCD. We examined carefully the controversy related to yo-yo dieting and support the notion that its effects in humans are not all that harmful. Ingestion of simple sugars in the high carbohydrate diet has negative effects both on carbohydrate and lipid metabolism.(ABSTRACT TRUNCATED AT 400 WORDS)


Microbiome-health interactions in older people.
Alterations in the composition and function of the gut microbiome have been implicated in a range of conditions and diseases. Culture-dependent and culture-independent studies both showed that older people harbour a gut microbiome that differs in composition from that of younger adults. Detailed analyses have identified discrete microbiota subtypes that characterize intermediates between a high diversity microbiota found in healthy community-dwelling subjects and a low diversity microbiota typical for elderly living in long-term residential care. There are also alterations in the microbiome composition associated with biological age, independent of health status. Even after adjusting for confounding factors such as age and medication, trends in microbiota composition correlate with gradients in clinical metadata particularly frailty and inflammatory status. There are few known mechanisms by which these associations might be causative rather than consequential, and this is a subject of intensive research. The strongest candidate effectors are microbial metabolites that could impact host energy balance, act as signalling molecules to modulate host metabolism or inflammation, and potentially also impact on the gut-brain axis.


A case of pseudoparacoccidioidomycosis: detection of the yeast phase of Mucor circinelloides in a clinical specimen.
A case is presented in which yeastlike cells resembling the yeast phase of Paracoccidioides brasiliensis were observed by microscopic examination of a urine sample. A mold identified as Mucor circinelloides was isolated from the specimen. It was converted to the yeast form by cultivation on Sabouraud agar incubated in a GasPak jar at 37 degrees C. The isolate was eventually shown not to be related to the patient's illness; however, the superficial resemblance of the yeast phase to P. brasiliensis caused some confusion in making a correct diagnosis.


A conceptual model to support curriculum review, revision, and design in an associate degree nursing program.
In light of declining NCLEX-RN first-write pass rates and the National League for Nursing call to transform nursing education, faculty at an associate degree nursing program in the southeastern United States began the process of intensive critique of its curriculum and program outcomes. Based on in-depth analysis and assessment of program outcomes, a conceptual framework was created to guide curriculum revision, development, and implementation processes. The framework serves as a guide for ongoing and systematic curriculum review and revision based upon evaluation findings. The basis of the conceptual framework was the result of an extensive literature review and an incorporation of the faculty-created philosophy.


When cost-effective interventions are unaffordable: Integrating cost-effectiveness and budget impact in priority setting for global health programs.
Potential cost-effective barriers in cost-effectiveness studies mean that budgetary impact analyses should also be included in post-2015 Sustainable Development Goal projects says Joshua Salomon and colleagues.


Trends in Breastfeeding Interventions, Skin-to-Skin Care, and Sudden Infant Death in the First 6 Days after Birth.
To determine if implementation of skin-to-skin care and the Baby-Friendly Hospital Initiative (BFHI) contributes to sudden unexpected infant death (SUID) and asphyxia in the first 6 days after birth. Survey data were used to determine a correlation between BFHI and deaths from SUID and asphyxia among infants <7 days in the US and Massachusetts. Using data from the Centers for Disease Control and Prevention, implementation of BFHI was tracked from 2004-2016 and skin-to-skin care was tracked from 2007-2015. Using data from Centers for Disease Control and Prevention WONDER and the Massachusetts Department of Public Health, SUID and asphyxia were tracked from 2004-2016. Nationally, births in Baby-Friendly facilities rose from 1.8% to 18.3% and the percentage of facilities in which most dyads experienced skin-to-skin care rose from 40% to 83%. SUID prevalence among infants <7 days was rare (0.72% of neonatal deaths) and decreased significantly from 2004-2009 compared with 2010-2016, from 0.033 per 1000 live births to 0.028, OR 0.85 (95% CI 0.77, 0.94). In Massachusetts, births in Baby-Friendly facilities rose from 2.8% to 13.9% and skin-to-skin care rose from 50% to 97.8%. SUID prevalence decreased from 2010-2016 compared with 2004-2009: OR 0.32 (95% CI 0.13, 0.82), with 0 asphyxia deaths during the 13-year period. Increasing rates of breastfeeding initiatives and skin-to-skin care are temporally associated with decreasing SUID prevalence in the first 6 days after birth in the US and Massachusetts.


Adherence to insulin therapy and associated factors among patients with diabetes mellitus in public hospitals of Central Zone of Tigray, Ethiopia, 2018: a cross-sectional study.
Adherence to insulin therapy is a critical factor for adequate control of diabetes mellitus. Despite the multiple well-known benefits of adherence to insulin therapy, poor adherence remains to be a common cause of diabetes mellitus-related complications. A better management of diabetes mellitus requires determining the level of patient adherence and identifying why non-adherence to insulin therapy occurs. Therefore, this study was designed to assess the level of adherence to insulin therapy and associated factors among diabetes mellitus patients. The study was conducted from May 1 to July 1, 2018, using a cross-sectional study design. Interviewer-administered questionnaire was employed for data collection and systematic random sampling technique was used to select study participants. The collected data were entered using Epi data version 3.1.1 and exported to SPSS version 22 for analysis. Logistic analysis was carried out to check the level of association between adherence to insulin therapy and the independent variables with significance level of 0.05 at 95% confidence interval. 273 respondents were selected with a 100% response rate. Near to one-fourth (24.2%) of the respondents were adherent to their insulin therapy. The study revealed that good knowledge of diabetes mellitus [AOR=6.51; 95% CI [1.58, 26.71], age [>30 years] [AOR=2.63; 95% CI [1.27, 5.42], knowledge regarding insulin self-injection [AOR=4.21; 95%CI [1.06,16.65], favorable attitude towards insulin injection [AOR=2.14; 95% CI [1.04,4.41], free-of-cost insulin therapy [AOR= 4.62, 95% CI [1.06,16.65], having of glucometer at home [AOR= 2.82, 95% CI [1.12,7.09], and being a member of Ethiopian diabetic association [AOR= 5.41, 95% CI [2.31,12.64] were found to significantly affect adherence to insulin therapy. Nearly one-fourth of the study participants were adherent to their insulin therapy. Good knowledge and favorable attitude towards insulin injection, good knowledge regarding diabetes mellitus, being a member of the Ethiopian Diabetes Association, age greater than thirty years old, free-of-cost insulin therapy and having glucometer at home were found to be significant predictors of adherence to insulin therapy.


Synthesis and evaluation of a photolyzable derivative of sphingosine 1-phosphate--caged SPP.
The synthesis of a photolyzable sphingosine 1-phosphate derivative is reported via the reaction of N-(tert-butoxycarbonyl)-2-N,3-O- isopropylidenesphingosine 7 and bis(alpha-methyl-o-nitrobenzyl) N,N-diisopropyl-phosphoramidite. Stimulation of DNA synthesis upon illumination of caged SPP-loaded cells is also described.


Glutathione disulfide production during arachidonic acid oxygenation in human platelets.
Washed human platelets stimulated with 50 microM sodium arachidonate rapidly accumulated glutathione disulfide to a peak concentration of 0.620 nmole per 10(9) cells, 200% of control (unstimulated) levels. Total glutathione remained unchanged. The rise in glutathione disulfide was transitory, returning to control values within 30 seconds in aggregating platelets. Similar findings were observed in washed platelets aggregated with 5 U/ml thrombin. Platelet aggregation was not necessary for the generation of glutathione disulfide. However, cyclooxygenase activity was necessary for the generation of glutathione disulfide. Aspirin treated platelets aggregated with thrombin demonstrated no thromboxane B2 production and no glutathione disulfide generation. Dose response studies with both agonists demonstrated a direct relationship between the amount of thromboxane B2 produced and the amount of glutathione disulfide generated by stimulated platelets. During the conversion of arachidonic acid to thromboxane B2, unesterified arachidonic acid is oxygenated to prostaglandin G2 which is subsequently reduced to prostaglandin H2. Both reactions are catalyzed by the enzyme prostaglandin H synthase. Our data support the hypothesis that glutathione is an important supplier of reducing equivalents to prostaglandin H synthase during the production of prostaglandin H2 in human platelets.


Effect of some abiotic factors on the biological activity of Gluconacetobacter diazotrophicus.
The effect of some abiotic factors, dryness, heat and salinity on the growth and biological activity of Gluconacetobacter diazotrophicus, and the influence of a salt stress on some enzymes involved in carbon metabolism of these bacteria is studied under laboratory conditions. Strain PAL-5 of G. diazotrophicus was incubated under different conditions of drying, heat and salinity. Cells showed tolerance to heat treatments and salt concentrations, and sensitivity to drying conditions. Higher NaCl dosage of 150 and 200 mmol l -1 limited its growth and drastically affected the nitrogenase activity and the enzymes glucose dehydrogenase, alcohol dehydrogenase, fumarase, isocitrate dehydrogenase and malate dehydrogenase. Gluconacetobacter diazotrophicus, despite its endophytic nature, tolerated heat treatments and salinity stress, but its nitrogenase activity and carbon metabolism enzymes were affected by high NaCl dosage. The investigation of the biological activity of G. diazotrophicus in response to different abiotic factors led to more knowledge of this endophyte and may help to clarify pathways involved in its transmission into the host plant.


Novel and improved Caenorhabditis briggsae gene models generated by community curation.
The nematode Caenorhabditis briggsae has been used as a model in comparative genomics studies with Caenorhabditis elegans because of their striking morphological and behavioral similarities. However, the potential of C. briggsae for comparative studies is limited by the quality of its genome resources. The genome resources for the C. briggsae laboratory strain AF16 have not been developed to the same extent as C. elegans. The recent publication of a new chromosome-level reference genome for QX1410, a C. briggsae wild strain closely related to AF16, has provided the first step to bridge the gap between C. elegans and C. briggsae genome resources. Currently, the QX1410 gene models consist of software-derived gene predictions that contain numerous errors in their structure and coding sequences. In this study, a team of researchers manually inspected over 21,000 gene models and underlying transcriptomic data to repair software-derived errors. We designed a detailed workflow to train a team of nine students to manually curate gene models using RNA read alignments. We manually inspected the gene models, proposed corrections to the coding sequences of over 8,000 genes, and modeled thousands of putative isoforms and untranslated regions. We exploited the conservation of protein sequence length between C. briggsae and C. elegans to quantify the improvement in protein-coding gene model quality and showed that manual curation led to substantial improvements in the protein sequence length accuracy of QX1410 genes. Additionally, collinear alignment analysis between the QX1410 and AF16 genomes revealed over 1,800 genes affected by spurious duplications and inversions in the AF16 genome that are now resolved in the QX1410 genome. Community-based, manual curation using transcriptome data is an effective approach to improve the quality of software-derived protein-coding genes. The detailed protocols provided in this work can be useful for future large-scale manual curation projects in other species. Our manual curation efforts have brought the QX1410 gene models to a comparable level of quality as the extensively curated AF16 gene models. The improved genome resources for C. briggsae provide reliable tools for the study of Caenorhabditis biology and other related nematodes.


Isolation of a potential anchoring motif based on proteome analysis of Escherichia coli and its use for cell surface display.
For bacterial cell surface display, the target protein needs to be linked to an anchoring motif, and it is essential to choose an appropriate anchoring motif for efficient and stable display of the protein on the cell surface. To isolate a potential anchoring motif that would allow a stable and enhanced display of target proteins on the surface of an Escherichia coli host, we analyzed the outer membrane proteome of E. coli. On the basis of this proteomic analysis, the outer membrane protein X (OmpX), which has a small, monomeric β-barrel structure and is highly expressed, was selected as a potential anchoring motif. The role of OmpX as an anchoring motif for cell surface display was demonstrated using three important industrial enzymes: endoxylanase, lipase, and alkaline phosphatase. Two different positions (Lys(122), Val(160)) in the extracellular loops of OmpX were examined for C-terminal fusion, and the biological activities and localization of the displayed enzymes were analyzed. All three enzymes examined were efficiently displayed on the E. coli cell surface with high activity. These results reveal that the use of OmpX as an anchoring motif is an efficient method to display functional enzymes on the surface of an E. coli host.


Continuous in vitro cultivation of a recently identified Babesia that infects small ruminants in China.
Babesia sp. Xinjiang was isolated from a splenectomised sheep infested by Rhipicephalus sanguineus and Hylomma anatolicum anatolicum, collected from sheep and cattle in Xinjiang province. It was considered to be a novel ovine Babesia species on the basis of its morphology, pathogenicity, vector tick species and alignments of 18S ribosomal RNA (18S rRNA) and internal transcribed spacers (ITS) gene sequences. Continuous in vitro cultures of the ovine parasite were established using infected sheep blood. In RPMI 1640 medium with 7.5% sheep red blood cells (RBCs) maintained in an incubator at 37 °C and 5% CO(2), the percentage of parasitized erythrocytes (PPE) peaked at 10% in 24- and 6-well plates. It increased to 20-50% with the same culture medium but with 2.5% RBC in 75 cm(2) flasks. Two clonal lines of Babesia sp. Xinjiang were screened using the limiting dilution method. Growth characteristics of these lines in vitro were measured by a microtiter-based spectrophotometric method and from the PPE. The generation time in sheep erythrocytes was between 15.20 h and 16.27 h. Furthermore, the host range of parasite was identified with in vitro culture and in vivo infection. Erythrocytes of sheep, cattle, sika deer and humans could be invaded into by lines in vitro, but the parasites could not propagate in human erythrocytes. The parasites could not enter erythrocytes from goats in vitro. However, in vivo, only sheep could be infected by lines. Finally, a Babesia sp. Xinjiang-like parasite (which shared 99.5% identity with the original strain of Babesia sp. Xinjiang) was isolated using this in vitro culture system from 1 of 19 sheep blood samples collected from western Gansu province, China.


When should Home-visit nurses initiate end-of-life discussions for patients with Organ failure and family caregivers? A qualitative study.
End-of-life (EOL) discussions for organ-failure patients with family caregivers are important factors for successful EOL care. However, identifying the appropriate time to initiate these discussions is difficult owing to the unpredictability of the disease trajectory. No practical tools or clinical indicators currently exist that can help identify non-cancer patients receiving home care who need EOL discussions. The survey was conducted from February 2020 to June 2021. To identify the appropriate time at which to initiate EOL discussions for patients with organ failure and their caregivers, we determined the time when home-visit nurses initiated EOL discussions. We interviewed 19 home-visit nurses (mean total home-visit nursing experience: 6.7 ± 5.9 years) and analyzed the data using Hsieh and Shannon's qualitative content approach. Three themes related to home-visit nurses' experiences of identifying the appropriate time to start EOL discussions were identified: symptomatic worsening, lack of patients' and family caregivers' EOL awareness, and decline in activities of daily living. It is necessary to develop a tool that will enable home-visit nurses to implement EOL discussions at the appropriate time.


Structure-activity profiles of macrolactam immunosuppressant FK-506 analogues.
The immunosuppressive agent FK-506 has received much attention due to its efficacy and potency in the areas of transplant rejection and autoimmune disease. Calcineurin, a Ca(2+)-calmodulin activated phosphatase, was recently implicated in the immunosuppressive mechanism of FK-506. In our ongoing search for superior immunosuppressive agents, we have synthesized several analogues of FK-506 and tested their mechanistic and immunosuppressive actions. It was found that C-18 hydroxyl analogues of ascomycin, an analogue of FK-506 also called FR900520, bound tightly to immunophilin FKBP-12, but do not show any immunosuppressive activity in vitro or in vivo despite good bioavailability. Further, they reverse the inhibition of calcineurin caused by FK-506/FKBP-12 complex.


Using patient-specific induced pluripotent stem cells to interrogate the pathogenicity of a novel retinal pigment epithelium-specific 65 kDa cryptic splice site mutation and confirm eligibility for enrollment into a clinical gene augmentation trial.
Retinal pigment epithelium-specific 65 kDa (RPE65)-associated Leber congenital amaurosis is an autosomal recessive disease that results in reduced visual acuity and night blindness beginning at birth. It is one of the few retinal degenerative disorders for which promising clinical gene transfer trials are currently underway. However, the ability to enroll patients in a gene augmentation trial is dependent on the identification of 2 bona fide disease-causing mutations, and there are some patients with the phenotype of RPE65-associated disease who might benefit from gene transfer but are ineligible because 2 disease-causing genetic variations have not yet been identified. Some such patients have novel mutations in RPE65 for which pathogenicity is difficult to confirm. The goal of this study was to determine if an intronic mutation identified in a 2-year-old patient with presumed RPE65-associated disease was truly pathogenic and grounds for inclusion in a clinical gene augmentation trial. Sequencing of the RPE65 gene revealed 2 mutations: (1) a previously identified disease-causing exonic leucine-to-proline mutation (L408P) and (2) a novel single point mutation in intron 3 (IVS3-11) resulting in an A>G change. RT-PCR analysis using RNA extracted from control human donor eye-derived primary RPE, control iPSC-RPE cells, and proband iPSC-RPE cells revealed that the identified IVS3-11 variation caused a splicing defect that resulted in a frameshift and insertion of a premature stop codon. In this study, we demonstrate how patient-specific iPSCs can be used to confirm pathogenicity of unknown mutations, which can enable positive clinical outcomes.


Selection and self-organization of self-reproducing macromolecules under the constraint of constant flux.
We investigate the dynamic behavior of a set of self-reproducing macromolecules (e.g., polynucleotides) under conditions such that the fluxes of all monomer units into the system are kept constant. Such conditions might prevail in an evolution reactor or in certain naturally occurring situations. A general set of equations is developed to describe the behavior of both the macromolecule and the monomer concentrations. The question of how the rate of macromolecule synthesis varies with the monomer levels is discussed briefly. With the help of several physically reasonable approximations, we obtain an exact solution for a simplified constant flux system. Comparison with the corresponding system under the constraint of constant overall organization reveals important similarities, most notably in the existance and composition of quasispecies. Given the same set of physical and chemical parameters, a system subject to constant flux will always evolve toward selective equilibrium more slowly than under the constraint of constant organization.


Prevention of early childhood caries (ECC)--review of literature published 1998-2007.
This was to examine the literature published during the last decade and review the effectiveness of methods used for the prevention of early childhood caries (ECC). A critical review of papers. A broad search of the PubMed database was conducted from 1998 through September 2007, using "early childhood caries", "baby bottle tooth decay", "nursing caries", "infant caries", "caries prevention" and "oral health education" as index terms. Relevant papers published in English between 1998 and 2007 were identified after a review of their abstracts. Papers were selected if they reported a prospective controlled design with preventive or non-invasive intervention directed to children under the age of 3 years. A defined endpoint measure of cavitated or non-cavitated clinical caries, expressed as incidence or prevalence, was required. The targeted publications were critically assessed by the author concerning design, methodology and performance. The initial search revealed 66 papers of which 22 met the inclusion criteria. The results reinforced the role of fluoride toothpaste as the most cost-effective home-care measure and semi-annual fluoride varnish applications as the best professional method for infants at risk. The evidence concerning the preventive effect of antibacterial agents, primary-primary prevention and dental health education were inconclusive but the included studies supported the importance of early start, outreach activities and motivational interviewing as key factors to overcome cultural and socioeconomic barriers. Although there is a body of evidence for the use of fluoride in preventing ECC, further high-quality studies are needed to further establish the best way to maintain oral health in infants.