CK1δ stimulates ubiquitination-dependent proteasomal degradation of ATF4 to promote chemoresistance in gastric Cancer.
Chemoresistance remains a major obstacle to successful cancer therapy, especially for advanced cancers. It used to be recognised as a stable outcome resulting from genetic changes. However, recent studies showed that chemoresistance can also be unstable and reversible with the involvement of non-genetic alterations. In the present study, we found that activating transcription factor 4 (ATF4) is downregulated in chemoresistant gastric cancer cells. The over-expression of ATF4 reversed chemoresistance by activating CHOP transcription to enhance drug-induced apoptosis, and vice versa. Moreover, casein kinase 1 delta (CK1δ) was identified as the kinase responsible for ATF4-S219 phosphorylation, which triggered βTrCP-mediated ATF4 polyubiquitination to promote its proteasomal degradation subsequently. Interestingly, drug withdrawal gradually restored chemosensitivity as well as ATF4 expression in chemoresistant cells, highlighting the dependence of dynamic drug resistance on ATF4 protein expression. In line with these findings, the inhibition of ATF4 protein degradation by CK1δ or proteasome inhibitors overcame chemoresistance both in vitro and in vivo. Taken together, these results indicate that CK1δ stimulates βTrCP-dependent ATF4 polyubiquitination and subsequent proteasomal degradation to promote chemoresistance in gastric cancer. Stabilisation of the ATF4 protein with bortezomib (BTZ), an anticancer drug that inhibits proteasomal degradation, might be a rational strategy to improve chemotherapeutic efficacy in gastric cancer.


Reinstatement of nicotine conditioned place preference in a transgenerational model of drug abuse vulnerability in psychosis: Impact of BDNF on the saliency of drug associations.
Psychotic disorders such as schizophrenia are often accompanied by high rates of cigarette smoking, reduced quit success, and high relapse rates, negatively affecting patient outcomes. However, the mechanisms underlying altered relapse-like behaviors in psychosis are poorly understood. The present study analyzed changes in extinction and reinstatement of nicotine conditioned place preference (CPP) and resulting changes in brain-derived neurotrophic factor (BDNF) in a novel heritable rodent model of psychosis, demonstrating increased dopamine D2 receptor sensitivity, to explore mechanisms contributing to changes in relapse-like behaviors. Male and female offspring of two neonatal quinpirole-treated (1 mg/kg quinpirole from postnatal day (P)1-21; QQ) and two neonatal saline-treated (SS) Sprague-Dawley rats (F1 generation) were tested on an extended CPP paradigm to analyze extinction and nicotine-primed reinstatement. Brain tissue was analyzed 60 min after the last nicotine injection for BDNF response in the ventral tegmental area (VTA), the infralimbic (IfL) and prelimbic (PrL) cortices. F1 generation QQ offspring demonstrated delayed extinction and more robust reinstatement compared to SS control animals. In addition, QQ animals demonstrated an enhanced BDNF response to nicotine in the VTA, IfL and Prl cortices compared to SS offspring. This study is the first to demonstrate altered relapse-like behavior in a heritable rodent model with relevance to comorbid drug abuse and psychosis. This altered pattern of behavior is hypothesized to be related to elevated activity-dependent BDNF in brain areas associated with drug reinforcement during conditioning that persists through the extinction phase, rendering aberrantly salient drug associations resistant to extinction and enhancing relapse vulnerability.


Childhood modifiable risk factors and later life chronic kidney disease: a systematic review.
Relationships between adulthood modifiable risk factors and chronic kidney disease (CKD) are well-established, but associations with childhood risk factors are unclear. This study systematically assesses the published evidence about childhood modifiable risk factors and adulthood CKD. We searched MEDLINE, EMBASE, and Web of Science to 6th May 2022. Articles were included if (1) they were population-based longitudinal studies, (2) exposures were potentially modifiable, for example through pharmacological or lifestyle modifications, including clinical conditions/measures (diabetes, blood pressure, adiposity, and dyslipidaemia); health behaviours (smoking, alcohol consumption, physical activity, fitness, and poor nutrition); and socio-economic factors (socio-economic position), and occurred during childhood (ages 2-19 years), and (3) outcome was CKD or surrogate markers of CKD in adulthood (ages 20 years or older). Three reviewers independently extracted the data. 15,232 articles were identified after deduplication; 17 articles met the inclusion criteria, reporting childhood blood pressure (n = 8), adiposity (n = 4), type 2 diabetes (n = 1), socio-economic position (n = 1), famine (n = 1), cardiorespiratory fitness (n = 1), and a healthy lifestyle score (n = 1). The results suggested positive associations of childhood adiposity, type 2 diabetes, and low socio-economic position and cardiorespiratory fitness in females with CKD in adulthood. Findings were inconsistent on associations between childhood BP and CKD in adulthood. Childhood healthy lifestyle score and exposure to famine were not associated with risk of CKD in adulthood. The limited evidence suggests childhood factors may contribute to the CKD risk in adulthood, particularly adiposity, type 2 diabetes, and low socio-economic position and cardiorespiratory fitness in females. Further high-quality community-based studies are needed with long-term follow-up and investigation of a broader range of modifiable risk factors.


Realistic and affordable lo-fidelity model for learning bronchoscopic transbronchial needle aspiration.
Transbronchial needle aspiration (TBNA) is used to sample mediastinal abnormalities and lymph node stations for diagnostic purposes and lung cancer staging. The procedure is underused, operator dependent, and reputed to have a steep learning curve. Other difficulties arise from a bronchoscopist's failure to insert the needle satisfactorily into the target node. The purpose of this study was to evaluate the realism and helpfulness of a lo-fidelity, easily constructed hybrid model used for learning and practicing TBNA. The model is constructed by attaching a porcine tracheobronchial tree to a Laerdal Airway Model mounted on polyvinyl chloride (PVC) piping. Twelve individuals with various levels of bronchoscopy training and experience were given a 15-min introductory PowerPoint presentation on TBNA strategy and planning, execution, and response to complications. Participants then practiced TBNA alone and with guidance, aided by an assistant, as many times as individually necessary to feel comfortable with the procedure. A five-point Likert scale 8-item questionnaire was then completed. Participants were unanimously positive about their experience (mean scores 4.25-4.91). The model was realistic, provided increased comfort with TBNA techniques, and allowed practice of communication skills. This realistic, affordable, and easily constructed hybrid lo-fidelity airway model allows beginner and experienced bronchoscopists opportunities to learn and practice basic TBNA techniques and team communication skills without placing patients at risk.


Effects of phytase-supplemented fermentation and household processing on the nutritional quality of Lathyrus sativus L. seeds.
Grass pea (Lathyrus sativus L.) is commonly consumed in cooked, fermented, and roasted forms in Ethiopia. However, the impacts of household processing practices on its nutrients, antinutrients, and toxic compounds have not been adequately studied. Therefore, the effects of household processing and fermentation in the presence and absence of a phytase on the contents of β-N-oxalyl-L-α,β-diaminopropionic acid (β-ODAP), myo-inositol phosphates, crude protein, minerals and the in vitro bioaccessibility were investigated. Fermentation exhibited a significant decline in β-ODAP (13.0-62.0%) and phytate (7.3-90.5%) irrespective of the presence of phytase. Pressure and pan cooking after discarding the soaking water resulted in a 27.0 and 16.2% reduction in β-ODAP. A 30% reduction in phytate was observed during germination followed by roasting. In addition, germination resulted in a significant (p < 0.05) increase in crude protein. Germination and germination followed by roasting resulted in the highest Fe bioaccessibilities (more than 25 fold higher compared to untreated samples) followed by pressure cooking and soaking. Processing also improved Zn bioaccessibilities by 50.0% (soaked seed without soaking water), 22.5% (soaked seed with soaking water), and 4.3% (germination). Thus, the processing technologies applied were capable of reducing the content of phytate (InsP6) and β-ODAP with a concomitant increase in mineral bioaccessibilities. Processing of grass peas could therefore contribute to their more widespread utilization.


Psychotic states associated with disorders of thyroid function.
Thyroid-related psychoses continue to pose diagnostic and treatment challenges for clinicians. Two case histories illustrate diverse clinical states associated with hyper- and hypo-thyroidism respectively and highlight the need to consider the possibility of thyroid disorder in all patients presenting with acute psychotic mental disorder. They also demonstrate treatment methods directed at control of psychotic symptoms and restoration of an euthyroid state.


The influence of aging and attentional demands on recovery from postural instability.
It is well known that the risk of a debilitating injury from a fall is much higher for elderly than for young individuals. In addition, it is well documented that healthy elderly subjects exhibit increased postural sway during normal stance tasks. In the present experiment, we explored the notion that control of minor postural instability in elderly subjects is attention demanding. Postural sway of eight elderly (mean age = 70.0 years) and eight young (mean age = 20.0 years) subjects was measured under two different secondary demands during stable and mildly unstable upright stance. There were two types of work loads. Either a cognitive (math task) or motor (hand-squeeze) task was performed during the second segment of a 50-second standing trial. The effect of these work loads on mean velocity, range, and variability of range of center of foot pressure was measured during the destabilizing activity of arm swinging and subsequent recovery period. Following seven seconds of 1 Hz arm-swinging activity, elderly subjects showed a marked increase in recovery time to normal stance when concurrently performing an arithmetic task. This result suggests that recovery from a posturally destabilizing activity, involving proprioceptive and vestibular information, places increased attentional demands on the postural support system of the elderly.


Continuous enzymatic biodiesel production from coconut oil in two-stage packed-bed reactor incorporating an extracting column to remove glycerol formed as by-product.
The transesterification of coconut oil with ethanol catalyzed by Burkholderia cepacia lipase immobilized on polysiloxane-polyvinyl alcohol was performed in a continuous flow. The experimental design consisted of a two-stage packed-bed reactor incorporating a column with cationic resin (Lewatit GF 202) to remove the glycerol formed as by-product and the reactor performance was quantified for three different flow rates corresponding to space-times from 10 to 14 h. The influence of space-time on the ethyl ester (FAEE) concentrations, yields and productivities was determined. The reactor operation was demonstrated for space-time of 14 h attaining FAEE concentrations of 58.5 ± 0.87 wt%, FAEE yields of 97.3 ± 1.9 % and productivities of 41.6 ± 1.0 mgester g medium (-1) h(-1). Biodiesel purified samples showed average kinematic viscosity values of 5.5 ± 0.3 mm(2) s(-1) that meet the criteria established by the American National Standard ASTM (D6751). The immobilized lipase was found to be stable regarding its morphological and catalytic characteristics, showing half-life time (t 1/2) around 1540 h. The continuous packed-bed reactor connected in series with simultaneous glycerol removal has a great potential to attain high level of transesterification yields, raising biodiesel productivity.


Dishevelled phase separation promotes Wnt signalosome assembly and destruction complex disassembly.
The amplitude of Wnt/β-catenin signaling is precisely controlled by the assembly of the cell surface-localized Wnt receptor signalosome and the cytosolic β-catenin destruction complex. How these two distinct complexes are coordinately controlled remains largely unknown. Here, we demonstrated that the signalosome scaffold protein Dishevelled 2 (Dvl2) undergoes liquid-liquid phase separation (LLPS). Dvl2 LLPS is mediated by an intrinsically disordered region and facilitated by components of the signalosome, such as the receptor Fzd5. Assembly of the signalosome is initiated by rapid recruitment of Dvl2 to the membrane, followed by slow and dynamic recruitment of Axin1. Axin LLPS mediates assembly of the β-catenin destruction complex, and Dvl2 attenuates LLPS of Axin. Compared with the destruction complex, Axin partitions into the signalosome at a lower concentration and exhibits a higher mobility. Together, our results revealed that Dvl2 LLPS is crucial for controlling the assembly of the Wnt receptor signalosome and disruption of the phase-separated β-catenin destruction complex.


Comparison of echocardiography at peak exercise and after bicycle exercise in evaluation of patients with known or suspected coronary artery disease.
To determine if echocardiography done immediately after bicycle exercise provides the same information as imaging at peak exercise, we evaluated 104 consecutive patients being studied for coronary artery disease. Interpretable exercise echocardiograms were obtained in 96 patients (92%), 29 of whom had a new wall motion abnormality detected with exercise echocardiography. Of these 29 patients, 10 had a wall motion abnormality detected in apical views obtained during peak exercise that resolved by the time apical imaging was performed after exercise. Three of these 10 patients, however, had wall motion abnormalities in parasternal views taken after exercise in areas adjacent to the wall motion abnormality imaged at peak exercise. The sensitivity of exercise-induced wall motion abnormality for the detection of significant coronary artery disease in those patients undergoing coronary arteriography was 70% for imaging done after exercise versus 100% for imaging done at peak exercise. Six patients' conditions would have been misclassified as normal if only imaging done after exercise had been performed. We conclude that the addition of echocardiographic imaging at peak exercise on a bicycle enhances the sensitivity for the detection of coronary artery disease with exercise echocardiography.


Expression of the SIN3 homologue from banana, MaSIN3, suppresses ABA responses globally during plant growth in Arabidopsis.
The SIN3 family of co-repressors is a family of highly conserved eukaryotic repressor proteins that regulates diverse functions in yeasts and animals but remains largely uncharacterized functionally even in plants like Arabidopsis. The sole SIN3 homologue in banana, MaSIN3, was identified as a 1408 amino acids, nuclear localized protein conserved to other SIN3s in the PAH, HID and HCR domains. Interestingly, MaSIN3 over-expression in Arabidopsis mimics a state of reduced ABA responses throughout plant development affecting growth processes such as germination, root growth, stomatal closure and water loss, flowering and senescence. The reduction in ABA responses is not due to reduced ABA levels but due to suppression of expression of several transcription factors mediating ABA responses. Transcript levels of negative regulators of germination (ABI3, ABI5, PIL5, RGL2 and RGL3) are reduced post-imbibition while those responsible for GA biosynthesis are up-regulated in transgenic MaSIN3 over-expressers. ABA-associated transcription factors are also down-regulated in response to ABA treatment. The HDAC inhibitors, SAHA and sodium butyrate, in combination with ABA differentially suppress germination in control and transgenic lines suggesting the recruitment by MaSIN3 of HDACs involved in suppression of ABA responses in different processes. The studies provide an insight into the ability of MaSIN3 to specifically affect a subset of developmental processes governed largely by ABA.


A randomised trial of an intervention to develop health promoting schools in Australia: the south western Sydney study.
This paper describes the evaluation of a health promoting schools intervention carried out in the south western region of Sydney, Australia. The evaluation sought to assess the project's impact on structures to support the health promoting school concept and changes in health-related policies and practices within 22 local schools. A randomised controlled study was employed. Intervention schools were offered seminars and training in the health promoting schools concept, encouraged to use a resource kit to help them establish their school as health promoting and invited to participate in a support network. Pre- and post measures of awareness, school structures and policies and practices to support the development of a health promoting school were taken and intervention and control schools compared. There was an increased level of awareness of the health promoting school concept among intervention schools. However, there were no significant changes in health-related policies and practices at the school level, among both intervention and control schools. A longer time frame and more structured support are required to influence school structures to support the development of a health promoting school. To facilitate organisational change in schools, health promoting school interventions need to provide structured support for schools and recognise that change takes time. A committee to initiate action, a plan to guide work and school support are likely to facilitate the adoption of the health promoting schools approach. Further work to refine the measurement of school-based action and its outcome and the development of indicators for a health promoting school is needed.


Chitosan nanoemulsion: Gleam into the futuristic approach for preserving the quality of muscle foods.
Trend for consumption of healthy meat without synthetic additives is blooming globally and has attracted the interest of consumers and research sphere to look for enhancement of quality and safety of food. Chitosan is multi-functional marine biopolymer with several befitting properties such as non-toxicity, ease of modification, antimicrobial activity, biodegradability and bio-compatibility, making it suitable for use in meat based food systems, which are highly prone to putrescence due to availability of high level protein, micronutrients and moisture. Bioactive components from plant extracts on account of their natural lineage are exquisite determinants for meat preservation in association with chitosan to replace synthetic molecules, which are considered to evince toxicological effects. Nanoemulsions are viable systems for integrating a myriad of active constituents framed by microfluidization, high-pressure homogenization, ultra-sonication, phase inversion (PIC and PIT) and spontaneous-emulsification with benefits of droplet size reduction, improved solubility, stability and their biological activity. This article summarizes the most important information on formulation, fabrication and advancements in chitosan-based nanoemulsions highlighting their potential benefit for applications in the muscle food system. Supervising the all-around executions of chitosan nanoemulsions for various food systems, the current review has been framed to lay down understandings regarding improvements made in the production and functionality of chitosan nanoemulsions for quality retention of meat products. Furthermore, it highlights the novel trends in chitosan-nanoemulsions application in meat based food systems from a preservation and shelf-life prolongation perspective.


Differentiation of compressive from glaucomatous optic neuropathy with spectral-domain optical coherence tomography.
To compare optic disc topography in eyes with compressive optic neuropathy (CON) and open-angle glaucoma (OAG) using spectral-domain (SD) optical coherence tomography (OCT) and Heidelberg retinal tomograph (HRT) (Heidelberg Engineering GmbH, Heidelberg, Germany). Cross-sectional, observational study. A total of 200 eyes from 123 patients with CON (69 eyes) or OAG (58 eyes) and controls (73 eyes). Univariate and multivariate analyses of HRT parameters, SD-OCT circumpapillary retinal nerve fiber layer (RNFL) thickness, and optic nerve head (ONH) parameters. Circumpapillary RNFL, OCT ONH parameters, and HRT parameters. The univariate analysis of OCT parameters demonstrated significant differences between the temporal and nasal quadrants; clock hours 3 (55 vs. 73 μm), 4, 8 (93.9 vs. 70.7 μm), 9, and 10; vertical cup-to-disc ratio (C:D) (0.6 vs. 0.8) and cup volume (0.2 vs. 0.5) (P<0.001) between patients with CON and OAG, respectively. The CON discs were significantly different from normal discs for all OCT parameters except cup volume. The CON discs were not significantly different from normal discs for HRT parameters, except for mean RNFL thickness and cup shape measure. The OAG discs were significantly different from normal discs in all HRT and OCT parameters (P<0.001). Multivariate analysis demonstrated that the OCT 3 o'clock temporal sector, average C:D ratio, vertical C:D ratio, and cup volume measurements were able to differentiate OAG from CON. Compressive optic neuropathy is associated with significantly thinner nasal and temporal sectors compared with OAG, whereas OAG results in larger cups and cup volume with OCT measurements. The Heidelberg retinal tomograph is not able to differentiate CON from normal discs.


Mechanistic and structural basis for activation of cardiac myosin force production by omecamtiv mecarbil.
Omecamtiv mecarbil is a selective, small-molecule activator of cardiac myosin that is being developed as a potential treatment for heart failure with reduced ejection fraction. Here we determine the crystal structure of cardiac myosin in the pre-powerstroke state, the most relevant state suggested by kinetic studies, both with (2.45 Å) and without (3.10 Å) omecamtiv mecarbil bound. Omecamtiv mecarbil does not change the motor mechanism nor does it influence myosin structure. Instead, omecamtiv mecarbil binds to an allosteric site that stabilizes the lever arm in a primed position resulting in accumulation of cardiac myosin in the primed state prior to onset of cardiac contraction, thus increasing the number of heads that can bind to the actin filament and undergo a powerstroke once the cardiac cycle starts. The mechanism of action of omecamtiv mecarbil also provides insights into uncovering how force is generated by molecular motors.Omecamtiv mecarbil (OM) is a cardiac myosin activator that is currently in clinical trials for heart failure treatment. Here, the authors give insights into its mode of action and present the crystal structure of OM bound to bovine cardiac myosin, which shows that OM stabilizes the pre-powerstroke state of myosin.


New guidance for self-harm: an opportunity not to be missed.
In this editorial we, as members of the 2022 NICE Guideline Committee, highlight and discuss what, in our view, are the key guideline recommendations (generated through evidence synthesis and consensus) for mental health professionals when caring for people after self-harm, and we consider some of the implementation challenges.


The innovators. Competitive pressures inspire creativity in clinical operations.
While medical practice administrators are expected to competently handle the day-to-day management of a practice, appropriately assessing and responding to the larger pressures may be the most important thing you do. Count innovation among the critical management skills for the profession. Learn the inspired solutions of three administrators to problems in their practices.


Extracellular Vesicles as Communicators of Senescence in Musculoskeletal Aging.
Extracellular vesicles (EVs), including exosomes and microvesicles, are released by numerous cell types. EVs are now acknowledged as playing a critical role in cell-cell communication in healthy aging as well as in age-related diseases. Recently it was shown that senescence, a key hallmark of aging, increases the secretion of EVs. Moreover, EVs can transport proteins and microRNAs (miRNAs) that are key components of the senescence-associated secretory phenotype (SASP). Here we review evidence that SASP-related miRNAs are involved in musculoskeletal degeneration with aging. Specifically, senescence-related miRNAs are elevated in EVs released by skeletal muscle myocytes and fibro-adipogenic progenitor cells with aging and disuse atrophy, respectively. Many of these same senescence-related miRNAs are detected in EVs from the synovial fluid of patients with osteoarthritis, and these miRNAs can contribute to cartilage degeneration. Finally, senescence-associated miRNAs are secreted from bone marrow-derived stem (stromal) cells impacting neighboring hematopoietic stem cells and circulating in the blood. The senescence-associated miRNA mir-34a, which is known to target Wnt and Notch pathways as well as the cell survival factors Sirt1 and Bcl2, is detected in EVs from human and animal subjects with muscle atrophy, bone loss, and osteoarthritis. These findings suggest that suppressing the secretion of EV-derived, senescence-related miRNAs, such as miR-34a, or increasing levels of competing endogenous long noncoding RNAs, such as MALAT1 that inhibit miR-34a, may help to improve musculoskeletal function with aging. © 2022 The Authors. JBMR Plus published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research.


Emergency laser vaporization and helium-oxygen administration for acute malignant tracheobronchial obstruction.
Two patients with malignant airway obstruction and acute respiratory insufficiency were given emergency treatment with yttrium aluminum garnet (YAG) laser tissue vaporization under local anesthesia only. A mixture of oxygen and helium was administered to reduce the respiratory distress. This case emphasizes the rapidity and effectiveness of YAG laser treatment via fiberoptic bronchoscopy under local anesthesia in the management of acute malignant airway obstructions.


Alpha-fodrin (brain spectrin) immunocytochemical localization in rat vestibular hair cells.
The presence of a spectrin-related protein in rat vestibular sensory receptors was demonstrated by immunocytochemistry and immunoblotting using affinity purified anti alpha-fodrin antibodies. Intense immunoreactivity was found in the apical pole of sensory hair cells where it seems to be concentrated in the cuticular plate. In contrast, alpha-fodrin immunoreactivity was absent from the stereocilia. We suggest that a spectrin-related protein participates in the organization of the cuticular plate of vestibular hair cells by cross-linking actin filaments as well as by anchoring the cuticular plate to the apical cell membrane.


Localization of GABAA-receptor gamma 2-subunit mRNA-containing neurons in the rat central nervous system.
The localization of neurons containing mRNA of the gamma 2-subunit of GABAA receptors was examined in the rat central nervous system with in situ hybridization histochemistry using an oligonucleotide probe to the sequence of the gamma 2-subunit. Neurons containing the gamma 2-subunit mRNA were widely but unevenly distributed in the brain. The location of gamma 2-subunit mRNA-containing neurons differed from those containing alpha- or beta-subunits. According to our results brain regions can be divided into three categories: one containing only gamma 2-subunit, one containing mRNA of at least one subunit other than gamma 2 and one containing more than one other subunit but not the gamma 2-subunit. The distribution of strongly labeled nuclei partly coincided with that of glutamate decarboxylase, suggesting that the GABAA receptor gamma 2-subunit would be involved in an autoreceptive mechanism of the GABAergic transmission.


Hydrogen peroxide-induced apoptosis mediated by p53 protein in glial cells.
It is now generally accepted that massive neuronal death due to oxidative stress is a regular feature of brains in neurodegenerative diseases. However, much less attention has been given to the death of glial cells. In this study, we examined p53-sensitive apoptosis of cells by using human glioblastoma A172 cells and p53-deficient mouse astrocytes. In human A172 cells, hydrogen peroxide (H2O2) caused cell death in a time- and concentration-dependent manner, accompanied by nucleosomal DNA fragmentation and chromatin condensation. After treatment with H2O2, p53 protein was highly expressed and protein levels of Bak, p21WAF1/CIP1 and GADD45 were also enhanced. However, the protein levels of Bcl-2 and Bax did not change. On the other hand, primary cultured astrocytes from p53-deficient mouse brain grew faster than wild-type and heterozygous astrocytes. In addition, p53-deficient astrocytes were more resistant to H2O2-induced apoptosis than wild-type and heterozygous astrocytes. These results suggest that glial proliferation and the repair of damaged DNA may be regulated by p53-induced p21WAF1/CIP1 and GADD45, and that glial apoptosis caused by oxidative stress may be mediated by p53-induced Bak.


Studies of the growth of human bone-derived cells in culture using aqueous two-phase partition.
Human bone cells, maintained in culture, have been subjected to partitioning in an aqueous two-phase system on a countercurrent distribution apparatus. A broad cell distribution was obtained indicating cell-surface heterogeneity. Two major cell populations were identified which appeared to be growing at different rates. The 'fast'-growing cells had a less hydrophobic cell surface than the 'slow'-growing cells. Possible relationships of these cell populations with osteoblast differentiation and the potential importance of this technique in studies of osteoblast differentiation are discussed.


Facile synthesis of Fe3O4@mesoporous TiO2 microspheres for selective enrichment of phosphopeptides for phosphoproteomics analysis.
Protein phosphorylation is one of the most important post-translational modifications. Due to the dynamic nature and low stoichiometry of the protein phosphorylation, enrichment of phosphopeptides from proteolytic mixtures is necessary prior to their characterization by mass spectrometry. In this work, we synthesized Fe3O4@mesoporous TiO2 magnetic microspheres with core-shell structure and large surface area for selective enrichment of phosphopeptides. To demonstrate its ability for selective enrichment of phosphopeptides, we applied Fe3O4@mesoporous TiO2 magnetic microspheres to isolation and enrichment of the phosphopeptides from tryptic digestion of standard proteins and real samples, and then the enriched peptides were analyzed by matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS) or liquid chromatography coupled to electrospray ionization mass spectrometry (LC-ESI-MS). Due to that the as-made Fe3O4@mesoporous TiO2 microspheres have large surface area, good dispersivity and biocompatibility, they have been demonstrated as a powerful tool for phosphoproteomics research.


Nucleoside-Nucleotide Analog Combination Therapy Is Effective in Preventing Recurrent Hepatitis B After Liver Transplantation.
Hepatitis B immune globulin (HBIg) in combination with a nucleos(t)ide analog is the mainstay of prophylactic regimen to prevent recurrence of hepatitis B following orthotopic liver transplantation (OLT). HBIg therapy is costly and inconvenient for the patients. There is a growing experience converting HBIg/nucleos(t)ide to combination nucleotide/nucleoside analogs from. Twenty-six patients that underwent OLT between March 2001 and July 2011 who had received at least 12 months of HBIg and single nucleos(t)ide were enrolled. HBsAg and HBV DNA were undetectable, and anti-HBs were detectable at the time of switch. HBV DNA and HBsAg were measured every 3 months following discontinuation of HBIg and addition of nucleos(t)ide. Patients included 23 Asians/3 Caucasian, 21 males/5 females. Mean time of conversion from HBIg/nucleos(t)ide to nucleoside/nucleotide combination was 77.5 (range 11-132) months after OLT. Mean duration of follow-up after conversion was 31.9 (range 14-70) months. All patients had undetectable HBV DNA, and 24 patients remained HBsAg negative during follow-up. Two patients recurred 7 and 9 months later, respectively, with detectable HBsAg. Both patients continued to have undetectable HBV DNA and normal ALT. HBsAg was neutralized by reinfusion of HBIg. Nucleoside/nucleotide combination is an effective alternative to HBIg/nucleos(t)ide to prevent recurrence of hepatitis B after OLT.


The cytoplasmic peptide:N-glycanase (NGLY1) - Structure, expression and cellular functions.
NGLY1/Ngly1 is a cytosolic peptide:N-glycanase, i.e. de-N-glycosylating enzyme acting on N-glycoproteins in mammals, generating free, unconjugated N-glycans and deglycosylated peptides in which the N-glycosylated asparagine residues are converted to aspartates. This enzyme is known to be involved in the quality control system for the newly synthesized glycoproteins in the endoplasmic reticulum (ER). In this system, misfolded (glyco)proteins are retrotranslocated to the cytosol, where the 26S proteasomes play a central role in degrading the proteins: a process referred to as ER-associated degradation or ERAD in short. PNGase-mediated deglycosylation is believed to facilitate the efficient degradation of some misfolded glycoproteins. Human patients harboring mutations of NGLY1 gene (NGLY1-deficiency) have recently been discovered, clearly indicating the functional importance of this enzyme. This review summarizes the current state of our knowledge on NGLY1 and its gene product in mammalian cells.


Repair of congenital malformations of the mitral valve: early and midterm results.
The aims of this study were to determine early and midterm survival and freedom from reoperation, and to identify the predictors for poor postoperative outcome in children undergoing mitral valve (MV) repair owing to congenital malformations of the mitral valve. Between January 1990 and February 2001, 94 consecutive children with congenital MV disease underwent valve repair. The mean age was 5.2+/-3.3 years (range 20 days to 15 years). Twenty-five (26.6%) children were less than 1 year old. Isolated MV disease was found in 21 (22.4%) patients. MV stenosis was the predominant lesion in 21 (22.4%) patients with a mean left atrial to left ventricle diastolic peak gradient of 24.5+/-9.2 mm Hg. MV regurgitation was the predominant pathophysiology in 73 (77.6%) patients with a mean regurgitation grade of 3.3+/-0.7. The hospital mortality was 8.5% (8 of 94). Three patients required permanent pacemaker implantation owing to complete atrioventricular block. Two patients underwent mediastinal exploration for significant bleeding. Postoperatively the echocardiography color Doppler study demonstrated a significantly lower mean end diastolic left atrium to left ventricle gradient 8.7+/-2.2 mm Hg (p < 0.001) in patients with MV stenosis and a mean regurgitation grade of 0.9+/-0.6 (p < 0.001) in patients with MV regurgitation. Actuarial survival and actuarial reoperation-free survival were 89.2% and 76.3%, respectively. Multivariate analysis demonstrated that age less than 1 year (p = 0.035), hammock MV (p = 0.0093), cardiothoracic ratio greater than 0.6 (p < 0.0001), and associated cardiac anomalies (p = 0.003) were strong predictors for poor overall freedom from reoperation and midterm survival. Mitral valve repair for congenital mitral valve disease yields acceptable early and midterm mortality and reoperation rates. Strong predictors for poor overall freedom from reoperation and midterm survival were age less than 1 year, hammock MV, cardiothoracic ratio greater than 0.6, and associated cardiac anomalies.


Culture and proliferation of highly purified adult Schwann cells from rat, dog, and man.
This chapter presents fast and easy protocols to obtain highly purified cultures of proliferating adult rat, canine, and human Schwann cells. Cell preparation from predegenerated adult sciatic nerves combined with the use of melanocyte growth medium supplemented with forskolin, fibroblast growth factor-2, pituitary extract, and heregulin as selective, serum-free culture medium and two methods for a consecutive cell-enrichment step are described. Our protocols result in approximately 90% pure Schwann cell cultures (or higher). The average time to obtain highly purified in vitro cultures of adult Schwann cells is 21 days.


Mechanical preparation of the large bowel for elective surgery. Comparison of whole-gut lavage with the conventional enema and purgative technique.
In this prospective, randomized study, 121 elective colorectal surgery patients had whole-gut lavage (n = 67) or enemas and purgatives (n = 54). Patient characteristics were well matched. Intravenous metronidazole and tobramycin were administered preoperatively initially in 53 patients, with the remaining 68 patients receiving the drugs perioperatively. Bowel preparation was satisfactory (minimal or no contents remaining) in 92.8 percent of patients with whole-gut lavage and 92.6 percent with enemas and purgatives (p = 0.72). Nasogastric tube insertion was poorly tolerated by 39 percent of the patients receiving whole-gut lavage, and enema tube insertion by 23 percent with enemas and purgatives. Fluid infusion tolerance was similar with both techniques. Abdominal wound sepsis occurred in 22 patients (18.8 percent), being unrelated to mechanical preparation or antimicrobial prophylaxis (p = 0.19). Colostomy closure was associated with a 42.8 percent sepsis rate. Excluding this group, wound sepsis with the remaining procedures was 13 percent (statistically significant, p = 0.03). Other complications included intraabdominal abscesses (3.3 percent), anastomotic leaks (2.5 percent), eviscerations (1.6 percent), and an operative mortality of 1.6 percent. We have concluded that whole-gut lavage and enemas and purgatives are equally efficacious mechanically with similar associated wound sepsis rates.


Effect of topical rosemary essential oil on Raynaud phenomenon in systemic sclerosis.
Raynaud's phenomenon is the earliest manifestation of systemic sclerosis. Nitroglycerin gel is the only proven topical therapy. A 53-year-old woman with systemic sclerosis had topical Rosamarinus officinalis (rosemary) oil, often used in anthroposophic medicine, applied to her hands over 3days and then, separately, olive oil. Thermography images showed significant warming of fingers after rosemary oil, but not after olive oil, coinciding with the patient's subjective experience. Topical Rosamarinus officinalis oil had a vasodilator and warming effect in a patient with systemic sclerosis and Raynaud's phenomenon.


Congenital triangular alopecia. Report of fourteen cases.
Fourteen patients affected by congenital triangular alopecia are presented. The clinical and histologic features of this condition are discussed. I suggest that the condition is considerably more common than hitherto has been thought.


Does serum nitrite concentration reflect gastric carcinogenesis in Japanese Helicobacter pylori-infected patients?
This study investigated whether the serum nitrite concentration reflects Helicobacter pylori-induced inflammation and atrophic changes of gastric mucosa. Ninety-seven patients underwent biopsy of both antrum and fundus. Samples were analyzed by the rapid urease test and histopathological examination according to the updated Sydney system. Fasting serum samples from each subject were analyzed for specific IgG Helicobacter pylori antibodies, pepsinogen I and II concentrations, and NO2-/NO3- content. Eleven patients had H. pylori eradicated with proton pump-based triple therapy. There was a strong positive correlation between the Helicobacter pylori density in the gastric mucosa and the serum nitrite concentration, but a negative correlation existed between the atrophic grade of the gastric mucosa and both serum nitrite concentration and Helicobacter pylori density in the gastric mucosa. Serum nitrite concentrations decreased significantly after successful eradication of Helicobacter pylori. Therefore, serum nitrite concentration may be a useful marker for oxidative DNA damage and apoptosis associated with Helicobacter pylori infection.


Nurses' readiness to prescribe under supervision in Saudi Arabia: A cross-sectional study.
To explore nurses' readiness to prescribe medications under supervision and identify associations between prescribing practices under supervision and demographic characteristics in Saudi Arabia. A cross-sectional study. Using convenience sampling, this study used a 32-item survey to collect data on nurses prescribing medications under supervision between December 2022 and March 2023. A total of 379 nurses were recruited from different regions in Saudi Arabia. Approximately 7% (n = 30) of the participants were prescribing medications independently, and 70% (n = 267) expressed their likelihood of becoming prescribers. The highest motivating factors to become prescribers were improvement of patient care (52.2%) and contribution to the multidisciplinary team (52.0%). Most participants (60%-81%) agreed that prescribing medications under supervision would improve potential outcomes at the system, nurse, and patient levels. Availability of appropriate mentors or supervisors (72.9%) was the highest rated facilitating factor, followed by support of nursing colleagues (72%). Based on demographic characteristics, findings revealed significant differences in the: a) likelihood and motivators of becoming prescribers; b) required minimum qualification, years of experience, and continuing professional education hours to become prescribers; and c) type of organizations delivering educational programs for nurse prescribing. Majority of nurses in Saudi Arabia favored becoming prescribers, and motivating factors were mostly relevant to optimizing patient care outcomes. Having the proper supervision was rated as the most facilitating factor for nurse prescribing. Nurses' views on potential outcomes, facilitating factors, and possible motivators varied based on nurses' demographical characteristics. Nurses favored prescribing under supervision to improve patient care outcomes, which is an opportunity to expand the benefits of health services, including easy access to healthcare. Results revealed that nurses support the implementation of prescribing practice under supervision. Thus, the findings may inform practice change in Saudi Arabia to allow prescribing under supervision, which was perceived to have a positive impact on patient care outcomes. This study adhered to STROBE guidelines.


Why some people prefer pickle juice: the research of Dr. Richard P. Lifton.
In this essay, the author interviews Dr. Richard P. Lifton and examines the use of genomics throughout his studies on the molecular pathophysiology of hypertensive and hypotensive diseases. To date, Dr. Lifton has identified 10 different gene mutations that result in severe hypertension and another 10 that result in severe hypotension.


Discovery of triazenyl triazoles as Nav1.1 channel blockers for treatment of epilepsy.
The voltage-gated sodium (Nav) channel is one of most important targets for treatment of epilepsy, and rufinamide is an approved third-generation anti-seizure drug as Nav1.1 channel blocker. Herein, by triazenylation of rufinamide, we reported the triazenyl triazoles as new Nav1.1 channel blocker for treatment of epilepsy. Through the electrophysiological activity assay, compound 6a and 6e were found to modulate the inactivation voltage of Nav 1.1 channel with shift of -10.07 mv and -11.28 mV, respectively. In the pentylenetetrazole (PTZ) mouse model, 6a and 6e reduced the seizure level, prolonged seizure latency and improved the survival rate of epileptic mice at an intragastric administration of 50 mg/kg dosage. In addition, 6a also exhibited promising effectiveness in the maximal electroshock (MES) mouse model and possessed moderate pharmacokinetic profiles. These results demonstrated that 6a was a novel Nav1.1 channel blocker for treatment of epilepsy.


Basilar skull fractures in children.
In this paper we are reporting a retrospective study of patients under 18 years of age managed at the Los Angeles County/University of Southern California Medical Center from January 1979 through December 1987 with the diagnosis of basilar skull fracture. Sixty-two patients with basilar skull fractures were admitted during that 7 1/2 year period. The most common etiology was pedestrain versus vehicle accidents (42%), followed by falls (27%), vehicle accidents (23%), and being hit by an object (8%). The most common physical findings were hemotympanum (58%) and bleeding in the ear canals (47%). Thirty-four percent of the patients complained of hearing loss. Cerebrospinal fluid otorrhea was noted in 16 patients (26%), while only 1 patient had cerebrospinal fluid rhinorrhea. Facial nerve paralysis was present in 8 patients (13%). Vestibular symptoms were rare. Sixty-three percent of the patients had the diagnosis confirmed by radiography. The clinical presentation, complications, management and outcome of basilar skull fractures in the pediatric population are discussed.


Raloxifene activates G protein-coupled estrogen receptor 1/Akt signaling to protect dopamine neurons in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine mice.
Raloxifene, used in the clinic, is reported to protect brain dopaminergic neurons in mice. Raloxifene was shown to mediate an effect through the G protein-coupled estrogen receptor 1 (GPER1). We investigated if raloxifene neuroprotective effect in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-treated male mice is mediated through GPER1 by using its antagonist G15. Striatal concentrations of dopamine, 3,4-dihydroxyphenylacetic acid, homovanillic acid to dopamine ratio as well as dopamine transporter and vesicular monoamine transporter 2 showed that raloxifene neuroprotection of dopaminergic neurons was blocked by G15. Protection by raloxifene was accompanied by activation of striatal Akt signaling (but not ERK1/2 signaling) and increased Bcl-2 and brain-derived neurotrophic factor levels; these effects were abolished by coadministration with G15. The effect of raloxifene was not mediated through increased levels of 17β-estradiol. MPTP mice had decreased plasma testosterone, dihydrotestosterone, and 3β-diol levels; this was prevented in raloxifene-treated MPTP mice. Our results suggest that raloxifene acted through GPER1 to mediate Akt activation, increase Bcl-2 and brain-derived neurotrophic factor levels, and protection of dopaminergic neurons and plasma androgens.


Biomarkers in Non-Schistosomiasis-related squamous cell carcinoma of the urinary bladder: A review.
Non-urothelial (NU) histologies represent less than 10% of bladder cancers, with squamous cell carcinoma (SCC) being the most common subtype (approximately 5%). Bladder SCCs are subdivided into Schistosoma-related or non-Schistosoma-related tumors, with the latter being the most frequent subtype in the western world. Typically, these patients have been excluded or under-represented in clinical trials and thus little is known about treatment efficacy in bladder SCC. To address this paucity of data, extrapolation from urothelial carcinoma (UC) trials can be performed but this approach has inherent limitations. In the era of precision medicine, efforts to characterize the genomic and molecular profiles of bladder tumors may yield evidence to support new targets for effective therapies. We reviewed the available data on biomarkers of bladder SCC and provide suggestions on how these may influence therapeutic testing and clinical trials in the future.


Insulin induces phosphatidylinositol-3-phosphate formation through TC10 activation.
Phosphatidylinositol-3-phosphate (PtdIns-3-P) is considered as a lipid constitutively present on endosomes; it does not seem to have a dynamic role in signalling. In contrast, phosphatidylinositol-3,4,5-trisphosphate (PtdIns-3,4,5-P(3)) plays a crucial role in different signalling pathways including translocation of the glucose transporter protein GLUT4 to the plasma membrane upon insulin receptor activation. GLUT4 translocation requires activation of two distinct pathways involving phosphatidylinositol 3-kinase (PI 3-K) and the small GTP-binding protein TC10, respectively. The contribution of each pathway remains to be elucidated. Here we show that insulin specifically induces the formation of PtdIns-3-P in insulin- responsive cells. The insulin-mediated formation of PtdIns-3-P occurs through the activation of TC10 at the lipid rafts subdomain of the plasma membrane. Exogenous PtdIns-3-P induces the plasma membrane translocation of both overexpressed and endogenous GLUT4. These data indicate that PtdIns-3-P is specifically produced downstream from insulin-mediated activation of TC10 to promote the plasma membrane translocation of GLUT4. These results give a new insight into the intracellular role of PtdIns-3-P and shed light on some aspects of insulin signalling so far not completely understood.


'Metabolic' factors in thermal injury: role of nucleic acids.
We believe that toxic events observed after thermal injury may be caused by the release of normally intracellular substances into the circulation. We define these substances as 'metabolic' factors. Analysis of extracts prepared from normal and burned mouse skin indicates that the burned skin extract contains increased clot-promoting (Thromboplastin-like) substances and, perhaps, less RNA than normal skin extracts. Injection of RNA or its breakdown products into the burned site significantly increases the acute mortality in burned mice. No increase in mortality is observed when these substances are injected into a non-burned site on burned mice. We suggest that 'Thromboplastin-like substances and RNA or RNA breakdown products may be some of the 'metabolic' factors involved in acute burn toxicity. Upon being released from their intracellular residence after thermal injury, their combined activity contributes to the acute mortality observed.


The yield and impact of pulmonologist-performed EUS-B-FNA of subdiaphragmatic lesions-an institutional experience.
Pulmonologists can biopsy structures below the diaphragm using the convex curvilinear ultrasound bronchoscope via the esophagus (EUS-B). The literature with respect to the value of EUS-B, rapid on-site evaluation, and final diagnostic yield for structures below the diaphragm is limited. We review our institutional experience. Our database was queried retrospectively for EUS-B fine needle aspirations (FNAs) from 2013 to 2021. All procedures involving EUS-B-FNA of subdiaphragmatic structures were selected for analysis. The following data elements were collected for each patient: age, gender, clinical indication, sample site, on-site adequacy (OSA), preliminary and final diagnoses, and sufficiency of cell block for ancillary studies. A total of 75 subdiaphragmatic sites were biopsied in 74 patients. Of which, 87% of samples subjected to rapid on-site evaluation were deemed to contain adequate material (OSA+). There were no false-positive OSAs. Six cases remained nondiagnostic at the final diagnosis. The final diagnostic yield (with cell block) was 92% (69/75 cases). Cell block was sufficient for immunohistochemistry or special stains in all applicable cases (n = 36). Molecular testing was requested for 11 cases and successful in 10 (91%). Sampling of subdiaphragmatic sites changed the stage in 67% (38/57) of lung cancer patients. Pulmonologists can perform EUS-B-FNA of subdiaphragmatic sites with high OSA and final diagnostic yield when assisted by cytopathologists. Strong correlations exist between OSA, cell block adequacy, and subsequent capacity to perform ancillary testing. EUS-B below the diaphragm can make an important contribution to the diagnosis of lung cancer, nonpulmonary malignancies, and other diseases.


[Study on natural infection of Plasmodium vivax in Anopheles sinensis in Korea]
On epidemiological grounds, Anopheles sinensis has been suspected for many years to be the malaria vector in Korea. Of 7,517 dissections of A. sinensis, one specimen was found for the first time in Korea naturally infected with sporozoites in August 1962. Experimental infection of A. sinensis with Plasmodium vivax had been successfully conducted at earlier date.


11C-Choline PET/CT vs. 99mTc-MIBI/123Iodide Subtraction SPECT/CT for Preoperative Detection of Abnormal Parathyroid Glands in Primary Hyperparathyroidism: A Prospective, Single-Centre Clinical Trial in 60 Patients.
In patients with primary hyperparathyroidism (PHPT) locating hyperfunctioning glands (HPGs) is crucial when planning minimally invasive surgery. Dual-isotope subtraction scintigraphy with 99mTc-MIBI/123Iodide using SPECT/CT and planar pinhole imaging (Method A) has previously shown a sensitivity >93%. However, the method is costly and time consuming and entails a high radiation dose. 11C-Choline PET/CT (Method B) is an appealing candidate method unencumbered by these disadvantages. Sixty patients with newly diagnosed PHPT participated and were scanned using both methods prior to parathyroidectomy. We investigated whether sensitivities of Method A and Method B are similar in a method-to-method comparison when using surgical findings as the true location. At the patient level, sensitivities were (A) 0.98 (95% CI: 0.90-1.00) and (B) 1.00 (95% CI: 0.93-1.00). At the gland level, sensitivities were (A) 0.88 (95% CI: 0.78-0.94) and (B) 0.87 (95% CI: 0.76-0.92). With a non-inferiority margin of ∆ = -0.1, we found a 1-sided p-value < 0.001. Our methods comparison study found that sensitivity of Method B was not inferior to Method A. We suggest that 11C-Choline PET/CT is a clinically relevant first-choice candidate for preoperative imaging of PHPT and that Method B can likely replace Method A in the near future.


Antimicrobial susceptibility of Pneumocystis carinii in culture.
Pneumocystis carinii from infected rats were cultured on infected WI-38 human lung fibroblasts. After cultures were established, medium was replaced by medium containing antimicrobials and supernate organisms counted at specified times. Results show that metronidazole had no effect, amphotericin B suppressed organisms at 5 but not 0.5 microgram/ml, and miconazole, chloroquine, trimethoprim-sulfamethoxazole, and pentamidine isethionate suppressed organisms at the concentrations tested.


Airway management by US and Canadian emergency medicine residents: a multicenter analysis of more than 6,000 endotracheal intubation attempts.
We determine success rates of endotracheal intubation performed in emergency departments (EDs) by North American emergency medicine residents. During 58 months, physicians performing intubations at 31 university-affiliated EDs in 3 nations completed a data form that was entered into the National Emergency Airway Registry 2 database. Included were all patients undergoing endotracheal intubation in the ED. The data form included patients' age, sex, weight, indication for intubation, technique of airway management, names and dosages of all medications used to facilitate intubation, level of training and specialty of the intubator, number of attempts, success or failure, and adverse events. We queried this prospectively gathered, observational data to analyze intubations done by US and Canadian emergency medicine residents. Enrollment was incomplete (eg, 85% at the main study center), so the study sample did not include all consecutive patients. Emergency medicine residents performed 77% (5768/7498; 95% confidence interval [CI] 76% to 78%) of all initial intubation attempts in the United States and Canada. The first intubator was successful in 90% (5,193/5,757; 95% CI 89% to 91%) of cases, including 83% (4,775/5,757; 95% CI 82% to 84%) on the first attempt. Success rates on the first attempt were as follows: postgraduate year 1 = 72% (498/692; 95% CI 68% to 75%), postgraduate year 2 = 82% (2,081/2,544; 95% CI 80% to 83%), postgraduate year 3 = 88% (1,963/2,238; 95% CI 86% to 89%), postgraduate year 4+ = 82% (233/283; 95% CI 77% to 87%), and attending physician = 89% (689/772; 95% CI 87% to 91%). Success rates by the first intubator were as follows: postgraduate year 1 = 80% (553/692; 95% CI 77% to 83%), postgraduate year 2 = 89% (2,272/2,544; 95% CI 88% to 90%), postgraduate year 3 = 94% (2,105/2,238; 95% CI 93% to 95%), postgraduate year 4+ = 93% (263/283; 95% CI 89% to 96%), and attending physician = 98% (755/772; 95% CI 96% to 99%). Rapid sequence intubation technique was used in 78% (4,513/5,768; 95% CI 77% to 79%) of initial attempts: it resulted in 85% (3,843/4,513; 95% CI 84% to 86%) success on the first attempt and 91% (4,117/4,513; 95% CI 90% to 92%) success by the first intubator. The overall rate of cricothyrotomy for all emergency resident intubations was 0.9% (50/5,757; 95% CI 0.6% to 1.1%). When an initial intubator failed, 40% (385/954; 95% CI 37% to 44%) of rescue attempts were performed by emergency medicine residents. Among emergency medicine residents, success on the first rescue attempt was 80% (297/371; 95% CI 76% to 84%), and success by the first rescue intubator was 88% (328/371; 95% CI 85% to 91%). Success of initial intubation attempts increased over the first 3 years of residency. This large multicenter study demonstrates the success of airway management by emergency medicine residents in North America. Using rapid-sequence intubation predominantly, emergency medicine residents achieved high levels of success.


External support of a polytetrafluoroethylene graft improves patency for bypass to below-knee arteries.
Patency and limb salvage after synthetic bypass to the arteries below the knee are inferior to that which can be achieved with autologous vein. The use of external support of synthetic polytetrafluoroethylene (PTFE) grafts to the below-knee position has been suggested to improve patency and limb salvage, a problem analyzed in this randomized clinical trial. We examined whether external graft support improves patency and/or limb salvage in patients undergoing reconstruction with synthetic PTFE grafts to the below-knee arteries. Three hundred thirty-four patients with critical limb ischemia undergoing PTFE bypass to below-knee arteries were randomly assigned to have an ordinary PTFE graft or one with external support. Follow-up was scheduled until amputation, death, or at most 5 years, whichever event occurred first. Patients in both the femoropopliteal and femorodistal groups were randomized to have an externally supported PTFE graft (101/195 patients in the femoropopliteal group and 72/139 patients in the femorodistal group). Follow-up information was available for 329 of 334 randomized patients (99%). At 1 year postprocedure, primary patency for below-knee bypass was 0.55 (95% confidence interval [CI], 0.47-0.64) with and 0.42 (95% CI, 0.34-0.50) without externally supported PTFE grafts, and secondary patency was 0.58 (95% CI, 0.51-0.67) and 0.47 (95% CI, 0.39-0.56), respectively. The corresponding figures for limb salvage were 0.75 (95% CI, 0.68-0.82) and 0.69 (95% CI, 0.62-0.77), respectively. The log rank test revealed statistically significant differences between patients with or without externally supported grafts for patency (primary patency: χ2=4.2 [degrees of freedom=1; P=0.041]; secondary patency: χ2=4.3 [degrees of freedom=1; P=0.037]) but not for limb salvage (limb salvage: χ2=0.2 [degrees of freedom=1; P=0.657]). External support to a PTFE graft used for bypass to below-knee arteries improves primary and secondary patency but not limb salvage.


Barrier properties and cost implications of a single versus a double wrap for storing sterile instrument packs.
Materials for wrapping sterile items continue to evolve, but evaluation of such products under clinical conditions is rare. The purpose of the current study was to test a new product before introducing it to the hospital's sterilizing processing unit. Four hundred packs containing 1199 items were prepared. Half were wrapped in linen and Kimguard sterile wrap (Kimberley-Clark Australia Pty, Ltd; Queensland, Australia), and half were wrapped in Kimguard One-Step sterile wrap (Kimberley-Clark). They were stored on shelves in 4 areas in the hospital. Items from the packs were periodically tested in the laboratory to evaluate shelf life. Time of wrapping was measured on a series of 50 packs (25 using each product), wrapped by 1 experienced person. These were unwrapped by an operating room nurse, and, again, the process was timed. Bacteria were cultured from 20 (1.7%) of the 1157 test items. There were no differences on this measure between the 2 products (P = .64). Coagulase-negative Staphylococcus was the most frequent isolate, accounting for 40% of the positive results. The average time taken to wrap the test tray with the double wrap was 56.4 seconds compared with 32.4 seconds with the single wrap (P < or = .000). Unwrapping the single pack (5.02 seconds) was also faster than unwrapping the double-wrap pack (6.92 seconds; P = .000). Wrapping sterile items using Kimguard one-step sterile wrap carries no greater risk of bacterial contamination than double-wrap methods and may lead to significant cost savings in both labor (time to wrap) and consumables (linen and recycling costs).


Changes of neuropeptides and their receptors in experimental stroke gerbil brains.
Eight kinds of neuropeptides and four kinds of neuropeptide receptors were examined in the right and left hemispheres of mongolian gerbils after unilateral carotid ligation-induced stroke and in normal controls. Five hours after ligation of the right common carotid artery, beta-endorphin concentration in the right hemisphere (ischemic side) of the stroke group was significantly increased compared with that in the contralateral hemisphere (non-ischemic side), but there were no differences between sides in other neuropeptides either with or without stroke. Furthermore, although there were no differences in [3H]naloxone binding, [3H]thyrotropin-releasing hormone binding or 125I-vasoactive intestinal polypeptide binding in the brain in this model of stroke, [3H]enkephalin binding was significantly lower on the ischemic side than on the non-ischemic side in the stroke group. These results suggest that increased activity in the beta-endorphinergic system in the brain might be partly caused by ischemic brain failure.


Correction: Inhibition of NAPDH Oxidase 2 (NOX2) Prevents Oxidative Stress and Mitochondrial Abnormalities Caused by Saturated Fat in Cardiomyocytes.
[This corrects the article DOI: 10.1371/journal.pone.0145750.].


Exclusion of linkage to 5q11-13 in families with schizophrenia and other psychiatric disorders.
Recently a linkage study on five Icelandic and two English pedigrees has provided evidence for a dominant gene for schizophrenia on 5q11-13 (ref. 1). In that study, families with bipolar illness were not included. Using the same probes, two similar but independent investigations on one Swedish pedigree and on fifteen Scottish families excluded linkage to schizophrenia. To evaluate whether the susceptibility gene on 5q11-13 is a common cause of schizophrenia in other populations, we examined five affected North American pedigrees using probes to the D5S39, D5S76 and dihydrofolate reductase loci. Two families in the present series had cases of bipolar disorder. We found that linkage can be excluded by multipoint analysis. These results, taken together, suggest that the disease gene on 5q11-13 does not account for most cases of familial schizophrenia.


The thermotropic behavior of dipalmitoyl phosphatidylcholine bilayers. A Fourier transform infrared study of specifically labeled lipids.
Fourier transform infrared spectroscopy has been used to study the thermotropic behavior of hydrated multibilayers of specifically deuterated derivatives of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine. It is shown that throughout the gel phase there is little or no conformational disorder introduced into the acyl chains. The pretransition effect is greatest in the central segments of the acyl chains, demonstrating that the interchain interactions are more pronounced in this region than in the center of the bilayer and suggesting the presence, in the gel phase, of a "plateau" in the strength of the interchain interactions. As the temperature is reduced, the rate of rotation of the terminal methyl group decreases steadily; below 0 degree C the conformation is rigid on the infrared time scale.


Negative inotropic mechanisms of β-cardiotoxin in cardiomyocytes by depression of myofilament ATPase activity without activation of the classical β-adrenergic pathway.
Beta-cardiotoxin (β-CTX) from the king cobra venom (Ophiophagus hannah) was previously proposed as a novel β-adrenergic blocker. However, the involvement of β-adrenergic signaling by this compound has never been elucidated. The objectives of this study were to investigate the underlying mechanisms of β-CTX as a β-blocker and its association with the β-adrenergic pathway. The effects of β-CTX on isolated cardiac myocyte functions, calcium homeostasis, the phosphorylation level of targeted proteins, and the myofibrillar ATPase activity were studied. Healthy Sprague Dawley rats were used for cardiomyocytes isolation. Like propranolol, β-CTX attenuated the cardiomyocyte inotropy and calcium transient alterations as induced by isoproterenol stimulation. In contrast, these effects were not observed in forskolin-treated cells. Interestingly, cardiomyocytes treated with β-CTX showed no changes in phosphorylation level at any PKA-targeted sites in the myofilaments as demonstrated in Western blot analysis. The skinned fibers study revealed no change in myofilament kinetics by β-CTX. However, this protein exhibited the direct inhibition of myofibrillar ATPase activity with calcium de-sensitization of the enzyme. In summary, the negative inotropic mechanism of β-CTX was discovered. β-CTX exhibits an atypical β-blocker mechanism. These properties of β-CTX may benefit in developing a novel agent aid to treat hypertrophic cardiomyopathy.


Primary fallopian tube carcinoma.
Primary fallopian tube carcinoma (PFTC) is a rare gynaecological tumour that accounts for 0.14-1.8% of genital malignancies. The most common age of occurrence is between 40 and 65 years, and the mean age is 55 years. The factors that contribute to its appearance are not well known. Population studies show that the mean incidence of PFTC is 3.6 per million women per annum. Overall survival percentages for patients with PFTC are generally low, in the range of 22-57%. Pre-operative diagnosis is rare and PFTC is usually confirmed by a pathologist, but earlier diagnosis with early clinical manifestation and prompt investigation improves the prognosis. Both PFTC and epithelial ovarian cancer (EOC) are treated with similar surgical and chemotherapy methods. Studies have shown that the prognosis for PFTC is worse than that for EOC or other primary gynaecological tumours. This article reviews and presents the current updates of this rare gynaecological malignancy.


Data on the survival times of breast cancer patients in a Teaching Hospital, Osogbo.
In a bid to assess the contribution of prognostic factors to survival times of breast cancer patients from day of presentation in Nigeria, a data was collected from Ladoke Akintola University of Technology Teaching Hospital Osogbo. This is a retrospective data culled from the case note files of the breast cancer patients. The survival time of the patients was recorded as the difference between the day of presentation and the day of last contact. The data is censored at 1 year. The prognostic factors considered are years of breastfeeding(In years), Age at menarche, stage at presentation, neoadjuvant treatment offered and use of contraceptives. Four survival models were applied to the data to assess the contribution of the prognostic factors to survival times.


Impairment of Quality of Life in Patients with Implanted Subcutaneous Cardioverter Defibrillator (S-ICD) Compared to Implanted Transvenous Cardioverter Defibrillator Therapy.
The subcutaneous cardioverter defibrillator (S-ICD) has been shown to be a viable alternative to transvenous ICDs (TV-ICD) in all patients at risk of sudden cardiac death (SCD) but without pacing indication. The aim of this study was to examine the impact of therapy with current S-ICD devices on quality of life (QoL) in comparison to patients with TV-ICD devices. In our single-centre study, 52 consecutive patients with S-ICD and 52 matched patients with TV-ICD were analysed. QoL has been assessed by a standardized questionnaire (EQ-5D-3L, modified). Additionally, clinical baseline and follow-up data were evaluated. Two-thirds of the total study population reported restrictions in daily routine compared to their life before ICD implantation. A total of 27.7% of S-ICD patients stated to expect an improvement of QoL by deactivation or explantation of their defibrillator compared to only 6.4% of patients with TV-ICD (p=0.006), which was mainly caused by discomfort and pain from the S-ICD pocket (relevant discomfort and pain in 32.6% vs 11.5%; p<0.01). Main limitation of the study is that quality of life was assessed for one single time point only and time since implantation differed significantly between S-ICD and TV-ICD. Furthermore our collective is younger, and, due to the high proportion of patients without cardiomyopathy, the mean EF is better than usual ICD collective. The absence of heart failure in about the half of our patients might have relevant impact on our QoL analysis. A relevant proportion of S-ICD patients expects an improvement of QoL by explantation of the device. Of note, this impression was not driven by the fear of receiving shocks but mainly by discomfort and pain caused by the pulse generator.


Probing the Architecture of a Multi-PDZ Domain Protein: Structure of PDZK1 in Solution.
The scaffolding protein PDZK1 has been associated with the regulation of membrane transporters. It contains four conserved PDZ domains, which typically recognize a 3-5-residue long motif at the C terminus of the binding partner. The atomic structures of the individual domains are available but their spatial arrangement in the full-length context influencing the binding properties remained elusive. Here we report a systematic study of full-length PDZK1 and deletion constructs using small-angle X-ray scattering, complemented with biochemical and functional studies on PDZK1 binding to known membrane protein partners. A hybrid modeling approach utilizing multiple scattering datasets yielded a well-defined, extended, asymmetric L-shaped domain organization of PDZK1 in contrast to a flexible "beads-on-string" model predicted by bioinformatics analysis. The linker regions of PDZK1 appear to play a central role in the arrangement of the four domains underlying the importance of studying scaffolding proteins in their full-length context.


Combined Application of Organic Fertilizer with Microbial Inoculum Improved Aggregate Formation and Salt Leaching in a Secondary Salinized Soil.
Greenhouse vegetable production provides significant quantities of vegetables throughout the year and improves farmers' income. However, over-fertilization with mineral fertilizer causes soil secondary salinization and decreases the stability of the soil structure. To improve aggregate formation and decrease salt accumulation in the soil profile, bio-organic fertilizers (Protaetia brevitarsis larvae frass with Bacillus amyloliticus and/or Trichoderma harziensis) were applied to partially substitute mineral fertilizer in a salinized vegetable soil. Soil nutrient condition, aggregate stability, and salt movement in the soil profile were measured in a greenhouse double-cucumber system. The results showed that soil organic matter (SOM), total nitrogen (TN), and available phosphorus (AP) increased significantly under bio-organic fertilizer treatments compared with control. Soil electrical conductivity (EC) and total salt content (TSC) decreased by 15.74-24.20% and 19.15-29.05%, respectively, with bio-organic fertilizers (p < 0.05). Cl-, NO3-, and SO42- content under double inoculation with B. amyloliticus and T. harziensis reduced by 31.19%, 26.30%, and 53.11%, respectively, compared to CK (p < 0.05). In addition, double inoculation was more efficient in reducing nitrate content in the soil profile than single inoculation. Soil microaggregates of 0.25-0.053 mm increased by 75.87-78.51% with bio-fertilizers compared with control, and double inoculation was the best for aggregate formation. In conclusion, the inoculation of plant-growth-promoting and salt-tolerant microorganisms with high humic acid larvae frass can alleviate salinization in vegetable soil, enhance soil nutrient content, and improve the soil structure.


[Seroprevalence and factors associated with viral hepatitis B among pregnant women in Parakou, Republic of Benin].
The purpose of this study is to determine the seroprevalence and the factors associated with viral hepatitis B among pregnant women at the Departmental University Hospital of Borgou (CHUD-B).We conducted a cross-sectional descriptive and analytical study with prospective collection of data. It focused on pregnant women treated and/or having given birth at the CHUD-B from 1 April 2017 to 30 June 2017. The study included patients who gave their consent to participate in the study. The variables studied were the sociodemographic data of pregnant women and the result of HBsAg test. The detectionof HBsAg in blood sampling of pregnant women was performed using a biological test for rapid diagnosis. Positive results were confirmed by ELISA technique. Two hundred and fourteen (214) pregnant women were included in the study. The average age of patients was 26.73±5.68 years. The mean gestation age was 31 weeks of amenorrhea (WA) ranging from 4 WA and 42 WA. Among them, 30 women were positive for HBsAg, reflecting a prevalence of 14.02%. The factors associated with this colonization were the practice of scarification, a personal history of jaundice and family history of viral hepatitis B. The seroprevalence of viral hepatitis B among pregnant women at the CHUD-B is high. This best reflect this major public health problem due to Hepatitis B virus.


Balaenoptera omurai is a newly discovered baleen whale that represents an ancient evolutionary lineage.
Balaenoptera omurai, formerly classified as a small form of Bryde's whale, was recently reclassified as a new baleen whale species of the family Balaenopteridae. Although researchers have investigated the evolutionary history of Balaenopteridae and their relatives using molecular phylogenetic methods, the taxonomy of the ordinary Bryde's whale (Balaenoptera brydei) and small-form Bryde's whales (Balaenoptera edeni and B. omurai) remains unclear. We have used complete mtDNA sequences and short interspersed repetitive element (SINE) insertion patterns to construct the evolutionary history of both B. omurai and the taxonomically redefined species, B. edeni. The combined results demonstrate that B. omurai forms a monophyletic lineage with B. musculus, B. brydei, B. edeni and B. borealis and that B. omurai and B. musculus successively diverged from their common ancestor. In addition, we also showed that B. edeni constitutes a sister taxon to B. brydei. Our data suggest that B. omurai evolved as an ancient independent lineage that diverged much earlier than B. borealis, B. brydei and B. edeni, which were previously believed to be closely related to B. omurai.


Measuring right-hemisphere dysfunction in children: validity of two new computer tests.
The validity of two new computer-mediated tests for the detection of right-cerebral hemisphere lesions in children--the Right-hemisphere Dysfunction Test and the Visual Perception Test--was evaluated. Normative data were drawn from a group of 91 children (aged five to 14 years) and 14 young adults. The tests were also administered to 14 children with acquired lesions of either right- or left-cerebral hemisphere. The results demonstrate that the Right-hemisphere Dysfunction Test and the Visual Perception Test, with predictive values of 71 per cent and 88 per cent, respectively, are useful in clinical practice for detection of right-hemisphere dysfunction in children.


Glioblastoma imitating a cavernoma radiologically: A unique image report.
Glioblastoma is the most common primary malignant brain tumor with characteristic radiological features in most cases. We highlight an unusual case of a 54-year-old woman, neurologically intact, with a diagnostically challenging lesion. The patient's magnetic resonance imaging revealed a left frontal lesion with surrounding edema and a hemosiderin ring, misleading it to be a cavernoma. Intraoperatively, the lesion was found to be a solid tumor with hematoma and was confirmed to be glioblastoma on histopathology. The dilemma associated with our patient's radiological findings and longstanding history of epilepsy is rare and a diagnostic challenge.


Inorganic Polyphosphate and Physiological Properties of Saccharomyces cerevisiae Yeast Overexpressing Ppn2.
The effect of the yeast endopolyphosphatase Ppn2 overproduction on the metabolism of inorganic polyphosphates in Saccharomyces cerevisiae yeast was studied. Expression of the PPN2 gene under control of the strong constitutive promoter of glyceraldehyde 3-phosphate dehydrogenase gene (PKG1) led to a significant increase in the endopolyphosphatase activity stimulated by cobalt/zinc ions. This activity was present in both soluble and membrane subcellular fractions; it was higher toward long-chain polyphosphates and could be stimulated by ADP. The content of short-chain polyphosphates in the cells of the overexpressing strain was ~2.5 times higher compared to the parent strain. The cells overexpressing Ppn2 were more resistant to peroxide and alkali. The role of short-chain polyphosphates in the adaptation to these stress factors is discussed.


Systematic review and meta-analysis of single-balloon enteroscopy-assisted ERCP in patients with surgically altered GI anatomy.
Surgically altered pancreaticobiliary anatomy increases the difficulty of performing ERCP. Single-balloon enteroscopy (SBE) is a relatively new technique that can be used for ERCP in patients with surgically altered anatomy. To evaluate the therapeutic and diagnostic success of SBE-ERCP among patients with surgically altered anatomy. Systematic review and meta-analysis of studies involving SBE-ERCP in patients with Roux-en-Y gastric bypass, hepaticojejunostomy, or Whipple procedure. Enteroscopy success was defined as success in reaching the papilla and/or biliary anastomosis by using SBE. Diagnostic success was defined as obtaining a cholangiogram. Procedural success was defined as the ability to provide successful intervention, if appropriate. A random-effects model was used. A total of 461 patients underwent SBE-ERCP from 15 trials. The pooled enteroscopy, diagnostic, and procedural success rates were 80.9% (95% confidence interval [CI], 75.3%-86.4%), 69.4% (95% CI, 61.0%-77.9%), and 61.7% (95% CI, 52.9%-70.5%), respectively. There was statistical large heterogeneity for enteroscopy, diagnostic, and therapeutic success (P < .001 for all). Adverse events occurred in 6.5% (95% CI, 4.7%-9.1%) of patients. There was no evidence of publication bias in this meta-analysis. Our findings and interpretations are limited by the quantity and heterogeneity of the studies included in the analysis. SBE-ERCP has high diagnostic and procedural success rates in this challenging patient population. It should be considered a first-line intervention when biliary access is required after Roux-en-Y gastric bypass, hepaticojejunostomy, or Whipple procedure.


High-resolution view of the type III secretion export apparatus in situ reveals membrane remodeling and a secretion pathway.
Type III protein secretion systems are essential virulence factors for many important pathogenic bacteria. The entire protein secretion machine is composed of several substructures that organize into a holostructure or injectisome. The core component of the injectisome is the needle complex, which houses the export apparatus that serves as a gate for the passage of the secreted proteins through the bacterial inner membrane. Here, we describe a high-resolution structure of the export apparatus of the Salmonella type III secretion system in association with the needle complex and the underlying bacterial membrane, both in isolation and in situ. We show the precise location of the core export apparatus components within the injectisome and bacterial envelope and demonstrate that their deployment results in major membrane remodeling and thinning, which may be central for the protein translocation process. We also show that InvA, a critical export apparatus component, forms a multiring cytoplasmic conduit that provides a pathway for the type III secretion substrates to reach the entrance of the export gate. Combined with structure-guided mutagenesis, our studies provide major insight into potential mechanisms of protein translocation and injectisome assembly.


Axonal and demyelinating forms of the MPZ Thr124Met mutation.
We report on a Japanese family with Charcot Marie Tooth disease (CMT) with the Thr124Met mutation in the peripheral myelin protein zero (MPZ) gene. Based on the clinical study, we investigated MPZ gene by direct sequence analysis and polymerase chain reaction restriction fragment length polymorphism analysis. Genotyping of four symptomatic family members showed that one family member with severe disease symptoms was homozygous, while the other three were heterozygous. The heterozygous cases were clinicopathologically determined to be the axonal type, which is characterized by late-onset and slow progression associated with Adie's pupil and deafness. The homozygous case was the demyelinating type, which showed earlier onset, rapid progression, sural nerve demyelination, and cranial nerve demyelination at autopsy. We suggest that axonal and demyelinating forms of CMT are not two distinct classes, but rather parts of a spectrum of genotypically related conditions, particularly with some MPZ mutations.


Near-infrared fluorescence imaging in immunotherapy.
Near-infrared (NIR) light possesses many suitable optophysical properties for medical imaging including low autofluorescence, deep tissue penetration, and minimal light scattering, which together allow for high-resolution imaging of biological tissue. NIR imaging has proven to be a noninvasive and effective real-time imaging methodology that provides a high signal-to-background ratio compared to other potential optical imaging modalities. In response to this, the use of NIR imaging has been extensively explored in the field of immunotherapy. To date, NIR fluorescence imaging has successfully offered reliable monitoring of the localization, dynamics, and function of immune responses, which are vital in assessing not only the efficacy but also the safety of treatments to design immunotherapies optimally. This review aims to provide an overview of the current research on NIR imaging of the immune response. We expect that the use of NIR imaging will expand further in response to the recent success in cancer immunotherapy. We will also offer our insights on how this technology will meet rapidly growing expectations in the future.


Guar gum and reduction of post-prandial glycaemia: effect of incorporation into solid food, liquid food, and both.
1. The influence of the dose and the form in which guar gum was given on the degree of "flattening" of blood glucose curves was studied in five subjects using meals of bread and soup containing 5 or 10 g guar gum. 2. When 5 g guar gum was added to bread the peak increase of blood glucose was reduced by 41% (P less than 0.002), with 5 g guar in soup, the reduction was 54% (P less than 0.001) while a reduction of 68% (P less than 0.001) was seen with 10 g guar gum (5 g in bread and 5 g in soup). The corresponding reduction in insulin peak increases were 37% (P less than 0.002), 50% (P less than 0.001) and 65% (P less than 0.001) respectively. 3. The difference between the two 5 g doses was significant with respect to the reduction of the peak increases in blood glucose and serum insulin; however the difference between the 5 g dose in bread and the 10 g dose was significantly different (P less than 0.02 for glucose, P less than 0.01 for insulin). 4. The results indicate that as little as 5 g guar gum may reduce the glycaemia following a 45 g carbohydrate meal, but perhaps due to earlier and more complete mixing, guar gum is most effective when added to the liquid phase of the meal.


Addition of Bevacizumab to Erlotinib as First-Line Treatment of Patients With EGFR-Mutated Advanced Nonsquamous NSCLC: The BEVERLY Multicenter Randomized Phase 3 Trial.
Adding bevacizumab to erlotinib prolonged progression-free survival (PFS) of patients with EGFR-mutated advanced NSCLC in the Japanese JO25567 trial, but limited data were available in non-Asian patients. BEVERLY is an Italian, multicenter, randomized, phase 3 investigating the addition of bevacizumab to erlotinib as first-line treatment of advanced EGFR-mutated NSCLC. Eligible patients were randomized 1:1 to erlotinib plus bevacizumab or erlotinib alone. Investigator-assessed PFS and blinded independent centrally reviewed PFS were coprimary end points. With 80% power in detecting a 0.60 hazard ratio and two-sided α error of 0.05, 126 events of 160 patients were needed. The trial was registered as NCT02633189 and EudraCT 2015-002235-17. From April 11, 2016, to February 27, 2019, a total of 160 patients were randomized to erlotinib plus bevacizumab (80) or erlotinib alone (80). At a median follow-up of 36.3 months, median investigator-assessed PFS was 15.4 months (95% confidence interval [CI]: 12.2-18.6) with erlotinib plus bevacizumab and 9.6 months (95% CI: 8.2-10.6) with erlotinib alone (hazard ratio = 0.66, 95% CI: 0.47-0.92). Blinded independent centrally reviewed PFS analysis confirmed this result. A statistically significant interaction with treatment effect was found for smoking habit (p = 0.0323), with PFS prolongation being clinically significant only among current or previous smokers. Hypertension (grade ≥3: 24% versus 5%), skin rash (grade ≥ 3: 31% versus 14%), thromboembolic events (any grade: 11% versus 4%), and proteinuria (any grade: 23% versus 6%) were more frequent with the combination. The addition of bevacizumab to first-line erlotinib prolonged PFS in Italian patients with EGFR-mutated NSCLC; toxicity was increased with the combination but without unexpected safety issues.


Reactive balance responses to a trip and slip during gait in people with multiple sclerosis.
To examine reactive balance responses to a trip and slip during gait in people with multiple sclerosis (MS). This cross-sectional laboratory study involved 29 participants with MS (50.6 ± 13.4 years) and 29 gender-and-aged-matched healthy controls (50.9 ± 19.2 years). Falls following an induced trip and slip along a 10 m walkway, approach (e.g. gait speed, step length, foot contact angle) and recovery strategies (e.g. response time, extrapolated centre of mass position, margin of stability) were compared between the two groups. The rate of falls was significantly higher in the participants with MS relative to healthy controls (rate ratio=2.82, 95% confidence interval [CI]=1.42, 5.61). Participants with MS also experienced more trip falls (odds ratio [OR]=3.90, 95% CI=1.16, 13.08) and more slip falls (OR=6.27, 95% CI=1.95, 20.22) than the heathy controls. Participants with MS had significantly slower gait speed, step length, cadence, and foot contact angle during approach (P < 0.05). Following slips, participants with MS had significantly greater stance limb knee flexion (P < 0.05), suggesting inadequate lower limb support to recover balance post-slip. Following trips, participants with MS had significantly delayed response initiation, lower toe clearance, shorter step length, and greater trunk sway (P < 0.05). Fewer participants with MS showed a hopping response to clear the obstacle (P < 0.05). Multiple sclerosis impairs reactive balance responses to a trip and slip associated with reduced lower limb function and delayed postural responses. Neurorehabilitation targeting reactive balance may facilitate fall prevention in people with multiple sclerosis.


Visceral fat-related systemic inflammation and the adolescent brain: a mediating role of circulating glycerophosphocholines.
Life-long maintenance of brain health is important for the prevention of cognitive impairment in older age. Low-grade peripheral inflammation associated with excess visceral fat (VF) may influence brain structure and function. Here we examined (i) if this type of inflammation is associated with altered white-matter (WM) microstructure and lower cognitive functioning in adolescents, and (ii) if recently identified circulating glycerophosphocholines (GPCs) can index this type of inflammation and associated variations in WM microstructure and cognitive functioning. We studied a community-based sample of 872 adolescents (12-18 years, 48% males) in whom we assessed VF and WM microstructure with magnetic resonance imaging, processing speed with cognitive testing, serum C-reactive protein (CRP, a common marker of peripheral inflammation) with a high-sensitivity assay, and serum levels of a panel of 64 GPCs with advanced mass spectrometry. VF was associated with CRP, and CRP in turn was associated with "altered" WM microstructure and lower processing speed (all p < 0.003). Further, "altered" WM microstructure was associated with lower processing speed (p < 0.0001). Of all 64 tested GPCs, 4 were associated with both VF and CRP (at Bonferroni corrected p < 0.0004). One of them, PC16:0/2:0, was also associated with WM microstructure (p < 0.0001) and processing speed (p = 0.0003), and mediated the directed associations between VF and both WM microstructure (p < 0.0001) and processing speed (p = 0.02). As a mediator, PC16:0/2:0 explained 21% of shared variance between VF and WM microstructure, and 22% of shared variance between VF and processing speed. Similar associations were observed in an auxiliary study of 80 middle-aged adults. Our results show that VF-related peripheral inflammation is associated with "altered" WM microstructure and lower cognitive functioning already in adolescents, and a specific circulating GPC may be a new molecule indexing this VF-related peripheral inflammation and its influences on brain structure and function.


Cellular copper content modulates differentiation and self-renewal in cultures of cord blood-derived CD34+ cells.
Several clinical observations have suggested that copper (Cu) plays a role in regulating haematopoietic progenitor cell (HPC) development. To further study this role we used an ex vivo system. Cord blood-derived CD34+ cells were cultured in liquid medium supplemented with Kit- ligand, FLt3, interleukin 6 (IL-6), thrombopoietin and IL-3. Under these conditions, Cu content, measured by atomic absorption, was 7 ng/10(7) cells. Modulation of intracellular Cu was achieved by supplementing the cultures with the Cu chelator tetraethylenepentamine, which reduced cellular Cu (4 ng/10(7) cells), or ceruloplasmin or Cu sulphate that elevated cellular Cu (18 and 14 ng/10(7) cells respectively). The results indicated that low Cu content delayed differentiation, as measured by the surface antigens CD34, CD14 and CD15, colony-forming unit (CFU) frequency and cell morphology, while high Cu accelerated differentiation compared with Cu unmanipulated cultures. As a result, expansion of total cells, CFU and CD34+ cells in low Cu was extended (12-16 weeks), and in high Cu was shortened (2-4 weeks), compared with control cultures (6-8 weeks). These effects required modulation of intracellular Cu only during the first 1-3 weeks of the culture; the long-term effects persisted thereafter, suggesting that the decision process for either self-renewal or differentiation is taken early during the culture. This novel method of controlling cell proliferation and differentiation by copper and copper chelators might be utilized for ex vivo manipulation of HPC for various clinical applications.


Somatosensory conditioning of sexual arousal and copulatory behavior in the male rat: a model of fetish development.
Previous studies have established that neutral olfactory cues associated with sexual reward or inhibition become conditioned stimuli that direct male or female rats toward or away from potential sex partners bearing the cue. Here we examined the ability of a somatosensory cue to exert stimulus control over sexual arousal and copulation in male rats. In the first experiment, two groups of sexually naïve male rats had their first copulatory experiences with receptive females in bilevel chambers with or without a rodent jacket. On a final copulatory test, half of the rats in each group were tested with the jacket on or off. Rats trained and tested without the jacket copulated normally, as did rats trained and tested with the jacket on, and rats trained without the jacket but tested with the jacket on. However, significantly fewer rats trained with the jacket on but tested with the jacket off copulated to ejaculation, and those that did made significantly fewer anticipatory level changes and had fewer significantly longer mount, intromission, and ejaculatory latencies, and fewer ejaculations. In the second experiment, one group of sexually naïve males was given differential conditioning trials in bilevel chambers to associate the jacket on with sexual reward (copulation to ejaculation with a sexually receptive female) and the jacket off with sexual inhibition (thwarted copulatory attempts with a sexually nonreceptive female). A second group had the opposite order of association. A final test with receptive females was made for all males with the jacket on. Males trained to associate the jacket with excitation displayed normal copulation whereas males trained to associate the jacket with inhibition displayed significantly fewer anticipatory level changes and ejaculations, and had significantly longer ejaculation latencies. Thus, somatosensory cues can signal sexual excitation or inhibition in male rats depending on the conditioning history.


Application of dairy manure as fertilizer in dry land in East China: field monitoring and model estimation of heavy metal accumulation in surface soil.
Manure-based fertilizer is usually applied to agricultural soils to increase soil fertility and improve soil quality. However, this practice has an impact on the soil environment, e.g., increasing heavy metal contents. The aim of this study was to evaluate and estimate the accumulation tendencies of eight heavy metals, including arsenic (As), cadmium (Cd), chromium (Cr), copper (Cu), mercury (Hg), lead (Pb), manganese (Mn), and zinc (Zn) in a soil fertilized continuously with dairy manure through a 5 years' field-scale experiment. Contents of the As, Cd, Cr, Cu, Mn, and Zn gradually increased with the fertilization time of dairy manure at the stable rate of around 326 t hm-2 year-1, leading to annual mean increases of 3.6%, 2.4%, 3.9%, 3.8%, 4.2%, and 6.1%, respectively. Based on the prediction of a dynamic mass balance model using the current practice, the contents of Cd and Zn in the fertilized soil would reach the Chinese standard values for agricultural soils in 48 and 35 years. The mitigation measures, such as lower application rates, for the environmental risk of heavy metal accumulation should be considered.


Blood plasma catecholamine concentration of poult embryos during the transition from diffusive to convective respiration.
1. Plasma catecholamines were determined in turkey embryos. 2. Decreased eggshell permeability depressed dopamine and epinephrine, but increased permeability also depressed epinephrine. 3. Norepinephrine (NE) was greatest before transition to convective respiration when hypoxia occurs. 4. NE was associated inversely with eggshell permeability. 5. Catecholamines appear to assist the embryo during hatching hypoxia.


Methylxanthines stimulate calcium transport and inhibit cyclic nucleotide phosphodiesterases in abalone sperm.
Experiments were designed to determine the mechanism by which methylxanthines elevate abalone sperm cAMP concentrations and induce the acrosome reaction (AR). Theophylline or, more effectively, 1-methyl-3-isobutylxanthine (MIX) inhibit the cyclic nucleotide phosphodiesterase activities of abalone sperm homogenates. 45Ca2+ uptake by sperm is also stimulated by theophylline, and more effectively by MIX, and this stimulatory effect is blocked by KCN. Verapamil, a compound known to antagonize Ca2+ conductance, has no effect on the Ca2+ or MIX-induced cAMP elevation at concentrations up to 200 microM. However, verapamil reduces the sperm cAMP elevation caused by the addition of Ca2+ plus MIX. This inhibition is not complete, even at 200 microM verapamil. The AR induced by Ca2+ plus MIX is completely inhibited by 200 microM verapamil. The data suggest that these methylxanthines elevate abalone sperm cyclic nucleotide concentrations by inhibiting cyclic nucleotide phosphodiesterase activities. Furthermore, since sperm cAMP metabolism is modulated by Ca2+ flux, methylxanthines also appear to elevate abalone sperm cAMP concentrations by their effects on Ca2+ transport. The Ca2+-induced cAMP elevation occurs through a verapamil-insensitive mechanism, whereas the potentiation by MIX of the Ca2+ effect to elevate cAMP occurs through both verapamil-insensitive and -sensitive mechanisms. The methylxanthine-induced AR is mediated by a primary effect on Ca2+ transport and occurs through a verapamil-sensitive mechanism. Cyclic AMP may play a role in the methylxanthine-induced AR, but does not appear to act as the primary mediator of this exocytotic event.