Optimal Dissociation Methods Differ for N- and O-Glycopeptides.
Site-specific characterization of glycosylation requires intact glycopeptide analysis, and recent efforts have focused on how to best interrogate glycopeptides using tandem mass spectrometry (MS/MS). Beam-type collisional activation, i.e., higher-energy collisional dissociation (HCD), has been a valuable approach, but stepped collision energy HCD (sceHCD) and electron transfer dissociation with HCD supplemental activation (EThcD) have emerged as potentially more suitable alternatives. Both sceHCD and EThcD have been used with success in large-scale glycoproteomic experiments, but they each incur some degree of compromise. Most progress has occurred in the area of N-glycoproteomics. There is growing interest in extending this progress to O-glycoproteomics, which necessitates comparisons of method performance for the two classes of glycopeptides. Here, we systematically explore the advantages and disadvantages of conventional HCD, sceHCD, ETD, and EThcD for intact glycopeptide analysis and determine their suitability for both N- and O-glycoproteomic applications. For N-glycopeptides, HCD and sceHCD generate similar numbers of identifications, although sceHCD generally provides higher quality spectra. Both significantly outperform EThcD methods in terms of identifications, indicating that ETD-based methods are not required for routine N-glycoproteomics even if they can generate higher quality spectra. Conversely, ETD-based methods, especially EThcD, are indispensable for site-specific analyses of O-glycopeptides. Our data show that O-glycopeptides cannot be robustly characterized with HCD-centric methods that are sufficient for N-glycopeptides, and glycoproteomic methods aiming to characterize O-glycopeptides must be constructed accordingly.


Nanometers-Thick Ferromagnetic Surface Produced by Laser Cutting of Diamond.
In this work, we demonstrate that cutting diamond crystals with a laser (532 nm wavelength, 0.5 mJ energy, 200 ns pulse duration at 15 kHz) produced a ≲20 nm thick surface layer with magnetic order at room temperature. We measured the magnetic moment of five natural and six CVD diamond crystals of different sizes, nitrogen contents and surface orientations with a SQUID magnetometer. A robust ferromagnetic response at 300 K was observed only for crystals that were cut with the laser along the (100) surface orientation. The magnetic signals were much weaker for the (110) and negligible for the (111) orientations. We attribute the magnetic order to the disordered graphite layer produced by the laser at the diamond surface. The ferromagnetic signal vanished after chemical etching or after moderate temperature annealing. The obtained results indicate that laser treatment of diamond may pave the way to create ferromagnetic spots at its surface.


His-Purkinje responses and refractory periods during atrial extrastimulation in children with heart defects.
During atrial extrastimulation, split His potentials, prolonged His-to-ventricular (H2V2) intervals and block distal to the His bundle deflection were observed in both preoperative and postoperative children with heart defects. His-Purkinje responses and refractoriness were identified in 31 of 78 (40%) pediatric patients (20 of 51 preoperative and 11 of 27 postoperative) during atrial extrastimulation coupled to sinus and/or paced cycle lengths. Split His potentials were found in 14 patients (eight preoperative and six postoperative) and His bundle relative refractory periods ranged from 250--490 msec. Prolonged H2V2 intervals were found in these and in an additional 16 patients (11 preoperative and five postoperative) and the relative refractory period of the His-Purkinje system ranged from 230--500 msec. Block distal to the His deflection occurred in seven patients (five preoperative and two postoperative) and the effective refractory period ranged from 230--510 msec. Split His potentials, long H2V2 intervals and block distal to the His bundle deflection produced by atrial extrastimulation were found in peroperative as well as postoperative children. These responses probably represent functional electrophysiologic characteristics of the pediatric cardiac conduction system.


Hyponatraemia and congestive heart failure refractory to diuretic treatment. Utility of tolvaptan.
Heart failure (HF) is currently one of the most significant healthcare problems in Spain and has a continuously increasing prevalence. Advances in our understanding of the various biological responses that promote cardiac remodelling and pulmonary venous congestion constitute the basis of current treatment. This article, prepared by members of the HF groups of the Spanish Society of Cardiology and the Spanish Society of Internal Medicine, discusses the current therapeutic strategies for patients with congestion refractory to diuretic treatment. The article includes our clinical experience with the use of tolvaptan as an additional treatment for congestion associated with hyponatraemia. To this end, we propose an algorithm for the use of tolvaptan in patients with congestive HF, natraemia <130mEq/l and poor response to conventional diuretic treatment.


Post-synthetic anion exchange in iron(ii) 1,2,4-triazole based spin crossover materials via mechanochemistry.
A facile method for post-synthetic exchange of anions in an iron(ii) spin crossover material using mechanochemistry is described. Dry grinding of the [Fe(atrz)3]Cl2 complex (atrz = 4-amino-1,2,4-triazole) in the presence of an excess of sodium halide salt results in the complete exchange of anions and formation of [Fe(atrz)3]Br2 and [Fe(atrz)3]I2 in a solid-state metathesis reaction. The method represents a new strategy for tuning active switching properties such as the transition temperature in spin crossover systems. Formation of stable by-products was identified as a major driving force for exchange and a straightforward method to predict the likely outcome of such reactions using simple thermodynamic considerations is presented.


Expression of a polygalacturonase associated with tomato seed germination.
Radicle protrusion from tomato (Lycopersicon esculentum Mill.) seeds to complete germination requires weakening of the endosperm tissue opposite the radicle tip. In common with other cell wall disassembly processes in plants, polygalacturonases (PGs) may be involved. Only calcium-dependent exo-PG activity was detected in tomato seed protein extracts. Chromatographic profiles of a partially acid-hydrolyzed fraction of polygalacturonic acid further digested with seed extract were consistent with the presence of only calcium-dependent exo-PG activity. In addition, a transcript encoding a previously unknown PG was detected prior to the completion of germination. The mRNA, produced from a gene (LeXPG1) estimated by Southern analysis to be represented once in the genome, was also present in flowers (anthers) and in lower amounts in roots and stems. LeXPG1 mRNA abundance was low during seed development, increased during imbibition, and was even greater in seeds that had completed germination. Expression of LeXPG1 during germination predominates in the endosperm cap and radicle tip, and in the radicle appears as a distinct band possibly associated with vascular tissue differentiation. We suggest that PG is involved in cell wall loosening of the endosperm necessary for radicle protrusion from tomato seeds and in subsequent embryo and seedling growth.


Creating thought diversity: the antidote to group think.
The Management Moment is a regular column within the Journal of Public Health Management and Practice. Edward L. Baker, MD, MPH, MSc, serves as the Management Moment Editor. Dr Baker is Director of the North Carolina Institute for Public Health, School of Public Health, at the University of North Carolina at Chapel Hill. This column provides commentary and guidance on timely management issues commonly encountered in public health practice.


Multiple potential regulatory elements in the 5' flanking region of the human alpha 1a-adrenergic receptor.
In spite of their critical importance in myocardial hypertrophy and benign prostatic hyperplasia, nothing is known about mechanisms underlying transcriptional regulation of alpha 1a-adrenergic receptors (alpha 1aARs). Therefore we cloned 6.2 kb of novel sequence upstream of the initiator ATG in the human alpha 1aAR gene. Sequence analysis reveals a TATA-less promoter, the presence of several initiator (Inr) consensus sequences, multiple GC rich regions consistent with Sp-1 binding, and consensus sequences for AP-1 and AP-2 as well as putative cis transcriptional regulatory elements for binding of CREB (cyclic-AMP response element binding protein), glucocorticoids, estrogen, and insulin. Compared to the alpha 1bAR, the alpha 1aAR has several more cis regulatory elements, suggesting more complex regulation. The importance of alpha 1aARs in human disease makes it imperative to determine mechanisms underlying transcription and ultimately expression of this receptor. These studies can now be undertaken with the availability of human alpha 1aAR 5'-flanking and 5'-untranslated sequence.


Development of Visible Spectrophotometric Methods for the Determination of Tricyclic Antidepressants Based on Formation of Molecular Complexes with p-Benzoquinones.
Tricyclic antidepressants are commonly employed in the management of major depressive disorders. The present work describes two visible (VIS) spectrophotometric techniques that utilize the formation of charge transfer complexes between four antidepressant compounds, namely, amitriptyline hydrochloride (AMI), imipramine hydrochloride (IMI), clomipramine hydrochloride (CLO), and trimipramine maleate (TRI) acting as electron donors and two p-benzoquinones, namely, p-chloranilic acid (pCA) and 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ), serving as electron acceptors. The stoichiometry of the compounds produced exhibited a consistent 1:1 ratio in all instances, as established by Job's method. Molar absorptivities, equilibrium association constants, and several other spectroscopic properties were determined for all complexes. The developed spectrophotometric techniques were validated intra-laboratory and successfully applied for quantitative assessment of the four antidepressant active ingredients in several commercial pharmaceutical formulations. The methods are relatively simple, fast, and use readily available laboratory instrumentation, making them easily applicable by most quality control laboratories worldwide.


The use of multiparametric monitoring during islet cell isolation and culture: a potential tool for in-process corrections of critical physiological factors.
Variables such as pH, pCO2, and PO2 have been established in the literature as critical factors that could affect the outcome of the islet cell processing and, therefore, the quality of the cells that could be transplanted. This report describes a highly accurate continuous multiparametric monitoring system and its evaluation for continuous monitoring of physiological variables during critical steps of the islet isolation procedure as well as during in vitro culture of the insulin-producing cells. Close monitoring of these variables could be of assistance to improve the outcome of islet cell processing, allowing to identify as soon as possible problems that could be corrected during the procedure, as well as during in vitro preservation, or shipment to remote sites.


Role of certain trace minerals in oxidative stress, inflammation, CD4/CD8 lymphocyte ratios and lung function in asthmatic patients.
Asthma is associated with increased inflammation, oxidative stress and abnormal immune system function. We determined the distributions of several essential trace minerals and assessed their relationships to factors that are associated with the pathophysiological status of patients with mild/moderate asthma. We enrolled 25 asthmatic patients and 25 healthy subjects. We measured: blood trace minerals, zinc (Zn), copper (Cu) and selenium (Se); oxidative stress markers thiobarbituric acid reactive substances (TBARS); antioxidant enzyme activities; percentages of CD4 and CD8 lymphocyte subsets; high-sensitivity C-reactive protein (hs-CRP); and a lung function index (FEV1/FVC%). Compared with healthy subjects, asthmatics had lower concentrations of Zn and Se; higher Cu concentrations, and Cu/Zn and Cu/Se ratios; and lower antioxidant enzyme glutathione peroxidase (GPx), glutathione reductase (GR) and catalase activities. Significantly increased concentrations of hs-CRP, TBARS and CD4/CD8 lymphocyte ratios were also observed. Furthermore, plasma TBARS or hs-CRP concentrations were negatively associated with Se concentrations, but were positively associated with Cu/Se ratios. CD4/CD8 lymphocyte ratios were inversely correlated with Se, while it was positively correlated with Cu/Se ratio. FEV1/FVC% was also significantly correlated with Se concentrations, and Cu/Se and Cu/Zn ratios. Abnormal distributions of these trace minerals may aggravate oxidative damage and inflammation, increased CD4/CD8 lymphocyte ratios and decreased lung function in asthma.


Enhancing capacity to make sexuality-related decisions in people with an intellectual disability.
The aim of the study was to apply an intervention to the area of sexual knowledge in order to determine if capacity to make sexuality-related decisions could be improved. The study adopted a single subject design using multiple baseline method with four adults with a moderate intellectual disability. The intervention consisted of individually tailored sex education adapted from Living Your Life (Bustard 2003). Treatment was offered to each participant twice weekly for a 10-week period on a one-to-one basis. The Sexual Consent and Education Assessment (SCEA, Kennedy 1993) was used for measurement purposes. The SCEA K-Scale (knowledge) and the S-Scale (safety practices) were administered weekly throughout the baseline, treatment and post-treatment phases of the study. Staff concerns were also assessed using the SCEA Inappropriate Sexual Behaviour Scale. All four participants improved their decision-making capacity in all targeted areas as measured by improvements in K-Scale and S-Scale scores. Staff concerns were not increased as indicated by results on the Inappropriate Sexual Behaviour Scale. Six-month follow-up data for three of the participants showed maintenance of scores on the S-Scale and some decay in scores on the K-Scale from post-intervention performance. The results demonstrate that tailored sexuality education can improve capacity to make sexuality-related decisions.


L-arginine does not improve cortical perfusion or histopathological outcome in spontaneously hypertensive rats subjected to distal middle cerebral artery photothrombotic occlusion.
The potential of nitric oxide (NO) to influence positively or negatively the outcome of mechanically induced focal cerebral ischemia is still controversial. Recent evidence suggests that NO of vascular origin, whether synthesized from exogenously administered L-arginine (L-Arg) or from NO donor compounds, is beneficial but that of neuronal origin is not. However, the therapeutic potential of NO to ameliorate stroke induced by arterial thrombosis has not been reported. We assessed the therapeutic effect of L-Arg administration in spontaneously hypertensive rats (SHR) subjected to permanent photothrombotic occlusion of the distal middle cerebral artery (dMCA). The ipsilateral carotid artery was left unligated to enhance L-Arg delivery into the putative penumbral region. Local CBF (LCBF) was assessed at 30 min by the [14C]iodoantipyrine technique (n = 9), while histological infarct volumes and index of peripheral ischemic cell change were determined at 3 days (n = 7). Rats (n = 9) given 300 mg/kg L-Arg at 18 and 3 h before photothrombotic dMCA occlusion and at 5 min afterward displayed no significant differences in LCBF compared with animals (n = 8) injected with water (the carrier vehicle) and similarly irradiated. Infarct volumes were also similar, being 37.0 +/- 9.7 mm3 (SD) in the vehicle-treated and 49.1 +/- 17.2 mm3 (SD) in the L-Arg-treated groups (both n = 7), as were assessments of ischemic neuronal density in the penumbra. In contrast, L-Arg administered intravenously in a dose of 300 mg/kg to nonischemic SHR (n = 5) increased cortical CBF by approximately 75% during a 70-min observation period. We conclude that thrombotic processes superimposed upon cerebral ischemia may facilitate tissue reactions that offset the potentially beneficial effect of L-Arg, and this caveat must be considered when proposing L-Arg for clinical treatment of focal thrombotic stroke.


Outcomes Comparison of Robot-Assisted and Video-Assisted Thoracoscopic Cardiac Sympathetic Denervation.
Cardiac sympathetic denervation (CSD) is a surgical antiadrenergic procedure that can reduce sustained ventricular tachyarrhythmia (VT). Video-assisted thoracoscopic surgery (VATS) is currently the standard approach used in CSD, and the practicality for robot-assisted thoracoscopic surgery (RATS) has yet to be investigated. We conducted a single-center retrospective study of all adult patients (N = 67) who underwent CSD from 2015 to 2021. We compared short-term outcomes of those treated with RATS versus VATS thoracic sympathectomy. For patients with VT, we examined the effectiveness of a RATS approach in reducing implantable cardioverter defibrillator (ICD) shock burden. A total of 34 patients underwent RATS cardiac denervation, and 33 underwent VATS cardiac denervation. Those undergoing RATS denervation had a significantly shorter procedure duration with a median of 129 min (P = 0.008). Patients receiving the VATS approach were significantly more complicated by pneumothorax (P = 0.004) and overall complications (P = 0.01) when compared with the RATS approach. At 1 year after surgery, both groups had significant reductions in ICD shocks compared with before surgery, both decreasing from a median of 4 to 0 shocks (P < 0.001). In addition, at 1 year after surgery, the percentage of patients with persistent ICD shocks and the median of ICD shocks were similar between the groups. The RATS approach to cardiac denervation has similar 1-year follow-up outcomes in reducing recurrent VT as the VATS approach. However, patients undergoing RATS denervation experienced better perioperative outcomes. This shows promise for robotic CSD to be an effective and safe therapeutic option for patients with malignant arrhythmias.


Treatment of intramedullary spinal cord tumors: a modified Delphi technique of the North American Spine Society Section of Spine Oncology.
Intramedullary spinal cord tumors (IMSCTs) are rare tumors with heterogeneous presentations and natural histories that complicate their management. Standardized guidelines are lacking on when to surgically intervene and the appropriate aggressiveness of resection, especially given the risk of new neurological deficits following resection of infiltrative tumors. Here, the authors present the results of a modified Delphi method using input from surgeons experienced with IMSCT removal to construct a framework for the operative management of IMSCTs based on the clinical, radiographic, and tumor-specific characteristics. A modified Delphi technique was conducted using a group of 14 neurosurgeons experienced in IMSCT resection. Three rounds of written correspondence, surveys, and videoconferencing were carried out. Participants were queried about clinical and radiographic criteria used to determine operative candidacy and guide decision-making. Members then completed a final survey indicating their choice of observation or surgery, choice of resection strategy, and decision to perform duraplasty, in response to a set of patient- and tumor-specific characteristics. Consensus was defined as ≥ 80% agreement, while responses with 70%-79% agreement were defined as agreement. Thirty-six total characteristics were assessed. There was consensus favoring surgical intervention for patients with new-onset myelopathy (86% agreement), chronic myelopathy (86%), or progression from mild to disabling numbness (86%), but disagreement for patients with mild numbness or chronic paraplegia. Age was not a determinant of operative candidacy except among frail patients, who were deemed more suitable for observation (93%). Well-circumscribed (93%) or posteriorly located tumors reaching the surface (86%) were consensus surgical lesions, and participants agreed that the presence of syringomyelia (71%) and peritumoral T2 signal change (79%) were favorable indications for surgery. There was consensus that complete loss of transcranial motor evoked potentials with a 50% decrease in the D-wave amplitude should halt further resection (93%). Preoperative symptoms seldom influenced choice of resection strategy, while a distinct cleavage plane (100%) or visible tumor-cord margins (100%) strongly favored gross-total resection. The authors present a modified Delphi technique highlighting areas of consensus and agreement regarding surgical management of IMSCTs. Although not intended as a substitute for individual clinical decision-making, the results can help guide care of these patients. Additionally, areas of controversy meriting further investigation are highlighted.


[A physiological analysis of kidney ion-regulating function in children with enuresis].
Peculiarities of the excretion of ions (Na, K, Ca, Mg) and water were studied in healthy children and children with the nocturnal enuresis, aged 6-15 years. A greater diuresis in the enuretic children is due to an increased excretion of the osmotically active substances including Na and Mg; excretion of K and Ca does nor differ from the control. A new formula is proposed for the quantitative evaluation of the role of different substances in the osmolar clearance. A high correlation is found between the sodium and magnesium excretion and the osmotic free water reabsorption. A single intranasal administration of 1-deamino-8-D-arginine-vasopressin (DDAVP) to the children before their going to bed returned to the norm the sodium and magnesium excretion in the enuretic children. It is suggested that the defect peculiar to this particular pathology is associated with a decrease in the ion reabsorption in the thick ascending Henle loop. The normal level of the ion transport is restored after stimulation of V2-receptors by DDAVP. An explanation is suggested of the mechanism of the increase of diuresis with a simultaneous rise in the osmotically free water reabsorption in children with enuresis.


ATP synthase hexamer assemblies shape cristae of Toxoplasma mitochondria.
Mitochondrial ATP synthase plays a key role in inducing membrane curvature to establish cristae. In Apicomplexa causing diseases such as malaria and toxoplasmosis, an unusual cristae morphology has been observed, but its structural basis is unknown. Here, we report that the apicomplexan ATP synthase assembles into cyclic hexamers, essential to shape their distinct cristae. Cryo-EM was used to determine the structure of the hexamer, which is held together by interactions between parasite-specific subunits in the lumenal region. Overall, we identified 17 apicomplexan-specific subunits, and a minimal and nuclear-encoded subunit-a. The hexamer consists of three dimers with an extensive dimer interface that includes bound cardiolipins and the inhibitor IF1. Cryo-ET and subtomogram averaging revealed that hexamers arrange into ~20-megadalton pentagonal pyramids in the curved apical membrane regions. Knockout of the linker protein ATPTG11 resulted in the loss of pentagonal pyramids with concomitant aberrantly shaped cristae. Together, this demonstrates that the unique macromolecular arrangement is critical for the maintenance of cristae morphology in Apicomplexa.


Prospective assessment of quality of life of octogenarians after cardiac surgery: factors predicting long-term outcome.
Our objective was to assess the long-term prognosis and quality of life (QoL) of elderly patients after cardiac surgery. The Nottingham Health Profile (NHP) QoL data were recorded from 104 patients who were older than 70 years at the time of primary cardiac surgery in 1993. All living patients were controlled at 15+/-3.2 months and 8.2+/-0.27 years after discharge. The 1-year, 5-year and actual survival rates were 94%, 76%, and 59%, respectively. Risk factors for death were urgency of the operation (relative risk ratio, 2.0; 95% confidence interval, 1.2-3.6), ejection fraction below 50% (2.1; 1.1-3.9), and preoperative renal failure (2.1; 1.0-4.0). Cardiac operated octogenarians took advantage from age and sex matched reference populations at 15 months in the NHP dimension of pain (P=0.001). The QoL decreased gradually during the follow-up similarly in both groups in dimensions describing energy (P=0.001), pain (P=0.003), and mobility (P=0.042). Diabetes, low energy score and high pain score at 15 months, treatment in intensive care unit >3 days, and duration of symptoms >120 days preoperatively were associated with impaired QoL. Survival and QoL were similar for cardiac operated octogenarians and age and sex matched controls at 15 months and 8.2 years after cardiac surgery.


Caring for People With Young Onset Dementia: An Interpretative Phenomenological Analysis of Family Caregivers' Experiences.
People with young onset dementia (YOD) experience many unique challenges. Similarly, family caregivers experience changing personal, familial, and societal roles. However, YOD is under recognized and under resourced with a lack of dedicated models of care. The current study explored family members' experience of caregiving for persons with YOD to identify barriers and opportunities to accessing support. Qualitative interviews were conducted with six family caregivers, using interpretative phenomenological analysis. Findings show that caregivers experience significant uncertainty within their role, difficulty accessing timely diagnosis, and face unclear care pathways and age-related restrictions to accessing support. These findings also reflect elements of ambiguous loss and anticipatory grief: caregivers experienced sudden changes to relationships and family structure, along with the anticipation of a changed future. Optimum models of care should target timely diagnosis and clear post-diagnosis pathways and services that are flexible, relevant, and accessible. Services should offer psychosocial support for family members as they adjust to their new role as caregivers. [Journal of Psychosocial Nursing and Mental Health Services, 57(11), 37-44.].


Primary anterior cruciate ligament reconstruction using fresh-frozen, nonirradiated patellar tendon allograft: minimum 2-year follow-up.
There are conflicting reports of allograft performance, immune response, tissue incorporation, and rerupture rates when used for anterior cruciate ligament reconstruction. To evaluate the clinical outcome of a fresh-frozen, nonirradiated, patellar tendon allograft for primary anterior cruciate ligament reconstruction surgery. Case series; Level of evidence, 4. Patients who underwent endoscopic primary anterior cruciate ligament reconstruction with allograft tissue a minimum of 2 years ago were evaluated with physical examinations, the KT-1000 arthrometer, functional testing, radiographic evaluation, subjective assessment, and outcomes tools. Fifty-nine patients (60 knees) were evaluated at an average of 51 months after surgery. Ninety-four percent of patients were mostly or completely satisfied. A negative pivot shift test result was noted in 90% of subjects. The KT-1000 arthrometer side-to-side differences were < or =3 mm in 95% of patients, and no patient exceeded 5 mm. The mean International Knee Documentation Committee score was 78 (SD = 19), and the mean Lysholm score was 82 (SD = 17). There were no clinical symptoms consistent with graft rejection or infection. Radiographic evaluation demonstrated infrequent significant tunnel widening. Use of a fresh-frozen, nonirradiated allograft for primary reconstruction of the anterior cruciate ligament is a successful procedure both subjectively and functionally for restoring stability in patients selected for allograft reconstruction. In the patients selected for this surgical procedure, clinical, arthrometric stability testing, and subjective satisfaction were comparable to our previously published cohort studies using patellar tendon autograft at similar postoperative follow-up.


Quantification of Atmospheric Mercury Deposition to and Legacy Re-emission from a Subtropical Forest Floor by Mercury Isotopes.
Air-soil exchange of elemental mercury vapor (Hg0) is an important component in the budget of the global mercury cycle. However, its mechanistic detail is poorly understood. In this study, stable Hg isotopes in air, soil, and pore gases are characterized in a subtropical evergreen forest to understand the mechanical features of the air-soil Hg0 exchange. Strong HgII reduction in soil releases Hg0 to pore gas during spring-autumn but diminishes in winter, limiting the evasion in cold seasons. Δ199Hg in air modified by the Hg0 efflux during flux chamber measurement exhibit seasonality, from -0.33 ± 0.05‰ in summer to -0.08 ± 0.05‰ in winter. The observed seasonal variation is caused by a strong pore-gas driven soil efflux caused by photoreduction in summer, which weakens significantly in winter. The annual Hg0 gross deposition is 42 ± 33 μg m-2 yr-1, and the corresponding Hg0 evasion from the forest floor is 50 ± 41 μg m-2 yr-1. The results of this study, although still with uncertainty, offer new insights into the complexity of the air-surface exchange of Hg0 over the forest land for model implementation in future global assessments.


[Molecular organization of the barley (Hordeum vulgare) genome].
Studies in peculiarities of the DNA secondary structure in barley by means of thermal denaturation and renaturation shows that there are three types of the nucleotide sequences organization in DNA. More than 95% of the genome composition contain distributed repetitive sequences, in one part of the concentration of the repetitive sequences being higher as compared to bulk of them. About 3.5% of DNA is enriched with A-T pairs and contains no repetitive sequences. There is no "unique" part in the barley genome, which is natural for animals. Slowly renaturation sequences repeat 4 times.


Vibration of the organ of Corti within the cochlear apex in mice.
The tonotopic map of the mammalian cochlea is commonly thought to be determined by the passive mechanical properties of the basilar membrane. The other tissues and cells that make up the organ of Corti also have passive mechanical properties; however, their roles are less well understood. In addition, active forces produced by outer hair cells (OHCs) enhance the vibration of the basilar membrane, termed cochlear amplification. Here, we studied how these biomechanical components interact using optical coherence tomography, which permits vibratory measurements within tissue. We measured not only classical basilar membrane tuning curves, but also vibratory responses from the rest of the organ of Corti within the mouse cochlear apex in vivo. As expected, basilar membrane tuning was sharp in live mice and broad in dead mice. Interestingly, the vibratory response of the region lateral to the OHCs, the "lateral compartment," demonstrated frequency-dependent phase differences relative to the basilar membrane. This was sharply tuned in both live and dead mice. We then measured basilar membrane and lateral compartment vibration in transgenic mice with targeted alterations in cochlear mechanics. Prestin(499/499), Prestin(-/-), and Tecta(C1509G/C1509G) mice demonstrated no cochlear amplification but maintained the lateral compartment phase difference. In contrast, Sfswap(Tg/Tg) mice maintained cochlear amplification but did not demonstrate the lateral compartment phase difference. These data indicate that the organ of Corti has complex micromechanical vibratory characteristics, with passive, yet sharply tuned, vibratory characteristics associated with the supporting cells. These characteristics may tune OHC force generation to produce the sharp frequency selectivity of mammalian hearing.


Multiparametric analysis of preoperative lymphatic mapping with the use of combined deep plus superficial radiotracer injection technique in early breast cancer.
According to literature, combined tracer injection techniques seem to be the more powerful choice to eliminate technical and patient-related limitations. In this study, we analysed the quantitative performance parameters of combined deep plus superficial radiotracer injection and their correlation with a set of clinical, pathological and technical factors. One hundred and sixteen women who underwent preoperative sentinel lymph node (SLN) mapping were studied prospectively. All patients received the simultaneous deep and superficial injection of 99mTc-nanocolloid. Mapping success rate, mean number of SLNs per patient and radiotracer uptake of SLNs were determined. The possible effects of age, tumour stage, laterality and location, type and time of previous biopsy, and SLN status on the quantitative parameters were analysed. Axillary SLNs were visualized in all cases. Mean number of axillary SLNs was 2.15 and advanced age (>50) significantly decreased the number of SLNs. Radiotracer uptake of SLNs was also significantly decreased by advanced age and a shorter time interval between biopsy and mapping (<10 days). Extra-axillary SLN visualization rates for medial, lateral and periareolar injection sites were 32%, 16% and 8%. Although SLN gamma probe counts were significantly higher in the single-day protocol, the 2-day protocol gave better contrast values which was also an important parameter in lesion detection. The combined radiotracer injection technique successfully demonstrated axillary and extra-axillary SLNs. Advanced age and previous biopsy time can lower the accuracy and reliability of SLN biopsy. Although periareolar injection gave the best results for the axilla, it was still insensitive for extra-axillary SLNs in spite of deep injection.


Neonatal Anthropometrics and Obesity Treatment Response in Children and Adolescents.
To investigate the relationship between in utero growth conditions, as indicated by neonatal anthropometric measures, and childhood obesity treatment response, to examine the potential usefulness of neonatal anthropometrics as a potential childhood obesity treatment stratification tool. The study included 2474 children and adolescents with obesity (mean age, 11.2 years; range, 5.0-18.9 years) treated at the Children's Obesity Clinic in Holbæk, Denmark. Treatment response was registered prospectively, and neonatal data were collected from national electronic registers. Birth weight, birth length, birth weight for gestational age, and large for gestational age status were positively associated with the degree of obesity at treatment initiation. After a mean (SD) of 1.27 (0.69) years of enrollment in obesity treatment, the children exhibited a mean reduction of -0.32 (0.50) in body mass index SD score. No significant associations between neonatal anthropometric measures and childhood obesity treatment response were detected. Neonatal anthropometric measures were positively associated with the degree of obesity at treatment initiation but not with response to multidisciplinary treatment of childhood obesity. Individualization of obesity treatment based on neonatal anthropometry does not seem warranted.


[Hansen's disease in children under fifteen years-old in Jequitinhonha Valley, Minas Gerais, Brazil].
Hansen's disease has a slow evolution, which mostly affects the adult population. Detecting cases in children shows that the endemic continues and there is early exposure of the population to the bacillus. This is an ecological study with the objective of analyzing the occurrence of Hansen's disease in children under fifteen in the Jequitinhonha Valley through epidemiological and operational indicators. The results show that the endemic continues in the region, demonstrating that the health services are not proactive and must implement specific prevention and control measures for this age group in order to reduce the transmission of the disease and the possible damage it can bring to the life of individuals, especially when it affects them in their childhood.


Superparamagnetic colloidal nanocrystal clusters coated with polyethylene glycol fumarate: a possible novel theranostic agent.
We report cell endocytosis, drug release, NMR relaxometry and in vitro MRI studies on a novel class of superparamagnetic colloidal nanocrystal clusters (CNCs) with various biocompatible coatings. It is shown that the transverse relaxivity r2, the parameter representing the MRI efficiency in negative contrast agents, for the PVA-coated, PEGF-coated, and crosslinked PEGF-coated CNCs, is high enough to contrast suitably the magnetic resonance images. The same samples have shown a good ability also in drug releasing (particularly the crosslinked PEGF-coated compound), thus finally allowing us to propose this class of compounds for future applications in theranostics.


Tissue proteomics for cancer biomarker development: laser microdissection and 2D-DIGE.
Novel cancer biomarkers are required to achieve early diagnosis and optimized therapy for individual patients. Cancer is a disease of the genome, and tumor tissues are a rich source of cancer biomarkers as they contain the functional translation of the genome, namely the proteome. Investigation of the tumor tissue proteome allows the identification of proteomic signatures corresponding to clinico-pathological parameters, and individual proteins in such signatures will be good biomarker candidates. Tumor tissues are also a rich source for plasma biomarkers, because proteins released from tumor tissues may be more cancer specific than those from non-tumor cells. Two-dimensional difference gel electrophoresis (2D-DIGE) with novel ultra high sensitive fluorescent dyes (CyDye DIGE Fluor satulation dye) enables the efficient protein expression profiling of laser-microdissected tissue samples. The combined use of laser microdissection allows accurate proteomic profiling of specific cells in tumor tissues. To develop clinical applications using the identified biomarkers, collaboration between research scientists, clinicians and diagnostic companies is essential, particularly in the early phases of the biomarker development projects. The proteomics modalities currently available have the potential to lead to the development of clinical applications, and channeling the wealth of produced information towards concrete and specific clinical purposes is urgent.


Serum lipid profiles, lipid ratios and Chronic Kidney Disease in a Chinese population.
To examine the association of serum lipids, lipid ratios with Chronic Kidney Disease (CKD) in a Chinese population. Data were drawn from a cross-sectional survey in China. CKD was defined as estimated glomerular filtration rate (eGFR) < 60 mL/min/1.73m(2) or albuminuria-to-creatinine ratio (ACR) > 30 mg/g. Multivariable logistic regressions and multivariate regression models were used. Serum lipids and lipid ratios included total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), TG/HDL-C ratio, TC/HDL-C ratio and LDL-C/HDL-C ratio. In men, only logarithm-transformed (log) TG was associated with CKD. The odds ratio (every SD increment) was 1.39 (95% CI 1.03-1.87, P = 0.03). In women, none of the serum lipids and lipid ratios was associated with CKD. Using multivariate regression models, it was shown that log TG and log TG/HDL-C were negatively correlated with eGFR (P < 0.05) in men and LDL-C and log LDL-C/HDL-C ratio were correlated with ACR in men. In female subjects, serum TC, log TG, log TG/HDL-C and log TC/HDL-C were negatively correlated with eGFR (P < 0.05). All of serum lipid profiles and lipid related ratio were not correlated with ACR in women. Serum TG is the only suitable predictor for CKD in men. However, in women, none of serum lipids and lipid ratio can be used as a predictor for CKD. Log TG and log TG/HDL-C are negatively correlated with eGFR in both genders.


Isolation of a novel mutated strain of Xanthomonas campestris for xanthan production using whey as the sole substrate.
This study was conducted to isolate novel lactose utilizing Xanthomonas campestris mutants. Such a mutant will assist the utilization of whey as the sole carbon source for xanthan gum production, lower costs of fermentation process and set a precise application for whey as a waste. In this study, a mutant strain (NA1) was isolated from Xanthomonas campestris cells exposed to nitrous acid mutagenesis Environmental conditions were optimized and maximum activity of the beta-galactosidase enzyme was obtained at pH 5.5 and 38 degrees C following which the beta-galactosidase activity in NA1 culture was increased 9.5 folds, compared to that of the wild type culture (336.1 U vs. 35.4 U). Xanthan gum production by NA1 using whey as carbon source was also studied. Using the experimental design of Plackett-Burman and statistical analysis, whey, as the main substrate and pH were the first factors affecting gum production among the seven parameters tested. Gum production using significant factors (such as substrate concentration and pH) was carried out in a lab-scale fermentor and 10 g L(-l) xanthan was obtained.


The sigma-1 receptor modulates methamphetamine dysregulation of dopamine neurotransmission.
Dopamine neurotransmission is highly dysregulated by the psychostimulant methamphetamine, a substrate for the dopamine transporter (DAT). Through interactions with DAT, methamphetamine increases extracellular dopamine levels in the brain, leading to its rewarding and addictive properties. Methamphetamine also interacts with the sigma-1 receptor (σ1R), an inter-organelle signaling modulator. Using complementary strategies, we identified a novel mechanism for σ1R regulation of dopamine neurotransmission in response to methamphetamine. We found that σ1R activation prevents methamphetamine-induced, DAT-mediated increases in firing activity of dopamine neurons. In vitro and in vivo amperometric measurements revealed that σ1R activation decreases methamphetamine-stimulated dopamine efflux without affecting basal dopamine neurotransmission. Consistent with these findings, σ1R activation decreases methamphetamine-induced locomotion, motivated behavior, and enhancement of brain reward function. Notably, we revealed that the σ1R interacts with DAT at or near the plasma membrane and decreases methamphetamine-induced Ca2+ signaling, providing potential mechanisms. Broadly, these data provide evidence for σ1R regulation of dopamine neurotransmission and support the σ1R as a putative target for the treatment of methamphetamine addiction.


Effect of ethanol on lipid metabolism in cultured hepatocytes.
Isolated rat hepatocytes were cultured in a modified HI-WO/BA medium for 16 h. In the following 24 h oleate or oleate plus ethanol was added to the medium. After this period the medium was changed again and the cultures were further incubated with [1-14C]oleate alone or with [1-14C]oleate plus ethanol for 6 h. This allowed a comparison of effects of short-term (6 h) and long-term (24 + 6 h) exposure to ethanol on fatty acid metabolism. The increased intracellular accumulation of triacylglycerol in the presence of ethanol was quantitatively accounted for by increased fatty acid uptake, by decreased fatty acid oxidation in the tricarboxylic acid cycle and by decreased VLDL (very-low-density lipoprotein)-triacylglycerol secretion. Ketone-body production was not affected. After short-term exposure the rate of accumulation of triacylglycerol was increased by 50%. This increase was accounted for by increased fatty acid uptake (44%), decreased tricarboxylic acid-cycle activity (49%) and decreased VLDL-triacylglycerol secretion (7%). After long-term exposure, the rate of accumulation of triacylglycerol was increased by 74%. This increase was accounted for by increased fatty acid uptake (34%), decreased tricarboxylic acid-cycle activity (34%) and decreased VLDL-triacylglycerol secretion (32%). The larger increase in accumulation of triacylglycerol after long-term exposure to ethanol was entirely accounted for by increased inhibition of secretion of VLDL-triacylglycerol. The biochemical mechanisms underlying the observations are discussed.


Prevalence of self-reported noncommunicable diseases in grassroot-level health worker of Petlad taluka: A cross-sectional study.
Noncommunicable diseases (NCDs) kill 41 million people each year globally. The financial burden of NCDs, including lifelong and expensive treatment with loss of income, forces many people into poverty. Healthcare workers (HCWs) are an essential group of the workforce in building a healthier society. They must choose a healthy lifestyle to have better health for themselves. To estimate the prevalence of self-reported NCDs in grassroot-level HCWs of Petlad taluka along with the study of various healthcare and lifestyle practices among them. In this cross-sectional study, grassroot-level HCWs of Petlad taluka of Anand district were given a self-administered questionnaire, which included their demographic details, their current health status, and health practices followed by them. The data collected were entered in Microsoft Excel-2019 and analysed by SPSS version 15. Descriptive analysis and univariate analysis were performed. NCDs' prevalence was 10.2%. Hypertension and diabetes were the commonest. Nearly 50% HCWs have their body mass index in a normal range. The majority of them were aware of healthy diet practices and exercise. Our study reported a lower prevalence of NCDs than other studies (50%). Dietary practices appear to be healthy, but a significant proportion of them are overweight. The majority of them prefer to consult at a government hospital. Healthy behaviour and practices should be encouraged and maintained in HCWs.


Caveolin-1 expression in odontogenic cysts and ameloblastomas.
The aim of this study is to evaluate the caveolin-1 expression in a group of odontogenic cysts and tumors. In this cross-sectional study, the expression of caveolin-1 was evaluated immunohistochemically in 75 samples including 18 cases of dentigerous cyst, 18 odontogenic keratocysts, 3 orthokeratinized odontogenic cysts, 2 calcifying odontogenic cysts and 34 ameloblastomas (solid and unicystic). Positive immunohistochemical reaction was found in 100% of odontogenic cysts and this was significantly more than both unicystic (65%) and solid (55%) ameloblastomas. The present study showed the expression of caveolin-1 in all odontogenic cysts and more than ameloblastomas. The results suggested that absence of caveolin-1 might enhance aggressiveness of odontogenic lesions and could be a useful marker for distinguishing ameloblastomas from other odontogenic lesions.


Ex vivo systems to study host-microbiota interactions in the gastrointestinal tract.
It is increasingly apparent that the microbial ecosystems in the mammalian gastrointestinal tract play an intricate role in health and disease. There is a growing interest in the development of targeted strategies for modulating health through the modification of these microbiota. Ecologists are faced with the challenge of understanding the structure and function of ecosystems, the component parts of which interact with each other in complex and diffuse ways. The human gut microbiota, with its high species richness and diversity (up to 1000 bacterial species per individual) including members of all three domains of life, situated in the dynamic environment of the gastrointestinal tract, is probably among the most complex ecosystems on this planet. In order to elucidate the mechanistic foundations, and physiological significance, of beneficial or pathogenic relationships between the gut microbiota and their hosts, researchers require tractable model ecosystems that allow to recapitulate and investigate host-microbe and microbe-microbe interactions. This review discusses ex vivo gastrointestinal models systems that can be used to gain mechanistic insights into the emergent properties of the host-microbial superorganism.


[Effects of bromocriptine on catecholamine metabolism in subjects with borderline arterial hypertension].
Effects of bromocriptine on hemodynamics and catecholamine exchange were evaluated in patients with borderline arterial hypertension with respect to their family history. The agent was shown to be able to bring down blood pressure and decrease plasma catecholamine levels. With this, platelet monoamine oxidase activity was increased in persons with a family history of hypertensive disease.


The role of Vitellogenin in the transfer of immune elicitors from gut to hypopharyngeal glands in honey bees (Apis mellifera).
Female insects that survive a pathogen attack can produce more pathogen-resistant offspring in a process called trans-generational immune priming. In the honey bee (Apis mellifera), the egg-yolk precursor protein Vitellogenin transports fragments of pathogen cells into the egg, thereby setting the stage for a recruitment of immunological defenses prior to hatching. Honey bees live in complex societies where reproduction and communal tasks are divided between a queen and her sterile female workers. Worker bees metabolize Vitellogenin to synthesize royal jelly, a protein-rich glandular secretion fed to the queen and young larvae. We ask if workers can participate in trans-generational immune priming by transferring pathogen fragments to the queen or larvae via royal jelly. As a first step toward answering this question, we tested whether worker-ingested bacterial fragments can be transported to jelly-producing glands, and what role Vitellogenin plays in this transport. To do this, we fed fluorescently labelled Escherichia coli to workers with experimentally manipulated levels of Vitellogenin. We found that bacterial fragments were transported to the glands of control workers, while they were not detected at the glands of workers subjected to RNA interference-mediated Vitellogenin gene knockdown, suggesting that Vitellogenin plays a role in this transport. Our results provide initial evidence that trans-generational immune priming may operate at a colony-wide level in honey bees.


Videofetoscopically assisted fetal tissue engineering: skin replacement.
Treatment of several congenital anomalies is frequently hindered by lack of enough tissue for surgical reconstruction in the neonatal period. The purposes of this study were (1) introduction of a novel concept in perinatal surgery, involving minimally invasive harvest of fetal tissue, which is then processed through tissue engineering techniques in vitro while pregnancy is allowed to continue, so that, at delivery, the newborn can benefit from having autologous, expanded tissue promptly available for surgical implantation at birth; (2) analysis of the progress of an engineered fetal skin graft with time, after implantation in the neonate; and (3) study of the effects of current tissue engineering techniques on fetal keratinocytes and fetal dermal fibroblasts. Ten 90- to 95-day-gestation fetal lambs underwent surgical creation of two large paramedian excisional skin defects on the posterior body wall. Subsequently, fetal skin specimens no larger than 1.5 x 1.5 cm were videofetoscopically harvested. Fetal keratinocytes and dermal fibroblasts were then separately cultivated and expanded in vitro for 45 to 50 days, resulting in a total of approximately 250 to 300 million cells. Seven to 10 days before fetal delivery, all cells were seeded in two layers on a 16 to 20-cm2, 3-mm thick biodegradable polyglycolic acid polymer matrix. One to 4 days after delivery, the autologous engineered skin was implanted over one of two previously created skin defects. The second skin defect region received an absorbable polymer scaffold without cells as a control. If necessary, the original skin wounds were further amplified before implantation. Each animal provided at least one time-point for histological analysis of both types of repair through excisional biopsies performed at weekly intervals, up to 8 weeks postimplantation. Normal skin specimens were also used as controls. Fetal and neonatal survival rates were 100%. Based on previous postnatal skin engineering studies, fetal dermal fibroblasts multiplied significantly faster in vitro (approximately fivefold) than expected. Fetal keratinocytes multiplied at expected postnatal rates. The engineered grafts induced faster epithelization of the wound (partial at 1 week and complete between 2 and 3 weeks postoperatively) than did the acellular ones (partial at 3 weeks and complete between 3 and 4 weeks postoperatively). Analysis of skin architecture showed a higher level of epidermal organization and less dermal scarring in the wounds that received the engineered, cell-implanted polymer scaffold. (1) Videofetoscopically assisted fetal tissue engineering is a viable method for obtaining expanded autologous tissue for prompt surgical reconstruction at birth. (2) Fetal skin can be expanded and engineered in vitro at faster rates than expected postnatally, with current tissue engineering techniques. (3) Engineered autologous fetal skin induces a faster and more organized healing of neonatal skin defects than that observed with second intention. This concept may prove useful for the treatment of certain human neonatal conditions such as giant neoplasias, ectopia cordis, and other body wall defects.


Talin and kindlin use integrin tail allostery and direct binding to activate integrins.
Integrin affinity regulation, also termed integrin activation, is essential for metazoan life. Although talin and kindlin binding to the β-integrin cytoplasmic tail is indispensable for integrin activation, it is unknown how they achieve this function. By combining NMR, biochemistry and cell biology techniques, we found that talin and kindlin binding to the β-tail can induce a conformational change that increases talin affinity and decreases kindlin affinity toward it. We also discovered that this asymmetric affinity regulation is accompanied by a direct interaction between talin and kindlin, which promotes simultaneous binding of talin and kindlin to β-tails. Disrupting allosteric communication between the β-tail-binding sites of talin and kindlin or their direct interaction in cells severely compromised integrin functions. These data show how talin and kindlin cooperate to generate a small but critical population of ternary talin-β-integrin-kindlin complexes with high talin-integrin affinity and high dynamics.


Reduced self-control leads to disregard of an unfamiliar behavioral option: an experimental approach to the study of neuroenhancement.
Neuroenhancement (NE), the use of psychoactive substances in order to enhance a healthy individual's cognitive functioning from a proficient to an even higher level, is prevalent in student populations. According to the strength model of self-control, people fail to self-regulate and fall back on their dominant behavioral response when finite self-control resources are depleted. An experiment was conducted to test the hypothesis that ego-depletion will prevent students who are unfamiliar with NE from trying it. 130 undergraduates, who denied having tried NE before (43% female, mean age = 22.76 ± 4.15 years old), were randomly assigned to either an ego-depletion or a control condition. The dependent variable was taking an "energy-stick" (a legal nutritional supplement, containing low doses of caffeine, taurine and vitamin B), offered as a potential means of enhancing performance on the bogus concentration task that followed. Logistic regression analysis showed that ego-depleted participants were three times less likely to take the substance, OR = 0.37, p = .01. This experiment found that trying NE for the first time was more likely if an individual's cognitive capacities were not depleted. This means that mental exhaustion is not predictive for NE in students for whom NE is not the dominant response. Trying NE for the first time is therefore more likely to occur as a thoughtful attempt at self-regulation than as an automatic behavioral response in stressful situations. We therefore recommend targeting interventions at this inter-individual difference. Students without previous reinforcing NE experience should be provided with information about the possible negative health outcomes of NE. Reconfiguring structural aspects in the academic environment (e.g. lessening workloads) might help to deter current users.


Factors influencing internet addiction tendency among middle school students in Gyeong-buk area.
This study was conducted to understand the degree of internet addiction tendency and to find out the factors influencing this addiction tendency among middle school students in Gyeong-buk area. A total of 450 middle school students in the Daegu and Gyeong-buk area were surveyed in this study. Data collection was conducted through the use of questionnaires. Internet addiction among middle school students was relatively low (Average user). In the overall ratio distribution, however, students who were classified as either addicted or at risk of addiction accounted for a high percentage, 27%. A positive correlation was found between Internet addiction and Internet expectation, depression and parent control over Internet use. A negative correlation was found between Internet addiction and interpersonal relationship, parent support and self-control. Multiple regression analysis revealed that the most powerful predictor of Internet addiction tendency was depression. Through the above results, it would be necessary to develop an Internet addiction prevention program for adolescents taking into account for the psychological factors such as depression and Internet use habits. In the future study, the need assessment will be useful for developing this prevention program.


Ceramide mimics tumour necrosis factor-alpha in the induction of cell cycle arrest in endothelial cells. Induction of the tumour suppressor p53 with decrease in retinoblastoma/protein levels.
Tumour necrosis factor (TNF)-alpha induces a transient increase in N-octanoylsphingosine (C8-ceramide) which has been postulated as an intracellular mediator in TNF-alpha signalling. We tested the ability of C8-ceramide to reproduce the TNF-alpha-mediated interference with endothelial cell proliferation and DNA synthesis. TNF-alpha (10 ng.mL-1) and C8-ceramide (20 microM) inhibited the incorporation of [3H]thymidine into DNA and led to an accumulation of cells in the G1 phase of the cell cycle. When the responses of the tumour suppressors p53 and RB were analysed, it was found that TNF-alpha and C8-ceramide induced increased expression of p53. Treatment with TNF-alpha or C8-ceramide lead to a significant decrease in total retinoblastoma protein (RB) content that correlated with high levels of p53. These results suggest that p53 and RB may complement each other in their contribution to cell cycle arrest. TNF-alpha prevented RB phosphorylation whereas C8-ceramide did not interfere with this process, suggesting that it follows a ceramide-independent pathway.


Tumor Mutation Burden and Differentially Mutated Genes Among Immune Phenotypes in Patients with Lung Adenocarcinoma.
Nowadays, immune checkpoint blockades (ICBs) have been extensively applied in non-small cell lung cancer (NSCLC) treatment. However, the outcome of anti-program death-1/program death ligand-1 (anti-PD-1/PD-L1) therapy is not satisfying in EGFR-mutant lung adenocarcinoma (LUAD) patients and its exact mechanisms have not been fully understood. Since tumor mutation burden (TMB) and tumor immune phenotype had been thought as potential predictors for efficacy of ICBs, we further studied the TMB and immune phenotype in LUAD patients to explore potential mechanisms for poor efficacy of ICBs in EGFR positive mutated patients and to find possible factors that could impact the tumor immune phenotype which might uncover some new therapeutic strategies or combination therapies. We enrolled 223 LUAD patients who underwent surgery in our hospital. We evaluated TMB through targeted panel sequencing. The tumor immune phenotype, which could be divided into non-inflamed, intermediate and inflamed, was determined through immunohistochemistry using formalin-fixed paraffin-embedded samples. Enumeration data were analyzed by Chi-square test or Fisher exact test and shown as number (proportion). Logistic regression model was employed for univariate and multivariate analysis of the association between TMB levels and clinical characteristics. The median TMB level was 4.0445 mutations/Mb. Multivariate analysis showed the TMB level was significantly associated with age (P=0.026), gender (P=0.041) and EGFR mutation status (P=0.015), and in EGFR-mutant patients we found a lower proportion of patients with mutated KRAS and BRCA2. Furthermore, we found patients with or without metastatic lesions would have different immune phenotype (P=0.007). And the mutational frequencies of ALK, CDKN2A, MAP2K1, IDH2 and PTEN were significantly different among three immune phenotypes. Low TMB level could be the reason for the poor efficacy of ICBs in patients having EGFR mutation. And mutational frequencies of KRAS and BRCA2 were lower in EGFR-mutant patients. Furthermore, ALK, CDKN2A, MAP2K1, IDH2 and PTEN might involve in the formation of immune phenotypes.


Bioinspired, graphene-enabled Ni composites with high strength and toughness.
Nature's wisdom resides in achieving a joint enhancement of strength and toughness by constructing intelligent, hierarchical architectures from extremely limited resources. A representative example is nacre, in which a brick-and-mortar structure enables a confluence of toughening mechanisms on multiple length scales. The result is an outstanding combination of strength and toughness which is hardly achieved by engineering materials. Here, a bioinspired Ni/Ni3C composite with nacre-like, brick-and-mortar structure was constructed from Ni powders and graphene sheets. This composite achieved a 73% increase in strength with only a 28% compromise on ductility, leading to a notable improvement in toughness. The graphene-derived Ni-Ti-Al/Ni3C composite retained high hardness up to 1000°C. The present study unveiled a method to smartly use 2D materials to fabricate high-performance metal matrix composites with brick-and-mortar structure through interfacial reactions and, furthermore, created an opportunity of developing advanced Ni-C-based alloys for high-temperature environments.


The auriculotemporal nerve in etiology of migraine headaches: compression points and anatomical variations.
The auriculotemporal nerve has been identified as one of the peripheral trigger sites for migraine headaches. However, its distal course is poorly mapped following emergence from the parotid gland. In addition, a reliable anatomical landmark for locating the potential compression points along the course of the nerve during surgery has not been sufficiently described. Twenty hemifaces on 10 fresh cadavers were dissected to trace the course of the auriculotemporal nerve from the inferior border of the zygomatic arch to its termination in the temporal scalp. The compression points were mapped and the distances were measured from the most anterosuperior point of the external auditory meatus, which was used as a fixed anatomical landmark. Three potential compression points along the course of the auriculotemporal nerve were identified. Compression points 1 and 2 corresponded to preauricular fascial bands. Compression point 1 was centered 13.1±5.9 mm anterior and 5.0±7.0 mm superior to the most anterosuperior point of the external auditory meatus, whereas compression point 2 was centered at 11.9±6.0 mm anterior and 17.2±10.4 mm superior to the most anterosuperior point of the external auditory meatus. A significant relationship was found between the auriculotemporal nerve and superficial temporal artery (compression point 3) in 80 percent of hemifaces, with three patterns of interaction: a single site of artery crossing over the nerve (62.5 percent), a helical intertwining relationship (18.8 percent), and nerve crossing over the artery (18.8 percent). Findings from this cadaver study provide information relevant to the operative localization of potential compression points along the auriculotemporal nerve.


Detecting the movement and spawning activity of bigheaded carps with environmental DNA.
Bigheaded carps are invasive fishes threatening to invade the Great Lakes basin and establish spawning populations, and have been monitored using environmental DNA (eDNA). Not only does eDNA hold potential for detecting the presence of species, but may also allow for quantitative comparisons like relative abundance of species across time or space. We examined the relationships among bigheaded carp movement, hydrography, spawning and eDNA on the Wabash River, IN, USA. We found positive relationships between eDNA and movement and eDNA and hydrography. We did not find a relationship between eDNA and spawning activity in the form of drifting eggs. Our first finding demonstrates how eDNA may be used to monitor species abundance, whereas our second finding illustrates the need for additional research into eDNA methodologies. Current applications of eDNA are widespread, but the relatively new technology requires further refinement.


Serum-induced differentiation of human meibomian gland epithelial cells.
We hypothesize that culturing immortalized human meibomian gland epithelial cells in serum-containing medium will induce their differentiation. The purpose of this investigation was to begin to test our hypothesis, and explore the impact of serum on gene expression and lipid accumulation in human meibomian gland epithelial cells. Immortalized and primary human meibomian gland epithelial cells were cultured in the presence or absence of serum. Cells were evaluated for lysosome and lipid accumulation, polar and neutral lipid profiles, and gene expression. Our results support our hypothesis that serum stimulates the differentiation of human meibomian gland epithelial cells. This serum-induced effect is associated with a significant increase in the expression of genes linked to cell differentiation, epithelium development, the endoplasmic reticulum, Golgi apparatus, vesicles, and lysosomes, and a significant decrease in gene activity related to the cell cycle, mitochondria, ribosomes, and translation. These cellular responses are accompanied by an accumulation of lipids within lysosomes, as well as alterations in the fatty acid content of polar and nonpolar lipids. Of particular importance, our results show that the molecular and biochemical changes of immortalized human meibomian gland epithelial cells during differentiation are analogous to those of primary cells. Overall, our findings indicate that immortalized human meibomian gland epithelial cells may serve as an ideal preclinical model to identify factors that control cellular differentiation in the meibomian gland.


Emission tunable, cyto/hemocompatible, near-IR-emitting Ag2S quantum dots by aqueous decomposition of DMSA.
Size tunable aqueous Ag2S quantum dots emitting in the near-infrared region were synthesized through decomposition of meso-2,3-dimercaptosuccinic acid (DMSA) in water. The resulting NIR QDs are highly cyto- and hemocompatible, have quantum yields as high as 6.5% and are effective optical imaging agents based on in vitro evaluation.


Impaired glycerophosphorylcholine synthesis in murine muscular dystrophy.
A test of some of the tenets of a proposed hypothesis on muscle phospholipid synthesis, and its possible defect in murine muscular dystrophy, shows that the cytidine pathways for the synthesis of phosphatidylcholine and phosphatidylethanolamine have a negligible flux in differentiated mouse gastrocnemius, while that of the respective proposed de novo glycerophosphodiester pathways is normally high in this muscle. Evidence is presented that de novo glycerophosphorylcholine synthesis in dystrophic mouse gastrocnemius is about half that of the wild type homozygotes, while that of the heterozygotes is near the mean of the two homozygous groups. No significant differences in rates of glycerophosphorylcholine or glycerophosphorylethanolamine synthesis were observed in brain and liver tissues among the three genotypes. These results suggest that defective de novo synthesis of glycerophosphorylcholine may be the primary biochemical lesion in murine muscular dystrophy.


Biliary-bronchial fistula after right hepatic lobectomy.
Biliary-bronchial fistula is an extremely unusual complication of thoracoabdominal trauma or suppurative hepatobiliary disease. We treated a patient with a biliary-bronchial fistula that developed after right hepatectomy for an infected traumatic intrahepatic hematoma. The diagnosis of biliary-bronchial fistula was confirmed by percutaneous transhepatic cholangiography. Surgical therapy included a primary transabdominal choledochostomy, with the insertion of a T-tube, sphincteroplasty, and open marsupialization of the chronic subphrenic abscess.


The prognostic value of chromosome studies in patients with the preleukemic syndrome (hemopoietic dysplasia).
Banded chromosome studies obtained at the time of diagnosis from 42 patients with the preleukemic syndrome had clonal abnormalities in fifteen. Using Kaplan-Meier product limit survival analysis, it was found that patients with chromosome abnormalities developed acute leukemia more frequently than those without (p less than 0.05). However, we noted that in a subset of patients with complex abnormalities and in a subset previously exposed to alkylating agents, almost all developed leukemia. When either subset was excluded from the larger group analysis there were no differences in leukemia-free survival between cytogenetically abnormal and normal groups. Thus, in the prognostic evaluation of our patients with the preleukemic syndrome a chromosome abnormality per se did not predict rapid evolution to acute leukemia. However, almost all preleukemic patients with complex chromosome abnormalities and almost all alkylator-exposed preleukemic patients can be expected to develop overt leukemia within 1 yr.


Survival following a diagnosis of heart failure in primary care.
Heart failure is a common long term condition affecting around 900 000 people in the UK and patients commonly present to primary care. The prognosis of patients with a code of heart failure in their primary care record is unknown. The study sought to determine the overall survival rates for patients with heart failure in a primary care population from the time of diagnosis. Survival analysis was carried out using UK primary care records from The Health Improvement Network (THIN) between 1 January 1998 and 31 December 2012. Patients age 45 or over with a first diagnostic label of heart failure were matched by age, sex and practice to people without heart failure. Outcome was death in the heart failure and no heart failure cohorts. Kaplan-Meier curves were used to compare survival. Age-specific survival rates at 1, 5 and 10 years were determined for men and women with heart failure. Survival rates by year of diagnosis and case definition were also calculated. During the study period, 54313 patients had a first diagnostic code of heart failure. Overall survival rates for the heart failure group were 81.3% (95%CI 80.9-81.6), 51.5% (95%CI 51.0-52.0) and 29.5% (95%CI 28.9-30.2) at 1, 5 and 10 years respectively and did not change over time. In a primary care population, the survival of patients diagnosed with heart failure did not improved over time. Further research is needed to explain these trends and to find strategies to improve outlook.


[Medical informatics in obstetrics and gynecology - the actual challenge?].
There is common agreement about the importance of information management systems in obstetrics and gynecology. Those systems are necessary tools for medical quality management and are essential for the actual preparation for the age of the "diagnosis related groups" that will be introduced in Germany next year. Nevertheless there are only small scientifically activities to improve information management systems and to evaluate their performance. Great efforts are necessary to develop new features and not to loose the conflict between the needs of the physicians and their patients and the needs and demands of hospital administrative authorities.


Synthesis and self-assembly of thermoresponsive amphiphilic biodegradable polypeptide/poly(ethyl ethylene phosphate) block copolymers.
We report the design and synthesis of new fully biodegradable thermoresponsive amphiphilic poly(γ-benzyl L-glutamate)/poly(ethyl ethylene phosphate) (PBLG-b-PEEP) block copolymers by ring-opening polymerization of N-carboxy-γ-benzyl L-glutamate anhydride (BLG-NCA) with amine-terminated poly(ethyl ethylene phosphate) (H2N-PEEP) as a macroinitiator. The fluorescence technique demonstrated that the block copolymers could form micelles composed of a hydrophobic core and a hydrophilic shell in aqueous solution. The morphology of the micelles as determined by transmission electron microscopy (TEM) was spherical. The size and critical micelle concentration (CMC) values of the micelles showed a decreasing trend as the PBLG segment increased. However, UV/Vis measurements showed that these block copolymers exhibited a reproducible temperature-responsive behavior with a lower critical solution temperature (LCST) that could be tuned by the block composition and the concentration.


Community-based treatment of venous thromboembolism with a low-molecular-weight heparin and warfarin.
This multicenter, prospective, open label, observational study evaluated practice patterns of physicians using tinzaparin, a low-molecular-weight heparin (LMWH), and warfarin for the treatment of deep venous thrombosis (DVT) with or without pulmonary embolism (PE). Short-term recurrence of venous thromboembolism (VTE) and safety were also evaluated. Patients with an objective diagnosis of DVT, with or without PE, were invited by their physician to participate in this study. Treatment was given according to the approved U.S. package inserts for tinzaparin (175 IU/kg SQ QD) and warfarin and the clinical judgment of the prescribing physician. Baseline patient history including demographic information and the results of tests to confirm the diagnosis of DVT, with or without PE, were collected. Follow-up information included the treatment setting in which each dose of tinzaparin was administered, medical training of the person administering tinzaparin doses, timing of initiation of warfarin with respect to that of tinzaparin, length of overlap of tinzaparin and warfarin therapy, and adverse experiences. A total of 334 patients were enrolled at 65 sites. Patients across a wide age (range 18-93 years old) and body weight (range 40-261 kg) were included. Overall, 27.3% of patients had cancer, and 50% of the overall study population reported more than one VTE risk factor. Mean duration of tinzaparin treatment was 7.61 days. Therapy at home was more common in suburban and rural settings than in urban settings. High proportions of patient, even among the small group with concurrent PE, were treated at home with self-injection. Severity of disease was the primary reason for hospitalization. Home treatment of DVT, with or without PE, with self administration of tinzaparin at 175 IU SQ once-daily was safe and resulted in an acceptably low rate of recurrent venous thromboembolism and adverse events. Home therapy in the usual practice setting should achieve substantial overall cost savings in the treatment of DVT.


Pharmacologic approach to aggression in neuropsychiatric disorders.
Aggressive behavior has been associated with numerous neurologic conditions including traumatic brain injury, mental retardation, developmental disorders, Huntington disease, and several dementias. Preclinical and human studies suggest that dysfunction of neural systems involving the brainstem, hypothalamus, amygdala, or prefrontal cortex can give rise to aggression. Several neurochemicals are felt to be relevant to modulation of aggression, including serotonin, dopamine, norepinephrine, GABA, acetylcholine, and androgens. Pharmacologic intervention studies have targeted these systems but have been limited by inconsistent definitions of aggression and a relative paucity of controlled trials. This article briefly reviews studies of neural systems and medication trials relevant to aggression and propose a clinical approach to treatment of patients manifesting aggressive behavior.


Common Mental Disorders among Occupational Groups: Contributions of the Latent Class Model.
Background. The Self-Reporting Questionnaire (SRQ-20) is widely used for evaluating common mental disorders. However, few studies have evaluated the SRQ-20 measurements performance in occupational groups. This study aimed to describe manifestation patterns of common mental disorders symptoms among workers populations, by using latent class analysis. Methods. Data derived from 9,959 Brazilian workers, obtained from four cross-sectional studies that used similar methodology, among groups of informal workers, teachers, healthcare workers, and urban workers. Common mental disorders were measured by using SRQ-20. Latent class analysis was performed on each database separately. Results. Three classes of symptoms were confirmed in the occupational categories investigated. In all studies, class I met better criteria for suspicion of common mental disorders. Class II discriminated workers with intermediate probability of answers to the items belonging to anxiety, sadness, and energy decrease that configure common mental disorders. Class III was composed of subgroups of workers with low probability to respond positively to questions for screening common mental disorders. Conclusions. Three patterns of symptoms of common mental disorders were identified in the occupational groups investigated, ranging from distinctive features to low probabilities of occurrence. The SRQ-20 measurements showed stability in capturing nonpsychotic symptoms.


The crystal structure of paramagnetic copper(II) oxalate (CuC₂O₄): formation and thermal decomposition of randomly stacked anisotropic nano-sized crystallites.
Synthetic copper(II) oxalate, CuC2O4, was obtained in a precipitation reaction between a copper(II) solution and an aqueous solution of oxalic acid. The product was identified from its conventional X-ray powder patterns which match that of the copper mineral Moolooite reported to have the composition CuC2O4·0.44H2O. Time resolved in situ investigations of the thermal decomposition of copper(II) oxalate using synchrotron X-ray powder diffraction showed that in air the compound converts to Cu2O at 215 °C and oxidizes to CuO at 345 °C. Thermo gravimetric analysis performed in an inert Ar-gas reveals that the material contains no crystal water and reduces to pure Cu at 295 °C. Magnetic susceptibility measurements in the temperature range from 2 K to 300 K show intriguing paramagnetic behaviour with no sign of magnetic order down to 2 K. A crystal structure investigation is made based on powder diffraction data using one neutron diffraction pattern obtained at 5 K (λ = 1.5949(1) Å) combined with one conventional and two synchrotron X-ray diffraction patterns obtained at ambient temperature using λ = 1.54056, 1.0981 and λ = 0.50483(1) Å, respectively. Based on the X-ray synchrotron data the resulting crystal structure is described in the monoclinic space group P2₁/c (#14) in the P12₁/n1 setting with unit cell parameters a = 5.9598(1) Å, b = 5.6089(1) Å, c = 5.1138 (1) Å, β = 115.320(1)°. The composition is CuC2O4 with atomic coordinates determined by FullProf refinement of the neutron diffraction data. The crystal structure consists of a random stacking of CuC2O4 micro-crystallites where half the Cu-atoms are placed at (2a) and the other half at (2b) positions with the corresponding oxalate molecules centred around the corresponding (2b) and (2a) site positions, respectively. The diffraction patterns obtained for both kinds of radiation show considerable broadening of several Bragg peaks caused by highly anisotropic microstructural size and strain effects. In contrast to the water reported to be present in Moolooite, neither thermogravimetric nor the in situ thermal decomposition investigations and crystal structure analysis of the neutron diffraction data revealed any trace of water. An appendix contains details about the profile parameters for the diffractometers used at the European Synchrotron Radiation Facility and the Institute Max von Laue-Paul Langevin.


Three genes of Citrus tristeza virus are dispensable for infection and movement throughout some varieties of citrus trees.
Citrus tristeza virus (CTV), a member of the Closteroviridae, possesses a 19.3-kb positive-stranded RNA genome that is organized into twelve open reading frames (ORFs). The CTV genome contains two sets of conserved genes, which are characteristic of this virus group, the replication gene block (ORF 1a and 1b) and the quintuple gene block (p6, HSP70 h, p61, CPm, and CP). With the exception of the p6 gene, they are required for replication and virion assembly. CTV contains five additional genes, p33, p18, p13, p20 and p23, in the 3' half of the genome, some of which (p33, p18 and p13) are not conserved among other members of this virus group, and have been proposed to have evolved for specific interactions with the citrus host. In the present study, the requirements for systemic infection of citrus trees of p33, p6, p18, p13 and p20 were examined. Viral mutants with a deletion in the p6 or the p20 ORF failed to infect citrus plants systemically, suggesting their possible roles in virus translocation/systemic infection. However, we found that deletions within the p33, p18 or p13 ORF individually resulted in no significant loss of ability of the virus to infect, multiply, and spread throughout citrus trees. Furthermore, deletions in the p33, p18 and p13 genes in all possible combinations including deletions in all three genes allowed the virus to systemically invade citrus trees. Green fluorescent protein-tagged CTV variants with deletions in the p33 ORF or the p33, p18 and p13 ORFs demonstrated that the movement and distribution of these deletion mutants were similar to that of the wild-type virus.


Increased serum ferritin levels are independently associated with carotid atherosclerosis in women.
Previous studies have supported the theory that there is a positive association between ferritin and carotid atherosclerosis in Western people. Diet plays an important role in determining serum ferritin concentration. Asian dietary patterns are different from Western dietary patterns, implying that there may be a difference in the association of ferritin with carotid atherosclerosis between Asian and Western people. However, few studies focus on the association between ferritin and carotid atherosclerosis among Asians. The aim of this study was to investigate how serum ferritin levels are associated with carotid atherosclerosis in an Asian adult population. A cross-sectional assessment was performed in 8302 adults in Tianjin, China. Carotid intima-media thickness (IMT) and plaques were assessed using ultrasonography, and serum ferritin was measured using the protein chip-chemiluminescence method. Multiple logistic regression analysis was used to examine the association between quartiles of serum ferritin concentration and carotid atherosclerosis. In the present study, the overall prevalence of IMT and carotid plaques in participants is 29·2 and 22·7 %, respectively. In women, after adjustments for potentially confounding factors, the OR of IMT and carotid plaques by increasing serum ferritin quartiles were 1·00, 1·39 (95 % CI 0·98-1·99), 1·39 (95 % CI 0·99-1·97), 1·81 (95 % CI 1·30-2·55) (P for trend<0·001) and 1·00, 1·24 (95 % CI 0·89-1·73), 1·18 (95 % CI 0·85-1·65), 1·59 (95 % CI 1·15-2·20) (P for trend<0·01), respectively. However, no association was found between serum ferritin and carotid atherosclerosis in men. The study demonstrated that increased serum ferritin levels are independently associated with IMT and carotid plaques in Asian women but not in Asian men.


Innovations in applications and prospects of non-isocyanate polyurethane bioplastics.
Currently, conventional plastics are necessary for a variety of aspects of modern daily life, including applications in the fields of healthcare, technology, and construction. However, they could also contain potentially hazardous compounds like isocyanates, whose degradation has a negative impact on both the environment and human health. Therefore, researchers are exploring alternatives to plastic which is sustainable and environmentally friendly without compromising its mechanical and physical features. This review study highlights the production of highly eco-friendly bioplastic as an efficient alternative to non-biodegradable conventional plastic. Bioplastics are produced from various renewable biomass sources such as plant debris, fatty acids, and oils. Poly-addition of di-isocyanates and polyols is a technique employed over decades to produce polyurethanes (PUs) bioplastics from renewable biomass feedstock. The toxicity of isocyanates is a major concern with the above-mentioned approach. Novel green synthetic approaches for polyurethanes without using isocyanates have been attracting greater interest in recent years to overcome the toxicity of isocyanate-containing raw materials. The polyaddition of cyclic carbonates (CCs) and polyfunctional amines appears to be the most promising method to obtain non-isocyanate polyurethanes (NIPUs). This method results in the creation of polymeric materials with distinctive and adaptable features with the elimination of harmful compounds. Consequently, non-isocyanate polyurethanes represent a new class of green polymeric materials. In this review study, we have discussed the possibility of creating novel NIPUs from renewable feedstocks in the context of the growing demand for efficient and ecologically friendly plastic products.


A Genotype-Phenotype Analysis of the Bardet-Biedl Syndrome in Puerto Rico.
Bardet-Biedl syndrome is a complex heterogeneous ciliopathy caused by genetic mutations. Although establishing genotype-phenotype correlations has been challenging, some regional variations have been previously reported. Due to its relative geographic isolation, Puerto Rico has a greater prevalence of Bardet-Biedl syndrome than do other regions. We sought to characterize the most frequent genotypic variations in a local cohort of Bardet-Biedl syndrome patients and report any genotypic-phenotypic trends. Twenty-seven patients from an ophthalmology clinic in Puerto Rico with genetically confirmed Bardet-Biedl syndrome took a questionnaire inquiring about their most common symptoms. Ophthalmological information was obtained from patient records. The frequencies of the genotypic variations and symptoms were calculated. In the study population, BBS1 was the most prevalent mutated gene, followed by BBS7. In the BBS1 group, we found homozygotes for c.1169T>G (p.Met390Arg) and c.1645G>T (p.Glu549*), and compound heterozygotes for c.1169T>G (p.Met390Arg) and c.1645G>T (p.Glu549*), with one patient having c.1645G>T (p.Glu549*) and c.432+1G>A (splice donor). All the BBS7 patients were homozygous for c.632C>T (p.Thr211Ile). Compared to BBS7, we found that BBS1 patients generally had a milder ocular and systemic phenotype. However, when analyzing different BBS1 variants, patients with mutations in c.1645G>T (p.Glu549*), both compound heterozygous and homozygous, had more severe systemic phenotypes, overall. Our study was the first detailed genotype-phenotype analysis of the Bardet-Biedl syndrome in Puerto Rico. Genetic mutations in BBS1 and BBS7 seem to be the most common culprits behind Bardet-Biedl syndrome in this population. Although patients diagnosed with BBS1 are likely to display milder systemic features, this was not the case with our BBS1 patients having the c.1645G>T (p.Glu549*) mutation. Further studies should focus on the c.1645G>T (p.Glu549*) mutation's impact on the BBS1 gene and protein product.


Ultrastructural changes in skeletal muscle of a patient with familial intrahepatic cholestasis associated with vitamin E deficiency.
The skeletal muscle from a patient with familial intrahepatic cholestasis associated with vitamin E deficiency was studied by electron microscopy. The muscle fibers showed a variety of pathologic features including degenerative, necrotic and regenerative changes. Granule-like inclusions found in our biopsy specimens were similar in structure to those observed in vitamin E deficient animals. These inclusions were noted not only in the skeletal muscle fibers but also in endothelial cells of the intramuscular capillaries, Schwann cells and perineural cells. Disruption and disappearance of the plasma membrane, separation, disruption and pleats formation of the external lamina, and multilayered external laminae were observed in muscle fibers most frequently. The nerves among muscle fibers also showed degenerative features. These severely degenerative alterations of the muscle fiber have not so far been reported in vitamin E deficient patients. We discuss the process of cell damages caused by vitamin E deficiency.


A proposal for harmonizing clinical neurophysiology training in the Europe, Middle East and Africa Chapter of the International Federation of Clinical Neurophysiology.
There are significant differences in duration and intensity of clinical neurophysiology specialty training within the countries of the Europe, Middle East and Africa Chapter of the International Federation of Clinical Neurophysiology. We address these differences by proposing recommendations which may facilitate harmonisation of training and education within the Chapter. They arose from two workshops whose recommendations were then circulated widely within national societies in the Chapter for feedback and for consensus. The recommendations are applicable to clinical neurophysiology as a medical monospecialty and/or as a subspecialty (usually of neurology). We make a number of recommendations on governance and regulation of training, on the requirements for competence and the numbers of various examinations and tests performed by trainees, some under supervision. We also recommend a modular approach considering primary and complementary modules. Primary modules are electroencephalography, electromyography, nerve conduction studies and evoked potentials, while complementary ones include sleep analysis, intraoperative monitoring, small fibre testing, peripheral nerve and muscle ultrasound, intracortical recordings, and analysis of movement disorders. It is recommended that national examinations should include a variety of techniques to assess knowledge and judgement, practical skills, teamwork, communication skills, as well as safety and quality. The aim of the suggested recommendations is to harmonize clinical neurophysiology training in the member societies throughout the Chapter. It is realised that this may mean that the numbers for competence are aspirational for some, though ways to mitigate this, for instance through supranational training centres, are also discussed.


Changes in acute kidney injury epidemiology in critically ill patients: a population-based cohort study in Korea.
Although no specific treatment facilitates renal tubular regeneration in acute kidney injury (AKI), the rapid increase in aging populations with more comorbidities and advances in critical care management are expected to change the epidemiology of AKI. However, few recent studies dissected the current epidemiologic characteristics of critically ill patients with AKI. We investigated recent epidemiologic changes in severe AKI in critically ill patients. All adult admissions to intensive care units (ICUs) in Korea from 2008 to 2015 were screened using the national health insurance review and assessment database, and 1,744,235 patients were included. Clinical characteristics and changes in AKI incidence and mortality rate were analyzed. The incidence of AKI increased from 7.4% in 2008 to 8.3% in 2015 (p for trend < 0.001). Age-standardized AKI rate was 7018.6 per 100,000 person-years. In-hospital mortality significantly decreased from 39.1% in 2008 to 37.2% in 2015 (p for trend < 0.001) with 2427.6 deaths per 100,000 person-years. Patients with AKI showed higher in-hospital mortality, prolonged ICU length of stay, and higher total cost. Multivariable analysis showed increased risk of in-hospital mortality (adjusted odds ratio [OR] 3.74), mechanical ventilation (OR 2.87), ECMO (OR 6.99), and vasopressor requirement (OR 2.75) in patients with AKI. Recent advances in medical management for AKI have improved in-hospital mortality of critically ill patients with AKI despite increases in the elderly population and AKI incidence.


Isolation and characterisation of legumin promoter sequence from chickpea (Cicer arietinum L.).
Seed specific promoters are useful for expression of foreign genes in the seeds. We have isolated a Cicer arietinum legumin promoter from lambda EMBL genomic library and subcloned in pBluescript II KS (-) in Eco RV and Pst I site. The 2.762 kb Hind II Pst I fragment was sequenced completely by dideoxy chain termination method by creating a set of unidirectional deletions of the inserts in pAKKIII. The insert contains mainly upstream sequence (2240 bps) and only a part of structural gene (522 bps) sequence. The 522 bps of the structural gene shows approximately 80% homology with pea legumin A and this is almost the same as chickpea legumin in its sequence. The amino acid sequence derived from the part of the structural gene was similar to the chickpea 5' part of the legumin structural gene with a few variations. A 21 amino acid signal peptide was also deduced like many other legumes. Transcription start site (CAT) was located at 55 bp upstream of the initiation codon ATG. One codon downstream to ATG codon Hind III site was present. TATA box was observed at -30 position, with a consensus of CCTATAAATAACC. The consensus CATGCAAG, a part of legumin box was noticed at -110 bp position. At -295 to -265 bp upstream AGGA box like sequences were observed and a 56 bp perfect repeat was located between -913 bp and -972 bp. Strong homology with pea promoter sequence near the CAT sequence was noticed which gradually decreased towards the upstream region. Thus the cloned fragment contains a full length promoter which can be utilised for expression of foreign genes in seeds of chickpea.


Challenges in systematic reviews that evaluate drug efficacy or effectiveness.
Increasingly, consumers, clinicians, regulatory bodies, and insurers are using systematic reviews of drug interventions to select treatments and set policies. Although a systematic review cannot provide all the information a clinician needs to make an informed choice for therapy, it can help decision makers distinguish what claims about effectiveness are based on evidence, identify critical information gaps, describe features of the evidence that limit applicability in practice, and address whether drug effectiveness differs for particular subgroups of patients. To improve the relevance and validity of reviews of drug therapies, reviewers need to delineate clinically important subgroups, specific aims of therapy, and most important outcomes. They may need to find unpublished trials, studies other than direct comparator (head-to-head) trials, and additional details of published trials from pharmaceutical manufacturers and regulatory agencies. In this paper, we address ways to formulate questions relevant to specific clinical therapeutic aims; discuss types of studies to include in drug efficacy and effectiveness reviews and how to find them; and describe ways to assess applicability of studies to actual practice.


Associations of tissue transglutaminase antibody seropositivity with coronary heart disease: Findings from a prospective cohort study.
Clinical experience and observational studies suggest that individuals with coeliac disease are at increased risk of coronary heart disease (CHD), but the precise mechanism for this is unclear. Laboratory studies suggest that it may relate to tissue transglutaminase antibodies (tTGAs). Our aim was to examine whether seropositivity for tTGA and endomysial antibodies (EMAs) are associated with incident CHD in humans. We used data from Mini-Finland Health Survey, a prospective cohort study of Finnish men and women aged 35-80 at study baseline 1978-80. TTGA and EMA seropositivities were ascertained from baseline blood samples and incident CHD events were identified from national hospitalisation and death registers. Cox regression was used to examine the associations between antibody seropositivity and incident CHD. Of 6887 men and women, 562 were seropositive for tTGAs and 72 for EMAs. During a median follow-up of 26 years, 2367 individuals experienced a CHD event. We found no clear evidence for an association between tTGA positivity (hazard ratio, HR: 1.04, 95% confidence interval, CI: 0.83, 1.30) or EMA positivity (HR: 1.16, 95% CI: 0.77, 1.74) and incident CHD, once pre-existing CVD and known CHD risk factors had been adjusted for. We found no clear evidence for an association of tTGA or EMA seropositivity with incident CHD outcomes, suggesting that tTG autoimmunity is unlikely to be the biological link between coeliac disease and CHD.