Metallated conjugated polymers as a new avenue towards high-efficiency polymer solar cells.
Bulk heterojunction solar cells have been extensively studied owing to their great potential for cost-effective photovoltaic devices. Although recent advances resulted in the fabrication of poly(3-hexylthiophene) (P3HT)/fullerene derivative based solar cells with efficiencies in the range 4.4-5.0%, theoretical calculations predict that the development of novel donor materials with a lower bandgap is required to exceed the power-conversion efficiency of 10%. However, all of the lower bandgap polymers developed so far have failed to reach the efficiency of P3HT-based cells. To address this issue, we synthesized a soluble, intensely coloured platinum metallopolyyne with a low bandgap of 1.85 eV. The solar cells, containing metallopolyyne/fullerene derivative blends as the photoactive material, showed a power-conversion efficiency with an average of 4.1%, without annealing or the use of spacer layers needed to achieve comparable efficiency with P3HT. This clearly demonstrates the potential of metallated conjugated polymers for efficient photovoltaic devices.


Association of beta2-glycoprotein I IgG and IgM antibodies with thrombosis and thrombocytopenia.
Antiphospholipid antibodies (APA) have been known for decades. Their relation to clinical manifestations, primarily thromboses and thrombocytopenia, was recognised in the 1980s. In this clinical study two cohorts of patients, a population-based (84 patients with systemic lupus erythematosus (SLE)) and a hospital-based (87 patients with SLE and 53 with other connective tissue diseases) were investigated for APA and associated clinical manifestations. Anticardiolipin antibodies (ACA) of IgG and IgM classes were found in 13 and 38% of the population-based patients and in 29 and 58% of the hospital-based patients, respectively. The corresponding figures for antibodies against beta2-glycoprotein I (anti-beta2GPI) were 15 and 10% in the population-based patients and 14 and 8% in the hospital-based cohort. Anti-beta2GPI antibodies were always found in association with the corresponding immunoglobulin class of ACA. In both cohorts anti-beta2GPI of the IgG class were associated with arterial/venous occlusion, a result concordant with other studies. A novel finding in both cohorts, however, was an association between thrombocytopenia and IgM anti-beta2GPI.


Root water potential integrates discrete soil physical properties to influence ABA signalling during partial rootzone drying.
To investigate the influence of different growing substrates (two mineral, two organic) on root xylem ABA concentration ([ABA](root)) and the contribution of the drying root system to total sap flow during partial rootzone drying (PRD), sunflower (Helianthus annuus L.) shoots were grafted onto the root systems of two plants grown in separate pots. Sap flow through each hypocotyl was measured below the graft union when one pot ('wet') was watered and other ('dry') was not. Each substrate gave unique relationships between dry pot matric potential (Psi(soil)), volumetric water content ((v)) or penetrometer resistance (Q) and either the fraction of photoperiod sap flow from roots in drying soil or [ABA](root). However, decreased relative sap flow, and increased [ABA](root), from roots in drying soil varied with root water potential (Psi(root)) more similarly across a range of substrates. The gradient between Psi(soil) and Psi(root) was greater in substrates with high sand or peat proportions, which may have contributed to a more sensitive response of [ABA](root) to Psi(soil) in these substrates. Whole plant transpiration was most closely correlated with the mean Psi(soil) of both pots, and then with detached leaf xylem ABA concentration. Although Psi(root) best predicted decreased relative sap flow, and increased [ABA](root), from roots in drying soil across a range of substrates, the inaccessibility of this variable in field studies requires a better understanding of how measurable soil variables (Psi(soil), (v), Q) affect Psi(root).


Intersubunit allosteric communication mediated by a conserved loop in the MCM helicase.
The minichromosome maintenance (MCM) helicase is the presumptive replicative helicase in archaea and eukaryotes. The archaeal homomultimeric MCM has a two-tier structure. One tier contains the AAA+ motor domains of the proteins, and these are the minimal functional helicase domains. The second tier is formed by the N-terminal domains. These domains are not essential for MCM helicase activity but act to enhance the processivity of the helicase. We reveal that a conserved loop facilitates communication between processivity and motor tiers. Interestingly, this allostery seems to be mediated by interactions between, rather than within, individual protomers in the MCM ring.


1H NMR-Based Metabolomics Study of the Toxicological Effects in Rats Induced by "Renqing Mangjue" Pill, a Traditional Tibetan Medicine.
"RenqingMangjue" pill (RMP), as an effective prescription of Traditional Tibetan Medicine (TTM), has been widely used in treating digestive diseases and ulcerative colitis for over a thousand years. In certain classical Tibetan Medicine, heavy metal may add as an active ingredient, but it may cause contamination unintentionally in some cases. Therefore, the toxicity and adverse effects of TTM became to draw public attention. In this study, 48 male Wistar rats were orally administrated with different dosages of RMP once a day for 15 consecutive days, then half of the rats were euthanized on the 15th day and the remaining were euthanized on the 30th day. Plasma, kidney and liver samples were acquired to 1H NMR metabolomics analysis. Histopathology and ICP-MS were applied to support the metabolomics findings. The metabolic signature of plasma from RMP-administrated rats exhibited increasing levels of glucose, betaine, and creatine, together with decreasing levels of lipids, 3-hydroxybutate, pyruvate, citrate, valine, leucine, isoleucine, glutamate, and glutamine. The metabolomics analysis results of liver showed that after RMP administration, the concentrations of valine, leucine, proline, tyrosine, and tryptophan elevated, while glucose, sarcosine and 3-hydroxybutyrate decreased. The levels of metabolites in kidney, such as, leucine, valine, isoleucine and tyrosine, were increased, while taurine, glutamate, and glutamine decreased. The study provides several potential biomarkers for the toxicity mechanism research of RMP and shows that RMP may cause injury in kidney and liver and disturbance of several pathways, such as energy metabolism, oxidative stress, glucose and amino acids metabolism.


Assessment of Impact of Long-Cassette Standing X-Rays on Surgical Planning for Cervical Pathology: An International Survey of Spine Surgeons.
Understanding the role of regional segments of the spine in maintaining global balance has garnered significant attention recently. Long-cassette radiographs (LCR) are necessary to evaluate global spinopelvic alignment. However, it is unclear how LCRs impact operative decision-making for cervical spine pathology. To evaluate whether the addition of LCRs results in changes to respondents' operative plans compared to standard imaging of the involved cervical spine in an international survey of spine surgeons. Fifteen cases (5 control cases with normal and 10 test cases with abnormal global alignment) of cervical pathology were presented online with a vignette and cervical imaging. Surgeons were asked to select a surgical plan from 6 options, ranging from the least (1 point) to most (6 points) extensive. Cases were then reordered and presented again with LCRs and the same surgical plan question. One hundred fifty-seven surgeons completed the survey, of which 79% were spine fellowship trained. The mean response scores for surgical plan increased from 3.28 to 4.0 (P = .003) for test cases with the addition of LCRs. However, no significant changes (P = .10) were identified for the control cases. In 4 of the test cases with significant mid thoracic kyphosis, 29% of participants opted for the more extensive surgical options of extension to the mid and lower thoracic spine when they were provided with cervical imaging only, which significantly increased to 58.3% upon addition of LCRs. In planning for cervical spine surgery, surgeons should maintain a low threshold for obtaining LCRs to assess global spinopelvic alignment.


A Cranial Mesoderm Origin for Esophagus Striated Muscles.
The esophagus links the oral cavity to the stomach and facilitates the transfer of bolus. Using genetic tracing and mouse mutants, we demonstrate that esophagus striated muscles (ESMs) are not derived from somites but are of cranial origin. Tbx1 and Isl1 act as key regulators of ESMs, which we now identify as a third derivative of cardiopharyngeal mesoderm that contributes to second heart field derivatives and head muscles. Isl1-derived ESM progenitors colonize the mouse esophagus in an anterior-posterior direction but are absent in the developing chick esophagus, thus providing evolutionary insight into the lack of ESMs in avians. Strikingly, different from other myogenic regions, in which embryonic myogenesis establishes a scaffold for fetal fiber formation, ESMs are established directly by fetal myofibers. We propose that ESM progenitors use smooth muscle as a scaffold, thereby bypassing the embryonic program. These findings have important implications in understanding esophageal dysfunctions, including dysphagia, and congenital disorders, such as DiGeorge syndrome.


Enduring impact of communication skills training: results of a 12-month follow-up.
The efficacy of a communication skills training programme was shown through a randomised trial. Oncologists (N=160) from 34 cancer centres were allocated to written feedback plus course; course alone; written feedback alone or control. Each clinician had 6 - 10 interviews with patients videotaped at baseline and 3 months postintervention. Analysis of videotapes revealed improvements in the communication skills of clinicians randomised to training (n=80) compared with others (n=80). A 12-month follow-up assessment is reported here. Robust Poisson conditional analyses of counts of changes in communication behaviours revealed no demonstrable attrition in those who had shown improvement previously, including fewer leading questions, appropriate use of focused and open-ended questions and responses to patient cues. Additional skills, not apparent at 3 months, were now evident; the estimated effect sizes corresponded to 81% fewer interruptions (P=0.001) and increased summarising of information to 38% (P=0.038). However, expressions of empathy (54%, P=0.001) declined. The overall results show that 12 - 15 months postintervention, clinicians had integrated key communication skills into clinical practice and were applying others. This is the first RCT to show an enduring effect of communication skills training with transfer into the clinic.


Dual mode switching of cholesteric liquid crystal device with three-terminal electrode structure.
We propose a cholesteric liquid crystal device with a three-terminal electrode structure that can be operated in both the dynamic and the bistable modes. Fast switching (less than 5 ms) between the planar and the in-plane-field-induced states can be realized by applying an in-plane electric field, and conventional bistable switching between the planar and focal conic states can be realized by applying a vertical electric field.


Analysis of a Biathlon Sprint Competition and Associated Laboratory Determinants of Performance.
Biathlon is an Olympic winter-sport where cross-country (XC) skiing in the skating technique is combined with rifle shooting. In the biathlon sprint competition for men, three laps of 3.3-km are interspersed with a 5-shot shooting sequence in the prone and standing position. Our purpose was to investigate the contribution from overall XC skiing performance, the performance in different terrain sections and shooting performance to the overall biathlon sprint race performance, as well as the relationship to laboratory-measured capacities obtained during treadmill roller ski skating. Eleven elite male biathletes were tracked by a Global Positioning System (GPS) device and a heart rate (HR) monitor during an international 10-km biathlon sprint competition. Within a period of 6 weeks prior to the competition, physiological responses, and performance during submaximal and maximal treadmill roller skiing were measured. Stepwise multiple regression analysis revealed that XC skiing time, shooting performance, shooting time and range time explained 84, 14, 1.8, and 0.2% of the overall sprint race performance (all p < 0.01). Time in uphill, varied, and downhill terrains were all significantly correlated to the total XC skiing time (r = 0.95, 0.82, 0.72, respectively, all p < 0.05). Percent of maximal HR (HRmax) and rating of perceived exertion (RPE) during submaximal roller skiing, and time-to-exhaustion during incremental roller skiing correlated significantly with overall biathlon sprint race performance and overall XC skiing time (r = 0.64-0.95, all p < 0.05). In conclusion, XC skiing performance provided greatest impact on biathlon sprint performance, with most of the variance determined by XC skiing performance in the uphill terrain sections. Furthermore, the ability to roller ski with a low RPE and %HRmax during submaximal speeds, as well as time-to-exhaustion during incremental roller skiing significantly predicted biathlon performance. Such laboratory-derived measures may therefore be validly used to distinguish biathletes of different performance levels and to track progress of their XC skiing capacity.


Serum apolipoproteins C-I and C-III are reduced in stomach cancer patients: results from MALDI-based peptidome and immuno-based clinical assays.
Finding new peptide biomarkers for stomach cancer in human sera that can be implemented into a clinically practicable prediction method for monitoring of stomach cancer. We studied the serum peptidome from two different biorepositories. We first employed a C8-reverse phase liquid chromatography approach for sample purification, followed by mass-spectrometry analysis. These were applied onto serum samples from cancer-free controls and stomach cancer patients at various clinical stages. We then created a bioinformatics analysis pipeline and identified peptide signature discriminating stomach adenocarcinoma patients from cancer-free controls. Matrix Assisted Laser Desorption/Ionization-Time of Flight (MALDI-TOF) results from 103 samples revealed 9 signature peptides; with prediction accuracy of 89% in the training set and 88% in the validation set. Three of the discriminating peptides discovered were fragments of Apolipoproteins C-I and C-III (apoC-I and C-III); we further quantified their serum levels, as well as CA19-9 and CRP, employing quantitative commercial-clinical assays in 142 samples. ApoC-I and apoC-III quantitative results correlated with the MS results. We then employed apoB-100-normalized apoC-I and apoC-III, CA19-9 and CRP levels to generate rules set for stomach cancer prediction. For training, we used sera from one repository, and for validation, we used sera from the second repository. Prediction accuracies of 88.4% and 74.4% were obtained in the training and validation sets, respectively. Serum levels of apoC-I and apoC-III combined with other clinical parameters can serve as a basis for the formulation of a diagnostic score for stomach cancer patients.


Study of the interaction of deoxynivalenol with human serum albumin by spectroscopic technique and molecular modelling.
The mechanism of interaction between deoxynivalenol (DON) and human serum albumin (HSA) was studied using spectroscopic methods including fluorescence spectra, UV-VIS, Fourier transform infrared (FT-IR) and circular dichroism (CD). The quenching mechanism was investigated in terms of the association constants, number of binding sites and basic thermodynamic parameters. The distance between the HSA donor and the acceptor DON was 2.80 nm as derived from fluorescence resonance energy transfer. The secondary structure compositions of free HSA and its DON complexes were estimated by the FT-IR spectra. Alteration of the secondary protein structure in the presence of DON was confirmed by UV-VIS and CD spectroscopy. Molecular modelling revealed that a DON-protein complex was stabilised by hydrophobic forces and hydrogen bonding. It was potentially useful for elucidating the toxigenicity of DON when combined with biomolecular function effect, transmembrane transport, toxicological testing and the other experiments.


Evidence for a potent antiinflammatory effect of rosiglitazone.
We have recently demonstrated a potent antiinflammatory effect of troglitazone, an agonist of peroxisome proliferator-activated receptor gamma (PPARgamma) and a partial agonist of PPARalpha in both the nondiabetic obese and diabetic obese subjects. We have now investigated the antiinflammatory actions of rosiglitazone, a selective PPARgamma agonist. Eleven nondiabetic obese subjects and 11 obese diabetic subjects were each given 4 mg of rosiglitazone daily for a period of 6 wk. Fasting blood samples were obtained at 0, 1, 2, 4, 6, and 12 wk (6 wk after the cessation of rosiglitazone). Eight obese subjects and five obese diabetic subjects were also included in the study as control groups. Fasting blood samples were obtained from the control groups at 0, 1, 2, 4, and 6 wk only. Nuclear factor kappaB (NFkappaB)-binding activity in mononuclear cells, plasma monocyte chemoattractant protein-1 (MCP-1), TNF-alpha, soluble intercellular adhesion molecule-1, C-reactive protein (CRP), and serum amyloid A (SAA) were measured. Blood glucose concentration changed significantly at 6 wk only in the obese diabetic subjects after rosiglitazone treatment for 6 wk, whereas insulin concentration decreased significantly at 6 wk in both groups. NFkappaB-binding activity in mononuclear cell nuclear extract fell in both obese and obese diabetic subjects (P < 0.02). Rosiglitazone treatment resulted in a reduction in plasma MCP-1 and CRP in both groups (P < 0.05). Plasma TNF-alpha and SAA concentrations were inhibited significantly in the obese group (P < 0.05) but not in the obese diabetic subjects. NFkappaB-binding activity and plasma MCP-1, CRP, SAA, and TNF-alpha did not change in the obese and obese diabetic control groups. We conclude that rosiglitazone, a selective PPARgamma agonist, exerts an antiinflammatory effect at the cellular and molecular level, and in plasma. These observations may have implications for atherogenesis in the long term in subjects treated with rosiglitazone and possibly other thiazolidinediones.


Photocatalytic paper from colloidal TiO(2)--fact or fantasy.
Photocatalytic paper encompasses a range of materials based on paper and nonwoven fabrics which performs a function based on the light-activated catalytic activity of colloidal TiO(2). The literature describing photocatalytic paper is surveyed, including mechanisms, applications, limitations and future opportunities. The technology is in its infancy with less than 10 patents and as many scientific publications appearing over the last decade. The main applications described are the destruction of organic molecules (mineralization) and photo-disinfection (sterilization). These disclosures build upon a much larger literature describing photochemical properties of TiO(2) both supported on non-cellulose substrates or simply as suspended particles in water or air. Current photocatalytic paper developments include methods to fix TiO(2) to cellulose substrates to minimize photochemical damage to the paper. Another theme is the use of multiple approaches, such as zeolites, for enhanced mineralization, and metals, such as silver and copper, for enhanced photocatalytic disinfection.


Resourcefulness, positive cognitions, relocation controllability and relocation adjustment among older people: a cross-sectional study of cultural differences.
The population of older people in both the United States and Egypt is expected to double by the year 2030. With ageing, chronic illnesses increase and many older people need to relocate to retirement communities. Research has shown that positive cognitions and resourcefulness are positively correlated with adaptive functioning and better adjustment. The purpose of this study was to examine and compare relocation controllability, positive cognitions, resourcefulness and relocation adjustment between American and Egyptian older people living in retirement communities. The purpose of this cultural comparison is to gain insight into influencing factors in each culture that might lead to interventions to help relocated older adults in both cultures adjust to their new surroundings. A cross-sectional, descriptive design was used to compare relocation controllability, positive cognitions, resourcefulness and relocation adjustment of a convenience sample of American older people (n = 104) and a convenience sample of Egyptian older people (n = 94). The study was a secondary analysis of two studies of older people residing in six retirement communities in Northeast Ohio and in five retirement communities in Alexandria, Egypt. Examination of mean scores and standard deviations on the measure of positive cognitions using independent sample t-tests indicated that on average, the American older people reported more positive cognitions (t (131.16) = 11.29, P < 0.001), more relocation controllability (t (196) = -6.78, P < 0.001) and more relocation adjustment (t (196) = 9.42, P < 0.001) than the Egyptian older people. However, there was no significant difference between Egyptians and Americans in resourcefulness (t (174.16) = -0.97, P > 0.05). The results provide direction for the development of positive cognition interventions and engaging older people in the decision-making process to help them to adjust to relocation. Implications for practice. Positive thinking and resourcefulness training interventions can be used by nurses to help relocated older people to adjust to the stress of relocation to retirement communities. These interventions can be used on primary, secondary, and tertiary levels. Primary interventions can help to prevent the stress of relocation before happening by helping older people to use their positive thinking and their resources and work with them before relocating to retirement communities. Secondary prevention can be used by nurses to help older people who have already relocated to retirement communities and have already experienced stress of relocation to help them out by decreasing the stress that they are suffering. Tertiary prevention can be used to prevent further stress and deterioration for those who have suffered physical and psychological symptoms as a result of relocation.


Finite element analysis on dental implant-supported prostheses without passive fit.
The purpose of this study is to use finite element analysis to investigate the effect of misfit prostheses, cantilever prostheses, and various occlusal forces on the stress distribution in the prostheses, implant components, and surrounding bone. Two 3-dimensional finite element models were constructed: (1) a 2-implant-supported, 2-unit fixed partial denture and (2) a 2-implant-supported, 2-unit fixed partial denture with a distal cantilever. Variations of the standard finite element models were made by placing a 111-microm gap between the gold cylinder on either the mesial or distal implant. The effects of load of 100 N were tested on all models. Subsequently, loads of 50 N, 200 N, and 300 N were evaluated on the cantilever model. When the gap was positioned near to the applied force, the stress in both models increased significantly in the implant components and surrounding bone. The stress increase in each component ranged from 8% to 64% in the non-cantilever models and 43% to 85% in the cantilever models. The greatest stress was found in the distal gold screw. The effect of the gap was clearly shown by the pattern of stress distribution in both models. Additionally, the presence of a cantilever and excessive occlusal force amplified the effect of prosthesis misfit. Prosthesis misfit influenced the pattern and magnitude of stress distribution in the prosthesis, implant components, and surrounding bone, and the presence of the cantilever or greater occlusal force amplified the effect of misfit.


Brain organoid formation on decellularized porcine brain ECM hydrogels.
Human brain tissue models such as cerebral organoids are essential tools for developmental and biomedical research. Current methods to generate cerebral organoids often utilize Matrigel as an external scaffold to provide structure and biologically relevant signals. Matrigel however is a nonspecific hydrogel of mouse tumor origin and does not represent the complexity of the brain protein environment. In this study, we investigated the application of a decellularized adult porcine brain extracellular matrix (B-ECM) which could be processed into a hydrogel (B-ECM hydrogel) to be used as a scaffold for human embryonic stem cell (hESC)-derived brain organoids. We decellularized pig brains with a novel detergent- and enzyme-based method and analyzed the biomaterial properties, including protein composition and content, DNA content, mechanical characteristics, surface structure, and antigen presence. Then, we compared the growth of human brain organoid models with the B-ECM hydrogel or Matrigel controls in vitro. We found that the native brain source material was successfully decellularized with little remaining DNA content, while Mass Spectrometry (MS) showed the loss of several brain-specific proteins, while mainly different collagen types remained in the B-ECM. Rheological results revealed stable hydrogel formation, starting from B-ECM hydrogel concentrations of 5 mg/mL. hESCs cultured in B-ECM hydrogels showed gene expression and differentiation outcomes similar to those grown in Matrigel. These results indicate that B-ECM hydrogels can be used as an alternative scaffold for human cerebral organoid formation, and may be further optimized for improved organoid growth by further improving protein retention other than collagen after decellularization.


All-optical 1st and 2nd order integration on a chip.
We demonstrate all-optical temporal integration of arbitrary optical waveforms with temporal features as short as ~1.9ps. By using a four-port micro-ring resonator based on CMOS compatible doped glass technology we perform the 1st- and 2nd-order cumulative time integral of optical signals over a bandwidth that exceeds 400GHz. This device has applications for a wide range of ultra-fast data processing and pulse shaping functions as well as in the field of optical computing for the real-time analysis of differential equations.


Antibiotic-resistant acne: getting under the skin.
Propionibacterium acnes is a key pathogenic factor in the development of acne. Antibiotics are the first choice of treatment for mild-to-moderate, mixed, papular/pustular, and moderate nodular acne, and an alternative choice in severe, nodular/conglobate acne. The emergence of resistance to the currently available antibiotics poses a serious set-back to this algorithm, and the reduced arsenal can diminish efficacy of treatment. This emerging situation should catalyze innovations in dermatology; for example, newer drugs and technologies such as next-generation antibiotics with excellent potency and low propensity to develop resistance, rapid diagnostic platforms to select responders and nonresponders, and delivery technologies that target the bacteria. Such innovations can dramatically expand the arsenal for dermatologists in the management of acne.


Synovial tissue cytokine profile in disc displacement of the temporomandibular joint.
Symptomatic disc displacement (DD) of the temporomandibular joint (TMJ) may cause pain and limited mouth opening. The aetiopathogenesis is obscure and probably complex, which makes the diagnostic classification crude and mainly based on clinical criteria rather than disease mechanisms, and tissue characteristics. The study aim was to characterise and quantify synovial tissue in DD, where specific cytokine patterns might serve as potential biomarkers. An observational cohort study was performed harvesting synovial tissue from 63 patients: 44 with DD without reduction (DDwoR) and 19 with DD with reduction (DDwR). DDwoR was subdivided depending on type of onset (sudden, n = 17; delayed, n = 27), and DDwR served as the control group. Proteins were extracted from tissue samples and investigated in a multi-analytic profiling system. DDwoR patients had significantly higher concentrations in 12 out of 28 analysed cytokines compared to DDwR. In the same statistical model, significantly lower concentrations of interferon gamma-induced protein (IP) 10, osteoprotegerin (OPG) and RANTES were detected in DDwoR patients. Women showed significantly higher concentrations of epidermal growth factor and interleukin (IL) 1ra compared to men. DDwoR with sudden onset had significant higher concentrations of bone morphogenetic protein 4, eotaxin and IL-8 compared to DDwoR with delayed onset. Characterising the biomarker panel for TMJ conditions may serve as suggestible targets for disease classification and novel treatment options. The significantly lower concentrations of IP-10, OPG and RANTES could be proposed as putative markers for the separation of the studied conditions to other TMJ diseases.


Cerebral blood flow changes during chanting meditation.
To examine changes in brain physiology during a chanting meditation practice using cerebral blood flow single-photon emission computed tomography. Single-photon emission computed tomography scans were acquired in 11 healthy individuals during either a resting state or meditation practice randomly performed on two separate days. Statistical parametric mapping analyses were conducted to identify significant changes in regional cerebral blood flow (rCBF) between the two conditions. When the meditation state was compared with the baseline condition, significant rCBF increases were observed in the right temporal lobe and posterior cingulate gyrus, and significant rCBF decreases were observed in the left parietotemporal and occipital gyri. The results offer evidence that this form of meditation practice is associated with changes in brain function in a way that is consistent with earlier studies of related types of meditation as well as with the positive clinical outcomes anecdotally reported by its users.


Preclinical Cushing's syndrome in adrenal "incidentalomas": comparison with adrenal Cushing's syndrome.
Adrenal tumors are usually diagnosed by clinical symptoms of hormone excess. The increasing use of ultrasound and computed tomography results in the detection of a substantial number of incidentally discovered adrenal tumors. Most of these tumors are nonfunctional adrenocortical adenomas, but a few cases of subclinical cortisol production in "incidentalomas" have been reported. We investigated prospectively the prevalence of autonomous cortisol production in 68 patients (44 females and 24 males, aged 25-90 yr) with adrenal incidentalomas at our institution. As a screening procedure all patients with incidentalomas underwent an overnight dexamethasone suppression test (1 mg). Patients who failed to suppress serum cortisol below 140 nmol/L (5 micrograms/dL) underwent more comprehensive studies (prolonged dexamethasone suppression test, determination of the diurnal rhythm of cortisol secretion in saliva, and CRH stimulation test). Eight patients (12% of all patients with incidentalomas; 5 females and 3 males, aged 25-71 yr) were finally identified as having cortisol-producing tumors, and the findings in these patients were compared with those of overt Cushing's syndrome in 8 patients (8 females, aged 26-50 yr) suffering from cortisol-producing adrenal adenomas. The tumor size of patients with cortisol-producing incidentalomas ranged from 2-5 cm. No specific signs and symptoms of hypercortisolism were present, but arterial hypertension (seven of eight subjects), diffuse obesity (four of eight subjects), and noninsulin-dependent diabetes mellitus (NIDDM; two of eight subjects) were frequently observed. Baseline cortisol levels were in the normal to upper normal range, whereas baseline ACTH levels were suppressed in five of the eight patients. In none of the patients was serum cortisol suppressible by low dose or high dose dexamethasone. The ACTH and cortisol responses to CRH were normal in two, blunted in one, and suppressed in four patients. Unilateral adrenalectomy was performed in seven patients and resulted in temporary adrenal insufficiency in four of them. After surgery, improvement of arterial hypertension, a permanent weight loss in obese subjects, and a better metabolic control of NIDDM were noted in the majority of patients. The following conclusions were reached. Incidentally diagnosed adrenal tumors with pathological cortisol secretion in otherwise clinically asymptomatic patients are more frequently observed than previously assumed. Adrenocortical insufficiency is a major risk in these patients after adrenalectomy. After surgery, hypertension, obesity, and NIDDM may improve. Patients with asymptomatic adrenal incidentalomas, therefore, should be screened for cortisol production by means of an overnight dexamethasone suppression test.


Nucleosome-remodelling machines and other molecular motors observed at the single-molecule level.
Through its capability to transiently pack and unpack our genome, chromatin is a key player in the regulation of gene expression. Single-molecule approaches have recently complemented conventional biochemical and biophysical techniques to decipher the complex mechanisms ruling chromatin dynamics. Micromanipulations with tweezers (magnetic or optical) and imaging with molecular microscopy (electron or atomic force) have indeed provided opportunities to handle and visualize single molecules, and to measure the forces and torques produced by molecular motors, along with their effects on DNA or nucleosomal templates. By giving access to dynamic events that tend to be blurred in traditional biochemical bulk experiments, these techniques provide critical information regarding the mechanisms underlying the regulation of gene activation and deactivation by nucleosome and chromatin structural changes. This minireview describes some single-molecule approaches to the study of ATP-consuming molecular motors acting on DNA, with applications to the case of nucleosome-remodelling machines.


Correlation between orbital volume, body mass index, and eyeball position in healthy East asians.
The objectives of this study were measure the orbital volume of healthy Koreans and analyze the differences between orbital tissue volume with respect to age and sex and to assess any correlation between body mass index (BMI), eyeball position, and orbital volume. We retrospectively evaluated the scan results of patients who had undergone orbital computed tomography scans between November 2010 and November 2011. We assessed the scan results of 184 orbits in 92 adults who had no pathology of the orbit. The individuals were classified into 3 groups with respect to age. Orbital volume, effective orbital volume (defined as the difference between orbital and eyeball volume), extraocular muscle volume, orbital fat volume, and transverse globe protrusion were recorded and analyzed. The records of the subjects were reviewed retrospectively, and BMI was calculated. A correlation analysis was performed to investigate the correlation between BMI, eyeball position, and orbital volume. Orbital tissue volume, with the exception of orbital fat volume, was larger in men compared with women. In both sexes, orbital fat volume increased with increasing age, whereas the other volumes decreased. Orbital tissue volumes increased with increasing BMI, but transverse globe protrusion was not significantly related to BMI. In addition, orbital volume and effective orbital volume were positively correlated with transverse globe protrusion. These results provide basic information about the effects of age, sex, and BMI on orbital volume and eyeball position in healthy Koreans. Furthermore, these results will be helpful in the diagnosis of orbital diseases and in planning orbital surgeries.


[Analysis of social and environmental risk for pleural involvement in severe pneumonia in children younger than 5 years of age].
To determine social and environmental risk factors associated with pleural involvement among children from 3 to 59 months old who were hospitalized with severe pneumonia in northeastern Brazil. This was an observational and descriptive cross-sectional study with an analytical component. We evaluated 154 patients hospitalized due to severe pneumonia, with or without pleural involvement. Pleural involvement was determined based on radiological findings. The following variables were analyzed: age, sex, place of residence, housing conditions, day care attendance, passive smoking, family income, presence of consumer goods in the home (e.g., television, radio, refrigerator, automobile), and schooling and occupation of the mother or other caregiver. The information was obtained by interviewing the mother or other caregiver or by consulting the inpatient medical chart. The frequency of pleural involvement among the 154 patients evaluated was 25.3%. The following factors were associated with pleural involvement: living in a rural area, being in a household with two or fewer rooms, family income below 170 U.S. dollars per month, and birthweight < 2 500 g. These findings indicate the need to improve the socioeconomic and living conditions of the less-privileged population, especially in rural areas. The health sector should emphasize primary care, focusing on a preventive approach beginning in the prenatal period.


High-throughput sequencing of SARS-CoV-2 in wastewater provides insights into circulating variants.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged from a zoonotic spill-over event and has led to a global pandemic. The public health response has been predominantly informed by surveillance of symptomatic individuals and contact tracing, with quarantine, and other preventive measures have then been applied to mitigate further spread. Non-traditional methods of surveillance such as genomic epidemiology and wastewater-based epidemiology (WBE) have also been leveraged during this pandemic. Genomic epidemiology uses high-throughput sequencing of SARS-CoV-2 genomes to inform local and international transmission events, as well as the diversity of circulating variants. WBE uses wastewater to analyse community spread, as it is known that SARS-CoV-2 is shed through bodily excretions. Since both symptomatic and asymptomatic individuals contribute to wastewater inputs, we hypothesized that the resultant pooled sample of population-wide excreta can provide a more comprehensive picture of SARS-CoV-2 genomic diversity circulating in a community than clinical testing and sequencing alone. In this study, we analysed 91 wastewater samples from 11 states in the USA, where the majority of samples represent Maricopa County, Arizona (USA). With the objective of assessing the viral diversity at a population scale, we undertook a single-nucleotide variant (SNV) analysis on data from 52 samples with >90% SARS-CoV-2 genome coverage of sequence reads, and compared these SNVs with those detected in genomes sequenced from clinical patients. We identified 7973 SNVs, of which 5680 were novel SNVs that had not yet been identified in the global clinical-derived data as of 17th June 2020 (the day after our last wastewater sampling date). However, between 17th of June 2020 and 20th November 2020, almost half of the SNVs have since been detected in clinical-derived data. Using the combination of SNVs present in each sample, we identified the more probable lineages present in that sample and compared them to lineages observed in North America prior to our sampling dates. The wastewater-derived SARS-CoV-2 sequence data indicates there were more lineages circulating across the sampled communities than represented in the clinical-derived data. Principal coordinate analyses identified patterns in population structure based on genetic variation within the sequenced samples, with clear trends associated with increased diversity likely due to a higher number of infected individuals relative to the sampling dates. We demonstrate that genetic correlation analysis combined with SNVs analysis using wastewater sampling can provide a comprehensive snapshot of the SARS-CoV-2 genetic population structure circulating within a community, which might not be observed if relying solely on clinical cases.


Introducing intentional rounding: a pilot project.
To report on the introduction of intentional rounding in an NHS trust. A two-stage pilot project with participatory methods underpinned by core practice development principles was undertaken to introduce intentional rounding. Patients and staff found intentional rounding to be a positive experience, although nurses and midwives felt they were inadequately prepared to implement the practice. Leaders and managers who engaged in learning opportunities before the pilot study tended to achieve more successful and sustained practice change. There is disagreement among nurses about the contribution of intentional rounding to patient care, particularly in light of the weak evidence base, time involved and associated documentation. Therefore, further research is needed in this area.


Practical pituitary pathology: what does the pathologist need to know?
The sellar region is the site of frequent pathology. The pituitary is affected by a large number of pathologic entities arising from the gland itself and from adjacent anatomical structures including brain, blood vessels, nerves, and meninges. The surgical pathology of this area requires the accurate characterization of primary adenohypophysial tumors, craniopharyngiomas, neurologic neoplasms, germ cell tumors, hematologic malignancies, and metastases as well as nonneoplastic lesions such as cysts, hyperplasias, and inflammatory disorders. To provide a practical approach to the diagnosis of pituitary specimens. Literature review and primary material from the University of Toronto. The initial examination requires routine hematoxylin-eosin to establish whether the lesion is a primary adenohypophysial proliferation or one of the many other types of pathology that occur in this area. The most common lesions resected surgically are pituitary adenomas. These are evaluated with a number of special stains and immunohistochemical markers that are now available to accurately classify these tumors. The complex subclassification of pituitary adenomas is now recognized to reflect specific clinical features and genetic alterations that predict targeted therapies for patients with pituitary disorders.


Molecular nature of spontaneous mutations at the hypoxanthine-guanine phosphoribosyltransferase (hprt) locus in Chinese hamster ovary cells.
The hypoxanthine-guanine phosphoribosyltransferase (hprt) locus has been widely used as a selectable genetic marker for studies of mammalian cell mutagenesis. We report here the spontaneous mutation spectrum at the hprt locus in 64 independently isolated mutants of Chinese hamster ovary (CHO) cells. All nine hprt exons were simultaneously analyzed via multiplex polymerase chain reaction (PCR) for rapid detection of gene deletions or insertions. Structural point mutations were identified by direct sequence analysis of the PCR amplified cDNA. The molecular nature of RNA splicing errors and insertions was analyzed by solid-phase direct exon sequencing. Single base substitutions were found in 24 mutants (38%), of which 21 were missense and 3 were nonsense mutations. Transversions were about twice as frequent as transitions. Fifteen mutants (23%) had deletions involving either intragenic small fragments (2), single exons (9), or multiple exons (4). The majority of deletion breakpoints (71%) were located in regions surrounding exons 4, 5, and 6. RNA splicing mutations were observed in 15 mutants (23%) and affected exons 3-8; most (6/15) resulted in the loss of exon 7. Two insertion mutants, one with a 209 bp insert in exon 4 and the other with a 88 bp insert accompanied by a 24 bp deletion in exon 6, represent novel mutations reported for the first time in spontaneous mutants of the mammalian hprt gene.


A comprehensive benchmarking study of protocols and sequencing platforms for 16S rRNA community profiling.
In the last 5 years, the rapid pace of innovations and improvements in sequencing technologies has completely changed the landscape of metagenomic and metagenetic experiments. Therefore, it is critical to benchmark the various methodologies for interrogating the composition of microbial communities, so that we can assess their strengths and limitations. The most common phylogenetic marker for microbial community diversity studies is the 16S ribosomal RNA gene and in the last 10 years the field has moved from sequencing a small number of amplicons and samples to more complex studies where thousands of samples and multiple different gene regions are interrogated. We assembled 2 synthetic communities with an even (EM) and uneven (UM) distribution of archaeal and bacterial strains and species, as metagenomic control material, to assess performance of different experimental strategies. The 2 synthetic communities were used in this study, to highlight the limitations and the advantages of the leading sequencing platforms: MiSeq (Illumina), The Pacific Biosciences RSII, 454 GS-FLX/+ (Roche), and IonTorrent (Life Technologies). We describe an extensive survey based on synthetic communities using 3 experimental designs (fusion primers, universal tailed tag, ligated adaptors) across the 9 hypervariable 16S rDNA regions. We demonstrate that library preparation methodology can affect data interpretation due to different error and chimera rates generated during the procedure. The observed community composition was always biased, to a degree that depended on the platform, sequenced region and primer choice. However, crucially, our analysis suggests that 16S rRNA sequencing is still quantitative, in that relative changes in abundance of taxa between samples can be recovered, despite these biases. We have assessed a range of experimental conditions across several next generation sequencing platforms using the most up-to-date configurations. We propose that the choice of sequencing platform and experimental design needs to be taken into consideration in the early stage of a project by running a small trial consisting of several hypervariable regions to quantify the discriminatory power of each region. We also suggest that the use of a synthetic community as a positive control would be beneficial to identify the potential biases and procedural drawbacks that may lead to data misinterpretation. The results of this study will serve as a guideline for making decisions on which experimental condition and sequencing platform to consider to achieve the best microbial profiling.


Microelectrodes for the measurement of catecholamines in biological systems.
Many of the molecules involved in biological signaling processes are easily oxidized and have been monitored by electrochemical methods. Temporal response, spatial considerations, and sensitivity of the electrodes must be optimized for the specific biological application. To monitor exocytosis from single cells in culture, constant potential amperometry offers the best temporal resolution, and a low-noise picoammeter improves the detection limits. Smaller electrodes, with 1-micron diameters, provided spatial resolution sufficient to identify the locations of release sites on the surface of single cells. For the study of neurotransmitter release in vivo, larger cylindrical microelectrodes are advantageous because the secreted molecules come from multiple terminals near the electrode, and the greater amounts lead to a larger signal that emerges from the Johnson noise of the current amplifier. With this approach, dopamine release elicited by two electrical stimulus pulses at 10 Hz was detected with fastscan cyclic voltammetry in vivo. Nafion-coated elliptical electrodes have previously been shown to be incapable of detecting such concentration changes without extensive signal averaging. In addition, we demonstrate that high-pass filtering (200 Hz) of cyclic voltammograms recorded at 300 V/s decreases the background current and digitization noise at these microelectrodes, leading to an improved signal. Also, high-pass filtering discriminated against ascorbic acid, DOPAC, and acidic pH changes, three common interferences in vivo.


The Impact of Fine-Scale Disturbances on the Predictability of Vegetation Dynamics and Carbon Flux.
Predictions from forest ecosystem models are limited in part by large uncertainties in the current state of the land surface, as previous disturbances have important and lasting influences on ecosystem structure and fluxes that can be difficult to detect. Likewise, future disturbances also present a challenge to prediction as their dynamics are episodic and complex and occur across a range of spatial and temporal scales. While large extreme events such as tropical cyclones, fires, or pest outbreaks can produce dramatic consequences, small fine-scale disturbance events are typically much more common and may be as or even more important. This study focuses on the impacts of these smaller disturbance events on the predictability of vegetation dynamics and carbon flux. Using data on vegetation structure collected for the same domain at two different times, i.e. "repeat lidar data", we test high-resolution model predictions of vegetation dynamics and carbon flux across a range of spatial scales at an important tropical forest site at La Selva Biological Station, Costa Rica. We found that predicted height change from a height-structured ecosystem model compared well to lidar measured height change at the domain scale (~150 ha), but that the model-data mismatch increased exponentially as the spatial scale of evaluation decreased below 20 ha. We demonstrate that such scale-dependent errors can be attributed to errors predicting the pattern of fine-scale forest disturbances. The results of this study illustrate the strong impact fine-scale forest disturbances have on forest dynamics, ultimately limiting the spatial resolution of accurate model predictions.


Plant growth-promotion and biofortification of chickpea and pigeonpea through inoculation of biocontrol potential bacteria, isolated from organic soils.
Seven strains of bacteria [Pseudomonas plecoglossicida SRI-156, Brevibacterium antiquum SRI-158, Bacillus altitudinis SRI-178, Enterobacter ludwigii SRI-211, E. ludwigii SRI-229, Acinetobacter tandoii SRI-305 and Pseudomonas monteilii SRI-360; demonstrated previously for control of charcoal rot disease in sorghum and plant growth-promotion (PGP) in rice] were evaluated for their PGP and biofortification traits in chickpea and pigeonpea under field conditions. When treated on seed, the seven selected bacteria significantly enhanced the shoot height and root length of both chickpea and pigeonpea over the un-inoculated control. Under field conditions, in both chickpea and pigeonpea, the plots inoculated with test bacteria enhanced the nodule number, nodule weight, root and shoot weights, pod number, pod weight, leaf weight, leaf area and grain yield over the un-inoculated control plots. Among the seven bacteria, SRI-229 was found to significantly and consistently enhance all the studied PGP and yield traits including nodule number (24 and 36%), nodule weight (11 and 44%), shoot weight (22 and 20%), root weight (23 and 16%) and grain yield (19 and 26%) for both chickpea and pigeonpea, respectively. When the harvested grains were evaluated for their mineral contents, iron (up to 18 and 12%), zinc (up to 23 and 5%), copper (up to 19 and 8%), manganese (up to 2 and 39%) and calcium (up to 22 and 11%) contents in chickpea and pigeonpea, respectively, were found enhanced in test bacteria inoculated plots over the un-inoculated control plots. This study further confirms that the selected bacterial isolates not only have the potential for PGP in cereals and legumes but also have the potential for biofortification of mineral nutrients.


Crystal Structure of Staphylococcus aureus Cas9.
The RNA-guided DNA endonuclease Cas9 cleaves double-stranded DNA targets with a protospacer adjacent motif (PAM) and complementarity to the guide RNA. Recently, we harnessed Staphylococcus aureus Cas9 (SaCas9), which is significantly smaller than Streptococcus pyogenes Cas9 (SpCas9), to facilitate efficient in vivo genome editing. Here, we report the crystal structures of SaCas9 in complex with a single guide RNA (sgRNA) and its double-stranded DNA targets, containing the 5'-TTGAAT-3' PAM and the 5'-TTGGGT-3' PAM, at 2.6 and 2.7 Å resolutions, respectively. The structures revealed the mechanism of the relaxed recognition of the 5'-NNGRRT-3' PAM by SaCas9. A structural comparison of SaCas9 with SpCas9 highlighted both structural conservation and divergence, explaining their distinct PAM specificities and orthologous sgRNA recognition. Finally, we applied the structural information about this minimal Cas9 to rationally design compact transcriptional activators and inducible nucleases, to further expand the CRISPR-Cas9 genome editing toolbox.


Airborne stability of simian virus 40.
The influence of relative humidity on the airborne survival of simian virus 40 (SV40) was studied by allowing virus aerosols to age in rotating drums at 21 or 32 C and at a relative humidity (RH) value ranging from 22 to 88%. Airborne SV40 virus was stable at every RH tested at 21 C, but aerosols maintained at 32 C were inactivated within 60 min at mid-range RH values. The unusual stability at 21 C over a broad RH range indicates that potentially biohazardous situations may occur under laboratory conditions if this virus becomes accidentally airborne.


Distinguishing true coagulase-negative Staphylococcus infections from contaminants in the neonatal intensive care unit.
To characterize true coagulase-negative Staphylococcus (CoNS) infections in infants receiving neonatal intensive care. Retrospective cohort study of neonatal intensive care unit (NICU) infants with clinical sepsis and CoNS isolated from ≥ 2 blood cultures (BCs) or one BC and a sterile site (proved infection) or CoNS isolated from one BC and deemed significant after blinded data review (probable infection). In all, 98% of 40 proved and 96% of 55 probable infections occurred in infants with birth weight (BW) <2000 g and gestation <34 weeks. Total central lines (CLs) placed, but not CL duration or presence in situ, predicted proved (odds ratio (OR) 3.5, 95% confidence interval (CI) 1.4 to 8.3; P=0.005) and probable infection (OR 2.7, 95% CI 1.3 to 5.6; P=0.007) by multivariate analysis as did lethargy and gastric residuals. True CoNS infection is unlikely in infants with BW >2000 g and gestation >34 weeks. Total CL required for care, lethargy and gastric residuals predicted true CoNS infection.


Genetics and mapping of the R₁₁ gene conferring resistance to recently emerged rust races, tightly linked to male fertility restoration, in sunflower (Helianthus annuus L.).
Sunflower oil is one of the major sources of edible oil. As the second largest hybrid crop in the world, hybrid sunflowers are developed by using the PET1 cytoplasmic male sterility system that contributes to a 20 % yield advantage over the open-pollinated varieties. However, sunflower production in North America has recently been threatened by the evolution of new virulent pathotypes of sunflower rust caused by the fungus Puccinia helianthi Schwein. Rf ANN-1742, an 'HA 89' backcross restorer line derived from wild annual sunflower (Helianthus annuus L.), was identified as resistant to the newly emerged rust races. The aim of this study was to elucidate the inheritance of rust resistance and male fertility restoration and identify the chromosome location of the underlying genes in Rf ANN-1742. Chi-squared analysis of the segregation of rust response and male fertility in F(2) and F(3) populations revealed that both traits are controlled by single dominant genes, and that the rust resistance gene is closely linked to the restorer gene in the coupling phase. The two genes were designated as R ( 11 ) and Rf5, respectively. A set of 723 mapped SSR markers of sunflower was used to screen the polymorphism between HA 89 and the resistant plant. Bulked segregant analysis subsequently located R ( 11 ) on linkage group (LG) 13 of sunflower. Based on the SSR analyses of 192 F(2) individuals, R ( 11 ) and Rf5 both mapped to the lower end of LG13 at a genetic distance of 1.6 cM, and shared a common marker, ORS728, which was mapped 1.3 cM proximal to Rf5 and 0.3 cM distal to R ( 11 ) (Rf5/ORS728/R ( 11 )). Two additional SSRs were linked to Rf5 and R ( 11 ): ORS995 was 4.5 cM distal to Rf5 and ORS45 was 1.0 cM proximal to R ( 11 ). The advantage of such an introduced alien segment harboring two genes is its large phenotypic effect and simple inheritance, thereby facilitating their rapid deployment in sunflower breeding programs. Suppressed recombination was observed in LGs 2, 9, and 11 as it was evident that no recombination occurred in the introgressed regions of LGs 2, 9, and 11 detected by 5, 9, and 22 SSR markers, respectively. R ( 11 ) is genetically independent from the rust R-genes R ( 1 ), R ( 2 ), and R ( 5 ), but may be closely linked to the rust R-gene R ( adv ) derived from wild Helianthus argophyllus, forming a large rust R-gene cluster of R ( adv )/R ( 11 )/R ( 4 ) in the lower end of LG13. The relationship of Rf5 with Rf1 is discussed based on the marker association analysis.


Two cases of rarely recognized infection with Entamoeba moshkovskii.
We report two cases (a 36-year-old woman and 2-year-old girl) infected with Entamoeba moshkovskii in Turkey. Entamoeba moshkovskii was identified and distinguished from the morphologically identical parasites E. histolytica and E. dispar by a nested polymerase chain reaction and restriction fragment length polymorphism analysis of the small subunit ribosomal DNA gene.


Increased authenticity in practical assessment using emergency case OSCE stations.
In case of an emergency, a fast and structured patient management is crucial for patient's outcome. The competencies needed should be acquired and assessed during medical education. The objective structured clinical examination (OSCE) is a valid and reliable assessment format to evaluate practical skills. However, traditional OSCE stations examine isolated skills or components of a clinical algorithm and thereby lack a valid representation of clinical reality. We developed emergency case OSCE stations (ECOS), where students have to manage complete emergency situations from initial assessment to medical treatment and consideration of further procedures. Our aim was to increase the authenticity and validity in the assessment of students' capability to cope with emergency patients. 45 students participated in a 10-station OSCE with 6 ECOS and 4 traditional OSCE stations. They were assessed using a case-specific checklist. An inter-station and post-OSCE-questionnaire was completed by each student to evaluate both ECOS and traditional OSCE. In this study, we were able to demonstrate that ECOS are feasible as time-limited OSCE stations. There was a high acceptance on both students and examiners side. They rated ECOS to be more realistic in comparison to the traditional OSCE scenarios. The reliability estimated via Crohnbach's alpha for the 6 ECOS is high (0.793). ECOS offer a feasible alternative to the traditional OSCE stations with adequate reliability to assess students' capabilities to cope with an acute emergency in a realistic encounter.


A prospective study of the modified Atkins diet for intractable epilepsy in adults.
The ketogenic diet is not typically offered to adults with epilepsy due to the significant lifestyle alterations needed for its use. The modified Atkins diet has been recently demonstrated to be therapeutic for children without the need for an admission, fasting period, weighing of foods, or fluid, calorie, and protein restriction. A prospective, open-label study was performed of adults over 18 years of age, having at least weekly seizures and prior use of at least two anticonvulsants. Carbohydrates were initially restricted to 15 g/day, fats were encouraged, and fluids, protein, and calories were allowed ad lib. Thirty patients, with age ranging from 18 to 53 years, were enrolled. Using an intent-to-treat analysis, 47% had a >50% seizure reduction after 1 and 3 months on the diet; 33% after 6 months. In those with seizure reduction, the median time to improvement was 2 weeks (range: 1-8 weeks). The mean weight loss was 6.8 kg, p < 0.001. Body-mass index (BMI) decrease correlated with efficacy at 3 months, p = 0.03. Ten subjects (30%) discontinued the diet prior to 3 months. Side effects included increased cholesterol (mean 187 to 201 mg/dL), blood urea nitrogen (BUN; 13 to 16 mg/dL), and urine calcium to creatinine ratio (0.14 to 0.19). A modified Atkins diet appears to demonstrate preliminary efficacy for adults with intractable epilepsy, especially in those who lost weight. Considering the rapid response in those who improved, but somewhat high discontinuation rate, a 2-month trial period may be adequate to assess for efficacy.


Relationship between body mass index with dietary fiber intake and skinfolds--differences among bodybuilders who train during morning and nocturne period.
The prevalence of overweight individuals has increased in recent years. Moreover, the importance of a healthy diet is associated with the practice of physical activity and attempt to verify the achievement of physical exercise influences on food choice. However, it relationship between food intake and physical activity have not been studied. To evaluate if the period in which the trainings are conducted, morning and nocturne, interfere qualitatively and quantitatively in food consumption as well as verify possible associations between anthropometric profiles and dietary habits. We collected data from 33 adult volunteers, between men and women, practitioners of bodybuilding. A total of 33 volunteers were interviewed (18 (54.5%) were men and 15 (45.5%) were women). Regarding anthropometric data, it turns out that that the volunteers of the two periods had similar characteristics, differentiating only weight. The consumption of nutritional supplements was observed in 30.77% of the practitioners in the morning period vs. 35% for the nocturne. Considering macronutrient intake, there was a significant difference in the consumption of protein between the periods. The consumption during nocturne period was greater (126 ± 5% of the daily requirement) than the morning period (115.7 ± 2%). As for micro-nutrients, calcium intake was greater among men when compared to women. There was a positive correlation between the BMI, and arm circumference for practitioners of the morning period. This study show that the practitioners who train in the morning have quietly better eating habits than those in the nocturne period, however both are inappropriate.


ThermoEnergy Ammonia Recovery Process for municipal and agricultural wastes.
The Ammonia Recovery Process (ARP) is an award-winning, low-cost, environmentally responsible method of recovering nitrogen, in the form of ammonia, from various dilute waste streams and converting it into concentrated ammonium sulfate. The ThermoEnergy Biogas System utilizes the new chemisorption-based ARP to recover ammonia from anaerobically digested wastes. The process provides for optimal biogas production and significantly reduced nitrogen levels in the treated water discharge. Process flows for the ammonia recovery and ThermoEnergy biogas processes are presented and discussed. A comparison with other techniques such as biological nitrogen removal is made. The ARP technology uses reversible chemisorption and double salt crystal precipitation to recover and concentrate the ammonia. The ARP technology was successfully proven in a recent large-scale field demonstration at New York City's Oakwood Beach Wastewater Treatment Plant, located on Staten Island. This project was a joint effort with Foster Wheeler Environmental Corporation, the Civil Engineering Research Foundation, and New York City Department of Environmental Protection. Independent validated plant data show that ARP consistently recovers up to 99.9% of the ammonia from the city's centrate waste stream (derived from dewatering of sewage sludge), as ammonium sulfate. ARP technology can reduce the nitrogen (ammonia) discharged daily into local bodies of water by municipalities, concentrated animal farming operations, and industry. Recent advances to ARP enhance its performance and economic competitiveness in comparison to stripping or ammonia destruction technologies.


Electricity generation from young landfill leachate in a microbial fuel cell with a new electrode material.
This study aims at evaluating the performance of a two-chambered continuously fed microbial fuel cell with new Ti-TiO₂ electrodes for bioelectricity generation from young landfill leachate at varying strength of wastewater (1-50 COD g/L) and hydraulic retention time (HRT, 0.25-2 days). The COD removal efficiency in the MFC increased with time and reached 45 % at full-strength leachate (50 g/L COD) feeding. The current generation increased with increasing leachate strength and decreasing HRT up to organic loading rate of 100 g COD/L/day. The maximum current density throughout the study was 11 A/m² at HRT of 0.5 day and organic loading rate of 67 g COD/L/day. Coulombic efficiency (CE) decreased from 57 % at feed COD concentration of 1 g/L to less than 1 % when feed COD concentration was 50 g/L. Increase in OLR resulted in increase in power output but decrease in CE.


Metabolic fate of L-arginine in relation to microbiostatic capability of murine macrophages.
L-arginine is required for the fungistatic action of murine macrophages in vitro. To further investigate this requirement, L-arginine metabolism by macrophages was measured under conditions where fungistasis either succeeded or failed. Macrophage fungistasis correlated with metabolism of L-arginine to citrulline, nitrite, and nitrate. The metabolic rate was dependent on extracellular L-arginine concentration, reaching a maximum of 67 nmol nitrite/h per mg protein. It accounted for one-third of arginine consumed by fungistatic macrophages. Equimolar amounts of citrulline and total nitrite plus nitrate accumulated in medium. This was consistent with the hypothesis that one of the equivalent guanidino nitrogens of L-arginine was oxidized to both nitrite and nitrate leaving L-citrulline as the amino acid reaction product. The analogue, NG-mono-methyl-L-arginine, selectively inhibited nitrogen oxidation and it was shown previously that it inhibited fungistatic capability. Resident macrophages were not fungistatic and their nitrogen oxidation was low. Once macrophages began producing nitrite/nitrate, protein synthesis was not required during the next 8 h for either fungistasis or nitrogen oxidation. Two-thirds of L-arginine consumption was due to macrophage arginase yielding L-ornithine and urea, which accumulated in medium. This activity was dissociated from macrophage fungistasis. Nitrogen oxidation metabolism by macrophages is linked to a mechanism that inhibits proliferation of fungi. This may involve synthesis of an intermediate compound(s) that has antimicrobial properties.


The relationship between psychosocial stress and allergic disease among children and adolescents in Gwangyang Bay, Korea.
Stress is considered a causal factor in many diseases, allergic disease being one of them. The prevalence of allergic disease is increasing in Korea, but the relationship between allergic symptoms and stress is not empirically well known. We aimed to evaluate the relationship between allergy-related symptoms and stress in children and adolescents. We investigated 698 children and adolescents living in Gwangyang Bay, Korea, using a multi-stage cluster sampling method. Using the International Study of Asthma and Allergies in Childhood and the Psychosocial Well-being Index, these subjects were surveyed on allergy-related symptoms and psychosocial stressors in their lives, respectively. We used a multivariate logistic analysis for odds ratios for the complaint rate of allergic symptoms, after adjusting for age, gender, household income, body mass index, and residence. After adjustments, lifetime rhinitis (odds ratio [OR], 1.024), rhinoconjunctivitis (OR, 1.090), diagnosis of itchy eczema (OR, 1.040), treatment of itchy eczema (OR, 1.049), 12-month allergic conjunctivitis (OR, 1.026), diagnosis of allergic conjunctivitis (OR, 1.031), and treatment of allergic conjunctivitis (OR, 1.034) were found to be significantly associated with stress. Our results support the notion that there is a relationship between stress and allergic symptoms in children and adolescents. Further research into any causal relationship between stress and allergies, as well as preventative public health plans for decreasing stress in children and adolescents are needed.


Balancing cancer immunotherapy and immune-related adverse events: The emerging role of regulatory T cells.
Advances in our understanding οf tumor immunity have prompted a paradigm shift in oncology, with the emergence of immunotherapy, where therapeutic agents are used to target immune cells rather than cancer cells. A real breakthrough in the field of immunotherapy came with the use of immune checkpoint inhibitors (ICI), namely antagonistic antibodies that block key immune regulatory molecules (checkpoint molecules), such as cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4), programmed cell death protein (PD-1) and its ligand PD-L1, that under physiologic conditions suppress T cell effector function. However, despite the enormous success, a significant proportion of patients do not respond, while responses are frequently accompanied by life-threatening autoimmune related adverse events (irAEs). A major impediment in the effectiveness of ICI immunotherapy is the tumoral resistance, which is dependent on the immunosuppressive nature of tumor microenvironment (TME). Regulatory T cells (Tregs) are among the most abundant suppressive cells in the TME and their presence has been correlated with tumor progression, invasiveness as well as metastasis. Tregs are characterized by the expression of the transcription factor Foxp3 and various mechanisms ranging from cell-to-cell contact to secretion of inhibitory molecules have been implicated in their function. Notably, Tregs amply express most of the checkpoint molecules such as CTLA4, PD1 and LAG3 and therefore represent a direct target of ICI immunotherapy. Taking into consideration the critical role of Tregs in maintenance of immune homeostasis and avoidance of autoimmunity it is plausible that targeting of Tregs by ICI immunotherapy results in the development of irAEs. Since the use of ICI becomes common, and new immune checkpoint molecules are currently under clinical trials for the treatment of cancer, the occurrence of irAEs is expected to dramatically rise. Herein we review the current literature focusing on the role of Tregs in cancer evolution, ICI response and development of irAEs. Unraveling the complex mechanisms that hinder the tumor immune surveillance and in particular how ICI immunotherapy imprint on Treg activities to promote cancer regression while avoid development of irAEs, will empower the design of novel immunotherapeutic modalities in cancer with increased efficacy and diminished adverse events.


Levofloxacin for NIH Category II Chronic Bacterial Prostatitis: A Real-Life Study.
Mounting worldwide resistance trends make the use of fluoroquinolone (FQ) antibacterial agents increasingly difficult. This is felt more acutely in the case of urogenital infections, which are mainly caused by Gram-negative pathogens. For years, levofloxacin and other FQs have been the first-line drugs for treating National Institutes of Health (NIH) category II chronic bacteria prostatitis (CBP). Eradication rates achieved by levofloxacin in the frame of randomized trials vary greatly, ranging between 71 and 86%. This was a retrospective observational study to investigate the efficacy of levofloxacin against CBP in a real-life setting (urological outpatient wards). A database including the clinical records of >2,500 CBP patients was reviewed. Patients were selected based on strict inclusion criteria. They were treated for 4 weeks with 500 mg levofloxacin per day, alone or combined with other antibacterials. Besides standard urological procedures including the 4-glass test for pathogen isolation, international symptom questionnaires (the NIH Chronic Prostatitis Symptom Index [NIH-CPSI] and International Prostate Symptom Score [IPSS]) were administered. Pathogen eradication was achieved in 79% of the cases treated with levofloxacin as a single agent and 87.8% of patients who received a combination of levofloxacin and azithromycin. The 11% increase in the eradication rate in the latter group is statistically significant. In addition, the levofloxacin-azithromycin combination caused a significant decrease in prostate volume and significantly increased the bladder-voided volume. IPSS and NIH-CPSI values and the urinary peak flow rate decreased to a similar extent in both treatment groups. No adverse effects were reported by patients belonging to either treatment group. Levofloxacin retained its therapeutic efficacy in patients assessed in a real-life setting, and high eradication rates were attained when it was administered as a single agent. A combination of an FQ with azithromycin induced a significant improvement of eradication rates. This strategy may be an interesting option in both first-referral and relapsing cases, although caution should be exercised when patients are at risk of developing arrhythmias, tendinitis, or other adverse effects.


[Gene therapy for castration resistant prostate cancer].
Prostate is an ideal target organ for the gene therapy as a translational research (TR). It has advantages as follows: prostate is not a life keeping organ, can be approached easily by ultrasound as a routine clinical technique and PSA is a potent tumor marked for the evaluation of clinical response. Many clinical gene therapy trials for castration resistant prostate cancer (CRPC) have been conducted in many institutes showing clinical safety and efficacy including Japanese trial. A phase I/II study of adenovirus-mediated REIC/Dkk-3 gene therapy for CRPC was initiated in Okayama university hospital and oncolytic Herpes virus therapy was also ongoing in Tokyo university hospital. In spite of limited cases, some clinically positive results are observed. In this paper ongoing development of gene therapy for CRPC is reported and legal issues which are essential for the management of clinical gene therapy are also explained.


Deciphering drift time measurements from travelling wave ion mobility spectrometry-mass spectrometry studies.
Detailed knowledge of the tertiary and quaternary structure of proteins and protein complexes is of immense importance in understanding their functionality. Similarly, variations in the conformational states of proteins form the underlying mechanisms behind many biomolecular processes, numerous of which are disease-related. Thus, the availability of reliable and accurate biophysical techniques that can provide detailed information concerning these issues is of paramount importance. Ion mobility spectrometry (IMS) coupled to mass spectrometry (MS) offers a unique opportunity to separate multi-component biomolecular entities and to measure the molecular mass and collision cross-section of individual components in a single, rapid (</= 2 min) experiment, providing 3D- architectural information directly. Here we report a method of calibrating a commercially available electrospray ionisation (ESI)-travelling wave ion mobility spectrometry (TWIMS)-mass spectrometer using known cross-sectional areas determined for a range of biomolecules by conventional IMS-MS. Using this method of calibration, we have analysed a range of proteins of differing mass and 3D architecture in their native conformations by ESI-TWIMS-MS and found that the cross-sectional areas measured in this way compare extremely favourably with cross-sectional areas calculated using an in-house computing method based on Protein Data Bank NMR-derived co-ordinates. This not only provides a high degree of confidence in the calibration method, but also suggests that the gas phase ESI- TWIMS-MS measurements relate well to solution-based measurements derived from other biophysical techniques. In order to determine which instrumental parameters affect the ESI-TWIMS-MS cross-sectional area calibration, a systematic study of the parameters used to optimise TWIMS drift time separations has been carried out, observing the effect each parameter has on drift times and IMS resolution. Finally, the ESI-TWIMS-MS cross-sectional area calibration has been applied to the analysis of the amyloidogenic protein beta(2)-microglobulin and measurements for three co-populated conformational families, present under denaturing conditions, have been made: the folded, partially unfolded and unfolded states.


Undeclared animal species in dry and wet novel and hydrolyzed protein diets for dogs and cats detected by microarray analysis.
Although the European Pet Food Industry Federation (FEDIAF) stated that labels must be accurate and provide detailed information on the ingredients, mislabeling of pet food has been documented by several authors. This phenomenon is of particular concern when related to products used as elimination diets for the diagnosis of adverse food reaction (AFR) in dogs and cats because the presence of undeclared ingredients may negatively interfere with the trial and prevent the veterinarian from making an appropriate diagnosis. The aim of this study was to shed light upon the problem of contamination and mislabeling in both dry and wet novel protein diets (NPDs) and hydrolyzed protein diets (HPDs) using a microarray-based commercial kit which tests for the presence of 19 animal species. Of the 40 analyzed products (9 dry NPDs, 22 wet NPDs, 6 dry HPDs and 3 wet HPDs), ten presented a content that correctly matched the label, while five did not contain the declared animal species, twenty-three revealed the presence of undeclared animal species, and two had a vague label that did not allow the evaluation of its accuracy. The most frequently contaminants identified in both dry and wet pet foods were pork, chicken and turkey. The presence of undeclared animal species was higher in dry than wet pet foods; furthermore, a lower number of contaminating animal species was identified in HPDs than NPDs (4 vs 10), and a lower number of contaminated HPDs (6 out of 9, 67%) than contaminated NPDs was detected (24 out of 31, 77%). Thirteen out of 14 brands tested presented at least one mislabeled product. Mislabeling seems to be a widespread issue in pet foods used as elimination diets. Contamination can occur in all types of products used for the purpose, although dry NPDs are the main issue. Due to the high risk of contamination, particular attention should be given to both the selection of raw material suppliers and the production process.


Conducting Online Behavioral Research Using Crowdsourcing Services in Japan.
Recent research on human behavior has often collected empirical data from the online labor market, through a process known as crowdsourcing. As well as the United States and the major European countries, there are several crowdsourcing services in Japan. For research purpose, Amazon's Mechanical Turk (MTurk) is the widely used platform among those services. Previous validation studies have shown many commonalities between MTurk workers and participants from traditional samples based on not only personality but also performance on reasoning tasks. The present study aims to extend these findings to non-MTurk (i.e., Japanese) crowdsourcing samples in which workers have different ethnic backgrounds from those of MTurk. We conducted three surveys (N = 426, 453, 167, respectively) designed to compare Japanese crowdsourcing workers and university students in terms of their demographics, personality traits, reasoning skills, and attention to instructions. The results generally align with previous studies and suggest that non-MTurk participants are also eligible for behavioral research. Furthermore, small screen devices are found to impair participants' attention to instructions. Several recommendations concerning this sample are presented.


Early changes in auditory function as a result of platinum chemotherapy: use of extended high-frequency audiometry and evoked distortion product otoacoustic emissions.
The objective is to describe progressive changes in hearing and cochlear function in children and adolescents treated with platinum-based chemotherapy and to begin preliminary evaluation of the feasibility of extended high-frequency audiometry and distortion product otoacoustic emissions for ototoxicity monitoring in children. Baseline and serial measurement of conventional pure-tone audiometry (0.5 to 8 kHz) and evoked distortion product otoacoustic emissions (DPOAEs) were conducted for 32 patients age 8 months to 20 years who were treated with cisplatin and/or carboplatin chemotherapy. Seventeen children also had baseline and serial measurement of extended high-frequency (EHF) audiometry (9 to 16 kHz). Audiologic data were analyzed to determine the incidence of ototoxicity using the American Speech-Language-Hearing Association criteria, and the relationships between the different measures of ototoxicity. Of the 32 children, 20 (62.5%) acquired bilateral ototoxicity in the conventional frequency range during chemotherapy treatment, and 26 (81.3%) had bilateral decreases in DPOAE amplitudes and dynamic range. Of the 17 children with EHF audiometry results, 16 (94.1%) had bilateral ototoxicity in the EHF range. Pilot data suggest that EHF thresholds and DPOAEs show ototoxic changes before hearing loss is detected by conventional audiometry. EHF audiometry and DPOAEs have the potential to reveal earlier changes in auditory function than conventional frequency audiometry during platinum chemotherapy in children.


Ipilimumab and radiation therapy for melanoma brain metastases.
Ipilimumab, an antibody that enhances T-cell activation, may augment immunogenicity of tumor cells that are injured by radiation therapy. We hypothesized that patients with melanoma brain metastasis treated with both ipilimumab and radiotherapy would have improved overall survival, and that the sequence of treatments may affect disease control in the brain. We analyzed the clinical and radiographic records of melanoma patients with brain metastases who were treated with whole brain radiation therapy or stereotactic radiosurgery between 2005 and 2012. The hazard ratios for survival were estimated to assess outcomes as a function of ipilimumab use and radiation type. Seventy patients were identified, 33 of whom received ipilimumab and 37 who did not. The patients who received ipilimumab had a censored median survival of 18.3 months (95% confidence interval 8.1-25.5), compared with 5.3 months (95% confidence interval 4.0-7.6) for patients who did not receive ipilimumab. Ipilimumab and stereotactic radiosurgery were each significant predictors of improved overall survival (hazard ratio = 0.43 and 0.45, with P = 0.005 and 0.008, respectively). Four of 10 evaluable patients (40.0%) who received ipilimumab prior to radiotherapy demonstrated a partial response to radiotherapy, compared with two of 22 evaluable patients (9.1%) who did not receive ipilimumab. Ipilimumab is associated with a significantly reduced risk of death in patients with melanoma brain metastases who underwent radiotherapy, and this finding supports the need for multimodality therapy to optimize patient outcomes. Prospective studies are needed and are underway.


Longitudinal Relationships between Perceived Social Support and Symptom Outcomes: Findings from a sample of Adult Survivors of Childhood Sexual Abuse.
Cross-sectional studies have consistently reported an inverse association between perceived social support and the severity of mental health symptoms among adult survivors of childhood sexual abuse (CSA). However, there is a lack of longitudinal studies investigating the bidirectional association between social support and the severity of symptoms among adult CSA-survivors, as well as the role of relational problems in predicting perceived social support and symptom levels over time. The present study addressed these questions in a sample of primarily female CSA-survivors. In a three-wave, four-year longitudinal study of 506 CSA-survivors (94.9% women, 5.1% men) recruited from support centers for sexual abuse survivors in Norway, we used cross-lagged panel structural equation modeling to examine the directionality of the longitudinal associations between perceived social support and symptoms of posttraumatic stress, anxiety, depression and insomnia. Cross-lagged panel analyses revealed significant weak reciprocal associations between perceived social support and depression, posttraumatic stress symptoms and anxiety symptoms, but not with insomnia symptoms. The observed effects were partly overlapping and partly inconsistent across the different symptom domains. Relational problems predicted social support cross-sectionally and longitudinally, whereas only cross-sectional associations were found between the relational problems variable and mental health symptoms. Theoretical and clinical implications of the findings are discussed, alongside methodological limitations of the study.


Staging of locally advanced breast cancer and the prediction of response to neoadjuvant chemotherapy: complementary role of scintimammography and 18F-FDG PET/CT.
The primary endpoint of the study was to established the role of sestamibi scintimammography and PET/CT findings in locally advanced breast cancer (LABC) before neoadjuvant systemic therapy (NST) in different histological subtypes. The secondary endpoint was to determine the role of FDG PET/CT as multi-drug resistance marker. From January 2012, we prospectively enrolled 51 consecutive women (median age: 49 years; range: 27-76 yrs) with a biopsy-proven LABC. All patients underwent both sestamibi scintimammography and FDG PET/CT within one week before to start NST. Both examinations were qualitatively and semiquantitatively analysed. For scintimammography we calculated the tumor to background ratio (T/B) and the most intense uptake of the tumor to background ratio (I/B) according the following formula: T/B=[cntsT-cntsB]/ [cntsB] and I/B [cntsI-cntsB]/[cntsB]. Furthermore, the percentage washout index (WO) for T and I were obtained, according to: WOT,I= [cntsT,I]early image-[cntsT,I]delayed image/[cntsT,I]early image. Maximum and average (avg) standardized uptake value (SUV) was computed by PET/CT, using a region of interest. Patients who had an evidence of systemic metastases or a second active cancer at imaging scans, were excluded. At the end of pre-operative therapy, the response to therapy was assessed by the analysis of surgical specimen and then correlated with both scintimammographic and PET/CT data. Based on the inclusion criteria, the final analysis was performed in 49 patients. Scintimammography and PET/CT showed a sensitivity of 100% for the evaluation of primary cancer, while PET/CT showed a slightly higher detection rate for axillary lymph node than scintimammography. According to the biological pattern, SUVmax and SUVavg resulted significantly different among histological subtypes, whereas scintimammographic data did not. At the end of neo-adjuvant therapy, pathological complete response was obtained in 12 (24.4%) patients, while 37 had a partial or no response to NST (identified as no-responders). On the basis of histopathological response to NST, median WOI resulted significantly lower in responders than non-responders (30.5% vs. 44%; P=0.027). Conversely, SUVmax and SUVavg were significantly higher in responders than non-responders (all P<0.05). In this latter subset of patients, high WOTs were associated with low SUVs. On the contrary, in responder group, high SUVs were reported particularly for high WOT values. Scintimammography with sestamibi did not accurately determine the responsiveness to therapy. FDG PET/CT is more accurate in the prediction of response to therapy, particularly in the aggressive LABC subtype. Moreover, semiquantitative data by FDG PET seems to be linked with the chemosensitivity to NST.


Origins of Enterovirus Replication Organelles Established by Whole-Cell Electron Microscopy.
Enterovirus genome replication occurs at virus-induced structures derived from cellular membranes and lipids. However, the origin of these replication organelles (ROs) remains uncertain. Ultrastructural evidence of the membrane donor is lacking, suggesting that the sites of its transition into ROs are rare or fleeting. To overcome this challenge, we combined live-cell imaging and serial block-face scanning electron microscopy of whole cells to capture emerging enterovirus ROs. The first foci of fluorescently labeled viral protein correlated with ROs connected to the endoplasmic reticulum (ER) and preceded the appearance of ROs stemming from the trans-Golgi network. Whole-cell data sets further revealed striking contact regions between ROs and lipid droplets that may represent a route for lipid shuttling to facilitate RO proliferation and genome replication. Our data provide direct evidence that enteroviruses use ER and then Golgi membranes to initiate RO formation, demonstrating the remarkable flexibility with which enteroviruses usurp cellular organelles.IMPORTANCE Enteroviruses are causative agents of a range of human diseases. The replication of these viruses within cells relies on specialized membranous structures termed replication organelles (ROs) that form during infection but whose origin remains elusive. To capture the emergence of enterovirus ROs, we use correlative light and serial block-face scanning electron microscopy, a powerful method to pinpoint rare events in their whole-cell ultrastructural context. RO biogenesis was found to occur first at ER and then at Golgi membranes. Extensive contacts were found between early ROs and lipid droplets (LDs), which likely serve to provide LD-derived lipids required for replication. Together, these data establish the dual origin of enterovirus ROs and the chronology of their biogenesis at different supporting cellular membranes.


Identification of tryptic peptides from large databases using multiplexed tandem mass spectrometry: simulations and experimental results.
Multiplexed tandem mass spectrometry (MS/MS) has recently been demonstrated as a means to increase the throughput of peptide identification in liquid chromatography (LC) MS/MS experiments. In this approach, a set of parent species is dissociated simultaneously and measured in a single spectrum (in the same manner that a single parent ion is conventionally studied), providing a gain in sensitivity and throughput proportional to the number of species that can be simultaneously addressed. In the present work, simulations performed using the Caenorhabditis elegans predicted proteins database show that multiplexed MS/MS data allow the identification of tryptic peptides from mixtures of up to ten peptides from a single dataset with only three "y" or "b" fragments per peptide and a mass accuracy of 2.5 to 5 ppm. At this level of database and data complexity, 98% of the 500 peptides considered in the simulation were correctly identified. This compares favorably with the rates obtained for classical MS/MS at more modest mass measurement accuracy. LC multiplexed Fourier transform-ion cyclotron resonance MS/MS data obtained from a 66 kDa protein (bovine serum albumin) tryptic digest sample are presented to illustrate the approach, and confirm that peptides can be effectively identified from the C. elegans database to which the protein sequence had been appended.


Characterization of cytochrome P450 monooxygenase CYP154H1 from the thermophilic soil bacterium Thermobifida fusca.
Cytochrome P450 monooxygenases are valuable biocatalysts due to their ability to hydroxylate unactivated carbon atoms using molecular oxygen. We have cloned the gene for a new cytochrome P450 monooxygenase, named CYP154H1, from the moderately thermophilic soil bacterium Thermobifida fusca. The enzyme was overexpressed in Escherichia coli at up to 14% of total soluble protein and purified to homogeneity in three steps. CYP154H1 activity was reconstituted using putidaredoxin reductase and putidaredoxin from Pseudomonas putida DSM 50198 as surrogate electron transfer partners. In biocatalytic reactions with different aliphatic and aromatic substrates of varying size, the enzyme converted small aromatic and arylaliphatic compounds like ethylbenzene, styrene, and indole. Furthermore, CYP154H1 also accepted different arylaliphatic sulfides as substrates chemoselectively forming the corresponding sulfoxides and sulfones. The enzyme is moderately thermostable with an apparent melting temperature of 67°C and exhibited still 90% of initial activity after incubation at 50°C.


Comparison of an elemental and polymeric enteral diet in patients with normal gastrointestinal function.
In a prospective controlled clinical trial, 70 patients with normal gastrointestinal function were randomised to receive either an elemental diet based on Vivonex HN or an isonitrogenous isocalorie polymeric diet based on Clinifeed 400, administered by continuous 24 hour nasogastric infusion. The two groups of patients were well matched for age, sex, diagnosis, prior starvation, duration of feeding, initial nutritional status, and metabolic status. Nitrogen losses were significantly less on the polymeric feed, despite similar intakes. Serum transferrin rose significantly (1.85 +/- 0.2 to 2.30 +/- 0.2 g/l, p less than 0.05) only in the Clinifeed group, but nutritional parameters were otherwise maintained in both groups. The incidence of diarrhoea (Vivonex, 23.5%; Clinifeed, 30.6%) was not significantly different and was attributable to antibiotics in most cases. Hypokalaemia, which occurred in nearly half the patients, was equally distributed in the two groups, but hypophosphataemia occurred more often in the Vivonex group (p less than 0.05). Liver enzyme disturbances were similar in both groups. The present findings, therefore, provide no evidence that chemically defined 'elemental' diets containing free amino acids as their nitrogen source are in any way superior to polymeric diets containing whole protein and fat when administered to patients with normal gastrointestinal function.


Bidirectional scalable motion for scalable video coding.
Motion information scalability is an important requirement for a fully scalable video codec, especially in low bit rate or small resolution decoding scenarios, for which the fully scalable motion model (SMM) has been proposed. SMM can collaborate flawlessly with other scalabilities, such as spatial, temporal and quality, in a scalable video codec. It performs better than the nonscalable motion model. To further improve the SMM, this paper extends the algorithm to support the hierarchical B frame structure and bidirectional or multidirectional motion estimation. Furthermore, the corresponding rate distortion optimized estimation for improved efficiency in several scenarios is discussed. Several simulation results based on the updated framework are presented to verify the advantage of this extension.