Continuous infusion in adult females dogs submitted to ovariohysterectomy with midazolam-xylazine and/or medetomidine pre-treated with methotrimeprazine and buprenorphine.
To compare, by continuous infusion of ketamine or medetomidine combined to methotrimeprazine and buprenorphine, ketamine and midazolam, the degree of hypnosis, myorelaxation, anesthetic quality and surgical feasibility through evaluation of possible parametric alterations and recovery quality. 20 healthy adult females dogs, aged 3 to 5 years, body weight between 7 and 15 kg, were assigned randomly and homogenously to 2 groups of 10 animals each (n=10), group 1 (G1) and group 2 (G2), respectively. Animals of G1 were subjected to a pre-treatment with intravenous 1.0 mg/kg methotrimeprazine and or 3ì/kg. After 15 minutes, a 5.0 mg/kg ketamine and 0.2 mg/kg midazolam were intravenously injected. Immediately after induction, an anesthetic combination of 0.4 mg/kg/h midazolam, 20 mg/kg/h ketamine and 1.0 mg/kg/h xylazine, was continuously and intravenously administered for 30 minutes. The same techniques were used in G2 except for the substitution of xylazine for 30ìg/kg/h medetomidine. In G1 there was a 1st and 2nd degree atrioventricular heart block, a longer recovery period and lower quality. In G2 a 1st degree atrioventricular heart block occurred but isolated and ephemeral. The continuous infusion method, besides reducing drugs utilization, prevented collateral effects allowing a more tranquil recovery with no excitations, both protocols permitted the surgical procedure (ovary-hysterectomy) bringing about a reduction in hypnosis and an accentuated myorelaxation. Xylazine and medetomidine showed a similar pharmacodynamic behavior but with different clinical aspects. The electrocardiographic alterations observed in G2 and in a lower degree in G1 must be well studied. Describers: dogs, ketamine, methotrimeprazine, medetomidine, midazolam and xylazine.


Histology of genital tract and breast tissue after long-term testosterone administration in a female-to-male transsexual population.
Growing evidence indicates that androgens play a positive role in follicle proliferation and growth. Hence, many authors have assumed that androgen supplementation in women with poor ovarian reserve might improve the number of antral follicles available for ovarian stimulation. As androgen administration may become more frequently used in reproductive medicine, this study aimed at describing the histological changes observed in the genital tract and the breast of female-to-male (FTM) transsexuals. A pathological analysis of the genital tract of 112 FTM subjects who were given androgen for at least 6 months before hystero-salpingo-oophorectomy was performed. In addition, 100 bilateral mastectomies were performed, allowing a study of the breast tissue. Mean ovarian volume was increased, with histological characteristics of polycystic ovaries (PCO), defined as >12 antral follicles per ovary, observed in 89 patients (79.5%). Endometrial atrophy was observed in 45%. Breast examination revealed marked reduction of glandular tissue and increase of fibrous connective tissue in 93%, without atypical hyperplasia or carcinoma. The present data confirms and expands the putative associations between long-term androgen administration and abnormalities in ovarian architecture with macroscopic and microscopic characteristics of PCO, increased risk of endometrial atrophy and fibrotic breast tissue with marked glandular reduction.


Arthroscopic treatment of posterior shoulder instability in patients with and without glenoid dysplasia: a comparative outcomes analysis.
The purpose of this study was to evaluate the influence of glenoid dysplasia on outcomes after isolated arthroscopic posterior labral repair in a young military population. Thirty-seven male patients who underwent arthroscopic posterior labral repair for symptomatic posterior shoulder instability were evaluated at a mean duration of 3.1 years. A comparative analysis was performed for those with glenoid dysplasia and without dysplasia. Additional factors analyzed included military occupational specialty (MOS), preoperative mental health clinical encounters and mental health medication use, and radiographic characteristics (version, posterior humeral head subluxation, and posterior capsular area) on a preoperative standard shoulder magnetic resonance arthrogram. The groups were analyzed with regard to shoulder outcome scores (subjective shoulder value [SSV], American Shoulder and Elbow Surgeons [ASES] rating scale, Western Ontario Shoulder Instability Index [WOSI]), need for revision surgery, and medical separation from the military. Of 37 patients, 3 (8.1%) underwent revision surgery and 6 (16%) underwent medical separation. Overall outcome assessment demonstrated a mean SSV of 67.9 (range, 25-100) ± 22.1, mean ASES of 65.6 (range, 15-100) ± 22, and mean WOSI of 822.6 (range, 5-1854) ± 538. There were no significant differences in clinical outcome scores between the glenoid dysplasia and no dysplasia groups (SSV, P = .55; ASES, P = .57; WOSI, P = .56). MOS (P = .02) and a history of mental health encounters (P = .04) were significantly associated with diminished outcomes. The presence or absence of glenoid dysplasia did not influence the outcome after arthroscopic posterior labral repair in a young military population. However, a history of mental health clinical encounters and an infantry MOS were significantly associated with poorer clinical outcomes.


Pharmacokinetics of fluconazole following intravenous and oral administration to koalas (Phascolarctos cinereus).
Clinically normal koalas (n = 12) received a single dose of 10 mg/kg fluconazole orally (p.o.; n = 6) or intravenously (i.v.; n = 6). Serial plasma samples were collected over 24 h, and fluconazole concentrations were determined using a validated HPLC assay. A noncompartmental pharmacokinetic analysis was performed. Following i.v. administration, median (range) plasma clearance (CL) and steady-state volume of distribution (Vss ) were 0.31 (0.11-0.55) L/h/kg and 0.92 (0.38-1.40) L/kg, respectively. The elimination half-life (t1/2 ) was much shorter than in many species (i.v.: median 2.25, range 0.98-6.51 h; p.o.: 4.69, range 2.47-8.01 h), and oral bioavailability was low and variable (median 0.53, range 0.20-0.97). Absorption rate-limited disposition was evident. Plasma protein binding was 39.5 ± 3.5%. Although fluconazole volume of distribution (Varea ) displayed an allometric relationship with other mammals, CL and t1/2 did not. Allometrically scaled values were approximately sevenfold lower (CL) and sixfold higher (t1/2 ) than observed values, highlighting flaws associated with this technique in physiologically distinct species. On the basis of fAUC/MIC pharmacodynamic targets, fluconazole is predicted to be ineffective against Cryptococcus gattii in the koala as a sole therapeutic agent administered at 10 mg/kg p.o. every 12 h.


Growth and paralytic shellfish poisoning toxin production by a Mexican dinoflagellate strain of Alexandrium tamiyavanichii Balech (1994) under different nutrient conditions.
Alexandrium tamiyavanichii is a marine dinoflagellate known to produce Paralytic Shellfish Poisoning (PSP) toxin. Thus, a strain was isolated from La Paz Bay, Baja California Sur, Mexico and used to explore whether stress conditions, such as phosphorus limitation (PL) and nitrogen enrichment (NE) modulate population growth and PSP toxin production in the GSe medium. Growth kinetics showed that the PL treatment produced a 3.4-fold increase in cell density versus control at day 30 of the culture cycle. The highest PSP concentration was found in the control culture (309 fmol cell-1) on day 21. Saxitoxin (STX) was the main analog in all the treatments (> 40 % mol). In conclusion, PL and NE treatments promoted growth kinetics in the species studied but did not affect the PSP toxin production. For the first time, the present research describes A. tamiyavanichii high toxicity strain isolated from Mexican coasts relative to the South-Atlantic strains.


Shewanella oneidensis: a new and efficient system for expression and maturation of heterologous [Fe-Fe] hydrogenase from Chlamydomonas reinhardtii.
The eukaryotic green alga, Chlamydomonas reinhardtii, produces H2 under anaerobic conditions, in a reaction catalysed by a [Fe-Fe] hydrogenase HydA1. For further biochemical and biophysical studies a suitable expression system of this enzyme should be found to overcome its weak expression in the host organism. Two heterologous expression systems used up to now have several advantages. However they are not free from some drawbacks. In this work we use bacterium Shewanella oneidensis as a new and efficient system for expression and maturation of HydA1 from Chlamydomonas reinhardtii. Based on codon usage bias and hydrogenase maturation ability, the bacterium S. oneidensis, which possesses putative [Fe-Fe] and [Ni-Fe] hydrogenase operons, was selected as the best potential host for C. reinhardtii [Fe-Fe] hydrogenase expression. Hydrogen formation by S. oneidensis strain AS52 (Delta hydA Delta hyaB) transformed with a plasmid bearing CrHydA1 and grown in the presence of six different substrates for anaerobic respiration was determined. A significant increase in hydrogen evolution was observed for cells grown in the presence of trimethylamine oxide, dimethylsulfoxide and disodium thiosulfate, showing that the system of S. oneidensis is efficient for heterologous expression of algal [Fe-Fe] hydrogenase. In the present work a new efficient system for heterologous expression and maturation of C. reinhardtii hydrogenase has been developed. HydA1 of C. reinhardtii was purified and shown to contain 6 Fe atoms/molecule of protein, as expected. Using DMSO, TMAO or thiosulfate as substrates for anaerobic respiration during the cell growth, 0.4 - 0.5 mg l(-1)(OD600 = 1) of catalytically active HydA1 was obtained with hydrogen evolution rate of approximately 700 micromol H2 mg(-1) min(-1).


The maximal amount of dietary alpha-tocopherol intake in U.S. adults (NHANES 2001-2002).
The current study was designed to determine the maximal amount of alpha-tocopherol intake obtained from food in the U.S. diet, and to examine the effect of different food group intakes on this amount. Data from 2138 men and 2213 women aged >18 y were obtained from the National Health and Nutrition Examination Survey (NHANES) 2001-2002. Linear programming was used to generate diets with maximal alpha-tocopherol intake, with the conditions of meeting the recommended daily allowances or adequate intakes for a set of nutrients, sodium and fat recommendations, and energy limits, and that were compatible with the observed dietary patterns in the population. With food use and energy constraints in models, diets formulated by linear programming provided 19.3-24.9 mg alpha-tocopherol for men and women aged 19-50 or >50 y. These amounts decreased to 15.4-19.9 mg with the addition of the sodium, dietary reference intake, and fat constraints. The relations between maximal alpha-tocopherol intake and food group intakes were influenced by total fat restrictions. Although meeting current recommendations (15 mg/d) appears feasible for individuals, dramatic dietary changes that include greater intakes of nuts and seeds, and fruit and vegetables, are needed. Careful selection of the highest vitamin E source foods within these groups could further increase the likelihood of meeting the current recommended daily allowance.


Prognostic factors for the outcome of surgical and conservative treatment of symptomatic spinal cord cavernous malformations: a review of a series of 20 patients.
In this study, the authors present a review of a series of 20 intramedullary spinal cord cavernous malformations (SCCMs) with particular focus on MR imaging and prognostic factors. Between 1994 and 2009, 20 patients with SCCM were treated under the care of the senior author. The diagnosis was made in all patients after the onset of clinical symptoms. The age of the 9 men and 11 women ranged between 26 and 71 years (median 38.5 years). The duration of symptoms prior to referral ranged from 1 week to 9 years (median 6.5 months). At the time of referral, 4 patients had no significant neurological deficits, 10 patients suffered significant functional restrictions, and 6 patients presented with severe paraparesis and loss of functional strength. None of the patients had complete paraplegia. Seventeen patients underwent microsurgical removal, while 3 patients opted for conservative therapy. For the present analysis, the medical records and MR images and/or reports were reviewed. Classification of length of history, pretreatment status, MR imaging pattern, and treatment modality was done and correlated with outcome. The cavernoma was located at the cervical level in 8 patients and between T-1 and L-1 in 12 patients. The cavernoma appeared as mainly T2 hyperintense on MR images in 7 patients, mainly T2 hypointense in 2 patients, and mixed in the remaining 10 patients. The craniocaudal extension of the core varied between 5 and 45 mm. In 2 patients with cervical cavernomas, a distinct T2 signal of the spinal cord cranial and distal to the cavernoma was seen, and in a patient with a large thoracic cavernoma, T2 extinction cranial and caudal to the cavernoma was seen as a sign of hemosiderosis. Neurological deficits improved postoperatively in 12 of the surgically treated patients, remained stable in 2, and deteriorated in 3. The 3 patients who were conservatively treated remained stable over a follow-up of 3-9 years. Postoperative improvement was seen in 5 of 7 surgical patients with a history of symptoms of 2 months or less, 5 of 6 patients with a history of 2-24 months, and in 2 of 4 patients with a history of more than 2 years. Two of the 3 patients with postoperative deterioration had a history of more than 2 years and the third a short history of 1 month. Although a satisfactory outcome can be achieved through surgical treatment of SCCMs, some patients worsen after surgery or during the postoperative course. Long-term stability is possible in oligosymptomatic conservatively treated patients. The prevalence and pathophysiological importance of segmental spinal cord edema and hemosiderosis is incompletely understood at the present time.


Why the resistance to statistical innovations? Bridging the communication gap.
While quantitative methodologists advance statistical theory and refine statistical methods, substantive researchers resist adopting many of these statistical innovations. Traditional explanations for this resistance are reviewed, specifically a lack of awareness of statistical developments, the failure of journal editors to mandate change, publish or perish pressures, the unavailability of user friendly software, inadequate education in statistics, and psychological factors. Resistance is reconsidered in light of the complexity of modern statistical methods and a communication gap between substantive researchers and quantitative methodologists. The concept of a Maven is introduced as a means to bridge the communication gap. On the basis of this review and reconsideration, recommendations are made to improve communication of statistical innovations.


Prognostic variables in patients with diffuse large-cell lymphoma treated with MACOP-B.
One hundred twenty-six patients with diffuse large-cell lymphoma were treated with methotrexate with leucovorin, doxorubicin, cyclophosphamide, vincristine, prednisone, and bleomycin (MACOP-B) between April 1981 and June 1986. Univariate and multivariate analyses were performed using overall survival as of September 1989 as the end point. Four independent negative predictors of survival were identified: presence of B symptoms; more than two involved lymph node sites; more than one extranodal site (variables related to tumor burden), and age older than 60, a variable related to the patient's ability to tolerate treatment. Each variable contributed the same relative risk of dying and, accordingly, this simple predictive formula was developed empirically: (4-N) x 30 = the approximate percentage of chance of survival at 5 years. "N" is the number of predictive variables present. The same four predictors were also found to be significant by multivariate analysis when only those patients achieving a complete response were analyzed.


Analysis of historical epidemiological data to guide intervention policy in the face of an infectious disease outbreak: poliomyelitis in Israel in 1988.
During the 1988 outbreak of 15 cases of Type 1 poliomyelitis mainly among previously vaccinated young adults in Israel, there were conflicting views on the need for and timing of mass vaccination. Since there was a possibility that the initially localized outbreak may spread more widely, it was felt that an early decision on vaccination policy should be made. Public health officials were faced with varied interpretations of the existing data on the extent of the immune status of the population; some indicating that as many as 15% of young adults lacked immunity to the Type 1 virus. Two major questions posed were whether or not the outbreak was a purely localized event in a single subdistrict, and whether the seasonality of the disease would limit its spread. Based on historical data, probability analysis of three cases occurring outside the subdistrict at the time of the outbreak suggested more widespread dispersion of the virus. Further analysis indicated very limited seasonality of the disease in this region, suggesting that the onset of colder weather would have little impact on the spread of the virus. Careful analysis of the local epidemiology of the disease is an important tool for guiding urgent decisions on mass intervention policy in outbreak situations.


Speech recognition for the disabled.
A simple and inexpensive system for telephone communication that includes a modular speech recognition device developed for use by a disabled person is presented. User considerations are reviewed, and hardware details are given. The complete automatic speech recognition (ASR) system consists of five main blocks: a TCM29C18 codec (analog interface for the digital signal processor), an SRS-1 speech recognizer, an LED display, a dual-tone multifrequency generator, and tone enable plus data input circuitry. Results of a semiqualitative assessment in terms of recognition success for the limited vocabulary of numbers and commands from the user are reported. The results are described in chronological order, beginning with testing of the SRS-1 and the codec working as a stand-alone recognizer system, and leading to the testing of the recognition system combined with the tone generator. Finally, results were obtained when the whole ASR system was interfaced to the telephone line. Some further work is suggested.


Was low CO2 a driving force of C4 evolution: Arabidopsis responses to long-term low CO2 stress.
The responses of long-term growth of plants under elevated CO2 have been studied extensively. Comparatively, the responses of plants to subambient CO2 concentrations have not been well studied. This study aims to investigate the responses of the model C3 plant, Arabidopsis thaliana, to low CO2 at the molecular level. Results showed that low CO2 dramatically decreased biomass productivity, together with delayed flowering and increased stomatal density. Furthermore, alteration of thylakoid stacking in both bundle sheath and mesophyll cells, upregulation of PEPC and PEPC-K together with altered expression of a number of regulators known involved in photosynthesis development were observed. These responses to low CO2 are discussed with regard to the fitness of C3 plants under low CO2. This work also briefly discusses the relevance of the data to C4 photosynthesis evolution.


Fusion of white and gray matter geometry: a framework for investigating brain development.
Current neuroimaging investigation of the white matter typically focuses on measurements derived from diffusion tensor imaging, such as fractional anisotropy (FA). In contrast, imaging studies of the gray matter oftentimes focus on morphological features such as cortical thickness, folding and surface curvature. As a result, it is not clear how to combine findings from these two types of approaches in order to obtain a consistent picture of morphological changes in both gray and white matter. In this paper, we propose a joint investigation of gray and white matter morphology by combining geometrical information from white and the gray matter. To achieve this, we first introduce a novel method for computing multi-scale white matter tract geometry. Its formulation is based on the differential geometry of curve sets and is easily incorporated into a continuous scale-space framework. We then incorporate this method into a novel framework for "fusing" white and gray matter geometrical information. Given a set of fiber tracts originating in a particular cortical region, the key idea is to compute two scalar fields that represent geometrical characteristics of the white matter and of the surface of the cortical region. A quantitative marker is created by combining the distributions of these scalar values using Mutual Information. This marker can be then used in the study of normal and pathological brain structure and development. We apply this framework to a study on autism spectrum disorder in children. Our preliminary results support the view that autism may be characterized by early brain overgrowth, followed by reduced or arrested growth (Courchesne, 2004).


Engineering a cyanobacterium as the catalyst for the photosynthetic conversion of CO2 to 1,2-propanediol.
The modern society primarily relies on petroleum and natural gas for the production of fuels and chemicals. One of the major commodity chemicals 1,2-propanediol (1,2-PDO), which has an annual production of more than 0.5 million tons in the United States, is currently produced by chemical processes from petroleum derived propylene oxide, which is energy intensive and not sustainable. In this study, we sought to achieve photosynthetic production of 1,2-PDO from CO2 using a genetically engineered cyanobacterium Synechococcus elongatus PCC 7942. Compared to the previously reported biological 1,2-PDO production processes which used sugar or glycerol as the substrates, direct chemical production from CO2 in photosynthetic organisms recycles the atmospheric CO2 and will not compete with food crops for arable land. In this study, we reported photosynthetic production of 1,2-PDO from CO2 using a genetically engineered cyanobacterium Synechococcus elongatus PCC 7942. Introduction of the genes encoding methylglyoxal synthase (mgsA), glycerol dehydrogenase (gldA), and aldehyde reductase (yqhD) resulted in the production of ~22 mg/L 1,2-PDO from CO2. However, a comparable amount of the pathway intermediate acetol was also produced, especially during the stationary phase. The production of 1,2-PDO requires a robust input of reducing equivalents from cellular metabolism. To take advantage of cyanobacteria's NADPH pool, the synthetic pathway of 1,2-PDO was engineered to be NADPH-dependent by exploiting the NADPH-specific secondary alcohol dehydrogenases which have not been reported for 1,2-PDO production previously. This optimization strategy resulted in the production of ~150 mg/L 1,2-PDO and minimized the accumulation of the incomplete reduction product, acetol. This work demonstrated that cyanobacteria can be engineered as a catalyst for the photosynthetic conversion of CO2 to 1,2-PDO. This work also characterized two NADPH-dependent sADHs for their catalytic capacity in 1,2-PDO formation, and suggested that they may be useful tools for renewable production of reduced chemicals in photosynthetic organisms.


The Raman spectrum of CaCO3 polymorphs calcite and aragonite: a combined experimental and computational study.
Powder and single crystal Raman spectra of the two most common phases of calcium carbonate are calculated with ab initio techniques (using a "hybrid" functional and a Gaussian-type basis set) and measured both at 80 K and room temperature. Frequencies of the Raman modes are in very good agreement between calculations and experiments: the mean absolute deviation at 80 K is 4 and 8 cm(-1) for calcite and aragonite, respectively. As regards intensities, the agreement is in general good, although the computed values overestimate the measured ones in many cases. The combined analysis permits to identify almost all the fundamental experimental Raman peaks of the two compounds, with the exception of either modes with zero computed intensity or modes overlapping with more intense peaks. Additional peaks have been identified in both calcite and aragonite, which have been assigned to (18)O satellite modes or overtones. The agreement between the computed and measured spectra is quite satisfactory; in particular, simulation permits to clearly distinguish between calcite and aragonite in the case of powder spectra, and among different polarization directions of each compound in the case of single crystal spectra.


Development of immunogenic recombinant Oka varicella vaccine expressing hepatitis B virus surface antigen.
Recombinant Oka varicella vaccine expressing hepatitis B virus (HBV) surface antigen (HBs) was constructed by inserting the HBs gene into the viral thymidine kinase (TK) gene and was examined for its immunogenicity in guinea-pigs. The HBs gene encoding 25 amino acids of preS2 and the whole of the S region was inserted into the TK gene of the cloned plasmid. The chimeric plasmid DNA and Oka varicella vaccine DNA were cotransfected and recombinant virus was isolated after immunofluorescence screening using a monoclonal antibody to HBs and a fluorescein-conjugated anti-mouse antibody. Expression of viral HBs was detected in the cytoplasm of infected cells and was stable over several repeated passages in vitro. The recombinant virus expressed 26K and 30K HBs molecules in infected cells and the culture supernatant contained 30K and 35K HBs molecules. HBs was purified at a density of 1.20 g/ml from the culture supernatants. The recombinant virus induced an antibody response to HBs as well as to varicella-zoster virus (VZV) in guinea-pigs, and the antibody titre to HBs was comparable to that induced by a recombinant HBs subunit vaccine produced in yeast. Thus a single dose of live recombinant Oka varicella vaccine could induce good immunity to VZV and HBs. The recombinant Oka varicella vaccine expressing HBs may be a good candidate for a combined HBV and VZV vaccine.


Expression of p210 and p190 BCR-ABL due to alternative splicing in chronic myelogenous leukaemia.
The hallmark of chronic myelogenous leukaemia (CML) is the presence of the Philadelphia chromosome and its resultant fusion message, BCR-ABL, and fusion protein, p210. Patients with CML in blast crisis, or with Philadelphia positive acute lymphoblastic leukaemia (ALL), can have a smaller BCR-ABL fusion transcript possessing only the first exon of BCR fused to ABL. This smaller transcript encodes a 190 kD protein which is more strongly transforming than the p210 protein derived from the larger CML-associated transcript. We performed RT-PCR on samples from CML patients in chronic phase to determine the frequency and mechanism of p190 and p210 co-expression and to see if this correlated with clinical indices. We examined the peripheral blood or marrow of 67 patients with CML and found that 35 of them expressed both transcripts whereas the remainder expressed the p210-encoding transcript exclusively. Additional PCR products of an intermediate size were also frequently detected and have been isolated and sequenced. Data from two of these products indicate that they are the result of alternative splicing and include variable combinations of BCR exons. We believe that the expression of the p190-encoding transcript in the chronic phase of CML is also due to alternative splicing. A comparison of patients co-expressing the p190- and p210-encoding transcripts with those patients who expressed only the p210-encoding transcript detected significantly higher white blood cell (WBC) counts and blast cell counts at time of testing as well as significantly higher white blood cell counts at diagnosis.


Anisotropically Conductive Biodegradable Scaffold with Coaxially Aligned Carbon Nanotubes for Directional Regeneration of Peripheral Nerves.
Fascicular rearrangement of an injured peripheral nerve requires reconnection of nerve sprouts from anterior and Büngner bands from distal sides of the lesion, failing to which leads to inefficient regeneration of the injured nerve. However, existing neural scaffolds have limited neuroregeneration efficiency because of either the lack of alignment of fibers and a conductive second phase, leading to compromised electrical conductivity, or the lack of extracellular matrix components and in vivo validation. The present study reports a biocompatible, multiwall carbon nanotube (MWCNT)-reinforced, anisotropically conductive, electrospun, aligned nanofibrous scaffold, ensuring maximal peripheral nerve regeneration. Electrospinning parameters were modulated to deposit random and parallel fibers in separate scaffolds for comparative analysis on the effect of fiber alignment on regeneration. Both types of scaffolds were reinforced with MWCNTs to impart electrical conductivity. Nonreinforced scaffolds were nonconductive. In this comparative study, MWCNT-reinforced, aligned scaffolds showed better tensile property with increased conductivity along the direction of alignment, thereby ensuring an escalated neural-regeneration rate. Collectively, in vitro studies established the scaffolds to be highly biocompatible, promoting cell growth and proliferation. With 85% more anisotropic conductivity in the direction of the alignment and the degradation kinetics tuned to the regeneration regime, the MWCNT-reinforced, aligned scaffold efficiently healed injured sciatic nerves in rats within 30 days. Rigorous revivification of the tissue was due to coordinated Wallerian degeneration and expedited guided axonal regeneration. Structural and functional analysis of nerves in vivo showed the aligned, MWCNT-reinforced scaffold to be very efficient in peripheral sciatic nerve regeneration. This study notes the efficacy of the coaxially aligned, MWCNT-reinforced neural scaffold, with a capability of establishing remarkable advancement in the field of peripheral neural regeneration.


A randomised trial of pre-exercise meal composition on performance and muscle damage in well-trained basketball players.
Attenuating muscle damage is important to subsequent sports performance. It is possible that pre-exercise protein intake could influence markers of muscle damage and benefit performance, however, published research provides conflicting results. At present no study has investigated protein and carbohydrate (PRO/CHO) co-ingestion solely pre-exercise, nor prior to basketball-specific exercise. The purpose of this study was to answer the research question; would pre-exercise protein intake enhance performance or attenuate muscle damage during a basketball simulation test? Ten well-trained male basketball players consumed either carbohydrate (1 g · kg(-1) body mass) with protein (1 g · kg(-1) body mass), or carbohydrate alone (2 g · kg(-1) body mass) in a randomised cross- over design, 90 minutes before completing an 87-minute exercise protocol. The rise in creatine kinase (CK) from baseline to post-exercise was attenuated following PRO/CHO (56 ± 13U · L(-1)) compared to carbohydrate (100 ± 10 U · L(-1)), (p = 0.018). Blood glucose was also higher during and post-exercise following PRO/CHO (p < 0.050), as was free throw shooting accuracy in the fourth quarter (p = 0.027). Nausea during (p = 0.007) and post-(p = 0.039) exercise increased following PRO/CHO, as did cortisol post-exercise (p = 0.038). Results suggest that in well-trained basketball players, pre-exercise PRO/CHO may attenuate the rise in CK, indicative of a decrease in muscle damage during exercise. However, unfamiliarity with the protein amount provided may have increased nausea during exercise, and this may have limited the ability to see an improvement in more performance measures.


A surgical technique for hip disarticulation.
Hip disarticulation is usually elected for malignant bony and soft tissue tumors below the lesser trochanter of the femur. The operation is performed with the patient in a posterolateral position; in the first phase of the procedure the surgeon stands anterior to the patient. After incision of the skin and division of the femoral vessels and nerve, muscles of the anterior thigh are transected off the pelvic bone from lateral to medial starting with the sartorius and finishing with the adductor magnus. Muscles are divided at their origin except for the iliopsoas and obturator externus which are divided at their insertion on the lesser trochanter of the femur. The quadratus femoris muscle is identified and preserved, then the flexor muscles are transected at their site of origin from the ischial tuberosity. During the next phase the surgeon is posterior to the patient, and the pelvis is rotated from the posterolateral to the anterolateral position. After completion of the skin incision, the gluteal fascia, tensor fascia lata, and the gluteus maximus muscles are divided and dissected free of their posterior attachments to expose the muscles inserting by way of a common tendon onto the greater trochanter. These muscles are then transected at their insertion on the bone. The posterior aspect of the joint capsule is then exposed and transected. Finally, the sciatic nerve is divided and allowed to retract beneath the piriformis muscle. To close the wound the preserved muscles are approximated over the joint capsule and the gluteal fascia secured to the inguinal ligament over suction drains. The skin is closed with interrupted sutures.


Gastric infarction after therapeutic embolization.
A case of extensive gastric necrosis after therapeutic transcatheter embolization of the left gastric artery with fragments of gelatin sponge for recurrent massive upper gastrointestinal hemorrhage is reported. Although future modifications in technique and in choice of embolic agents undoubtedly will be forthcoming, postembolic ischemic necrosis may prove to be a major limitation of this technique. Until more widespread experience is accumulated and the frequency of postembolic ischemic necrosis is known, it would seem prudent to reserve this procedure for those patients presenting prohibitive surgical risks.


Gating of Na channels. Inactivation modifiers discriminate among models.
Macroscopic Na currents were recorded from N18 neuroblastoma cells by the whole-cell voltage-clamp technique. Inactivation of the Na currents was removed by intracellular application of proteolytic enzymes, trypsin, alpha-chymotrypsin, papain, or ficin, or bath application of N-bromoacetamide. Unlike what has been reported in squid giant axons and frog skeletal muscle fibers, these treatments often increased Na currents at all test pulse potentials. In addition, removal of inactivation gating shifted the midpoint of the peak Na conductance-voltage curve in the negative direction by 26 mV on average and greatly prolonged the rising phase of Na currents for small depolarizations. Polypeptide toxins from Leiurus quinquestriatus scorpion and Goniopora coral, which slow inactivation in adult nerve and muscle cells, also increase the peak Na conductance and shift the peak conductance curve in the negative direction by 7-10 mV in neuroblastoma cells. Control experiments argue against ascribing the shifts to series resistance artifacts or to spontaneous changes of the voltage dependence of Na channel kinetics. The negative shift of the peak conductance curve, the increase of peak Na currents, and the prolongation of the rise at small depolarization after removal of inactivation are consistent with gating kinetic models for neuroblastoma cell Na channels, where inactivation follows nearly irreversible activation with a relatively high, voltage-independent rate constant and Na channels open only once in a depolarization. As the same kind of experiment does not give apparent shifting of activation and prolongation of the rising phase of Na currents in adult axon and muscle membranes, the Na channels of these other membranes probably open more than once in a depolarization.


Feeding and digestion of the marine isopod Idotea emarginata challenged by poor food quality and microplastics.
Ingestion of microplastics can impair nutrition of marine invertebrates. In a laboratory study, we tested whether microplastics affect ingestion rates and gastrointestinal enzyme activities in the marine isopod Idotea emarginata. Isopods were fed for eight days with one out of four different food formulations: natural food (the brown alga Fucus vesiculosus) or synthetic diet consisting of freeze-dried algal powder embedded in agarose, both, with or without microplastic particles (fluorescent polymethyl methacrylate, 10-100 μm) at a concentration of 40 items per mg of food. The isopods accepted both types of food but consumed significantly more (average 3.1-fold) of the agar based synthetic food. I. emarginata responded to the reduced content of digestible organic matter in the synthetic food by a compensatory adjustment of the ingestion rates. Addition of microplastics had no effect on ingestion rates in natural food whereas the feeding rates for synthetic food varied in response to microplastics. Similarly, activity patterns of digestive enzymes, particularly those of esterases, changed significantly in the treatment with synthetic food. Isopods fed with synthetic food alone showed elevated esterase activities in the gut while those isopods fed with synthetic food and microplastics showed elevated esterase activities in the midgut gland but not in the gut. Apparently, not the exposure to microplastic alone, but the combined effects of reduced nutrient availability and microplastic ingestion caused considerable biochemical reactions in the digestive organs of the isopods.


Glomerular basement membrane discontinuities. Scanning electron microscopic study of acellular glomeruli.
Glomerulonephritides which develop necrotizing and crescentic lesions usually have glomerular basement membrane (GBM) disruptions when carefully examined by light microscopy or transmission electron microscopy. Despite numerous excellent and detailed ultrastructural investigations of GBM discontinuities, a complete appreciation of their actual number, appearance, and distribution within a glomerulus has been difficult to achieve by reconstruction of two-dimensional light or transmission electron microscopic images. Selective removal of podocytes by a sequence of lytic and solubilization procedures has been developed which exposes any structural alteration of the GBM to direct examination by scanning electron microscopy. A case of idiopathic, immune-complex-negative, focal-segmental necrotizing glomerulonephritis has been studied by this technique, permitting three-dimensional visualization of the GBM defects which result in free communication between the vascular and urinary spaces. These disruptions were distinctive by their frequency within an affected lobule, variable size, and sharply demarcated edges. Application of this technique to human renal biopsies is capable of enhancing our understanding of the morphologic alterations occurring in human glomerulonephritis.


The effect of a transforming care initiative on patient outcomes in acute surgical units: a time series study.
To examine the relationship between the implementation of a transforming care initiative and two patient outcomes, inpatient falls and hospital acquired pressure ulcers. Patients continue to experience harm in hospitals from adverse events such as inpatient falls and hospital acquired pressure ulcers. These and other patient safety concerns led to transforming care initiatives. This cohort study used historical controls and a time series design. The setting was two surgical units of an acute care hospital; 7125 patients discharged from these units between July 2008-December 2010 were included. Those patients discharged during the initial 3 months of implementation were excluded. Several interventions were adopted as part of the transforming care initiative such as bedside handover and bedside whiteboards. Coded administrative data were accessed to identify patients who had experienced a fall or acquired a pressure ulcer during their hospital stay. Statistical process control was used to identify changes in outcomes over time. The findings demonstrated variation in the proportion of patients experiencing a fall in both units and for the proportion of patients acquiring a pressure ulcer in one unit, following implementation. These results demonstrate that implementing a transforming care initiative may have some influence on the quality and safety of patient care as measured by patient falls and pressure ulcers but findings varied. Further research is required to gain an understanding of the inconsistency of the impact of these initiatives across all clinical contexts.


Highly scalable, atomically thin WSe2 grown via metal-organic chemical vapor deposition.
Tungsten diselenide (WSe2) is a two-dimensional material that is of interest for next-generation electronic and optoelectronic devices due to its direct bandgap of 1.65 eV in the monolayer form and excellent transport properties. However, technologies based on this 2D material cannot be realized without a scalable synthesis process. Here, we demonstrate the first scalable synthesis of large-area, mono and few-layer WSe2 via metal-organic chemical vapor deposition using tungsten hexacarbonyl (W(CO)6) and dimethylselenium ((CH3)2Se). In addition to being intrinsically scalable, this technique allows for the precise control of the vapor-phase chemistry, which is unobtainable using more traditional oxide vaporization routes. We show that temperature, pressure, Se:W ratio, and substrate choice have a strong impact on the ensuing atomic layer structure, with optimized conditions yielding >8 μm size domains. Raman spectroscopy, atomic force microscopy (AFM), and cross-sectional transmission electron microscopy (TEM) confirm crystalline monoto-multilayer WSe2 is achievable. Finally, TEM and vertical current/voltage transport provide evidence that a pristine van der Waals gap exists in WSe2/graphene heterostructures.


Immunology of multiple sclerosis.
Multiple sclerosis (MS) develops in young adults with a complex predisposing genetic trait and probably requires an inciting environmental insult such as a viral infection to trigger the disease. The activation of CD4+ autoreactive T cells and their differentiation into a Th1 phenotype are a crucial events in the initial steps, and these cells are probably also important players in the long-term evolution of the disease. Damage of the target tissue, the central nervous system, is, however, most likely mediated by other components of the immune system, such as antibodies, complement, CD8+ T cells, and factors produced by innate immune cells. Perturbations in immunomodulatory networks that include Th2 cells, regulatory CD4+ T cells, NK cells, and others may in part be responsible for the relapsing-remitting or chronic progressive nature of the disease. However, an important paradigmatic shift in the study of MS has occurred in the past decade. It is now clear that MS is not just a disease of the immune system, but that factors contributed by the central nervous system are equally important and must be considered in the future.


Iodine-125 plaque radiotherapy of choroidal hemangioma.
Circumscribed choroidal hemangioma (CCH) is a benign vascular tumor that is not associated with systemic abnormalities and its pathogenesis is unknown. Most choroidal hemangiomas remain stable in size, and treatment is periodic observation unless complicated by an overlying serous retinal detachment. A 43-year-old white man came to our clinic reporting decreased vision in the left eye for 1 year. On funduscopic evaluation, a large elevated nonpigmented red-orange lesion, just temporal to the fovea of the left eye, was observed with surrounding areas of subretinal fluid. On the basis of findings obtained through ophthalmoscopy, fluorescein angiography, and ultrasonography, the patient was diagnosed with a circumscribed choroidal hemangioma. Treatment of the CCH was with iodine-125 plaque radiotherapy, which significantly reduced the tumor size and improved vision. CCH is a benign vascular tumor that is characteristically nonprogressive, but in cases of vision loss caused by serous detachment of the fovea, argon laser photocoagulation or radiotherapy is the treatment of choice. Recent studies indicate that iodine-125 plaque radiotherapy is indicated for large choroidal hemangiomas involving the macula or with associated retinal detachment. The main goal of treating CCH is to preserve vision and to prevent the progression of vision loss.


Cerebral mitochondrial metabolism in early Parkinson's disease.
Abnormal cerebral energy metabolism owing to dysfunction of mitochondrial electron transport has been implicated in the pathogenesis of Parkinson's disease (PD). However, in vivo data of mitochondrial dysfunction have been inconsistent. We directly investigated mitochondrial oxidative metabolism in vivo in 12 patients with early, never-medicated PD and 12 age-matched normal controls by combined measurements of the cerebral metabolic rate of oxygen (CMRO(2)) and the cerebral metabolic rate of glucose (CMRglc) with positron emission tomography. The primary analysis showed a statistically significant 24% increase in bihemispheric CMRO(2) and no change in CMRO(2)/CMRglc. These findings are inconsistent with a defect in mitochondrial oxidative phosphorylation owing to reduced activity of the mitochondrial electron transport system (ETS). Because PD symptoms were already manifest, deficient energy production owing to a reduced activity of the mitochondrial ETS cannot be a primary mechanism of neuronal death in early PD. Alternatively, this general increase in CMRO(2) could be due not to an increased metabolic demand but to an uncoupling of ATP production from oxidation in the terminal stage of oxidative phosphorylation. Whether this is the case in early PD and whether it is important in the pathogenesis of PD will require further study.


Tradeoffs of managing cod as a sustainable resource in fluctuating environments.
Sustainable human exploitation of living marine resources stems from a delicate balance between yield stability and population persistence to achieve socioeconomic and conservation goals. But our imperfect knowledge of how oceanic oscillations regulate temporal variation in an exploited species can obscure the risk of missing management targets. We illustrate how applying a management policy to suppress fluctuations in fishery yield in variable environments (prey density and regional climate) can present unintended outcomes in harvested predators and the sustainability of harvesting. Using Atlantic cod (Gadus morhua, an apex predatory fish) in the Barents Sea as a case study we simulate age-structured population and harvest dynamics through time-varying, density-dependent and density-independent processes with a stochastic, process-based model informed by 27-year monitoring data. In this model, capelin (Mallotus villosus, a pelagic forage fish), a primary prey of cod, fluctuations modulate the strength of density-dependent regulation primarily through cannibalistic pressure on juvenile cod survival; sea temperature fluctuations modulate thermal regulation of cod feeding, growth, maturation, and reproduction. We first explore how capelin and temperature fluctuations filtered through cod intrinsic dynamics modify catch stability and then evaluate how management to suppress short-term variability in catch targets alters overharvest risk. Analyses revealed that suppressing year-to-year catch variability impedes management responses to adjust fishing pressure, which becomes progressively out of sync with variations in cod abundance. This asynchrony becomes amplified in fluctuating environments, magnifying the amplitudes of both fishing pressure and cod abundance and then intensifying the density-dependent regulation of juvenile survival through cannibalism. Although these transient dynamics theoretically give higher average catches, emergent, quasicyclic behaviors of the population would increase long-term yield variability and elevate overharvest risk. Management strategies that overlook the interplay of extrinsic (fishing and environment) and intrinsic (life history and demography) fluctuations thus can inadvertently destabilize fish stocks, thereby jeopardizing the sustainability of harvesting. These policy implications underscore the value of ecosystem approaches to designing management measures to sustainably harvest ecologically connected resources while achieving socioeconomic security.


Connecting the Micro to the Macro: An Exploration of Micro-Behaviors of Individuals Who Drive CSR Initiatives at the Macro-Level.
Grounded on a case study on the formation of an inter-corporate CSR initiative in which four corporations from Chennai, India collaborate, this paper explores the micro-behaviors that individual actors engage in to create CSR solutions later adopted at the macro-organizational level. Based on the findings, the paper (1) identifies five categories of micro-behaviors, namely increasing stakeholder salience by turning attention to the ethical and social responsibilities to specific stakeholder groups, emerging as a self-appointed CSR champion by assuming personal responsibility for action, creating CSR initiative prototypes by leveraging personal skills, garnering support by leveraging personal networks and amassing operational resources by organizational resources; (2) explicates the characteristics of individual approach to CSR that makes it different from, but complementary to organizational approach to CSR.


Predicting Agent Behaviour and State for Applications in a Roundabout-Scenario Autonomous Driving.
As human drivers, we instinctively employ our understanding of other road users' behaviour for enhanced efficiency of our drive and safety of the traffic. In recent years, different aspects of assisted and autonomous driving have gotten a lot of attention from the research and industrial community, including the aspects of behaviour modelling and prediction of future state. In this paper, we address the problem of modelling and predicting agent behaviour and state in a roundabout traffic scenario. We present three ways of modelling traffic in a roundabout based on: (i) the roundabout geometry; (ii) mean path taken by vehicles inside the roundabout; and (iii) a set of reference trajectories traversed by vehicles inside the roundabout. The roundabout models are compared in terms of exit-direction classification and state (i.e., position inside the roundabout) prediction of query vehicles inside the roundabout. The exit-direction classification and state prediction are based on a particle-filter classifier algorithm. The results show that the roundabout model based on set of reference trajectories is better suited for both the exit-direction and state prediction.


[Green coffee bean allergy and ultrafine particles exposure in Trieste dock workers].
The paper reviews allergy to green coffee bean and castor bean in dock workers and in coffee processing workers from '80 to nowadays in Trieste (NE of Italy). The avoidance of use of jute sacks contaminated with castor bean caused a decrease in sensitization to castor bean and the better work practices to handle jute sacks permitted to reduce airborne exposure to green coffee been powders, that resulted below occupational exposure limits. However, the measurement of ultrafine particles emitted during the handling of sacks showed exposure to high level of particles below 40 nm and permitted to identify some work tasks that can cause a more elevated exposure. Moreover, some sacks, coming from Tanzania, are still contaminated with castor bean, causing mild allergic symptoms. The work condition in dock workers in Trieste improved in years with a reduction of exposure to these allergens. However, the adoption of protective measures as well as periodical medical surveillance are needed to prevent sensitization or to detect the early onset of new cases.


pH-Controlled Resettable Modular DNA Strand-Displacement Circuits.
Sophisticated dynamic molecular systems with diverse functions have been fabricated by using the fundamental tool of toehold-mediated strand displacement (TMSD) in the field of dynamic DNA nanotechnology. However, simple approaches to reset these TMSD-based dynamic systems are lacking due to the difficulty in creating kinetically favored pathways to implement the backward resetting reactions. Here, we develop a facile proton-driven strategy to achieve complete resetting of a modular DNA circuit by integrating a pH-responsive intermolecular CG-C+ triplex DNA and an i-motif DNA into the conventional DNA substrate. The pH-programmed strategy allows modular DNA components to specifically associate/dissociate to promote the forward/backward TMSD reactions, thereby enabling the modular DNA circuit to be repeatedly operated at a constant temperature without generating any DNA waste products. Leveraging this tractable approach, we further constructed two resettable DNA logic gates used for logical computation and two resettable catalytic DNA systems with good performance in signal transduction and amplification.


Genome-wide analysis of ionotropic receptors provides insight into their evolution in Heliconius butterflies.
In a world of chemical cues, smell and taste are essential senses for survival. Here we focused on Heliconius, a diverse group of butterflies that exhibit variation in pre- and post-zygotic isolation and chemically-mediated behaviors across their phylogeny. Our study examined the ionotropic receptors, a recently discovered class of receptors that are some of the most ancient chemical receptors. We found more ionotropic receptors in Heliconius (31) than in Bombyx mori (25) or in Danaus plexippus (27). Sixteen genes in Lepidoptera were not present in Diptera. Only IR7d4 was exclusively found in butterflies and two expansions of IR60a were exclusive to Heliconius. A genome-wide comparison between 11 Heliconius species revealed instances of pseudogenization, gene gain, and signatures of positive selection across the phylogeny. IR60a2b and IR60a2d are unique to the H. melpomene, H. cydno, and H. timareta clade, a group where chemosensing is likely involved in pre-zygotic isolation. IR60a2b also displayed copy number variations (CNVs) in distinct populations of H. melpomene and was the only gene significantly higher expressed in legs and mouthparts than in antennae, which suggests a gustatory function. dN/dS analysis suggests more frequent positive selection in some intronless IR genes and in particular in the sara/sapho and melpomene/cydno/timareta clades. IR60a1 was the only gene with an elevated dN/dS along a major phylogenetic branch associated with pupal mating. Only IR93a was differentially expressed between sexes. All together these data make Heliconius butterflies one of the very few insects outside Drosophila where IRs have been characterized in detail. Our work outlines a dynamic pattern of IR gene evolution throughout the Heliconius radiation which could be the result of selective pressure to find potential mates or host-plants.


Effects of computerized provider order entry and nursing documentation on workflow.
The objective was to measure the effects of the implementation of computerized provider order entry (CPOE) and electronic nursing documentation on provider workflow in the emergency department (ED). The authors performed a before-and-after time-motion study of the activities of physicians and nurses. The percentages of time spent in task categories were calculated by provider session and averaged across provider sessions. There was a shift in physician time from working with paper alone, 13.1% to 9.6% (p = 0.05), to working with paper while using a computer, 1.6% to 4.3% (p = 0.02), and an increase in time spent working on computer and/or paper from 30.0% to 38.9% (p = 0.02). For nurses, the increase in time spent on computer from 9.5% to 25.7% (p < 0.01) was offset by a decrease in time spent working with paper from 16.5% to 1.8% (p < 0.01). Direct care decreased minimally for nurses from 56.9% to 55.3% (p = 0.69), but from 36.8% to 29.1% (p = 0.07) for physicians, approaching statistical significance. Care planning decreased for nurses from 9.4% to 6.4% (p = 0.04) and from 21.7% to 19.5% (p = 0.60) for physicians. The net effects of an implementation on provider workflow depend not only on changes in tasks directly related to the provider-computer interface, but also on changes in underlying patient care processes and information flows. The authors observed an unanticipated shift in physician time from interacting with nurses and patients toward retrieving information from the electronic patient record. Implementers should carefully consider how implementations will affect information flow and then expect the unexpected.


AIM2 inhibits colorectal cancer cell proliferation and migration through suppression of Gli1.
Colorectal cancer (CRC) is a common malignant tumor and is one of the leading causes of cancer-related deaths worldwide. Absent in melanoma 2 (AIM2), as a member of the pyrin-HIN family proteins, plays contentious roles in different types of cancers. In the present work, we provide evidence that AIM2 was commonly downregulated in human CRC and loss of AIM2 significantly correlated with tumor size, depth of invasion, lymph node metastasis (LNM) and TNM (Tumor, Node, Metastases) stage in patients suffering from CRC. AIM2 knockdown promoted CRC cell proliferation, migration and epithelial-mesenchymal transition (EMT) progress, whereas AIM2 overexpression did the opposite. AIM2 inhibited glioma-associated oncogene-1 (Gli1) expression through Smoothened homolog (SMO)-independent pathway and regulated CRC cell proliferation and migration in a Gli1-dependent manner. Moreover, AIM2 could modulate Protein kinase B (AKT)/mechanistic target of rapamycin (mTOR) signaling pathway and the increased Gli1 expression and EMT progress induced by AIM2 depletion was reversed after incubation with AKT inhibitor Ly294002 in CRC cells. In conclusion, our results define AIM2 as a novel regulator of Gli1 in CRC cell growth and metastasis, and suggest that the AIM2/AKT/mTOR/Gli1 signaling axis may serve as a potential target for treatment of CRC.


Breeding system of diploid sexuals within the Ranunculus auricomus complex and its role in a geographical parthenogenesis scenario.
The larger distribution area of asexuals compared with their sexual relatives in geographical parthenogenesis (GP) scenarios has been widely attributed to the advantages of uniparental reproduction and polyploidy. However, potential disadvantages of sexuals due to their breeding system have received little attention so far. Here, we study the breeding system of five narrowly distributed sexual lineages of Ranunculus notabilis s.l. (R. auricomus complex) and its effects on outcrossing, inbreeding, female fitness, and heterozygosity. We performed selfing and intra- and interlineage crossings by bagging 481 flowers (59 garden individuals) followed by germination experiments. We compared seed set and germination rates, and related them to genetic distance and genome-wide heterozygosity (thousands of RADseq loci). Selfings (2.5%) unveiled a significantly lower seed set compared with intra- (69.0%) and interlineage crossings (69.5%). Seed set of intra- (65%) compared to interpopulation crossings (78%) was significantly lower. In contrast, all treatments showed comparable germination rates (32%-43%). Generalized linear regressions between seed set and genetic distance revealed positive relationships in general and between lineages, and a negative one within lineages. Seed set was the main decisive factor for female fitness. Germination rates were not related to genetic distance at any level, but were positively associated with heterozygosity in interlineage crossings. Experiments confirmed full crossability and predominant outcrossing among sexual R. notabilis s.l. lineages. However, up to 5% (outliers 15%-31%) of seeds were formed by selfing, probably due to semi-self-compatibility in a multi-locus gametophytic SI system. Less seed set in intrapopulation crossings, and higher seed set and germination rates from crossings of genetically more distant and heterozygous lineages (interlineage) indicate negative inbreeding and positive outbreeding effects. In GP scenarios, sexual species with small and/or isolated populations can suffer from decreased female fitness due to their breeding system. This factor, among others, probably limits range expansion of sexuals.


Extraction of 3,4,4'-Trichlorocarbanilide from Rat Fecal Samples for Determination by High Pressure Liquid Chromatography-Tandem Mass Spectrometry.
Triclocarban (3,4,4'-Trichlorocarbanilide; TCC) in the environment has been well documented. Methods have been developed to monitor TCC levels from various matrices including water, sediment, biosolids, plants, blood and urine; however, no method has been developed to document the concentration of TCC in fecal content after oral exposure in animal studies. In the present study, we developed and validated a method that uses liquid extraction coupled with HPLC-MS/MS determination to measure TCC in feces. The limit of detection and limit of quantitation in control rats without TCC exposure was 69.0 ng/g and 92.9 ng/g of feces, respectively. The base levels of TCC in feces were lower than LOD. At 12 days of treatment, the fecal TCC concentration increased to 2220 µg/g among 0.2% w/w exposed animals. The concentration in fecal samples decreased over the washout period in 0.2% w/w treated animals to 0.399 µ/g feces after exposure was removed for 28 days. This method required a small amount of sample (0.1 g) with simple sample preparation. Given its sensitivity and efficiency, this method may be useful for monitoring TCC exposure in toxicological studies of animals.


Cervical cancer screening in a sexually transmitted disease clinic: screening adoption experiences from a midwestern clinic.
We examined whether a sexually transmitted disease (STD) clinic could reach women who had not received a Papanicolau (Pap) test in the past 3 years. We also explored staff attitudes and implementation of cervical cancer screening. Women (n = 123) aged 30 to 50 years were offered cervical cancer screening in an Indiana STD clinic. We measured effectiveness by the patients' self-reported last Pap test. We explored adoption of screening through focus groups with 34 staff members by documenting their attitudes about cervical cancer screening and screening strategy adaptation. We also documented recruitment and screening implementation. Almost half (47.9%) of participants reported a last Pap test 3 or more years previously; 30% had reported a last Pap more than 5 years ago, and 11.4% had a high-risk test outcome that required referral to colposcopy. Staff supported screening because of mission alignment and perceived patient benefit. Screening adaptations included eligibility, results provision, and follow-up. Cervical cancer screening was possible and potentially beneficial in STD clinics. Future effectiveness-implementation studies should expand to include all female patients, and should examine the degree to which adaptation of selected adoption frameworks is feasible.


Association between use of humidifier disinfectant and allergic rhinitis in Korean children: a cross-sectional study based on the eighth Panel Study on Korean Children (PSKC).
Rhinitis was the most frequently diagnosed disorder among users of humidifier disinfectants (HDs). The aim of our study was to investigate on the relationship between allergic rhinitis (AR) and HD uses. Our study used the data from the eighth Panel Study on Korean Children; a total of 1,540 participants were enrolled. The χ2 test and multiple logistic regression analyses were conducted to debunk the association between AR and HD uses. In our analysis, odds ratios (ORs) of doctor-diagnosed AR increased significantly when simply the response of whether HDs were used in the past was considered. When the brands of HD were considered, ORs of doctor-diagnosed AR were found to be increased significantly for those who have used polyhexamethylene guanidine phosphate/oligo(2-[2-ethoxy]-ethoxyethyl)guanidinium-containing HDs (model 3: 1.41, 95% confidence interval [CI]: 1.02-1.95). However, once duration of usage was additionally considered, ORs of doctor-diagnosed AR increased significantly only for those who have used chloromethylisothiazolinone/methylisothiazolinone-containing HDs for more than or equal to 3 months (model 3: 2.08, 95% CI: 1.17-3.69). Further, past use of HD was associated with significantly increased ORs of AR diagnosed before 2013 (model 3: 1.35, 95% CI: 1.02-1.79). Results of our study suggest that past uses of HDs may be associated with an increased risk of AR.


The diagnostic and economic impact of contrast imaging techniques in the diagnosis of small hepatocellular carcinoma in cirrhosis.
Contrast-enhanced ultrasound (CE-US), contrast CT scan and gadolinium dynamic MRI are recommended for the characterisation of liver nodules detected during surveillance of patients with cirrhosis with US. To assess the sensitivity, specificity, diagnostic accuracy and economic impact of all possible sequential combinations of contrast imaging techniques in patients with cirrhosis with 1-2 cm liver nodules undergoing US surveillance. 64 patients with 67 de novo liver nodules (55 with a size of 1-2 cm) were consecutively examined by CE-US, CT, MRI, and a fine-needle biopsy (FNB) as diagnostic standard. Undiagnosed nodules were re-biopsied; non-malignant nodules underwent enhanced imaging follow-up. The typical radiological feature of hepatocellular carcinoma (HCC) was arterial phase hypervascularisation followed by portal/venous phase washout. HCC was diagnosed in 44 (66%) nodules (2, <1 cm; 34, 1-2 cm; 8, >2 cm). The sensitivity of CE-US, CT and MRI for 1-2 cm HCC was 26, 44 and 44%, with 100% specificity, the typical vascular pattern of HCC being identified in 22 (65%) by a single technique versus 12 (35%) by at least two techniques carried out at the same time point (p=0.028). Compared with the cheapest dual examination (CE-US+CT), the cheapest single technique of stepwise imaging diagnosis of HCC was equally expensive (euro 26 440 vs euro 28 667), but led to a 23% reduction of FNB procedures (p=0.031). In patients with cirrhosis with a 1-2 cm nodule detected during surveillance, a single imaging technique showing a typical contrast pattern confidently permits the diagnosis of HCC, thereby reducing the need for FNB examinations.


Equine anti-hapten antibody. IX. IgM anti-lactose antibodies.
The immune response to a bacterial vaccine of Streptococcus faecalis (strain N) was characterized in all of the seven horses studied by the sustained production of about 90% IgM anti-lactose antibody over a period of 44 weeks with maximum values of the total antibody ranging from 4 mg/ml of serum to 12 mg/ml of serum. With respect to the binding of a lactose-containing ligand the association constants of the antibodies purified from sera obtained between 5 and 44 weeks fell in the range of 1 times 10-5 M-1 to 2 times 10-5 M-1. Not only was there no significant indication of maturation of a-finity in this period but there was a selective limitation of affinity compared to that of 7S antibodies. It was inferred that the synthesis of IgM antibody involves the selective utilization of V-H and/or V-L genes.


Make them blink: probes for super-resolution microscopy.
In recent years, a number of approaches have emerged that enable far-field fluorescence imaging beyond the diffraction limit of light, namely super-resolution microscopy. These techniques are beginning to profoundly alter our abilities to look at biological structures and dynamics and are bound to spread into conventional biological laboratories. Nowadays these approaches can be divided into two categories, one based on targeted switching and readout, and the other based on stochastic switching and readout of the fluorescence information. The main prerequisite for a successful implementation of both categories is the ability to prepare the fluorescent emitters in two distinct states, a bright and a dark state. Herein, we provide an overview of recent developments in super-resolution microscopy techniques and outline the special requirements for the fluorescent probes used. In combination with the advances in understanding the photophysics and photochemistry of single fluorophores, we demonstrate how essentially any single-molecule compatible fluorophore can be used for super-resolution microscopy. We present examples for super-resolution microscopy with standard organic fluorophores, discuss factors that influence resolution and present approaches for calibration samples for super-resolution microscopes including AFM-based single-molecule assembly and DNA origami.


Uncovering system-specific stress signatures in primate teeth with multimodal imaging.
Early life stress can disrupt development and negatively impact long-term health trajectories. Reconstructing histories of early life exposure to external stressors is hampered by the absence of retrospective time-specific biomarkers. Defects in tooth enamel have been used to reconstruct stress but the methods used are subjective and do not identify the specific biological systems impacted by external stressors. Here we show that external physical and social stressors impart biochemical signatures in primate teeth that can be retrieved to objectively reconstruct the timing of early life developmental disruptions. Using teeth from captive macaques, we uncovered elemental imprints specific to disruptions of skeletal growth, including major disruptions in body weight trajectory and moderate to severe illnesses. Discrete increases in heat shock protein-70 expression in dentine coincided with elemental signatures, confirming that elemental signals were associated with activation of stress-related pathways. To overcome limitations of conventional light-microscopic analysis, we used high resolution Raman microspectral imaging to identify structural and compositional alterations in enamel and dentine that coincided with elemental signatures and with detailed medical and behavioural data. Integrating these objective biochemical markers with temporal mapping of teeth enables the retrospective study of early life developmental disruptions and their ensuing health sequelae.


Retinal degeneration and local oxygen metabolism.
Vision loss due to various forms of outer retinal degeneration remains a major problem in clinical ophthalmology. Most retinal degenerations are precipitated by genetic mutations affecting the retinal pigment epithelium and sensory retina, but it is becoming increasingly evident that resultant metabolic changes within the retina may also contribute to the further progression of photoreceptor cell loss. In particular, a role for the local oxygen environment within the retina has been proposed. The correct balance between retinal oxygen supply and oxygen consumption in the retina is essential for retinal homeostasis, and disruption of this balance is a factor in many retinal diseases. In animal models of photoreceptor degeneration, manipulation of environmental oxygen levels has been reported to be able to modulate the rate of photoreceptor degeneration. Clinically, hyperbaric oxygen therapy has already been used in retinitis pigmentosa patients and other types of oxygen therapy have been proposed. It therefore seems appropriate to review our current understanding of the oxygen environment in the normal and degenerating retina, and to build a clearer picture of how the retinal oxygen environment can be modulated. We focus on techniques that have been, or may be, applied clinically, such as modulation of systemic oxygen levels and modulation of retinal oxygen metabolism by light deprivation. Data from direct measurements of intraretinal oxygen distribution in rat models at different stages of photoreceptor degeneration will be reviewed. These models include the Royal College of Surgeons (RCS) rat, and the P23H rat model of outer retinal degeneration. Microelectrode based techniques have allowed the intraretinal oxygen distribution to be measured as a function of retinal depth under well-controlled systemic conditions at different stages of the degeneration process. Both models showed changes in the intraretinal oxygen distribution during the degenerative period, with the changes reflecting the gradual loss of oxygen metabolism of the degenerating photoreceptors. This results in higher than normal oxygen levels in the remaining outer retina and a significant alteration in the oxygen flux from the choroid to the inner retina. The maintenance of normal oxygen levels in the inner retina implies that inner retinal oxygen uptake is well preserved, and that there is also reduced oxygen input from the deeper capillary layer of the retinal circulation. Choroidal oxygen tension and the oxygen tension in the pre-retinal vitreous were unaffected at any of the time periods studied prior to, and during, the degeneration process. It is well known that both hypoxia and hyperoxia can cause neural cell stress and damage. Logically, any therapeutic intervention based on oxygen therapy should attempt to restore the oxygen environment of the remaining retinal cells to within the physiological range. Before any oxygen based therapies for the treatment of retinal degeneration should be seriously considered, the oxygen environment in the degenerating retina should be determined, along with clinically usable methods to restore the oxygen environment to the critical cell layers.


The clinical side of bipolar disorders.
This article summarizes key facts on the epidemiology, diagnosis and clinical treatment of bipolar disorder. Bipolar disorder is a common mental disorder with a high disease burden, but still does not get the attention it deserves in research and clinical training. The nature of the disorder is complex, but it is apparent that biological factors are decisive. Thus, understanding the biological systems and cycles affected will become crucial for developing more targeted interventions. Currently, standard treatments seem to have a low specificity for Bipolar Disorder, and only few experimental interventions target directly potential underlying disturbances as HPA axis or circadian clock dysregulation. Systemic analysing and modelling of bipolar disorder is a novel approach which might open up new ways in developing more selective therapies.


Dysregulated FOXO transcription factors in articular cartilage in aging and osteoarthritis.
Aging is a major risk factor for osteoarthritis (OA). Forkhead-box class O (FoxO) transcription factors regulate mechanisms of cellular aging, including protein quality control, autophagy and defenses against oxidative stress. The objective of this study was to analyze FoxO transcription factors in normal, aging and OA cartilage. Knee joints from humans ages 23-90 and from mice at the age of 4-24 months and following surgically induced OA were analyzed for expression of FoxO proteins. Regulation of FoxO protein expression and activation was analyzed in cultured chondrocytes. Human cartilage expressed FOXO1 and FOXO3 but not FOXO4 proteins. FOXO1 and FOXO3 were more strongly expressed the superficial and mid zone as compared to the deep zone and were mainly localized in nuclei. During human joint aging, expression of FOXO1 and FOXO3 was markedly reduced in the superficial zone of cartilage regions exposed to maximal weight bearing. In OA cartilage, chondrocyte clusters showed strong FOXO phosphorylation and cytoplasmic localization. Similar patterns of FOXO expression in normal joints and changes in aging and OA were observed in mouse models. In cultured chondrocytes, IL-1β and TNF-α suppressed FOXO1, while TGF-β and PDGF increased FOXO1 and FOXO3 expression. FOXO1 and FOXO3 phosphorylation was increased by IL-1β, PDGF, bFGF, IGF-1, and the oxidant t-BHP. Normal articular cartilage has a tissue specific signature of FoxO expression and activation and this is profoundly altered in aging and OA in humans and mice. Changes in FoxO expression and activation may be involved in cartilage aging and OA.


Random parameter sampling of a generic three-tier MAPK cascade model reveals major factors affecting its versatile dynamics.
The mitogen-activated protein kinase (MAPK) pathway is considered to be a central block in many biological signaling networks. Despite the common core cascade structure, the activation of MAPK in different biological systems can exhibit different types of dynamic behaviors. Computer modeling may help to reveal the mechanisms underlying such variations. However, so far most computational models of the MAPK cascade have been system-specific, or to reflect a particular type among the wide spectrum of possible dynamics. To obtain a general and integrated view of the relationship between the dynamics of MAPK activation and the structures and parameters of the MAPK cascade, we constructed a generic model by comparing previous models covering different specific biological systems. We assumed that reliable qualitative results could be predicted through a qualitative model with pseudo parameters. We used randomly sampled parameters instead of a specific set of "best-fit" parameters to avoid biases towards any particular systems. A range of dynamics behaviors for MAPK activation, including ultrasensitivity, bistability, transient activation and oscillation, were successfully predicted by the generic model. The results indicated that the steady state dynamics (ultrasensitivity and bistability) was jointly determined by the three-tiered structure of the MAPK cascade and the competitive substrate binding in the dual-phosphorylation processes of the central components, while the temporal dynamics (transient activation and oscillation) was mainly affected by the upstream signaling pathway and feedbacks. Moreover, MAPK kinase (MAPKK) played more important roles than the other two components in determining the dynamics of MAPK activation. We hypothesize that this is an important and advantageous property for the regulation and for the functional diversity of MAPK pathways in real cells. Finally, to assist developing generic models for signaling motifs through model comparisons, we proposed a reaction-based database to make the model data more flexible and interoperable.


Monocyte-mediated defense against microbial pathogens.
Circulating blood monocytes supply peripheral tissues with macrophage and dendritic cell (DC) precursors and, in the setting of infection, also contribute directly to immune defense against microbial pathogens. In humans and mice, monocytes are divided into two major subsets that either specifically traffic into inflamed tissues or, in the absence of overt inflammation, constitutively maintain tissue macrophage/DC populations. Inflammatory monocytes respond rapidly to microbial stimuli by secreting cytokines and antimicrobial factors, express the CCR2 chemokine receptor, and traffic to sites of microbial infection in response to monocyte chemoattractant protein (MCP)-1 (CCL2) secretion. In murine models, CCR2-mediated monocyte recruitment is essential for defense against Listeria monocytogenes, Mycobacterium tuberculosis, Toxoplasma gondii, and Cryptococcus neoformans infection, implicating inflammatory monocytes in defense against bacterial, protozoal, and fungal pathogens. Recent studies indicate that inflammatory monocyte recruitment to sites of infection is complex, involving CCR2-mediated emigration of monocytes from the bone marrow into the bloodstream, followed by trafficking into infected tissues. The in vivo mechanisms that promote chemokine secretion, monocyte differentiation and trafficking, and finally monocyte-mediated microbial killing remain active and important areas of investigation.


Frequency of EGFR mutations in non-small cell lung cancer patients: screening data from West Siberia.
Incorporation of molecular analysis of the epidermal growth factor receptor (EGFR) gene into routine clinical practice has shown great promise to provide personalized therapy of the non-small cell lung cancer (NSCLC) in the developed world. However, the genetic testing of EGFR mutations has not yet become routine clinical practice in territories remote from the central regions of Russia. Therefore, we aimed to study the frequency of major types of activating mutations of the EGFR gene in NSCLC patients residing in West Siberia. We examined EGFR mutations in exons 19 and 21 in 147 NSCLC patients (excluding squamous cell lung carcinomas) by real time polymerase chain reaction. EGFR mutations were detected in 28 of the 147 (19%) patients. There were 19 (13%) cases with mutations in exon 19 and 9 cases (6%) in exon 21. Mutations were more frequently observed in women (42%, p=0.000) than in men (1%). A significantly higher incidence of EGFR mutations was observed in bronchioloalveolar carcinomas (28%, p=0.019) and in adenocarcinomas (21%, p=0.024) than in large cell carcinomas, mixed adenocarcinomas, and NOS (4%). The EGFR mutation rate was much higher in never-smokers than in smokers: 38% vs. 3% (p=0.000). The frequency of EGFR mutations in the Kemerovo and Tomsk regions was 19%. The incorporation of molecular analysis of the EGFR gene into routine clinical practice will allow clinicians to provide personalised therapy, resulting in a significant increase in survival rates and improvement in life quality of advanced NSCLC patients.


Adverse childhood experiences, adult trauma, and depressive symptom trajectories.
Associations among adverse childhood experiences prior to age 18 (ACEs), subjective reports of trauma during Hurricane Sandy, and trajectories of depressive symptoms reported by community-dwelling older people were examined. We analyzed 6 waves of data from 5,688 people aged 50-74 recruited in 2006 and followed for 12-years using multilevel mixed effects models. We found that: (1) people who experienced ACEs had trajectories of depressive symptoms in late life that were higher than people not having these experiences, (2) people experiencing two or more ACEs were more likely to report fear and distress when Hurricane Sandy hit than people experiencing either one or no adverse childhood experiences, and (3) while both ACE exposure and peri-traumatic stress were associated with trajectories having higher levels of depressive symptoms, the risk associated with ACEs (especially multiple ACEs) was greater. Findings support life course stress theories including the cumulative inequality theory and stress proliferation theory, suggesting that inequalities are manifested over the life course and that people experiencing adversity during childhood are at increased risk of experiencing adversity in late life. By studying the relationship between adverse childhood experiences and response to Hurricane Sandy our findings demonstrate that adverse childhood experiences can alter the way traumatic events in adulthood are experienced. This finding in turn, has important implications for clinical practice, as it identifies a group of people likely to be at risk for adult trauma.


Deinococcus mumbaiensis Shashidhar and Bandekar 2006 is a later heterotypic synonym of Deinococcus ficus Lai et al. 2006.
Deinococcus mumbaiensis DSM 17424(T) was compared with Deinococcus ficus CC-FR2-10(T) to clarify the taxonomic relationship between the two species. 16S rRNA gene sequence comparisons demonstrated that these strains share 99.5 % sequence similarity. Investigation of fatty acid patterns and substrate utilization profiles displayed no striking differences between the type strains of the two species. DNA-DNA hybridizations between the two strains showed 75.4 % (76.6 %) relatedness. Therefore, the reclassification of Deinococcus mumbaiensis as a later heterotypic synonym of Deinococcus ficus is proposed, based upon the estimated phylogenetic position derived from 16S rRNA gene sequence data, fatty acid and biochemical data and DNA-DNA hybridization results.


Purification of a Novel Bacteriocin-Like Inhibitory Substance Produced by Enterococcus faecium ICIS 8 and Characterization of Its Mode of Action.
The aim of this work was to purify and characterize a bacteriocin-like antimicrobial substance produced by an antagonistic active strain of Enterococcus faecium. A novel bacteriocin-like inhibitory substance (BLIS) produced by the E. faecium ICIS 8 strain was purified and characterized using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and N-terminal amino acid sequencing revealed the following partial sequence: NH2-APKEKCFPKYCV. The proteinaceous nature of purified BLIS was assessed by treatment with proteolytic enzyme. Studies of the action of BLIS using bacteriological and bioluminescence assays revealed a dose-dependent inhibition of Listeria monocytogenes 88BK and Escherichia coli K12 TG1 lac::lux viability. The interaction of the BLIS with the bacterial surface led to the compensation of a negative charge value, as shown by zeta-potential measurements. Assessments of membrane integrity using fluorescent probes and atomic force microscopy revealed the permeabilization of the cellular barrier structures in both L. monocytogenes and E. coli. The novel BLIS from E. faecium ICIS 8 was characterized by a unique primary peptide sequence and exerted bactericidal activity against L. monocytogenes and E. coli by disrupting membrane integrity.


Auditory perception skills in children receiving simultaneous bilateral cochlear implants: early speech-discrimination results.
The objective of the present study is to evaluate early speech-perception abilities using VRISD in toddlers who received simultaneous bilateral CIs between 12 and 36 months of age and to compare them with the findings in NH infants and to monitor the development of speech-perception skills within 6 months after cochlear implantation. The VRISD test was performed using video visual reinforcement to assess speech-discrimination ability in the CI and NH groups. Four stimuli were used for testing in the present study: /a/, /i/, /ba/, and /da/. The two contrasts used for the research were /a-i/ and /ba-da/. Auditory and listening skills in the CI group were assessed using the IT-MAIS. The responses to the /a-i/ and /ba-da/ phoneme contrast were found to be similar in the NH and CI groups. No statistically significant difference was found between the groups (p > 0.05). VRISD test result and the IT-MAIS score were highly correlated in CI group (p = 0.001, r = 0.822). The VRISD test can be effectively used to evaluate the development of speech-discrimination skills in hearing-impaired babies before and after CI. This research suggests that the development of speech-perception ability with CI is seriously influenced by environmental exposure and sound access. To the best of our knowledge, this is the first study to evaluate the speech-perception skills in toddlers with simultaneous bilateral CI.


TFEB Expression Profiling in Renal Cell Carcinomas: Clinicopathologic Correlations.
TFEB is overexpressed in TFEB-rearranged renal cell carcinomas as well as in renal tumors with amplifications of TFEB at 6p21.1. As recent literature suggests that renal tumors with 6p21.1 amplification behave more aggressively than those with rearrangements of TFEB, we compared relative TFEB gene expression in these tumors. This study included 37 TFEB-altered tumors: 15 6p21.1-amplified and 22 TFEB-rearranged (including 5 cases from The Cancer Genome Atlas data set). TFEB status was verified using a combination of fluorescent in situ hybridization (n=27) or comprehensive molecular profiling (n=13) and digital droplet polymerase chain reaction was used to quantify TFEB mRNA expression in 6p21.1-amplified (n=9) and TFEB-rearranged renal tumors (n=19). These results were correlated with TFEB immunohistochemistry. TFEB-altered tumors had higher TFEB expression when normalized to B2M (mean: 168.9%, n=28), compared with non-TFEB-altered controls (mean: 7%, n=18, P=0.005). Interestingly, TFEB expression in tumors with rearrangements (mean: 224.7%, n=19) was higher compared with 6p21.1-amplified tumors (mean: 51.2%, n=9; P=0.06). Of note, classic biphasic morphology was only seen in TFEB-rearranged tumors and when present correlated with 6.8-fold higher TFEB expression (P=0.00004). Our results suggest that 6p21.1 amplified renal tumors show increased TFEB gene expression but not as much as t(6;11) renal tumors. These findings correlate with the less consistent/diffuse expression of downstream markers of TFEB activation (cathepsin K, melan A, HMB45) seen in the amplified neoplasms. This suggests that the aggressive biological behavior of 6p21.1 amplified renal tumors might be secondary to other genes at the 6p21.1 locus that are co-amplified, such as VEGFA and CCND3, or other genetic alterations.


The effect of extracellular acidosis on the behaviour of mesenchymal stem cells in vitro.
The stem cell fraction of a cell population is finely tuned by stimuli from the external microenvironment. Among these stimuli, a decrease of extracellular pH (pHe) may occur in a variety of physiological and pathological conditions, including hypoxia and inflammation. In this study, by using bone marrow stem cells and dental pulp stem cells, we provided evidence that extracellular acidosis endows the maintenance of stemness in mesenchymal cells. Indeed, continuous exposure for 21 d to low pHe (6.5-6.8) conditions impaired the osteogenic differentiation of both cell types. Moreover, the exposure to low pHe, for 1 and up to 7 d, induced the expression of stemness-related genes and proteins, drove cells to reside in the quiescent G0 alert state and enhanced their ability to form floating spheres. The pre-conditioning with extracellular acidosis for 7 d did not affect the differentiation potential of dental pulp stem cells since, when the cells were cultured again at physiological pHe, their multilineage potential was almost unmodified. Our data provided evidence of the role of extracellular acidosis as a modulator of the stemness of mesenchymal cells. This condition is commonly found both in systemic and local bone conditions, hence underlining the relevance of this phenomenon for a better comprehension of bone healing and regeneration.


New concepts in the use of axillofemoral bypass grafts.
Over a four-year period, 52 patients underwent axillofemoral bypass operations. Indications for operation were divided into three groups: those that were performed emergently (aortoenteric fistula, graft infection, and leaking aortic aneurysm), those performed electively for aneurysm, and those for ischemia. Operative procedures were categorized as follows: axillofemoral bypass alone, axillofemorofemoral bypass with the proximal femorofemoral anastomosis being graft to graft, or axillofemorofemoral bypass with the femorofemoral anastomosis to either a transected proximal common femoral artery or superficial femoral artery distal to the axillofemoral anastomosis. Thirty-three percent of the unilateral axillofemoral grafts failed, while only 14% of the axillobifemoral grafts failed. There was a further difference between the two methods of femorofemoral grafting with 22% failure in the former group but no failures in the latter.


The Impact of COVID-19 on Neurosurgical Services in Africa.
COVID-19 has affected the global provision of neurosurgical services. We sought to review the impact of COVID-19 on the neurosurgical services in Africa. A cross-sectional survey was distributed to African neurosurgeons seeking to review demographics, national and neurosurgical preparedness, and change in clinical services in April 2020. A total of 316 responses from 42 countries were received. Of these, 81.6% of respondents were male and 79.11% were under the age of 45 years. In our sample, 123 (38.92%) respondents were in training. Most (94.3%) respondents stated they had COVID-19 cases reported in their country as of April 2020. Only 31 (41.50%) had received training on managing COVID-19. A total of 173 (54.70%) respondents were not performing elective surgery. There was a deficit in the provision of personal protective equipment (PPE): surgical masks (90.80%), gloves (84.80%), N95 masks (50.80%), and shoe covers (49.10%). Health ministry (80.40%), World Health Organization (74.50%), and journal papers (41.40%) were the most common sources of information on COVID-19. A total of 43.60% had a neurosurgeon in the COVID-19 preparedness team; 59.8% were concerned they may contract COVID-19 at work with a further 25.90% worried they may infect their family. Mental stress as a result of COVID-19 was reported by 14.20% of respondents. As of April 2020, 73.40% had no change in their income. Most African countries have a national COVID-19 policy response plan that is not always fully suited to the local neurosurgery services. There is an ongoing need for PPE and training for COVID-19 preparedness. There has been a reduction in clinical activities both in clinic and surgeries undertaken.


Psychiatric comorbidity in the evolution from migraine to medication overuse headache.
We set out to study the role of psychiatric comorbidity in the evolution of migraine to medication overuse headache (MOH) by a comparative study of 41 migraineurs (MIG) and 41 patients suffering from MOH deriving from migraine. There was an excess risk of suffering from mood disorders [odds ratio (OR) = 4.5, 95% confidence interval (CI) 1.5, 13.5], anxiety (OR = 5, 95% CI 1.2, 10.7) and disorders associated with the use of psychoactive substances other than analgesics (OR = 7.6, 95% CI 2.2, 26.0) in MOH compared with MIG. Retrospective study of the order of occurrence of disorders showed that in the MOH group, psychiatric disorders occurred significantly more often before the transformation from migraine into MOH than after. There was no crossed-family transmission between MOH and psychiatric disorders, except for substance-related disorders. MOH patients have a greater risk of suffering from anxiety and depression, and these disorders may be a risk factor for the evolution of migraine into MOH. Moreover, MOH patients have a greater risk of suffering from substance-related disorders than MIG sufferers. This could be due to the fact that MOH is part of the spectrum of addictive disorders.


Gas Hydrate Combustion in Five Method of Combustion Organization.
Experiments on the dissociation of a mixed gas hydrate in various combustion methods are performed. The simultaneous influence of two determining parameters (the powder layer thickness and the external air velocity) on the efficiency of dissociation is studied. It has been shown that for the mixed hydrate, the dissociation rate under induction heating is 10-15 times higher than during the burning of a thick layer of powder, when the combustion is realized above the layer surface. The minimum temperature required for the initiation of combustion for different combustion methods was studied. As the height of the sample layer increases, the rate of dissociation decreases. The emissions of NOx and CO for the composite hydrate are higher than for methane hydrate at the same temperature in a muffle furnace. A comparison of harmful emissions during the combustion of gas hydrates with various types of coal fuels is presented. NOx concentration as a result of the combustion of gas hydrates is tens of times lower than when burning coal fuels. Increasing the temperature in the muffle furnace reduces the concentration of combustion products of gas hydrates.


Methotrexate levels in the interstitial space and seminiferous tubule of rat testis.
The ability of methotrexate (MTX) to pass from the blood into the interstitial space and seminiferous tubule of the rat was investigated using testicular micropuncture. MTX was administered to anesthetized adult Wister rats via a femoral vein cannula. Constant plasma levels of MTX were achieved by giving a priming dose followed by a constant infusion of 1, 10, or 100 mg/kg/hr with 6 to 27 rats studied at each dose. Blood (via a jugular vein cannula), testicular interstitial fluid, and seminiferous tubule fluid (via direct micropuncture) were periodically sampled during the 4 hr of drug infusion. Under steady-state conditions, when compared to corresponding plasma values, MTX levels were 2- to 4-fold lower in the testicular interstitial fluid and 18- to 50-fold lower in the seminiferous tubule. These results indicate that, in the rat, a significant blood-testis barrier to MTX exists at the tubular but probably not at the capillary-interstitial level. If these results can be extrapolated to humans, they do not provide a pharmacological explantation for the frequent occurrence of leukemic relapse in the interstitium of the testes in boys with acute lymphocytic leukemia.


Pharmacological treatment of ADHD and the short and long term effects on sleep.
There is growing research focusing on the sleep problems of children with attention-deficit/hyperactivity disorder (ADHD) in recent years. High incidence of sleep disorders in children with ADHD may be associated with a substantial impact on their quality of life and exacerbation of ADHD symptoms. The core symptoms of ADHD can be effectively treated by various medications, including methylphenidate (MPH), amphetamine, pemoline, and the newly FDA-approved extended-release α2 adrenergic agonists. However, most of them are known to affect patients' sleep because of their pharmacological actions on dopaminergic and/or noradrenergic release in the central nervous system. Previous studies have found increased incidence of insomnia and sleep disturbances in ADHD children treated with CNS (central nervous system) stimulants. In contrast, recent prospective, double-blind, placebo-controlled trials concluded that MPH, by objective polysomnographic or actigraphic measurements, did not cause significant sleep problems in children or adolescents with ADHD. Given the fact that sleep quality and core symptoms of ADHD are highly correlated, it is imperative that we understand the effects of ADHD medications on sleep while prescribing either CNS stimulants or non-CNS stimulants. Here we will concisely review the pharmacological treatments of ADHD, and provide the relevant data discussing their short- and long-term effects on sleep.