Noise perturbation in functional principal component analysis filtering for two-dimensional correlation spectroscopy: its theory and application to infrared spectra of a poly(3-hydroxybutyrate) thin film.
A method based on noise perturbation in functional principal component analysis (NPFPCA) is for the first time introduced to overcome the noise interference problem in two-dimensional correlation spectroscopy (2D-COS). By the systematic addition of synthetic noise to the dynamic multivariate spectral data, the functional principal component analysis (FPCA) described in this report is able to accurately determine which eigenvectors are representing significant signals instead of noise in the original data. This feature is especially useful for the data reconstruction and noise filtering. Reconstructed data resulted from the smooth eigenvectors can produce much more reliable 2D correlation spectra by removing the correlation artifacts from noise, which in turn enable more accurate interpretation of the spectral variations. The usefulness of this method is demonstrated with a theoretical framework and applications to the 2D correlation analyses of both simulated data and temperature-dependent reflection-absorption infrared spectra of a poly(3-hydroxybutyrate) (PHB) thin film.


Simple and Rapid Dual-Dispersive Liquid-Liquid Microextraction as an Innovative Extraction Method for Uranium in Real Water Samples Prior to the Determination of Uranium by a Spectrophotometric Technique.
An innovative, rapid, and simple dual-dispersive liquid-liquid microextraction (DDLL-ME) approach was used to extract uranium from real samples for the first time. The main objective of this study was to disperse extraction solvent by using an air-agitated syringe system to overcome matrix effects and avoid dispersion of hazardous dispersive organic solvents by using heat. The DDLL-ME method consisted of two dispersive liquid-liquid extraction steps with chloroform as the extracting solvent. Uranium formed complexes with 4-(2-thiazolylazo) resorcinol in the aqueous phase and was extracted in extracting solvent (chloroform) after the first dispersive liquid-liquid process. Uranium was then back-extracted in the acidic aqueous phase in a second dispersive liquid-liquid process. Finally, uranium was determined by a spectrophotometric detection technique. The variables that played a key role in the proposed method were studied and optimized. The LOD and sensitivity enhancement factor for uranium were found to be 0.60 µg/L and 45, respectively, under optimized conditions. Calibration graphs were found to be linear in the range of 5.0-600 µg/L. The RSD was 2.5%. Reliability of the proposed method was verified by analyzing certified reference material TM-28.3.


Social Regulation of Negative Valence Systems During Development.
The ability to sense, perceive, and respond appropriately to aversive cues is critical for survival. Conversely, dysfunction in any of these pathway components can lead to heightened avoidance of neutral or rewarding cues, such as social partners. The underlying circuitry mediating both negative valence processing and social behavior is particularly sensitive to early life experience, but mechanisms linking experience to pathology remain elusive. Previous research in humans, rodents, and non-human primates has highlighted the unique neurobiology of the developing infant and the role of the caregiver in mediating the infant's negative valence circuitry, and the importance of this early social relationship for scaffolding lasting social behavior. In this review, we summarize the current literature on the development of negative valence circuits in the infant and their social regulation by the caregiver following both typical and adversity-rearing. We focus on clinically-relevant research using infant rodents which highlights the amygdala and its interface with the mesolimbic dopamine system through innervation from the ventral tegmental area (VTA) as a locus of dysfunction following early-life adversity. We then describe how these circuits are recruited to perturb life-long social behavior following adversity and propose additional therapeutic targets in these circuits with an eye toward developing age-appropriate interventions.


Reconstruction of massive midfoot bone and soft tissue loss as a result of blast injury.
Lower extremity blast injuries represent a unique challenge to surgeons and often involve complex, limb-threatening wounds with extensive soft tissue and bone loss. Surgical treatment of these injuries can be difficult because of limited autogenous resources for reconstruction of the defect. In this article, we describe a technique for medial column reconstruction using iliac crest bone graft and soft tissue coverage with an abductor hallucis rotational flap combined with a split-thickness skin graft. This method addresses the extensive bone and soft tissue defects that frequently characterize blast injuries to the foot, and may be applicable in other situations where trauma or infection has caused extensive destruction of the medial column.


IP3R1 underlies diastolic ANO1 activation and pressure-dependent chronotropy in lymphatic collecting vessels.
Pressure-dependent chronotropy of murine lymphatic collecting vessels relies on the activation of the Ca2+-activated chloride channel encoded by Anoctamin 1 (Ano1) in lymphatic muscle cells. Genetic ablation or pharmacological inhibition of ANO1 results in a significant reduction in basal contraction frequency and essentially complete loss of pressure-dependent frequency modulation by decreasing the rate of the diastolic depolarization phase of the ionic pacemaker in lymphatic muscle cells (LMCs). Oscillating Ca2+ release from sarcoendoplasmic reticulum Ca2+ channels has been hypothesized to drive ANO1 activity during diastole, but the source of Ca2+ for ANO1 activation in smooth muscle remains unclear. Here, we investigated the role of the inositol triphosphate receptor 1 (Itpr1; Ip3r1) in this process using pressure myography, Ca2+ imaging, and membrane potential recordings in LMCs of ex vivo pressurized inguinal-axillary lymphatic vessels from control or Myh11CreERT2;Ip3r1fl/fl (Ip3r1ismKO) mice. Ip3r1ismKO vessels had significant reductions in contraction frequency and tone but an increased contraction amplitude. Membrane potential recordings from LMCs of Ip3r1ismKO vessels revealed a depressed diastolic depolarization rate and an elongation of the plateau phase of the action potential (AP). Ca2+ imaging of LMCs using the genetically encoded Ca2+ sensor GCaMP6f demonstrated an elongation of the Ca2+ flash associated with an AP-driven contraction. Critically, diastolic subcellular Ca2+ transients were absent in LMCs of Ip3r1ismKO mice, demonstrating the necessity of IP3R1 activity in controlling ANO1-mediated diastolic depolarization. These findings indicate a critical role for IP3R1 in lymphatic vessel pressure-dependent chronotropy and contractile regulation.


[Effects of the zinc oxide and cod liver oil containing ointment Zincojecol in an animal model of wound healing].
The effects of Zincojecol an ointment containing zinc oxide and cod liver oil on wound healing were compared with ointments that either contained no active ingredients or zinc oxide or cod liver oil alone. All formulations enhanced the epidermal proliferation after mechanical irritation of the tail skin. The combination of zinc oxide and cod liver oil was found to be superior to the vehicle control and formulations containing only one active ingredient. This combination was also found to be most efficient in accelerating wound healing being retarded by repeated dexamethasone treatment.


An analysis of implantable cardiac device reliability. The case for improved postmarketing risk assessment and surveillance.
Implantable cardiac devices have become the mainstay of the treatment of patients with heart disease. However, data regarding their reliability and, inferentially, safety have been called into question. We reviewed annual reports submitted to the Food and Drug Administration Office of Device Evaluation by device manufacturers from 2003 to 2007. The annual number of implantable cardiac defibrillators (ICDs) and cardiac resynchronization therapy defibrillator (CRT-D) implants, explants, and returned devices were tabulated along with the cumulative (Cum) number of implants for each device. We derived an annual explantation rate (AER) defined as the ratio of the annual number of explants less the number of normal battery depletions/Cum (×1000). From 2003 to 2007, 256,392 CRT-D and 459,300 ICD devices were implanted in the United States. The overall mean (±SD) AERs for ICD and CRT-D devices were, respectively, 49.5 (15.6) per 1000 ICD devices and 82.6 (35.5) per 1000 CRT-D devices. The AER for each device type significantly decreased over the study period (P for trend <0.001) although the AER for CRT-D devices was 38% higher than that for ICD devices (P < 0.001). On average, 20.3% of CRT-D devices and 22.6% of ICD devices were returned to the manufacturer for analysis after explantation. The rates of explanted CRT-D and ICD devices decreased from 2003 to 2007. Notwithstanding this favorable trend, the AER for CRT-D devices was higher than that for ICD devices. Improved methods for tracking individual device histories are needed for more precise estimates of the risk of device explantation for suspected malfunction. The proportion of devices returned to the manufacturer is suboptimal and needs to be improved to better understand the mechanisms of device malfunction.


Classical master equation for excitonic transport under the influence of an environment.
In a previous paper [Phys. Rev. E 83, 051911 (2011)] we have shown that the results of a quantum-mechanical calculation of electronic energy transfer (EET) over aggregates of coupled monomers can be described also by a model of interacting classical electric dipoles in a weak-coupling approximation, which we referred to as the realistic coupling approximation (RCA). The method was illustrated by EET on a simple linear chain of molecules and also by energy transfer on an arrangement of monomers corresponding to that of the Fenna-Matthews-Olson (FMO) complex relevant for photosynthesis. The study was limited to electronic degrees of freedom, since this is the origin of coherent EET in the quantum case. Nevertheless, more realistic models of EET require the inclusion of the decohering effects of coupling to an environment, when the molecular aggregate becomes an open quantum system. Here we consider the quantum description of EET on a linear chain and on the FMO complex, incorporating environment coupling and constructing the classical version of the same systems in the density matrix formalism. The close agreement of the exact quantum and exact classical results in the RCA is demonstrated and justified analytically in the RCA. This lends further support to the conclusion that the coherence properties of EET in the FMO complex are evident at the classical level and should not be ascribed as solely due to quantum effects.


Bioactive capacity of some Romanian wild edible mushrooms consumed mainly by local communities.
Wild edible mushrooms are considered as a cheap food source, but rich in bioactive compounds, especially in phenolic compounds. The purpose of the study was to determine the antioxidant and antimicrobial activity and the content of polyphenolcarboxylic acids in 10 species of mushrooms commonly used (consumed) in Romania and two controls. The effect against free radicals, lipid peroxidation and reducing power were determined. The antimicrobial effect was revealed on some strains with pathogenic effect by disk diffusion assay. The antioxidant capacity expressed in vitro was correlated both with the presence of the main polyphenolcarboxylic acids, and also with the presence of other bioactive molecules (flavonoids, carotenoid compounds, etc.).


Physiological and brain alterations produced by high-fat diet in male and female rats can be modulated by increased levels of estradiol during critical periods of development.
Overnutrition due to a high-fat diet (HFD) can increase the vulnerability of the metabolic system to maladjustments. Estradiol has an inhibitory role on food intake and this hormone has demonstrated to be a crucial organizer during brain development. Our aim was to determine whether increased levels of estradiol in the early postnatal period modulate the alterations in metabolism and brain metabolic circuits produced by overnutrition. Twenty-four male and 24 female Wistar rats were submitted to a HFD (34.9% fat) or a control diet (5% fat) from gestational day 6. From postnatal (P) 6 to P13, both control and HFD groups were administered a s.c. injection of vehicle or estradiol benzoate (0.4 mg/kg), resulting in eight experimental groups (n = 6 in each group). Body weight, food intake and subcutaneous, visceral, and brown fat pads were measured. Agouti-related peptide, neuropeptide Y, orexin, and proopiomelanocortin (POMC) were analyzed by quantitative real-time polymerase chain reaction assay and plasma estradiol levels were measured by ELISA. Males fed a HFD showed an increase in body weight and the amount of visceral and subcutaneous fat, which was coincident with an increase in the number of kilocalories ingested. Neonatal estradiol treatment restored the body weight and subcutaneous fat of HFD males to control levels. Hypothalamic POMC mRNA levels in HFD females were increased with respect to control females. This increase was reverted with estradiol treatment during development. HFD and estradiol treatment have different effects on males and females. Overnutrition affects physiological parameters, such as body weight, visceral, and subcutaneous fat content, in males, while females present alterations in hypothalamic POMC mRNA levels. Hence, the increase in estradiol levels during a period that is critical for the programing of the feeding system can modulate some of the alterations produced by the continuous intake of high-fat content food.


Influence of template strandedness on in vitro replication of mutagen-damaged DNA.
We analyzed the ability of DNA polymerases to bypass damage on single- and double-stranded templates. In vitro DNA synthesis was studied on UV-irradiated and polyaromatic hydrocarbon reacted (benzo[a]pyrenediol epoxide and oxiranylpyrene) double-stranded templates by a protocol involving initiation on a uniquely nicked circular double-stranded template. The template was prepared by treating single-stranded (+)M13mp2 circular strands with mutagen and then hybridizing with restricted M13 RFmp2, followed by isolation of the nicked RFII forms. The protocol permits either (+), (-), or both strands to carry lesions. We found that the rules for termination and bypass of lesions previously observed with single-stranded DNA templates also hold for double-stranded templates. Termination of synthesis occurs primarily one nucleotide 3' to the lesion in the template strand. Bypass of UV-induced lesions can be followed in a series of three partial reactions in the presence of Mn2+ and dGMP, which relax the specificity of nucleotide insertion and 3'----5' exonuclease activity, respectively. There is no evidence for greater permissivity of bypass in double-as opposed to single-stranded templates. As with single-stranded templates, purines and preferentially deoxyadenosine (dA) are inserted opposite lesions. Lesions in the nontemplate strand elicit neither termination nor pausing. The addition of Rec A protein resulted in a measurable increase of bypass in this system.


Cognitive decline in old age: separating retest effects from the effects of growing older.
Repeated administration of cognitive tests improves test performance, making it difficult for researchers to gauge the true extent of age-related cognitive decline. The authors examined this issue using data from the Rush Religious Orders Study and linear mixed-effects models. At annual intervals for up to 8 years, more than 800 older Catholic clergy members completed the same set of 19 cognitive tests from which previously established composite measures of cognitive domains were derived. Retest effects on some measures were substantial and continued to accumulate even after 8 annual test readministrations, but effects on other measures were minimal. Across cognitive measures, retest effects were not related to age, sex, or education. Individual differences in retest effects were substantial but not consistent across cognitive measures. The results suggest that retest-based improvement in cognitive test performance can be substantial and persistent.


The use of a novel smartphone testing platform for the development of colorimetric sensor receptors for food spoilage.
This work presents a novel smartphone testing platform for the validation of colorimetric sensor receptors (CSRs) in the form of layers that enables reliable and straightforward determination of their color change in a closed system using a commercially available color sensor. The food-compatible model CSR used for the method development was made of black carrot extract and ethyl cellulose. The colorimetric responses were studied in detail for NH3, dimethylamine (DMA), and trimethylamine (TMA) by analyzing changes in the value of the total color difference (ΔE) with the increasing logarithm of the mass concentration (log γ) of the analytes. The method was partially validated for the detection limit (LOD), the limit of quantification, sensitivity, and linear γ range. The fastest reaction times were obtained for the NH3 analyte, while the calculated LOD values were quite similar (1.48 mg L-1 for NH3, 1.55 mg L-1 for DMA, and 1.58 mg L-1 for TMA). The applicability of CSRs was shown for different types of muscle food. Frozen (boneless and skinless) hake fillets were used for additional experimental work in which the color changes of the CSRs were correlated with the values of the total volatile basic nitrogen (TVB-N) and the total counts of aerobic and anaerobic microorganisms. The developed testing platform shows great promise for the development of CSRs that define the quality of a broad variety of muscle food.


Emotions and skin (II)-the conditioning of scratch responses in cases of lichen simplex.
Lichen simplex is generally regarded as a condition initiated and perpetuated by scratching and emotional tension. It was felt that the scratching might partly be a conditional response to itching and other signals, and that feelings of guilt, anxiety and hostility would be prominent features in these patients. Conditioning experiments designed to establish scratch responses to an itch stimulus (UCS) and a tone (CS) showed that lichen simplex patients conditioned more readily and extinguished more slowly than controls. These finding were more marked when the itch stimulus was applied to affected as compared with normal skin. It was not possible to distinguish differences in the psychological tests between patients and controls. The possible significance of these findings is discussed.


Deep cooling of optically trapped atoms implemented by magnetic levitation without transverse confinement.
We report a setup for the deep cooling of atoms in an optical trap. The deep cooling is implemented by eliminating the influence of gravity using specially constructed magnetic coils. Compared to the conventional method of generating a magnetic levitating force, the lower trap frequency achieved in our setup provides a lower limit of temperature and more freedoms to Bose gases with a simpler solution. A final temperature as low as ∼6nK is achieved in the optical trap, and the atomic density is decreased by nearly two orders of magnitude during the second stage of evaporative cooling. This deep cooling of optically trapped atoms holds promise for many applications, such as atomic interferometers, atomic gyroscopes, and magnetometers, as well as many basic scientific research directions, such as quantum simulations and atom optics.


[Increasing peripheral blood neutrophils after G-CSF treatment is a predictor of early response to immunosuppressive therapy in severe aplastic anemia].
To testify whether absolute neutrophil count (ANC) response to preimmunosuppressive-therapy (pre-IST) granulocyte-stimulating factor (G-CSF) treatment could predict early response to IST in severe aplastic anemia (SAA). Clinical data and hematologic response of 125 SAA patients treated with antithymocyte globulin (r-ATG) combined with cyclosporine were retrospectively analyzed. Correlation of ANC response to pre-IST G-CSF treatment and early response to IST were statistically analyzed, and receiver operating characteristic (ROC) curve was used to estimate the value of increased ANC (∆ANC) in predicting early IST response. The hematologic response (HR) rate to IST in ANC reponded patients was significantly higher than non-responded group (3-month HR 49.0% vs 28.9%, P=0.023; 6-month HR 61.2% vs 40.8%, P=0.026). With ∆ANC≥0.5×10⁹/L as cutoff level, the best point to predict early IST response was 10 days after G-CSF (d 10). Response of ANC to pre-IST G-CSF treatment at d 10 was among the independent factors of predicting 3-month (P=0.004), but not for 6-month response to IST. The overall 5-year survival rate was 92.8% and 69.5% in ANC responded and non-responed groups, respectively (P=0.025). Responding to pre-IST G-CSF treatment reflected the residual bone marrow hematopoiesis, and could act as a convenient and practical predictor to early IST response as well as long-term survival in SAA.


Iron-Catalyzed para-Selective C-H Allylation of Aniline Derivatives.
Transition-metal-catalyzed directed C-H allylation of arenes offers an efficient and straightforward approach to construct value-added allylic arenes. However, these reactions are often performed with precious transition-metal catalysts and mainly limited to ortho-C-H allylation of arenes. Herein, we disclose a novel iron-catalyzed para-C-H allylation of aniline derivatives with allyl alcohols via a chelation-induced strategy, providing various allylic arenes in good yields with excellent regio- and chemoselectivity. A simple FeCl3·6H2O is employed as a catalyst, serving a dual role in the reaction: (1) coordination with N-arylpicolinamide to alter the electronic property of the aromatic ring and (2) reaction with allyl alcohol to form allyl-Fe species.


Lipoxygenase activity in rat embryos and its potential for xenobiotic oxidation.
This paper reports dioxygenase activity in rat embryos and demonstrates for the first time the ability of rat embryo lipoxygenase (LO) to oxidize xenobiotics in vitro. Significant dioxygenase activity towards linoleic acid was found in cytosol isolated from rat embryos in different developmental stages on day 9 and 10 of gestation. All four xenobiotics tested were oxidized at significant rate by the LO in the presence of linoleic acid. Both dioxygenase activity and benzidine oxidation were inhibited by the known inhibitors of LO, i.e. nordihydroguaiaretic acid and eicosatriynoic acid. These findings suggest that LO may be an important pathway for xenobiotic metabolism in the rat embryo.


Bias in Evaluating Erectile Function in Lifelong Premature Ejaculation Patients with the International Index of Erectile Function-5.
Lifelong premature ejaculation (LPE) is the most important ejaculating dysfunction. Relatively little is known about erectile function in this population. We attempted to evaluate the erectile function of patients with LPE using the International Index of Erectile Function-5 (IIEF-5) to determine if it is sufficiently reliable and accurate to make such an assessment. A total of 406 patients with LPE were enrolled in our study. The participants voluntarily answered the Premature Ejaculation Diagnostic Tool (PEDT) and IIEF-5 questionnaires and underwent a full history evaluation and clinical examination by doctors. Their answers were converted into data analyzed by a statistic software. The patients were diagnosed with LPE based on the diagnostic criteria and PEDT scores. The intravaginal ejaculation latency time was recorded according to patient self-reports. The IIEF-5 was used to evaluate their erectile function. Thorough history and clinical examination helped doctors make more correct diagnoses of erectile dysfunction (ED). Of the 406 patients, 70 (17.24%) patients had ED, as confirmed by doctors. IIEF-5 was accurate for the assessment of the erectile function of LPE patients when the cutoff was decreased to 15.5. Question 5 (1.34 ± 0.53) was the main reason for the drop in the total IIEF-5 score. Questions 1 and 5 shared low consistency with the other three IIEF-5 items, thus they lowered the reliability of the IIEF-5 scores. These questions created a confounding bias that decreased the diagnostic threshold of IIEF-5. However, they could not be removed from the IIEF-5 because they did not reduce its diagnostic accuracy in patients with LPE. Bias from questions 1 and 5 lowered the reliability of the IIEF-5 scores; however, it did not reduce the diagnostic accuracy of the IIEF-5. The recommendation is to edit questions 1 and 5 when they are applied to populations with LPE.


Current gene therapy trials for vascular diseases.
In the previous gene therapy trials for vascular diseases, safety of the therapies has been demonstrated with some evidence for clinical benefits. In the future, it will be important to also test the potential clinical benefits of the treatments in randomized and controlled trials with sufficient numbers of patients. This review covers 15 currently ongoing cardiovascular gene therapy trials that aim to treat coronary artery disease, heart failure and peripheral artery disease. This review summarizes current trials and their main features in the cardiovascular field. In the gene therapy trials for vascular diseases, some limiting factors are still present. The trials have enrolled mainly elderly and severely affected patients who might not have the capacity to respond optimally to the therapies. Also, major cardiac adverse events, major amputations, mortality and other very demanding hard clinical end points have been used in relatively small patient populations. Therefore, there is an urgent need to enroll less severely affected patients and to use more informative surrogate end points in the forthcoming clinical trials.


Decrease in the Uptake of Seasonal Influenza Vaccine in Persons Aged 65 Years and Older In Ireland since the 2009 Influenza A (H1N1) Pdm09 Pandemic.
In Ireland, there is year-on-year variation in the seasonal influenza vaccine uptake for people aged 65 years and older. We obtained aggregate vaccine uptake data (2004/05-2014/15) by area and age group and applied the 2011 relative deprivation score and GP-density by area. Using Poisson regression model, we estimated the associations between those variables and uptake before and after the 2009 pandemic. After adjusting for age, deprivation status and GP-density, overall persons aged ?65 years were less likely to receive the vaccine after the pandemic compared to before (RR=0.89, 95%CI: 0.82-0.96). Uptake increased with age; both those in the 70-74 and ?75 year age groups were more likely to receive the vaccine compared to those aged 65-69 years (RR=1.18, 95%CI:1.14-1.24; RR=1.34, 95%CI:1.28-1.39 respectively). Qualitative studies should be undertaken to explore whether changes in public perception on risks of the disease and the vaccine and/or public funding and policy influenced the uptake.


Comparison of Outcomes after Device Closure with Transseptal Puncture and Standard Technique in Patients with Patent Foramen Ovale and Ischemic Events.
The purpose of this study was to compare the effectiveness of device closure with the transseptal puncture and standard technique in patients with patent foramen ovale (PFO) and ischemic events. Eighty-two consecutive patients (men: 60 patients, mean age: 45.2 years) who underwent PFO closure with the Amplatzer PFO Occluder were enrolled. PFO closure with the transseptal puncture was performed in 22 patients (transseptal puncture technique, group I). In the remaining patients (n = 60), PFO closure was performed with the standard technique (group II). The co-primary end points were the incidence of significant residual shunt on follow-up transesophageal echocardiography (TEE) and a composite of death, stroke, transient ischemic attack (TIA), and peripheral embolism. Baseline characteristics were similar between the two groups. On TEE, despite similar grade of interatrial right-to-left shunt, shunt at rest/septal hypermobility was less common in group I than in group II (40.9% vs. 72.9%, P < 0.010). The device was successfully implanted in all patients. On follow-up TEE, significant residual shunt was more common in group I than in group II (28.6% vs. 4.3%, P = 0.021). In addition, composite of death, stroke, TIA, or peripheral embolism was more common in group I than in group II (13.6% vs. 0%, P = 0.017) during the follow-up period (mean 25.4 months). Compared to the standard technique, PFO closure with the transseptal puncture technique showed higher incidence of residual shunt and ischemic events. Therefore, this technique might be considered in only highly selected patients as the last option.


[Trends in caesarean section rates and places of delivery in Burkina Faso].
Objective: The objective of this study was to evaluate the trends, mortality rates and places of caesarean section by level of care in Burkina Faso.Method: A retrospective study was conducted using data from Burkina Faso Ministry of Health annual health statistics and public hospital reports from 2000 to 2014. Linear regression was used to analyse caesarean section trends and mortality rates in District Hospitals (HD), Regional Hospitals (CHR) and University Hospitals (CHU).Results: From 2000 to 2014, the number of caesarean sections performed in Burkina Faso public hospitals increased almost tenfold from approximately 2,365 to 19,081, corresponding to a growth of the caesarean section rate from 0.5 to 2 per 100 deliveries. This growth was linear. Between 2000 and 2006, University Hospitals performed most of these caesarean sections. Starting from 2007, the majority of caesarean sections were performed by HD. The proportion of caesarean sections performed in Regional Hospitals remained almost constant, although the number of caesarean sections increased threefold. In 2014, HD performed 52% of caesarean sections, followed by University Hospitals (25%) and Regional Hospitals (23%). Analysis of mortality rates following caesarean section showed upward and downward trends, oscillating between 1 to 2% before 2008 and 0.15% in 2014. This same mortality trend was observed in each type of hospital.Conclusion: The results show an increase of better quality caesarean section rates, performed closer to the woman's home in Burkina Faso. The health care services decentralization policy and quality of care approaches have helped to achieve these results.


Challenges and future developments in liver transplantation.
Liver transplantation (LT) has become the treatment of choice for a wide range of liver diseases in both adult and pediatric patients. Until recently, the largest proportion of LT in adults, were performed in patients with hepatitis C (HCV) related cirrhosis. The recent availability of safe and effective direct antiviral agents to cure HCV infection in almost all patients whatever the HCV genotype and severity of liver disease, will reduce the need for LT in this category of recipients. Thus, it is presumed that in the next 1 to 2 decades HCV related liver disease will diminish substantially, whereas non-alcoholic steato-hepatitis (NASH) will correspondingly escalate as an indication for LT. The greatest challenges facing LT remain the limited supply of donor organs, and the need for chronic immunosuppression, which represent the true obstacles to the greater application and durable success of the LT procedure. This review aimed to highlight, in different sections, the main open issues and future developments in LT. These will be focused to explore current and future strategies to maximize the use of limited organs, to offer an update on potential new approaches to immunosuppression and to imagine new indications for LT when the number of patients awaiting transplants for HCV related liver disease is reduced.


An on-line investigation of lexical ambiguity processing in schizophrenia.
The processing of lexical ambiguity in context was investigated in eight individuals with schizophrenia and a matched control group. Participants made speeded lexical decisions on the third word in auditory word triplets representing concordant (coin-bank-money), discordant (river-bank-money), neutral (day-bank-money), and unrelated (river-day-money) conditions. When the interstimulus interval (ISI) between the words was 100 ms, individuals with schizophrenia demonstrated priming consistent with selective, context-based lexical activation. At 1250 ms ISI, a pattern of nonselective meaning facilitation was obtained. These results suggest an attentional breakdown in the sustained inhibition of meanings on the basis of lexical context.


Automated 3-D Printed Arrays to Evaluate Genotoxic Chemistry: E-Cigarettes and Water Samples.
A novel, automated, low cost, three-dimensional (3-D) printed microfluidic array was developed to detect DNA damage from metabolites of chemicals in environmental samples. The electrochemiluminescent (ECL) detection platform incorporates layer-by-layer (LbL) assembled films of microsomal enzymes, DNA and an ECL-emitting ruthenium metallopolymer in ∼10 nm deep microwells. Liquid samples are introduced into the array, metabolized by the human enzymes, products react with DNA if possible, and DNA damage is detected by ECL with a camera. Measurements of relative DNA damage by the array assess the genotoxic potential of the samples. The array analyzes three samples simultaneously in 5 min. Measurement of cigarette and e-cigarette smoke extracts and polluted water samples was used to establish proof of concept. Potentially genotoxic reactions from e-cigarette vapor similar to smoke from conventional cigarettes were demonstrated. Untreated wastewater showed a high genotoxic potential compared to negligible values for treated wastewater from a pollution control treatment plant. Reactivity of chemicals known to produce high rates of metabolite-related DNA damage were measured, and array results for environmental samples were expressed in terms of equivalent responses from these standards to assess severity of possible DNA damage. Genotoxic assessment of wastewater samples during processing also highlighted future on-site monitoring applications.


Within Session Sequence of Balance and Plyometric Exercises Does Not Affect Training Adaptations with Youth Soccer Athletes.
The integration of balance and plyometric training has been shown to provide significant improvements in sprint, jump, agility, and other performance measures in young athletes. It is not known if a specific within session balance and plyometric exercise sequence provides more effective training adaptations. The objective of the present study was to investigate the effects of using a sequence of alternating pairs of exercises versus a block (series) of all balance exercises followed by a block of plyometric exercises on components of physical fitness such as muscle strength, power, speed, agility, and balance. Twenty-six male adolescent soccer players (13.9 ± 0.3 years) participated in an 8-week training program that either alternated individual balance (e.g., exercises on unstable surfaces) and plyometric (e.g., jumps, hops, rebounds) exercises or performed a block of balance exercises prior to a block of plyometric exercises within each training session. Pre- and post-training measures included proxies of strength, power, agility, sprint, and balance such as countermovement jumps, isometric back and knee extension strength, standing long jump, 10 and 30-m sprints, agility, standing stork, and Y-balance tests. Both groups exhibited significant, generally large magnitude (effect sizes) training improvements for all measures with mean performance increases of approximately >30%. There were no significant differences between the training groups over time. The results demonstrate the effectiveness of combining balance and plyometric exercises within a training session on components of physical fitness with young adolescents. The improved performance outcomes were not significantly influenced by the within session exercise sequence.


COVID-19: A Geriatric Emergency.
The older Italian population is posing a challenge in the number of deaths for coronavirus disease 2019 (COVID-19). According to previous data from China, pre-existing health conditions dramatically increase the risk of dying from COVID-19. The presence of multiple diseases in older patients may be considered as a mark of frailty, which increases the person's vulnerability to stress and impairs the multisystemic compensatory effort to restore homeostasis. The clinical complexity associated with the management of frailty may increase the risk of complications during infection as well as the lack of the early recognition of atypical symptoms. There is an urgent need to share expertise and clinical management skills with geriatricians as well as the need for early diagnosis to start treatment at the earliest convenience in the community, with the aim to avoid the collapse of intensive care units.


Evaluation of the effects of nanoparticles on the therapeutic function of platelet: a review.
Nanotechnology and nanoparticles are used in different applications in disease monitoring and therapy in contact with blood. Nanoparticles showed different effects on blood components and reduced or improved the function of therapeutic platelet during the storage time. This review study was performed to evaluate the impacts of various sizes and charges of nanoparticles on platelet function and storage time. The present review contains the literature between 2010 and 2020. The data have been used from different sites such as PubMed, Wiley, ScienceDirect and online electronic journals. From the literature survey, it has been demonstrated that among various properties, size and charge of nanoparticles were critical on the function of therapeutic platelet during the storage and inhibition of their aggregation. Overall, this study described that nanoparticles with smaller size and negative charge were more effective in increasing the survival time, inhibition of aggregation and improving the function of therapeutic platelet. Based on the current review, it can be confirmed that nanoparticles such as dendrimer, Au, Ag and iron oxide nanoparticles with smaller size and negative charge have significant advantages for improving the efficacy of platelets during the storage chain and inhibition of their aggregation.


[Prosthetic Joint Infection after Total Hip Arthroplasty: Results of an Online Questionnaire about Current Diagnostic and Therapeutic Procedures in 107 German Hospitals].
Background Prosthetic hip joint infection is a common severe complication with a high socio-economic impact. The inconsistency of the available data and the absence of binding guidelines lead to a variety of diagnostic and therapeutic strategies. The aim of this study is to present the current diagnostic and therapeutic approach to treating infections after total hip arthroplasties in German hospitals, link it with current evidence, and evaluate the willingness of these hospitals to participate in prospective multicentre trials. Material and Methods An online questionnaire for digital processing was sent to hospitals performing joint replacement procedures. These institutions included district hospitals, private hospitals, non-university maximum care facilities, statutory accident insurance hospitals, and university hospitals. Results A total of 107 hospitals took part in the survey, corresponding to a response rate of 27.6%. These hospitals perform approximately 2,951 revisions of infected total hip arthroplasties per year. Two-stage revision arthroplasty is the preferred procedure after prosthetic hip infections. The algorithm proposed by Zimmerli et al. is widely recognised. There is a lack of uniformity in the key features for revision of prosthetic joint infections - long vs. short interval in case of two-stage revision, duration of intravenous and oral administration of antibiotics, cemented vs. cement-free implant procedures, and follow-up intervals after revision surgery. The willingness to participate in clinical trials is high. Conclusion The controversial data leads to multiple treatment approaches. The high willingness to participate in adequately funded clinical trials offers a potential for multicentre trials to be conducted. There is an urgent need for funding to make this research possible.


Hysteroscopy in the evaluation of patients with recurrent pregnancy loss: a cohort study in a primary care population.
The purpose of this study was to determine the prevalence of structural uterine defects (SUD) in patients with recurrent pregnancy loss (RPL) attending a large family medicine practice and to determine the effectiveness of the therapeutic intervention. This prospective cohort study from October 1995 to October 1998 included 23 patients aged 23 to 35 years (mean, 28.1) with an otherwise unexplained history of three or more 1st- or 2nd-trimester miscarriages and no live births. None of the couples were tobacco or alcohol users, all of them and had normal cytogenetic examinations and testing. Their jobs were unrelated to chemical handling. All patients had a complete history taken and underwent physical examination and hysteroscopy with directed biopsy. They had normal values for complete blood count, sedimentation rate, urinalysis and culture (gonorrhea, Chlamydia, syphilis, hepatitis B, HIV), lupus anticoagulant, anticardiolipin antibodies, spermiogram, progesterone, and pelvic ultrasound. Fourteen patients (60.9%) had a normal hysteroscopy (with biopsies). Nine patients (39.1 %) had SUD, as follows: Five patients (21.8%) had intrauterine adhesions, two patients (8.7%) had a septated uterus, one patient (4.3%) had submucosal myoma, and one patient (4.3%) had multiple factors. After appropriate therapy of the SUD, seven patients (77.8%) achieved successful pregnancy and two patients (22.2%) had recurrent miscarriage. In the normal hysteroscopy group, eight patients (57.1%) had recurrent miscarriages, two patients (14.3%) had infertility, and four patients (28.6%) achieved successful pregnancy with no further therapy. The prevalence of SUD in our studied population was 39.1 %. After appropriate treatment, the rate of live-birth pregnancies in these patients was 77.8%. Because SUD are the most treatable cause of RPL, these patients should be identified early after other potential causes of RPL are eliminated.


Toxicity assessment of combined fluoroquinolone and tetracycline exposure in zebrafish (Danio rerio).
Fluoroquinolones (FQs) and tetracyclines (TCs), the two β-diketone antibiotics (DKAs), are two frequently detected pollutants in the environment; however, little data are available on their combined toxicity to zebrafish (Danio rerio). This study reports that toxicologic effects of combined DKA (FQs-TCs) exposure on zebrafish were comparable with or slightly less than those of TCs alone, showing that TCs played a major toxicologic role in the mixtures. The effects of FQs, TCs, and DKAs on malformation rates of zebrafish were dose dependent, with EC50 values of 481.3, 16.4, and 135.1 mg/L, respectively. According to the combined effects of DKAs on zebrafish hatching, mortality, and malformation rates, the interaction between FQs and TCs was shown to be antagonistic based on three assessment methods: Toxic Unit, Additional Index, and Mixture Toxic Index. The 1.56 mg/L TC and 9.38 mg/L DKA treatments resulted in higher zebrafish basal swimming rate compared with the control group at 120 hours postfertilization (hpf). in both light and light-to-dark photoperiod experiments. Under conditions of no obvious abnormality in cardiac development, the heart beats were decreased significantly because of DKA exposure, such as decreasing by ∼20% at 150 mg/L DKAs. Transmission electron microscopy observation of myocytes from DKA-exposed hearts displayed prominent interruptions and myofibrillar disorganization of the normal parallel alignment of thick and thin filaments, and partial edematous and dissolved membranes of cell nuclear tissues. At 90 mg/L DKAs, the transcriptional levels of the acta1a, myl7, and gle1b genes, related to heart development and skeletal muscle formation, were significantly changed. This is consistent with the swimming behavior and histopathologic results obtained by transmission electron microscopy. In summary, the toxicity of the combined DKAs to zebrafish was comparable with or less than that of TCs alone and had the ability to impair individual behaviors that are of great importance in the assessment of their ecologic fitness. © 2014 Wiley Periodicals, Inc. Environ Toxicol 31: 736-750, 2016.


Two-dimensional transferred nuclear Overhauser effect spectroscopy (TRNOESY) studies of nucleotide conformations in arginine kinase complexes.
Two-dimensional proton transfer nuclear Overhauser spectroscopy (TRNOESY) studies of the conformations of the adenosine moieties in the nucleotides bound at the active site of the lobster (Homarus americanus) muscle arginine kinase are reported. TRNOESY measurements were made using a sample protocol chosen to minimize contributions from weak non-specific binding of the nucleotides to the observed NOE's. This was done by making the measurements as a function of ligand concentration while keeping the ligand to enzyme concentration ratio fixed at 10:1. The experiments were performed at 500 MHz and 10 degrees C for six different mixing times in the range 40-300 ms. The measurements were made on three complexes of the enzyme: E.MgATP, E.MgADP, and the long-lived transition-state-analog complex (E.MgADP.NO-3.arginine). All the complexes, including the transition-state-analog complex with an estimated lifetime of about 50 ms, satisfy the fast-exchange condition. The TRNOE buildup curves for all the nucleotide-proton pairs in each complex were analyzed using a complete relaxation matrix appropriate for fast exchange. The interproton distances obtained from the NOE analysis were used as constraints in obtaining an energy-minimized conformation on the basis of the program CHARMm. The glycosidic torsion angle (chi) for the adenosine moiety in all three complexes is about 50 degrees +/- 5 degrees. The glycosidic orientation agrees well with that determined for MgATP and MgADP complexes of creatine kinase (Murali et al., 1993), MgATP bound at the active and ancillary sites of pyruvate kinase (Jarori et al., 1994a), and PRPP synthetase (Jarori et al., 1994b).(ABSTRACT TRUNCATED AT 250 WORDS)


Verified the rapid evaluation of the edible safety of wild porcini mushrooms, using deep learning and PLS-DA.
How to quickly identify poisonous mushrooms is a worldwide problem, because poisonous mushrooms and edible mushrooms have very similar appearances. Even some edible mushrooms must be processed further before they can be eaten. In addition, mushrooms from different geographical origins contain different levels of heavy metals. Eating frequent mushrooms with excessive heavy metal content can also cause food poisoning. This information is very important and needs to be informed to consumers in advance. Through the demand for the safety of porcini mushrooms in the Yunnan area we propose a hierarchical identification system based on Fourier-transform near-infrared (FT-NIR) spectroscopy to evaluate the edible safety of porcini species. We found that deep learning is the most effective means to identify the edible safety of porcini, and the recognition accuracy was 100%, by comparing two pattern recognition tools, deep learning and partial least square discriminant analysis (PLS-DA). Although the accuracy of the PLS-DA test set is 96.10%, the poisonous porcini is not allowed to be wrongly judged. In addition, the cadmium (Cd) content of Leccinum rugosiceps in the Midu area exceeded the standard. Deep learning can trace Le. rugosiceps geographic origin with an accuracy of 100%. The overall results show that deep learning methods based on FT-NIR can identify porcini that is at risk of being eaten. This has useful application prospects in food safety. © 2021 Society of Chemical Industry.


Spatial resolution study and power calibration of the high-k scattering system on NSTX.
NSTX high-k scattering system has been extensively utilized in studying the microturbulence and coherent waves. An absolute calibration of the scattering system was performed employing a new millimeter-wave source and calibrated attenuators. One of the key parameters essential for the calibration of the multichannel scattering system is the interaction length. This interaction length is significantly different from the conventional one due to the curvature and magnetic shear effect.


Test-retest reliability of mandibular morphology measurements on cone-beam computed tomography-synthesized cephalograms with random head positioning errors.
Cephalometric radiography has been used for orthodontic and surgical treatment planning and assessment, and for quantifying mandibular growth. However, it remains unclear how head positioning errors and the level of examiner experience affect the reliability of such morphometric measurements. The current study aimed to bridge the gap by determining the intra-, inter-rater, and inter-session reliability of measurements of mandibular morphology with random head positioning errors as measured by a junior and a senior dentist. Cone-beam computed tomography data of twelve mandibles were obtained with each rotated randomly away from the neutral position within the range of +3 and -3° along each of the anatomical axes to simulate six imaging trials. A synthetic cephalogram for each trial was obtained via a digitally reconstructed radiography (DRR) technique and eleven landmarks for twelve morphological parameters on the cephalogram were identified manually six times by a junior and a senior dentist. The procedure was repeated on another day within 5 days. Test-retest reliability was assessed in terms of an intra-class correlation coefficient (ICC) using a two-way mixed-effects model. Good to very good intra-rater (senior: ICC > 0.92; junior: ICC > 0.78), inter-rater (ICC > 0.70 for most parameters) and inter-session reliability (senior: ICC > 0.84; junior: ICC > 0.62) were found. Bland & Altman plots of inter-rater comparisons show that there were systematical biases between the examiners on most parameters, except for the distance between Gonion and Pogonion. The current results suggest that good to very good intra-rater, inter-rater and inter-session reliability can be achieved for most parameters with randomized head positioning errors; higher inter-session reliability can be achieved by more experienced examiners; and that long-term monitoring of mandibular growth based on cephalographic measurements should be made by the same more experienced examiner. The current DRR-based approach can be used to evaluate individual factors that affect the morphological measurements.


A genealogy of tubercular diseases in Japan.
This paper examines the historical transformation of concepts of tubercular diseases in Japan from premodern to modern times. Its principal conclusion is that, between the eighteenth and the twentieth centuries, intellectual and cultural circumstances in Japan first facilitated and then hindered the adoption of Western concepts of tubercular diseases. The Japanese adopted Western concepts of tubercular diseases from the early nineteenth century not because they suggested more effective therapeutic measures but because they mirrored ideas that the Japanese had developed during the previous century. During the late nineteenth and early twentieth centuries concepts of tubercular disease based on Western medical thought conflicted with popular concepts, creating a point of cultural dissonance in Japanese society that was overcome only after effective medicinal cures for tuberculosis were invented. To demonstrate these points, this paper surveys representative Chinese, Japanese, and European medical texts and ideas from earliest times to the first half of the twentieth century. Although the focus is on one specific set of diseases, this example helps explain why the Japanese adopted Western medicine much more quickly than their cultural neighbours in China and Korea, although this conclusion is left implied rather than stated.


BiDil for heart failure in black patients: implications of the U.S. Food and Drug Administration approval.
In 2005, the combination of hydralazine hydrochloride and isosorbide dinitrate was approved by the U.S. Food and Drug Administration (FDA) for treating heart failure in black patients. In departing from its long history of approving drugs for general clinical indications without regard to demographic classification, the FDA cited the need to address racial disparities in health as an important contributor to their decision. The authors argue that this decision, although perhaps well-intentioned, was based on flawed scientific interpretation of trial results that claimed differential drug response by race and ignored the considerable literature on the cause of racial disparities in health and health care. Because of its potential impact on future drug approvals, the FDA's decision is a setback in the scientific and policy discourse on medical therapeutics and race and specifically hinders the efforts aimed at eliminating health and health care disparities.


Formation of high density lipoproteins containing both apolipoprotein A-I and A-II in the rabbit.
Human plasma HDLs are classified on the basis of apolipoprotein composition into those that contain apolipoprotein A-I (apoA-I) without apoA-II [(A-I)HDL] and those containing apoA-I and apoA-II [(A-I/A-II)HDL]. ApoA-I enters the plasma as a component of discoidal particles, which are remodeled into spherical (A-I)HDL by LCAT. ApoA-II is secreted into the plasma either in the lipid-free form or as a component of discoidal high density lipoproteins containing apoA-II without apoA-I [(A-II)HDL]. As discoidal (A-II)HDL are poor substrates for LCAT, they are not converted into spherical (A-II)HDL. This study investigates the fate of apoA-II when it enters the plasma. Lipid-free apoA-II and apoA-II-containing discoidal reconstituted HDL [(A-II)rHDL] were injected intravenously into New Zealand White rabbits, a species that is deficient in apoA-II. In both cases, the apoA-II was rapidly and quantitatively incorporated into spherical (A-I)HDL to form spherical (A-I/A-II)HDL. These particles were comparable in size and composition to the (A-I/A-II)HDL in human plasma. Injection of lipid-free apoA-II and discoidal (A-II)rHDL was also accompanied by triglyceride enrichment of the endogenous (A-I)HDL and VLDL as well as the newly formed (A-I/A-II)HDL. We conclude that, irrespective of the form in which apoA-II enters the plasma, it is rapidly incorporated into spherical HDLs that also contain apoA-I to form (A-I/A-II)HDL.


Helping Children with Obesity "Move Well" To Move More: An Applied Clinical Review.
Children with obesity experience musculoskeletal pain and reduced physical function and well-being, which collectively impact their fitness, strength, motor skills, and even their ability to undertake simple tasks, like walking and climbing stairs. Disrupting obesity-related disability may be critical to increasing children's physical activity. Thus, barriers to movement should be considered by health practitioners to improve the efficacy of prescribed physical activity. This applied clinical review highlights key subjective and objective findings from a hypothetical case scenario, linking those findings to the research evidence, before exploring strategies to enhance movement and increase physical activity.


The effect of short implants placed in the posterior region on tilted implants in the 'All-On-Four' treatment concept: a three-dimensional finite element stress analysis.
This study aims to investigate the impact of a short implant placed behind the mental foramen with the all-on-four treatment concept using the three-dimensional (3D) finite element stress analysis (FEA).Six different finite element analysis models were designed according to tilted implant angle (17° and 30°), presence of short implants, and short implant diameter (4.1 mm and 4.8 mm). A 100 N force was applied vertically from the central fossa of the lower right second premolar tooth. Maximum equivalent (von Mises) and Minimum/Maximum principal (Pmin/Pmax) stress values and distributions were evaluated by 3D-FEA.The highest stress value among tilted implants was in the T17 group. T30 was the group that caused the most stress in the cortical bone. Adding the short implant to the all-on-four design reduced von Mises stress on multi-unit abutments, abutment screws, and tilted implants in both the 17° and 30° groups. At the same time, it reduced Pmin/Pmax stresses in cortical bone. Similar behavior was observed in terms of stress values and distributions for the 4.1 and 4.8 mm short implant groups.The results show that short implant placement in the posterior region in the all-on-four concept reduces stress on the bone, implants, and prosthetic parts, regardless of the diameter of the short implant. In cases where biomechanical risks such as parafunctional habits and poor bone quality increase, we recommend increasing the number of implants with short implants.


Identification and characterization of the interactive proteins with cytotoxic T-lymphocyte antigen-2α.
Cytotoxic T-lymphocyte antigen-2α (CTLA-2α) is a potent inhibitor of cathepsin L-like cysteine proteases. Recombinant CTLA-2α is known to be a potent, competitive inhibitor of cathepsin L-like cysteine proteases. In this study, cathepsin L, cathepsin C, and tubulointerstitial nephritis antigen-related protein 1 (TINAGL1) were identified as novel interactive proteins of CTLA-2α by the yeast two-hybrid screening system. The direct interactions and co-localization of these proteins with CTLA-2α were confirmed using co-immunoprecipitation and immunofluorescence staining, respectively. The disulfide-bonded CTLA-2α/cathepsin L complex was isolated from mouse tissue. CTLA-2α was found to be specific and consistently expressed on the maternal side of the mouse placenta. Double immunofluorescence analysis showed that CTLA-2α was co-localized with cathepsin L, cathepsin C, and TINAGL1 in placenta. A simple cell-based fluorescence assay revealed that CTLA-2α exhibited inhibitory activity toward cathepsin C in live cells, which indicated that CTLA-2α is a novel endogenous inhibitor of cathepsin C.


Manganese superoxide dismutase deficiency enhances cell turnover via tumor promoter-induced alterations in AP-1 and p53-mediated pathways in a skin cancer model.
Previous studies in our laboratories demonstrated that overexpression of manganese superoxide dismutase (MnSOD) suppressed both the incidence and multiplicity of papillomas in a DMBA/TPA multi-stage skin carcinogenesis model. The activity of activator protein-1 (AP-1), which is associated with tumor promotion, was reduced in MnSOD transgenic mice overexpressing MnSOD in the skin, suggesting that MnSOD may reduce tumor incidence by suppressing AP-1 activation. In the present study, we report that reduction of MnSOD by heterozygous knockout of the MnSOD gene (Sod2 -/+, MnSOD KO) increased the levels of oxidative damage proteins and the activity of AP-1 following TPA treatment. RNA levels of ornithine decarboxylase (ODC) were also increased, suggesting an increase in cell proliferation in the KO mice. Histological examination confirmed that the number of proliferating cells in DMBA/TPA-treated mouse skin were higher in the KO mice. Interestingly, histological examination also demonstrated greater numbers of apoptotic cells in the KO mice after DMBA/TPA treatment. Evidence of apoptosis, including DNA fragmentation, cytochrome c release from mitochondria, and caspase 3 activation were also observed by biochemical assays of the skin tissues. Apoptosis was associated with an increase in nuclear levels of p53 as determined by Western analysis. Quantitative immunogold ultrastructural analysis confirmed that p53 immunoreactive protein levels were increased to a greater level in the nuclei of epidermal cells from MnSOD KO mice compared to epidermal nuclei from wild type mice similarly treated. Moreover, p53 levels further increased in the mitochondria of DMBA/TPA treated mice, and this increase was much greater in the MnSOD KO than in the wild type mice, suggesting a link between MnSOD deficiency and mitochondrial-mediated apoptosis. Pathological examination reveals no difference in the incidence and frequency of papillomas comparing the KO mice and their wild type littermates. Taken together, these results suggest that: (1) MnSOD deficiency enhanced TPA-induced oxidative stress and AP-1 and p53 levels, consistent with the increase in both proliferation and apoptosis events in the MnSOD KO mice, and (2) increased apoptosis may negate increased proliferation in the MnSOD deficient mice during an early stage of tumor development.


Genetic diversity and dynamic distribution of Mycobacterium tuberculosis isolates causing pulmonary and extrapulmonary tuberculosis in Thailand.
This study examined the genetic diversity and dynamicity of circulating Mycobacterium tuberculosis strains in Thailand using nearly neutral molecular markers. The single nucleotide polymorphism (SNP)-based genotypes of 1,414 culture-positive M. tuberculosis isolates from 1,282 pulmonary tuberculosis (PTB) and 132 extrapulmonary TB (EPTB) patients collected from 1995 to 2011 were characterized. Among the eight SNP cluster groups (SCG), SCG2 (44.1%), which included the Beijing (BJ) genotype, and SCG1 (39.4%), an East African Indian genotype, were dominant. Comparisons between the genotypes of M. tuberculosis isolates causing PTB and EPTB in HIV-negative cases revealed similar prevalence trends although genetic diversity was higher in the PTB patients. The identification of 10 reported sequence types (STs) and three novel STs was hypothesized to indicate preferential expansion of the SCG2 genotype, especially the modern BJ ST10 (15.6%) and ancestral BJ ST19 (13.1%). An association between SCG2 and SCG1 genotypes and particular patient age groups implies the existence of different genetic advantages among the bacterial populations. The results revealed that increasing numbers of young patients were infected with M. tuberculosis SCGs 2 and 5, which contrasts with the reduction of the SCG1 genotype. Our results indicate the selection and dissemination of potent M. tuberculosis genotypes in this population. The determination of heterogeneity and dynamic population changes of circulating M. tuberculosis strains in countries using the Mycobacterium bovis BCG (bacillus Calmette-Guérin) vaccine are beneficial for vaccine development and control strategies.


[Role of ICS/LABA on COPD treatment].
In the treatment of chronic obstructive pulmonary disease (COPD), bronchodilators such as long acting muscarinic antagonist (LAMA) and long acting β agonist(LABA) play key roles for improving respiratory function and symptoms, and reducing risk of exacerbation. However, inhaled corticosteroid (ICS), a key medicine for bronchial asthma, is limitedly used in COPD treatment. Japanese Respiratory Society recommends to use ICS for severe COPD patients who have been frequently exacerbated, because previous clinical studies indicated that ICS reduces exacerbation in moderate to severe COPD patients. Asthma sometimes overlaps with COPD, and symptoms of those patients are not well controlled by the bronchodilation therapy alone. Therefore, ICS/LABA or ICS/LAMA should be prescribed to those overlapped patients. Concentration of exhaled nitrogen oxide and percentage of peripheral eosinophil may be good biomarkers for discriminating the COPD patients who have good response to ICS treatment.


Effect of physical exercise prelabyrinthectomy on locomotor balance compensation in the squirrel monkey.
This study examines the effectiveness of physical exercise, during a prepathology state, on locomotor balance compensation after subsequent unilateral labyrinthectomy in squirrel monkeys. An experimental group underwent 3 hr. of daily running exercise on a treadmill for 3 mo. prior to the surgery, whereas a control group was not exercised. Postoperatively, the locomotor balance function of both groups was tested for 3 mo. There was no significant difference in gait deviation counts in the acute phase of compensation. However, in the chronic compensation maintenance phase, the number of gait deviation counts was fewer in the exercise group, which showed significantly better performance stability.


Biological effects of a new ultraviolet A1 prototype based on light-emitting diodes on the treatment of localized scleroderma.
Ultraviolet A1 (UVA1 ) phototherapy (spectral range 340-400 nm) is a well-established treatment option for various skin diseases such as localized scleroderma. Recent improvements of conventional UVA1 light sources (metal-halide or fluorescent lamps) have brought attention to a new light-emitting diode (LED) technology with remarkable advantages in handling and clinical routine. This study provides a preclinical histological and molecular evaluation of an LED-based UVA1 prototype with a narrower spectral range (360-400 nm) for treating localized scleroderma. Scleroderma mouse models and fibroblasts in vitro were exposed to LED-based UVA1 phototherapy or to irradiation with a commercially available metal-halide lamp emitting low-dose (20, 40 J/cm2 ), medium-dose (60 J/cm2 ) and high-dose (80, 100 J/cm2 ) UVA1 light. Both UVA1 light sources affected inflammatory genes (IL-1α and IL-6) and growth factors (TGFß-1 and TGFß-2). Increased collagen type 1 was reduced after UVA1 phototherapy. Matrix metalloproteinase-1 was more enhanced after a medium dose of LED-based UVA1 phototherapy than after conventional treatment. In vivo, dermal thickness and the amount of collagen were reduced after both treatment methods. Remarkably, myofibroblasts were more effectively reduced by a medium dose of LED-based UVA1 phototherapy. The study indicates that LED-based UVA1 phototherapy yields similar or even better results than conventional treatment. In terms of biosafety and patient comfort, LED-based UVA1 phototherapy offers clear advantages over conventional treatment because of the use of a narrower and less harmful UVA1 spectrum, less heat generation and shorter treatment times at the same irradiation intensity. Clinical studies are required to confirm these results in patients with localized scleroderma.


Effects of tooth preparation burs and luting cement types on the marginal fit of extracoronal restorations.
Although surface roughness of axial walls could contribute to precision of a cast restoration, it is unclear how the roughness of tooth preparation affects marginal fit of the restoration in clinical practice. The purpose of this study was to describe the morphologic features of dentin surfaces prepared by common rotary instruments of similar shapes and to determine their effects on the marginal fit for complete cast crowns. Ninety crowns were cast for standardized complete crown tooth preparations. Diamond, tungsten carbide finishing, and crosscut carbide burs of similar shape were used (N = 30). The crowns in each group were subdivided into three groups (n = 10) for use with different luting cements: zinc phosphate cement (Fleck's), glass ionomer cement (Ketac-Cem), and adhesive resin cement (Panavia 21). Marginal fit was measured with a light microscope in a plane parallel to the tooth surface before and after cementation between four pairs of index indentations placed at equal distances around the circumference of each specimen. Difference among groups was tested for statistical significance with analysis of variance (ANOVA) followed by Ryan-Einot-Gabriel-Welsch Multiple Range Test (alpha= 0.05). Analysis of measurements disclosed a statistically significant difference for burs used to finish tooth preparations (p < 0.001); however, luting cement measurements were not significantly different (p= 0.152). Also, the interaction effect was not significantly different (p= 0.685). For zinc phosphate cement, the highest marginal discrepancy value (100 +/- 106 microm) was for tooth preparations refined with carbide burs, and the lowest discrepancy value (36 +/- 30 microm) was for tooth preparations refined with finishing burs. For glass ionomer cement, the highest marginal discrepancy value (61 +/- 47 microm) was for tooth preparations refined with carbide burs, and the lowest discrepancy value (33 +/- 40 microm) was for tooth preparations refined with finishing burs. For adhesive resin cement, the highest marginal discrepancy value (88 +/- 81 microm) was for tooth preparations refined with carbide burs, and the lowest discrepancy value (19 +/- 17 microm) was for tooth preparations refined with finishing burs. Marginal fit of complete cast crowns is influenced by tooth preparation surface characteristics, regardless of the type of luting agent used for cementation. Tooth preparations refined with finishing burs may favor the placement of restorations with the smallest marginal discrepancies, regardless of the type of cement used.


Comparing problem-based learning with case-based learning: effects of a major curricular shift at two institutions.
Problem-based learning (PBL) is now used at many medical schools to promote lifelong learning, open inquiry, teamwork, and critical thinking. PBL has not been compared with other forms of discussion-based small-group learning. Case-based learning (CBL) uses a guided inquiry method and provides more structure during small-group sessions. In this study, we compared faculty and medical students' perceptions of traditional PBL with CBL after a curricular shift at two institutions. Over periods of three years, the medical schools at the University of California, Los Angeles (UCLA) and the University of California, Davis (UCD) changed first-, second-, and third-year Doctoring courses from PBL to CBL formats. Ten months after the shift (2001 at UCLA and 2004 at UCD), students and faculty who had participated in both curricula completed a 24-item questionnaire about their PBL and CBL perceptions and the perceived advantages of each format A total of 286 students (86%-97%) and 31 faculty (92%-100%) completed questionnaires. CBL was preferred by students (255; 89%) and faculty (26; 84%) across schools and learner levels. The few students preferring PBL (11%) felt it encouraged self-directed learning (26%) and valued its greater opportunities for participation (32%). From logistic regression, students preferred CBL because of fewer unfocused tangents (59%, odds ration [OR] 4.10, P = .01), less busy-work (80%, OR 3.97, P = .01), and more opportunities for clinical skills application (52%, OR 25.6, P = .002). Learners and faculty at two major academic medical centers overwhelmingly preferred CBL (guided inquiry) over PBL (open inquiry). Given the dense medical curriculum and need for efficient use of student and faculty time, CBL offers an alternative model to traditional PBL small-group teaching. This study could not assess which method produces better practicing physicians.


Reversible binding of heme to proteins in cellular signal transduction.
Heme plays critical roles in numerous biological phenomena. Recent evidence has uncovered a new role of heme in cellular signal transduction, and its mechanism involves reversible binding of heme to proteins. This Account highlights the novel function of heme as an intracellular messenger in the regulation of gene expression and ion channel function.


The effectiveness of a magnetized water oral irrigator (Hydro Floss) on plaque, calculus and gingival health.
The purpose of this study was to evaluate the effects of a magnetized water oral irrigator on plaque, calculus and gingival health. 29 patients completed this double-blind crossover study. Each patient was brought to baseline via an oral prophylaxis with a plaque index < or = 1 and a gingival index < or = 1. Subjects used the irrigator for a period of 3 months with the magnet and 3 months without the magnet. After each 3 month interval, data were collected using the plaque index, gingival index, and accretions index. The repeated measures analysis on plaque, gingival and calculus indices yielded a statistically-significant period effect for PlI (p=0.0343), GI (p=0.0091), and approached significance for calculus (p=0.0593). This meant that the effect of irrigation resulted in a decrease of all indices over time. Therefore, the treatment effect on each index was evaluated using only the measurements obtained at the end of the first period (i.e., assuming a parallel design). Irrigation with magnetized water resulted in 64% less calculus compared to the control group. The reduction was statistically significant (p< or =0.02). The reduction by 27% in gingival index was not statistically significant. The reduction in plaque was minimal (2.2%). A strong positive correlation between the plaque index and the Watt accretion index was observed. The magnetized water oral irrigator could be a useful adjunct in the prevention of calculus accumulation in periodontal patients, but appears to have minimal effect on plaque reduction. The results indicated a clinical improvement in the gingival index, but this was not a statistically significant finding.


[Genome-wide identification and analysis of heat shock protein 90 in tomato].
Heat shock proteins 90 (Hsp90) are a kind of specific proteins in plant which were produced under environmental stresses. By referring to the tomato genome database, we identified and analyzed Hsp90 gene family members using bioinformatics methods. Results indicated that the tomato genome contained at least 7 Hsp90 genes, which were distributed unevenly on 6 chromosomes. Amino acid sequence length of these proteins ranged from 267 to 794aa. Numbers of intron ranged from 2 to 19. Microsynteny analysis showed that two pairs of Hsp90 genes (Hsp90-1and Hsp90-3, Hsp90-5 and Hsp90-7) were identified by segment duplication. In addition, multiple conservation motifs were found in Hsp90 proteins. Phylogenetic analysis revealed that Hsp90 genes from tomato, rice and Arabidopsis can be divided into 5 groups. Three pair of orthologous genes and four pairs of homologous genes were found. Expression analysis based on RNA-seq showed that the expression of three genes (Hsp90-5, Hsp90-6 and Hsp90-7) was high in vegetable and reproductive organs, while the expression of other four genes (Hsp90-1, Hsp90-2, Hsp90-3 and Hsp90-4) was relatively low except for its expression at the breaking stage of fruit. Analysis of promoter regions of Hsp90 genes showed that multiple cis-elements were involved in plant responses to biotic and abiotic stresses. The expression of 7 genes under heat stress was also detected by qRT-PCR. Expression of all Hsp90 genes in tomato leaf was enhanced. The results indicated that these genes could be participated in tomato leaf response to heat stresses. Together, these results will lay a foundation for analyzing Hsp90 gene function and molecular evolution in the future.


[Levels of cytokeratin CK19 expression in mononuclear blood cells evaluated using a reverse PCR (RT-PCR)].
The sensitivity and specificity of a reverse transcription PCR method (RT-PCR) to detect cytokeratin K19 (CK19) expression was evaluated with the purpose of assessing its capability to detect the presence of breast cancer tumour cells in peripheral blood progenitor cell collection that had to be reinfused to breast cancer patients submitted to intensive chemotherapy as haematopoietic support. Two breast cancers as positive samples were used and 34 samples of mononucleated blood cells as negative controls: 18 peripheral blood samples from normal subjects, 14 from different types of leukaemias (M3, M4Eo, M2, etc.) and two from two patients with Hodgkin's lymphoma. The method studied is a nested RT-PCR that amplifies the CK19 expression from the sample RNA extracted following the method of phenol-chloroform. The right performance of the method is demonstrated by observing the detection of CK19 transcripts in the breast cancer RNA and by obtaining good blank results both with non transcribed RNA and with DNA. Moreover, the method has an excellent sensitivity such as to allow the detection of CK19 transcripts in a 10(-6) dilution of cDNA reverse transcribed from 1 microgram of breast cancer RNA. The CK19 transcripts were also detected in the 64% of RNA obtained from the mononucleated blood cells controls, although the percentage of positivities was lower (47%) in the RNA from peripheral blood samples. Nevertheless it should be remarked that the levels of CK19 expression in the blood mononucleated cells is almost negligible since it used to extinguish at 1:5 cDNA dilution. The method studied is specific and has a high sensitivity that explains the detection of CK19 illegitimate expression approximately a half in mononucleated blood cells negative controls. However, the levels of CK19 expression in mononucleated blood cells were almost negligible and it used to extinguish at 1:5 cDNA dilution, therefore it could be concluded that the method might be useful to detect breast cancer occult tumours cells in mononucleated blood cell collection, always provided that a lower amount of cDNA is taken, thus decreasing to nil almost the false positive samples and keeping always a good sensitivity.


Serum cardio-specific troponin T after open heart surgery in patients with and without perioperative myocardial infarction.
One hundred and sixteen consecutive patients undergoing open heart surgery were studied to evaluate the diagnostic use of cardiac specific troponin T in serum (S-TnT) measured preoperatively, at day 1 and day 4 postoperatively. The results were related to perioperative myocardial infarction (POMI), diagnostically based on ECG-changes, as well as to other perioperative variables. Cardiac surgery resulted in increased levels of S-TnT day 1 in all patients, and the level of this increase was dependent on the type of surgical procedure performed and the duration of cardiac perioperative ischaemia. Similar results were observed for serum creatine kinase isoenzyme (mass determination) (S-CKMB), but differences were generally less well correlated with other perioperative variables. At day 1, patients with POMI had higher levels of S-TnT as well as S-CKMB when compared to patients without POMI. At day 4, most patients still had elevated levels of S-TnT, but the difference in S-TnT levels between patients with POMI and patients without POMI was more pronounced. In contrast, the levels of S-CKMB were essentially normalized in both groups. Measurements of S-TnT at day 4 appears to be of significant value in diagnosing POMI. However, most of the patients without POMI had increased levels of S-TnT at day 4, suggesting that some irreversible operatively induced myocardial damage had occurred. Thus, even at a late postoperative stage the perioperative duration of ischaemia and type and extent of the surgical procedure should be taken into consideration.


Self-disclosure patterns in clinical and nonclinical couples.
Examined differences in the self-disclosing patterns of 10 clinical and 10 nonclinical couples in a structured interview using the Self-Disclosure Coding System (SDCS), a behavioral content analysis system designed to assess the basic parameters of disclosing behavior. A discriminant function analysis revealed that two SDCS variables, (1) time spent talking; and (2) judged degree of congruence between verbal content and affective manner of presentation, could discriminate the two groups and would classify correctly 70% of the couples. These results suggest that it is not what clinical couples say, but how they say it that will distinguish them from nonclinical couples.


Aortic Valve Reconstruction with Ozaki Technique.
Modern bioprostheses offer a complete and definitive solution to elderly patients who need aortic valve surgery. Nonetheless, the scenario is more demanding when dealing with younger and less fragile patients. In this setting, any prosthetic aortic valve replacement can provide only a suboptimal solution and its related issues have not been fixed yet. The answer to the needs of this special population is the enhancement and refinement of the surgical technique. The Ozaki technique relies on custom-tailored autologous aortic cusps individually sutured in the aortic position. This approach has been showing optimal results if performed after a dedicated training period.


Stability of common biochemical analytes in serum gel tubes subjected to various storage temperatures and times pre-centrifugation.
Blood samples collected in rural and remote areas of Australasia are often exposed to a range of environmental conditions prior to analysis in a laboratory. The aim of this study was to determine analyte stability of venous blood specimens in serum gel tubes exposed to a range of storage temperatures and times prior to centrifugation. Thirty healthy adult volunteers were enrolled in the study. Blood was collected into 11 serum gel separator tubes. All samples were allowed to clot at room temperature for 30 min. Two samples were centrifuged and analysed as controls. Nine samples were stored at 15, 25 or 35 degrees C for 4, 8 or 24 h, respectively, before centrifugation. Thirty-five biochemical analytes were measured on each sample. Most analytes remained stable in all storage conditions including sodium, total protein, albumin, bilirubin, alanine transferase, aspartate aminotransferase, alkaline phosphatase, gamma glutamyl transferase, creatinine kinase, lipase, cholesterol, triglycerides, transferrin, urate, C-reactive protein, vitamin B(12), thyroid-stimulating hormone, free thyroxine, free triiodothyronine, follicle-stimulating hormone, oestradiol, prostate-specific antigen, cortisol and vitamin D. Potassium, glucose, phosphate, creatinine, urea, ferritin, iron, lactate dehydrogenase, magnesium and calcium were not stable in at least one of the storage conditions. These results can be used to determine which analytes produce valid results despite exposure to variable storage conditions for up to 24 h prior to centrifugation. The majority of analytes were unaffected by a delay in centrifugation at a variety of temperatures, however, some important analytes were significantly affected.


Rightward directional bias in art produced by cultures without a written language.
In this study, we coded art painted on rocks located in southern Africa, which was painted with a mixture of ochre, blood, and clay by the San, a Neolithic culture with no written language. These images depict a mixture of humans and animals in a variety of contexts, including (but not limited to) hunts and dances. We calculated a laterality index for the collected available art from each region, finding that although there was variability across regions in the direction of the laterality scores, most regions contained a majority of figures facing rightward. This is in stark contrast with reports of artists drawing leftward facing animals and human profiles (an effect that is influenced by native language writing system direction, gender, and handedness), but interestingly our sample also contained regions with strong leftward biases. Our results are, however, in accord with studies that report people preferring images that depict left-to-right motion, as well as the left-to-right bias in depicting transitive actions, an effect that seems to result from greater right hemispheric activation in scene processing and interpretation. Thus, this study shows that in the absence of a writing system, right-lateralized neural architecture may guide the hands of artists.


Food and Agriculture Policy in Europe.
Diet includes many risk factors for the most common non-communicable diseases (NCDs), but diets consumed in Europe and in other parts of the developed world are not being modified sufficiently to take account of health priorities concerning, in particular, the prevention of NCDs, while much excess mortality and morbidity could be prevented by government actions to regulate appropriately both the agricultural and food industries, and to apply appropriate taxes and subsidies to promote healthier nutrition. In Europe, the Common Agricultural Policy (CAP) continues to promote production of saturated fat rich foods and sugar, with scarce attempts to promote increased production of fruit and vegetables. Meanwhile, the food industry continues to market secondary food products rich in sugar, salt and saturated fats. Powerful lobbies seek to block reform; however, necessary reforms are indicated in the interests of improved nutritional health.


Silent katydid females are at higher risk of bat predation than acoustically signalling katydid males.
Males that produce conspicuous mate attraction signals are often at high risk of predation from eavesdropping predators. Females of such species typically search for signalling males and their higher motility may also place them at risk. The relative predation risk faced by males and females in the context of mate-finding using long-distance signals has rarely been investigated. In this study, we show, using a combination of diet analysis and behavioural experiments, that katydid females, who do not produce acoustic signals, are at higher risk of predation from a major bat predator, Megaderma spasma, than calling males. Female katydids were represented in much higher numbers than males in the culled remains beneath roosts of M. spasma. Playback experiments using katydid calls revealed that male calls were approached in only about one-third of the trials overall, whereas tethered, flying katydids were always approached and attacked. Our results question the idea that necessary costs of mate-finding, including risk of predation, are higher in signalling males than in searching females.


Mechanism and Energetics of the Hydrolysis of Th+ To Form Th(OD)3+: Guided Ion Beam and Theoretical Studies of ThO+, ThO2+, and OThOD+ Reacting with D2O.
The kinetic energy dependences of the reactions of ThO+, ThO2+, and OThOD+ with D2O, ThO2+ with D2, and OThOD+ with Xe were studied using guided ion beam tandem mass spectrometry. Exothermic formation of OThOD+ is the dominant process observed in reactions of both ThO+ and ThO2+ with D2O. Minor products formed in endothermic reactions include ThO2+, DThO+, and ThO2D2+. OThOD+ is also formed in the reaction of ThO2+ with D2 but in an endothermic process. Collision-induced dissociation (CID) of OThOD+ with Xe leads to endothermic loss of the hydroxide ligand. OThOD+ reacts further with D2O to form the associative complex ThO3D3+, which is long-lived before dissociating back to the reactants. The OThOD+-D2O bond energy of the associative complex is measured to be 2.96 ± 0.05 eV by modeling the kinetic energy-dependent cross section for association using a phase space theory model that rigorously conserves angular momentum. By comparison with theory, this bond energy identifies the ThO3D3+ species as the trihydroxide cation, Th(OD)3+. From the endothermic reactions and CID of OThOD+ with Xe, the OTh+-D, OTh+-O, and OTh+-OD bond dissociation energies (BDEs) are measured to be 2.33 ± 0.24, 4.66 ± 0.15, and 6.00 ± 0.17 eV, respectively. All four of these BDEs are experimentally determined for the first time and agree reasonably well with values calculated at the B3LYP, B3PW91, and PBE0 levels of theory with cc-pVQZ basis sets. Complete potential energy surfaces for all reactions were calculated at the B3LYP/cc-pVTZ level and elucidate the mechanisms for all processes observed.


The different roles of selective autophagic protein degradation in mammalian cells.
Autophagy is an intracellular pathway for bulk protein degradation and the removal of damaged organelles by lysosomes. Autophagy was previously thought to be unselective; however, studies have increasingly confirmed that autophagy-mediated protein degradation is highly regulated. Abnormal autophagic protein degradation has been associated with multiple human diseases such as cancer, neurological disability and cardiovascular disease; therefore, further elucidation of protein degradation by autophagy may be beneficial for protein-based clinical therapies. Macroautophagy and chaperone-mediated autophagy (CMA) can both participate in selective protein degradation in mammalian cells, but the process is quite different in each case. Here, we summarize the various types of macroautophagy and CMA involved in determining protein degradation. For this summary, we divide the autophagic protein degradation pathways into four categories: the post-translational modification dependent and independent CMA pathways and the ubiquitin dependent and independent macroautophagy pathways, and describe how some non-canonical pathways and modifications such as phosphorylation, acetylation and arginylation can influence protein degradation by the autophagy lysosome system (ALS). Finally, we comment on why autophagy can serve as either diagnostics or therapeutic targets in different human diseases.


Development of a whole-cell screening system for evaluation of the human CYP1A2-mediated metabolism.
Cytochrome P450 1A2 (CYP1A2) is an important member of cytochrome P450 involved in drug metabolism. In this study, a cell line, Huh7-1A2-I-E, with high expression level of CYP1A2 is established based on Huh7 cells. To achieve this, we constructed a recombinant lentiviral vector, pLenti-1A2-I-E, containing a single promoter encoding CYP1A2 followed by an internal ribosome entry site (IRES) to permit the translation of enhanced green fluorescence protein (EGFP). Such a design has greatly facilitated the selection of stable cell lines because the translations of CYP1A2 and EGFP proteins would be based on a single bi-cistronic mRNA. The Huh7-1A2-I-E cells were evaluated as a cell-based model for identification of CYP1A2 inhibitors and for studies of cytotoxicity resulted from CYP-mediated drug metabolism. Treatment of Huh7-1A2-I-E cells and the Huh7-E control cells with aflatoxin B1 showed that cells with CYP1A2 expression are much more sensitive to aflatoxin B1 and the cellular toxicity of aflatoxin B1 in Huh7-1A2-I-E cells could be prevented by furafylline, a CYP1A2 inhibitor. A collection of approximately 200 drugs were screened using this system and results indicate that for most drugs the metabolism by CYP1A2 is unlikely to have made a major contribution to the in vitro cytotoxicity except for thimerosal and evoxine. Several previously unidentified CYP1A2 inhibitors such as evoxine and berberine were also identified in this study.


Compressed Hydrogen-Induced Synthesis of Quaternary Trimethyl Chitosan-Silver Nanoparticles with Dual Antibacterial and Antifungal Activities.
Quaternary Trimethyl Chitosan (QTMC) and QTMC-Silver Nanoparticles (QTMC-AgNPs) have been synthesized, characterized, and tested as antibacterial agents against Staphylococcus aureus, Escherichia coli, and two plant fungi (Sclerotium rolfsil and Fusarium oxysporum). The as-prepared water-soluble QTMC was in situ reacted with silver nitrate in the presence of clean compressed hydrogen gas (3 bar) as a reducing agent to produce QTMC-AgNPs. UV-vis, ATR-FTIR, HR-TEM/SEM, XPS, DLS, XRD, and TGA/DTG were employed to assess the optical response, morphology/size, surface chemistry, particle size distribution, crystal nature, and thermal stability of the synthesized QTMC-AgNPs, respectively. The as-prepared QTMC-AgNPs were quasi-spherical in shape with an average particle size of 12.5 nm, as determined by ImageJ software utilizing HR-TEM images and further validated by DLS analysis. The development of crystalline nanoparticles was confirmed by the presence of distinct and consistent lattice fringes with an approximate interplanar d-spacing of 2.04 nm in QTMC-AgNPs. The QTMC-AgNPs exhibited significant antibacterial activity with a clear zone of inhibition of 30 mm and 26 mm around the disks against E. coli and S. aureus, respectively. In addition, QTMC-AgNPs showed highly efficient antifungal activity with 100% and 76.67% growth inhibition against two plant pathogens, S. rolfsii and F. oxysporum, respectively, whereas QTMC revealed no impact. Overall, QTMC-AgNPs showed a promising therapeutic potential and,thus, can be considered for drug design rationale.


Elevated Microparticles, Thrombin-antithrombin and VEGF Levels in Colorectal Cancer Patients Undergoing Chemotherapy.
Hypercoagulable state and neoangiogenesis are common phenomena associated with malignancy. Cancer patients have increased levels of circulating endothelium-derived microparticles (EMPs), which have been hypothesized to be involved in numerous pathophysiological processes. Hemostasis and angiogenesis are also activated in colorectal cancer (CRC) patients. The study aimed to investigate potential influence of chemotherapy on EMPs, thrombin anti-thrombin complex (TAT) and vascular endothelial growth factor (VEGF) levels in CRC patients undergoing chemotherapy. The study group consisted of 18 CRC patients: 8 stage III colon cancer (CC) and 10 stage IV rectal cancer (RC) patients. EMPs, TAT and VEGF levels were assessed before chemotherapy and after the third course. Results were compared with 10 healthy subjects. EMP concentration was measured by flow cytometry, while TAT and VEGF concentrations were assayed employing ELISA. Compared to the control group, CC and RC patients had significantly higher levels of tissue factor (TF)-bearing and non-TF-bearing EMPs before and after three courses of chemotherapy. VEGF concentrations in CRC patients were higher than in the control groups and increased following chemotherapy. TAT levels were elevated in CRC patients before chemotherapy compared to healthy subjects and significantly increased after the third course of chemotherapy. No significant correlation was found either between EMP and TAT levels, or between EMP concentrations and VEGF levels in the study group. CRC patients have increased EMPs, and TAT as well as VEGF levels tend to increase during chemotherapy.


Long-term spaceflight composite stress induces depressive behaviors in model rats through disrupting hippocampus synaptic plasticity.
Long-term spaceflight composite stress (LSCS) can cause adverse effects on human systems, including the central nervous system, which could trigger anxiety and depression. This study aimed to identify changes in hippocampus synaptic plasticity under LSCS. The present study simulated the real long-term space station environment by conducting a 42-day experiment that involved simulating microgravity, isolation, noise, circadian rhythm disruptions, and low pressure. The mood and behavior of the rats were assessed by behavior test. Transmission electron microscopy and patch-clamp were used to detect the changes in synapse morphology and electrophysiology, and finally, the expression of NMDA receptor channel proteins was detected by western blotting. The results showed that significant weight loss, anxiety, and depressive behaviors in rats were observed after being exposed to LSCS environment for 42 days. The synaptic structure was severely damaged, manifested as an obvious decrease in postsynaptic density thickness and synaptic interface curvature (p < 0.05; p < 0.05, respectively). Meanwhile, LTP was significantly impaired (p < 0.0001), and currents in the NMDAR channel were also significantly reduced (p < 0.0001). Further analysis found that LSCS decreased the expression of two key subtype proteins on this channel. These results suggested that LSCS-induced depressive behaviors by impairing synaptic plasticity in rat hippocampus.


Parascedosporium and its relatives: phylogeny and ecological trends.
The genus Scedosporium and its relatives comprising microascalean anamorphs with slimy conidia were studied. Graphium and Parascedosporium also belong to this complex, while teleomorphs are found in Pseudallescheria, Petriella, Petriellopsis, and Lophotrichus. Species complexes were clearly resolved by rDNA ITS sequencing. Significantly different ecological trends were observed between resolved species aggregates. The Pseudallescheria and Scedosporium prolificans clades were the only lineages with a marked opportunistic potential to mammals, while Petriella species were associated primarily with soil enriched by, e.g. dung. A consistent association with bark beetles was observed in the Graphium clade. The ex-type strain of Rhinocladium lesnei, CBS 108.10 was incorrectly implicated by Vuillemin (1910) in a case of human mycetoma; its sequence was identical to that of the ex-type strain of Parascedosporium tectonae, CBS 127.84.


Specimen Identification Errors in Breast Biopsies: Age Matters. Report of Two Near-Miss Events and Review of the Literature.
The consequences of patient identification errors due to specimen mislabeling can be deleterious. We describe two near-miss events involving mislabeled breast specimens from two patients who sought treatment at our institution. In both cases, microscopic review of the slides identified inconsistencies between the histologic findings and patient age, unveiling specimen identification errors. By correlating the clinical information with the microscopic findings, we identified mistakes that had occurred at the time of specimen accessioning at the original laboratories. In both cases, thanks to a timely reassignment of the specimens, the patients suffered no harm. These cases highlight the importance of routine clinical and pathologic correlation as a critical component of quality assurance and patient safety. A review of possible specimen identification errors in the anatomic pathology setting is presented.