Prevalence and predictors of dyslipidemia among HAART treated and HAART naive HIV positive clients attending Debre Tabor Hospital, Debre Tabor, Ethiopia.
Highly active anti-retroviral therapy has been reported to be associated with a number of side effects in human immunodeficiency virus patients among which dyslipidemia isa common metabolic disorder. A Hospital based comparative cross-sectional study among 228 HIV positive patients was conducted from July to August 2020. Socio-demographic and clinical data were collected using structured questionnaires. Fasting venous blood sample was collected and analyzed for Lipid profiles. EDTA sample was analyzed for CD4+ T cell determination. Anthropometric measurement was done. Data were analyzed using SPSS version 22. Independent t-test was done. Logistic and binary regression was done. A total of 228 HIV patients were enrolled in the study. Prevalence of dyslipidemia in HAART naive and HAART treated patients was 61 (53.5%) and 84 (73.7%), respectively. The prevalence of Total Cholesterol ≥200 mg/dl was 50% and 30%; High density lipoprotein cholesterol <40 mg/dl was 43.8% and 36%; Low density lipoprotein cholesterol ≥130 mg/dl was 48.3% and 28.1%; and Triglyceride ≥ 150 mg/dl 59.6% and 39% among HAART treated and HAART naive, respectively. Age greater than 40 years (AOR = 3.27, 95% C.I: 1.47-7.25), blood pressure ≥140/90 (AOR = 16.13, 95% C.I: 5.81-44.75), being on HAART (AOR = 2.73, 95% C.I: 1.35-5.53) and body mass index >25 kg/m2 (AOR = 1.92, 95% C.I: 1.20-4.81) were identified as determinants of dyslipidemia. The mean value of lipid profile was significantly higher among HAART treated as compared to those HAART naive HIV positive clients.


The disparity between homologous chromosomes during DNA replication.
When the patterns of replicating chromosome bands of homologous chromosomes within a diploid cell during DNA synthesis phase are compared, the frequency of disparity (i.e. a band present on only one homologue) is less than expected on the basis of chance. This could be taken as implying some "link" between homologues which constrains their programmes of replication to keep in step. This paper develops a model showing that the observed disparities can be accommodated within a framework of homologue independence. Differences between cells in the time of appearance of bands lead in any sample to the summation of an infinitude of binomial distributions and hence to over-dispersion. The model fits observed data for bands replicating in euchromatic and heterochromatic chromosome regions obtained from Syrian hamster fibroblast cells growing in vitro.


Assessing responses of the Hiroshima Bay ecosystem to increasing or decreasing phosphorus and nitrogen inputs.
The Japanese Government is seeking an appropriate level of nutrient load from the land to maintain the highest possible estuarine fishery production and water transparency simultaneously. To provide a scientific basis for the governmental inquiry, we conducted sensitivity analyses using an ecosystem model of Hiroshima Bay in order to assess the ecosystem's responses to phosphorus and nitrogen inputs. Load levels of phosphorus (Case P), nitrogen (Case N) and both phosphorus and nitrogen (Case NP) that were different from the average loading recorded during 1991-2000 (±25%, ±50%, and ±75%) were applied. The results showed that phosphorus had a significantly greater impact on the primary production of the bay than nitrogen. Case P+25 increased the primary production but led to N-limitation. However, it was found that Case NP at the levels over +25% could bring the Hiroshima Bay ecosystem back to its eutrophic state of 30 years ago.


Mass transfer process and separation mechanism of four 5'-ribonucleotides on a strong acid cation exchange resin.
5'-ribonucleotides including adenosine 5'-monophosphate (AMP), cytidine 5'-monophsphate (CMP), guanosine 5'-monophosphate (GMP) and uridine 5'-monophosphate (UMP) have been widely used in the food and pharmaceutical industries. This work focused on the assessment of mass transfer process and separation mechanism of four 5'-ribonucleotides and counter-ion Na+ on the strong cation exchange resin NH-1. The intraparticle diffusion was determined as the rate-limiting step for the mass transfer of AMP, CMP, GMP, and Na+ on the resin NH-1 through the Boyd model. Meanwhile, a homogeneous surface diffusion model (HSDM) combing ion exchange and physical adsorption was proposed and tested against adsorption kinetic data in the batch adsorption systems. The fixed-bed film-surface diffusion model based on the HSDM was then developed and successfully predicted the concentration profiles of 5'-ribonucleotides and the change of pH at the outlet of the fixed-bed in the dynamic adsorption and separation process. Finally, the separation mechanism of 5'-ribonucleotides was presented combining model prediction and experimental results. The separation of UMP, GMP and CMP were mainly based on their differences in isoelectric points, while that of AMP and CMP were lied with the discrepancy of their physical adsorption binding capacity with the resin NH-1.


Is endobronchial ultrasound-transbronchial needle aspiration (EBUS-TBNA) reliable and safe procedure in geriatric patients?
Even though studies have indicated the usefulness and safety of endobronchial ultrasound-transbronchial needle aspiration (EBUS-TBNA), elderly patient data are limited due to the small sample sizes. We aimed to evaluate usage and safety of EBUS-TBNA in elderly population. This single-center retrospective study was conducted with patients who underwent an EBUS-TBNA procedure between September 2011 and December 2019. The patients were categorized into two groups: those aged 65 years or older (elderly group) and those younger than 65 years (younger group). 2444 patient data, 1069 of which were in the elderly group, were analyzed. The cytological examination of EBUS-TBNA identified specimen adequacy in 96.8% of patients. One hundred and thirty patients (5.3%) experienced complications, with similar complication rates recorded in both the elderly and younger groups (5.4% vs 5.2%, p: 0.836). Logistic regression analyses revealed that age, and presence of hypertension, diabetes mellitus, coronary artery disease and malignancy are associated significantly with complication-related EBUS-TBNA. For the lymph nodes with a final diagnosis of malignancy, the sensitivity, specificity, positive predictive value, negative predictive value and accuracy of EBUS-TBNA revealed a diagnostic performance in excess of 90% except for metastasis and lymphoma. EBUS-TBNA can be considered a safe and effective technique in patients aged 65 years and over.


Antiinflammatory Effect of N-Acetylcysteine Combined with Exogenous Surfactant in Meconium-Induced Lung Injury.
Neonatal meconium aspiration syndrome (MAS) can be treated by exogenous surfactant (S). However, aspirated meconium initiates local inflammation and oxidation which may inactivate surfactant and reduce its action. This experimental study estimated whether combined use of surfactant and the antioxidant N-acetylcysteine (NAC) can enhance effectiveness of therapy. Meconium-instilled rabbits were non-treated (M), treated with monotherapies (M + S, M + NAC), combined therapy (M + S + NAC), or received saline instead of meconium (controls, C). Surfactant therapy consisted of two lung lavages (BAL) with diluted Curosurf (5 mg phospholipids/ml, 10 ml/kg) followed by undiluted Curosurf (100 mg phospholipids/kg). N-acetylcysteine (Acc Injekt, 10 mg/kg) was given intravenously in M + S + NAC group 10 min after surfactant therapy. Animals were oxygen-ventilated for additional 5 h. Then, differential white cell count in the blood (WBC) was determined. Left lung was saline-lavaged and differential cell count in BAL was determined. In right lung tissue, wet/dry weight ratio, oxidation markers (TBARS, 3NT) and interleukines (IL-2, IL-6, IL-13, and TNFα) using ELISA and RT-PCR were estimated. Combined S + NAC therapy significantly decreased W/D ratio, TBARS, 3NT, and IL, whereas the effect of monotherapies (either S or NAC) was less obvious. In conclusion, addition of NAC to surfactant treatment may enhance the therapeutic outcome in MAS.


Peripheral nerve dysfunction in adult patients with cystic fibrosis.
The prevalence of peripheral nerve dysfunction was assessed in 24 randomly selected adult patients with cystic fibrosis. Median, peroneal and sural nerves were studied. In 15 patients (62%), one or more electrical abnormality were detected. In the median nerve, conduction velocity was slowed in 29% of sensory studies and 12.5% of motor studies; compound muscle action potential (CMAP) and sensory nerve action potential (SNAP) amplitudes were normal. The CAMP was low in 17% of peroneal studies with motor conduction velocity slow in 8%. The sural SNAP was low in 17% and sensory nerve conduction velocity slow in 25%. Using a multiple regression analysis only age at diagnosis was identified as a significant predictor of peripheral nerve dysfunction (multiple R = 0.55; p < 0.01). This study demonstrates that mild peripheral nerve dysfunction is common in patients with cystic fibrosis. With the improving long term survival of these patients we predict that the prevalence and severity of this complication will increase.


[Anatomy and adaptation to environment study of endangered alpine medical plant Neopicrorhiza scrophulariiflora].
To study the anatomical structure of endangered alpine medical plant Neopicrorhiza scrophulariiflora and the high altitude adaptability. The leaf epidermis character as well as section structure of leaf, aerial stem and rhizome were observed by light microscopical technique. The leaf surface of Neopicrorhiza scrophulariiflora was covered with two kinds of glandular hair, and the stommata was anomocytic type. Moreover, the leaf was isolateral and differed from most of alpine plant. The aerial stem had well-developed mechanical tissue. The rhizome was distributed by well-developed cork layers and collenchyma. Large numbers of aerenchymas distributed widely in leaf, aerial stem and rhizome. There existed characteristic traits in Neopicrorhiza scrophulariiflora that adapted the alpine environment, however, there still had some particular character different from other alpine plant. Thus, the adaptive style of alpine plant to high altitude environment was diversity.


Degradation of nonylphenol by anaerobic microorganisms from river sediment.
We investigated the degradation of nonylphenol monoethoxylate (NP1EO) and nonylphenol (NP) by anaerobic microbes in sediment samples collected at four sites along the Erren River in southern Taiwan. Anaerobic degradation rate constants (k1) and half-lives (t1/2) for NP (2 microg/g) ranged from 0.010 to 0.015 1/day and 46.2 to 69.3 days respectively. For NP1EO (2 microg/g), the ranges were 0.009-0.014 1/day and 49.5-77.0 days respectively. Degradation rates for NP and NP1EO were enhanced by increasing temperature and inhibited by the addition of acetate, pyruvate, lactate, manganese dioxide, ferric chloride, sodium chloride, heavy metals, and phthalic acid esters. Degradation was also measured under three anaerobic conditions. Results show the high-to-low order of degradation rates to be sulfate-reducing conditions > methanogenic conditions > nitrate-reducing conditions. The results show that sulfate-reducing bacteria, methanogen, and eubacteria are involved in the degradation of NP and NP1EO, with sulfate-reducing bacteria being a major component of the river sediment.


Unveiling the Activity Origin of Iron Nitride as Catalytic Material for Efficient Hydrogenation of CO2 to C2+ Hydrocarbons.
Developing efficient catalytic materials and unveiling the active species are significant for selective hydrogenation of CO2 to C2+ hydrocarbons. Fe2 N@C nanoparticles were reported to exhibit outstanding performance toward selective CO2 hydrogenation to C2+ hydrocarbons (C2+ selectivity: 53.96 %; C2 -C4 = selectivity, 31.03 %), outperforming corresponding Fe@C. In situ X-ray diffraction, ex situ Mössbauer and X-ray photoelectron spectra revealed that iron nitrides were in situ converted to highly active iron carbides, which acted as the real active species. Moreover, the combined results of in situ diffuse reflectance infrared Fourier transform spectroscopy and control experiments suggested an in situ formed carbonyl iron-mediated conversion mechanism from iron nitrides to iron carbides.


Clinical indicators that influence a clinician's decision to start L-thyroxine treatment in prematurity with transient hypothyroxinemia.
Transient hypothyroxinemia of prematurity (THOP) is defined as a low level of circulating thyroxine (T4), despite low or normal thyroid-stimulating hormone (TSH) levels. We aimed to evaluate the incidence of THOP, the clinical and laboratory findings of preterm infants with this condition and the levothyroxine (L-T4) treatment. Preterm infants (n = 181) delivered at 24-34 weeks of gestation were evaluated by their thyroid function tests that were performed between the 10th and 20th days of postnatal life and interpreted according to the gestational age (GA) references. Clinical and laboratory characteristics of the patients with THOP and normal thyroid function tests were compared. Patients with THOP and treated with L-T4 were compared with the ones who were not regarding laboratory, and clinical characteristics. Incidence of hypothyroxinemia of prematurity was 45.8% (n = 83). Euthyroidism, primary hypothyroidism, and subclinical hypothyroidism were diagnosed in 47.5% (n = 86), 5% (n = 9) and 1.7% (n = 3) of the patients, respectively. Mean birth weight (BW) and GA were significantly lower in the hypothyroxinemia group than in the euthyroid group (p < 0.001). L-T4 was started in 43% (n = 36) of the patients with THOP. Treatment initiation rate was 44.4% (n = 16) in 24-27 wk, 41.6% (n = 15) in 28-30 wk, and 13.8% (n = 5) in 31-34 wk. As the GA increased, the incidence of THOP and the rate of treatment initiation decreased (p < 0.001). The lowest free thyroxine (FT4) cut-off value was 0.72 ng/dl in the treated group. In addition, incidences of vancomycin + amikacin, caffeine, dopamine treatments, RDS, IVH, BPD, central catheter, FFP transfusion, and ventilator support were higher in the treated group (P < 0.05). This study revealed that prevalence of THOP increased as the GA and BW decreased. As the GA decreased, THOP patients requiring L-T4 treatment increased. Additionally, association with comorbid diseases increased the requirement of treatment.


Non-tuberculous mycobacterial infections in geriatric patients-A neglected and emerging problem.
The diseases caused by Non-tuberculous mycobacteria (NTM) has increased steadily in the last two decades. Increase in incidence of NTM infections are being reported in elderly people as they are more susceptible and often experiencing high morbidity. There is prediction that NTM infections will further rise because of expected increase in elderly population by 2050. Given the importance of NTM infection in the elderly, the interest in studying NTM characteristics in the aged population is increasing. In this review, we summarize the characteristics of NTM infection among elderly patients. We focus on epidemiology, clinical presentation, and treatment options of NTM in this age group. We highlight the differences in the diagnosis and treatment between rapid and slow growing mycobacterial infections. The current recommendations for treatment of NTM have been discussed. Finally, we have reviewed the prognosis of NTM disease in elderly patients.


Structural identification of novel glucoside and glucuronide metabolites of (-)-epigallocatechin-3-gallate in mouse urine using liquid chromatography/electrospray ionization tandem mass spectrometry.
(-)-Epigallocatechin-3-gallate (EGCG), the most abundant and most biologically active polyphenolic compound in tea, has been proposed to have many health beneficial effects. The metabolic fate of EGCG, however, is not well understood. In the present study, we identified a novel EGCG metabolite, 7-O-beta-D-glucopyranosyl-EGCG-4''-O-beta-D-glucupyranoside, in a mouse urine sample using liquid chromatography/electrospray ionization tandem mass spectrometry. The structure of this metabolite was confirmed by analyzing the MSn (n = 1-4) spectra as well as comparing the MS/MS spectra of its product ions with those from EGCG and EGCG-4''-O-beta-D-glucupyranoside standards. To our knowledge, this is the first report of the identification of a glucoside metabolite of EGCG in mammals. Our results indicate that glucosidation represents a novel pathway in the metabolism of EGCG in mice.


8-chloroadenosine 3',5'-monophosphate (8-Cl-cAMP) selectively eliminates protein kinase A type I to induce growth inhibition in c-ras-transformed fibroblasts.
8-Chloroadenosine 3',5'-monophosphate (8-Cl-cAMP), a site-selective cyclic adenosine 3',5'-monophosphate (cAMP) analogue, exhibits growth inhibition in a broad spectrum of cancer cell lines. We investigated the effect of 8-Cl-cAMP on c-ras-transformed mouse fibroblasts (MP3/3T3) which were established by transfection of Balb3T3 cells (Balb3T3) with the point-mutated c-ras gene [G12-->V12]. 8-Cl-cAMP (2-5 microM) exerted over 80% growth inhibition by day 4 on MP3/3T3, while inhibiting parental Balb3T3 cell growth less than 40%. In order to distinguish the effect of 8-Cl-cAMP from that of 8-chloroadenosine (8-Cl-adenosine), we examined the effect of 8-Cl-cAMP in serum-free medium. 8-Cl-cAMP demonstrated a potent growth inhibition of MP3/3T3 cells cultured in serum-free medium, suggesting that the growth inhibitory effect of 8-Cl-cAMP was not due to its hydrolysed product, 8-Cl-adenosine. In addition, both Balb3T3 and MP3/3T3 contained cAMP phosphodiesterases mainly composed of isozyme IV which has previously been reported to be insensitive towards the hydrolysis of 8-Cl-cAMP. Non-transformed Balb3T3 cells contained only type II cAMP-dependent protein kinase (PKA), whereas transformed MP3/3T3 exhibited a marked increase in type I PKA. The growth inhibition of MP3/3T3 by 8-Cl-cAMP accompanied almost complete elimination of type I PKA without affecting type II PKA. Moreover, 8-Cl-cAMP induced an arrest in the G0/G1-phase of the cell cycle in MP3/3T3. 8-Cl-adenosine had little or no effect on the cell cycle kinetics of MP3/3T3 cells. These results show that 8-Cl-cAMP is a novel cAMP analogue which selectively eliminates type I PKA to induce growth inhibition in transformed fibroblasts.


Stereotaxic Surgical Targeting of the Nonhuman Primate Caudate and Putamen: Gene Therapy for Huntington's Disease.
Stereotaxic surgery is an invaluable tool to deliver a variety of gene therapy constructs to the nonhuman primate caudate and putamen in preclinical studies for the genetic, neurodegenerative disorder, Huntington's disease (HD). Here we describe in detail how to perform this technique beginning with a pre-surgical magnetic resonance imaging scan to determine surgical coordinates followed by the stereotaxic surgical injection technique. In addition, we include methodology of a full necropsy including brain and peripheral tissue removal and a standard immunohistochemical technique to visualize the injected gene therapy agent.


A review: Biological technologies for nitrogen monoxide abatement.
Nitrogen oxides (NOx), including nitrogen monoxide (NO) and nitrogen dioxide (NO2), are among the most important global atmospheric pollutants because they have a negative impact on human respiratory health, animals, and the environment through the greenhouse effect and ozone layer destruction. NOx compounds are predominantly generated by anthropogenic activities, which involve combustion processes such as energy production, transportation, and industrial activities. The most widely used alternatives for NOx abatement on an industrial scale are selective catalytic and non-catalytic reductions; however, these alternatives have high costs when treating large air flows with low pollutant concentrations, and most of these methods generate residues that require further treatment. Therefore, biotechnologies that are normally used for wastewater treatment (based on nitrification, denitrification, anammox, microalgae, and combinations of these) are being investigated for flue gas treatment. Most of such investigations have focused on chemical absorption and biological reduction (CABR) systems using different equipment configurations, such as biofilters, rotating reactors, or membrane reactors. This review summarizes the current state of these biotechnologies available for NOx treatment, discusses and compares the use of different microorganisms, and analyzes the experimental performance of bioreactors used for NOx emission control, both at the laboratory scale and in industrial settings, to provide an overview of proven technical solutions and biotechnologies for NOx treatment. Additionally, a comparative assessment of the advantages and disadvantages is performed, and special challenges for biological technologies for NO abatement are presented.


Latent Profiles of Health and Reproductive Risk and Protective Factors among Women in Appalachia.
Women who use opiates and are involved in the criminal justice system in Appalachia may be prone to adverse health outcomes. In this study, we performed a latent class analysis of risk and protective factors on 400 drug-using women recruited from rural, Appalachian jails. A two-profile solution best fit the data. Both profiles evinced low levels of condom use, reproductive and physical health screens, and STD history. However, the primary substantive difference between the profiles was partner risk behavior: the higher risk class had main male partners with histories of injection drug use and incarceration. Results suggest that interventions need to be tailored to unique profiles of risk and protective factors, which should include taking partner risk into consideration.


Role of the NO/cGMP pathway and renin-angiotensin system in the hypotensive and diuretic effects of aqueous soluble fraction from Crataegus songarica K. Koch.
Fruits of Crataegus songarica K. Koch. (Rosaceae) are commonly used in folk medicine for their diuretic properties to treat hypertension and congestive heart failure. To date, no scientific data has been published to support the diuretic potential. The purpose of this study was to evaluate efficacy and mechanism underlying the hypotensive and diuretic action of C. songarica in normotensive rats and to determine the constituents from the extracts by LC-DAD-MS. Firstly, phytochemical profiling and antioxidant potential of C. songarica extracts was determined. Then, to evaluate changes in blood pressure, different groups of anesthetized normotensive rats were intravenously treated with crude extract (CS-Cr, 10-80 mg/kg), aqueous soluble (AS-CS, 0.1-20 mg/kg), and n-butanol soluble fractions of C. songarica (BS-CS, 1-80 mg/kg). The diuretic effects of CS-Cr (100-500 mg/kg, p.o), AS-CS (100-300 mg/kg, p.o) and BS-CS (100-300 mg/kg, p.o) were evaluated in comparison with hydrochlorothiazide (HCTZ, 10 mg/kg, p.o). The urinary volume, sodium, potassium and pH were estimated in the sample collected for 6 h from saline-loaded rats. Using pharmacological antagonists or inhibitors, we determine the involvement of acetylcholine, prostaglandins, and nitric oxide in C. songarica induced hypotensive and diuresis action. In addition, the activities of angiotensin converting enzyme, erythrocytary carbonic anhydrase and renal Na+/K+/ATPase were evaluated in vitro. From the LC-DAD-MS analyses, thirty-nine compounds were detected, showing a complex chemical profile and an expressive antioxidant activity "in vitro". Acute treatment with CS-Cr, AS-CS, and BS-CS exhibited significant hypotensive and diuretic potential in normotensive rats. However, AS-CS produced most potent and significant dose-dependent hypotension in normotensive rats, and also produced highly significant diuretic and saluretic effects. Despite the changes in urinary excretion of electrolytes, the plasmatic levels of sodium and potassium were not changed. Previous treatment with atropine and L-NAME significantly reduced the hypotensive and diuretic action of AS-CS in normotensive rats. Moreover, the 7-day treatment with AS-CS also resulted in significant ACE inhibitory activity. This research supports and extends the ethnomedicinal use of C. songarica as diuretic and hypotensive agent. The results showed that AS-CS from C. songarica could present compounds responsible for hypotensive and diuretic activities with no signs of toxicity, and these effects could involve nitric oxide pathway activated by muscarinic receptors or/and inhibition of angiotensin converting enzyme.


Diazinon dissipation in pesticide-contaminated paddy soil: kinetic modeling and isolation of a degrading mixed bacterial culture.
Dissipation kinetics of diazinon was investigated in soils culled from a paddy field with a long history of the pesticide application. Goodness of fit statistical indices derived from several fitted mono- and bi-exponential kinetic models revealed a bi-phasic pattern of the diazinon dissipation curve at 15 and 150 mg kg-1 spiking levels, which could be described best by the first-order double exponential decay (FODED) model. Parameters obtained from this model were able to describe the enhanced dissipation of diazinon as the result of repeated soil applications, where a larger fraction of the pesticide readily available in the solution phase was dissipated with a fast rate. Cluster and principal component analysis (PCA) of denaturing gradient gel electrophoresis (DGGE) obtained from soil bacterial populations revealed that they were only affected at the 150 mg kg-1 diazinon concentration. This was also supported by the phylogenetic tree obtained from sequences of the main gel bands. Accordingly, bacterial populations belonging to Proteobacteria were enriched in the soil following three treatments with diazinon at 150 mg kg-1. The Shannon's index revealed a nonsignificant increase (P ≤ 0.05) in overall diversity of soil bacteria following diazinon application. Diazinon-degrading bacteria were isolated from the paddy soils in a mineral salt medium. Results showed that the isolated mixed culture was able to remove 90% of the pesticide at two concentrations of 50 and 100 mg L-1 by 16.81 and 19.60 days, respectively. Sequencing the DGGE bands confirmed the role of Betaproteobacteria as the main components of the isolated mixed culture in the degradation of diazinon.


Construction, expression, and activity of a bivalent bispecific single-chain antibody.
This report describes the design, construction, and expression of a bivalent bispecific single-chain antibody (SCA) protein in Escherichia coli. The bispecificity of the bivalent protein was based on two previously constructed monovalent single-chain antibody molecules possessing distinct specificities, SCA 4-4-20 (anti-fluorescein) and SCA 04-01 (anti-single-stranded DNA). A flexible linker, modeled after a secreted fungal cellulase protein, was incorporated as the interdomain linker covalently joining the two active sites. Bivalent bispecific SCA protein that accumulated in bacteria as insoluble inclusion bodies was harvested, denatured, refolded, and affinity-purified in vitro. Affinity-purified bivalent bispecific SCA showed nearly identical ligand binding properties at each site relative to the individual monovalent single-chain antibody prototype molecules. In both solid and solution phase binding assays, the bivalent bispecific single-chain antibody simultaneously bound both ligands (fluorescein and (dT)6). Construction of a model bivalent bispecific molecule provides a foundation for future assembly of similar molecules designed to identify parameters involved in enhanced binding of antibodies due to avidity and dual specificity.


Trabecular microfracture.
Healing trabecular microfractures are a common feature in cancellous bone. These lesions, when observed in macerated cancellous bone slices, measure about 500 microns in diameter and surround fractures in trabeculae with microcallus. Whether microcallus is a structure acting primarily as a transient brace, preventing relative movement of the fragmented segments and enabling the trabecula to heal, or whether it is a permanent buttress reducing the stress on the fractured strut, preventing the healing process, is not known. Microfractures are the result of normal physical activity. Hence, the widespread occurrence of trabecular microfracture in cancellous bone implies that a reasonable rate of microfracture is physiologically tolerable. There are three putative effects for trabecular microfracture. One is that, in response to impulse loading, cancellous subchondral bone increases its rigidity due to osteosclerosis resulting from bone formed around microfractures. Another hypothesis is that, if sufficient trabecular microfractures occur, they will compromise the trabecular structure of the vertebra and the proximal femur leading to osteoporotic fracture. By inducing remodeling changes, microfractures have an effect on the maintenance of joint structure. There are two histological patterns for microfractures: an early stage, when actively forming woven bone is bridging the fracture; and a more common late stage, when woven bone is inactive. Femoral studies fail to demonstrate that an increasing number of healed or healing microfractures in osteoarthrosis causes the increase of bone in the head of femur. Only one study has reported a significant increase in the number of trabecular microfractures in osteoarthritic femoral heads compared with normal controls. This significant increase was in patients taking antiinflammatory drugs.(ABSTRACT TRUNCATED AT 250 WORDS)


Female turtles from hot nests: is it duration of incubation or proportion of development at high temperatures that matters?
Mean daily temperature in natural nests of freshwater turtles with temperature-dependent sex determination is known to be a poor predictor of hatchling sex ratios when nest temperatures fluctuate. To account for this, a model was developed on the assumption that females will emerge from eggs when more than half of embryonic development occurs above the threshold temperature for sex determination rather than from eggs that spend more than half their time above the threshold. The model is consistent with previously published data and in particular explains the phenomenon whereby the mean temperature that best distinguishes between male and female nests decreases with increasing variability in nest temperature. The model, if verified by controlled experiments, has important implications for our understanding of temperature-dependent sex determination in natural nests. Both mean nest temperature and "hours spent above the threshold" will be poor predictors of hatchling sex ratios. Studies designed to investigate latitudinal trends and inter-specific differences in the threshold temperature will need to consider latitudinal and inter-specific variation in the magnitude of diel fluctuations in nest temperature, and variation in factors influencing the magnitude of those fluctuations, such as nest depth. Furthermore, any factor that modifies the relationship between developmental rate and temperature can be expected to influence hatchling sex ratios in natural nests, especially when nest temperatures are close to the threshold.


Mouse organogenesis atlas at single-cell resolution.
The generation of spatial transcriptomes of whole embryo has been limited in scale and resolution due to various technological restrictions. In this issue of Cell, Chen et al. introduce a DNA nanoball-based sample-capture technology for spatial transcriptome analysis to generate a molecular atlas of mouse organogenesis at single-cell resolution.


Physiology and pathophysiology of the digestive organs in critical illness.
Prolongation of life in critical illness has led to the increasing involvement of the digestive organs by systemic hypoperfusion. The end result of profound ischemia is multiple systems organ failure. Digestive system failure, in most probability, becomes the terminal event. Understanding the physiology and pathophysiology of the digestive organs in critical illness allows for early detection and institution of prophylactic measures. Involvement of the liver, biliary tract, pancreas, stomach, and intestines is individually discussed.


Watching the grin fade: tracing the effects of polyploidy on different evolutionary time scales.
Polyploidy, or whole-genome duplication (WGD), is a recurrent mutation both in cell lineages and over evolutionary time. By globally changing the relationship between gene copy number and other cellular entities, it can induce dramatic changes at the cellular and phenotypic level. Perhaps surprisingly, then, the insights that these events can bring to understanding other cellular features are not as well appreciated as they could be. In this review, we draw on examples of polyploidy from animals, plants and yeast to explore how investigations of polyploid cells have improved our understanding of the cell cycle, biological network complexity, metabolic phenotypes and tumor biology. We argue that the study of polyploidy across organisms, cell types, and time scales serves not only as a window into basic cell biology, but also as a basis for a predictive biology with applications ranging from crop improvement to treating cancer.


Contribution of micelles to ionic strength of surfactant solution.
A new definition of the concentration-dependent "shielding factor" has been proposed, along with its particular value at a concentration assumed to be equal to critical micelle concentration (cmc). It has been shown that herein defined shielding factor enables one to calculate the values of monomer (a(x)) and counterion activities (a(y)) which are in better agreement with the measured ones than those calculated using Burchfield and Wooley's constant value of the shielding factor. In addition, it was shown that by using particular value of shielding factor at cmc, one is able to calculate the micellar charge radius which is dependent on aggregation number, counterion parameter and cmc, all of them having important implications for thermodynamic treatment of the micellization process.


Adenosine A2B receptor activation stimulates glucose uptake in the mouse forebrain.
ATP consumption during intense neuronal activity leads to peaks of both extracellular adenosine levels and increased glucose uptake in the brain. Here, we investigated the hypothesis that the activation of the low-affinity adenosine receptor, the A2B receptor (A(2B)R), promotes glucose uptake in neurons and astrocytes, thereby linking brain activity with energy metabolism. To this end, we mapped the spatiotemporal accumulation of the fluorescent-labelled deoxyglucose, 2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-2-deoxyglucose (2-NBDG), in superfused acute hippocampal slices of C57Bl/6j mice. Bath application of the A(2B)R agonist BAY606583 (300 nM) triggered an immediate and stable (>10 min) increase of the velocity of 2-NBDG accumulation throughout hippocampal slices. This was abolished with the pretreatment with the selective A(2B)R antagonist, MRS1754 (200 nM), and was also absent in A(2B)R null-mutant mice. In mouse primary astrocytic or neuronal cultures, BAY606583 similarly increased (3)H-deoxyglucose uptake in the following 20 min incubation period, which was again abolished by a pretreatment with MRS1754. Finally, incubation of hippocampal, frontocortical, or striatal slices of C57Bl/6j mice at 37 °C, with either MRS1754 (200 nM) or adenosine deaminase (3 U/mL) significantly reduced glucose uptake. Furthermore, A(2B)R blockade diminished newly synthesized glycogen content and at least in the striatum, increased lactate release. In conclusion, we report here that A(2B)R activation is associated with an instant and tonic increase of glucose transport into neurons and astrocytes in the mouse brain. These prompt further investigations to evaluate the clinical potential of this novel glucoregulator mechanism.


Draft Whole-Genome Sequence of Sphingobium sp. 22B, a Polycyclic Aromatic Hydrocarbon-Degrading Bacterium from Semiarid Patagonia, Argentina.
Sphingobium sp. 22B is a polycyclic aromatic hydrocarbon-degrading strain isolated from Patagonia, Argentina, with capabilities to withstand the environmental factors of that semiarid region. The draft genome shows the presence of genes related with responses to carbon starvation and drying environmental conditions.


Adult-derived Index for Mortality Prediction After Cardiac Transplantation (IMPACT) risk score predicts short-term mortality after pediatric heart transplantation.
We have previously derived and validated the Index for Mortality Prediction After Cardiac Transplantation (IMPACT), which predicts short-term mortality after adult orthotopic heart transplantation (OHT). This study evaluated the predictive accuracy of the IMPACT score in pediatric OHT. The United Network for Organ Sharing registry was used to identify pediatric (< 18 years) OHT patients from 2000 to 2008. The IMPACT score was calculated for each patient. The association of IMPACT score with 1-year mortality was evaluated with univariate and multivariable logistic regression analysis. The correlation coefficient between predicted and actual 1-year mortality was determined for each IMPACT score. Kaplan-Meier survival estimates were calculated and stratified by IMPACT score. We identified 2,518 eligible pediatric OHT patients (1,128 girls [44.8%]). Mean IMPACT score was 10.3 ± 6.3 (range, 0 to 38). A total of 297 patients (11.8%) died within the first year after OHT. Each point increase in the IMPACT score increased the odds of 1-year mortality by 13% (odds ratio, 1.13; 95% confidence interval, 1.11 to 1.15; p < 0.001). The correlation coefficient between predicted and actual 1-year mortality was 0.93 (p < 0.001). One-year survival by disjoint categories of the IMPACT score was 0 to 4 (96.7%), 5 to 9 (92.9%), 10 to 14 (87.6%), 15 to 19 (81.3%), and 20 or more (64.2%; p < 0.001). In this large-cohort analysis, the IMPACT score accurately predicted mortality following pediatric OHT. The IMPACT score could therefore be useful to providers for organ allocation and prognostication in this patient population.


Valproic acid impairs carnitine uptake in cultured human skin fibroblasts. An in vitro model for the pathogenesis of valproic acid-associated carnitine deficiency.
The mechanisms of valproate-associated carnitine deficiency are controversial. The urinary excretion of valproylcarnitine is insufficient to account for tissue carnitine depletion. To explore this mechanism, we studied the effects of valproic acid (VPA) on carnitine uptake in cultured human skin fibroblasts by the method of Tein et al. (Pediatr Res 28:247-255, 1990). Fibroblasts were preincubated with varying concentrations (0-2000 microM) of VPA for 1, 3, 5, 7, 10, 14, 21, and 28 d and then incubated with fixed carnitine concentrations of 50 microM (normal physiologic concentration), 20 microM (as seen in secondary carnitine deficiency disorders), or 5 microM (as seen in the plasma membrane carnitine transport defect). There was an exponential dose-dependent decrease in carnitine uptake with increasing VPA concentrations, and the relative inhibitory effect was the same for all three carnitine concentrations. The mean percentages +/- SD (n-1) of residual carnitine uptake for all combined preincubation periods (1-28 d) and combined carnitine concentrations (5, 20, and 50 mumol/L) with increasing concentrations of VPA varied from 83.4 +/- 2.6% (10 microM VPA) to 56.7 +/- 0.1% (500 microM) to 19.8 +/- 1.3% (2000 microM). The degree of inhibition was directly proportional to the time of VPA preincubation and parallel for all three carnitine concentrations; the longer the preincubation period, the lower the toxic dose of VPA (to a minimum of 450 microM), resulting in a 50% suppression of carnitine uptake (TD50).(ABSTRACT TRUNCATED AT 250 WORDS)


Characterization of insecticide resistance in Trinidadian strains of Aedes aegypti mosquitoes.
Bioassays and biochemical assays were conducted on eight Trinidadian strains of Aedes aegypti larvae to determine the involvement of biochemical mechanisms in resistance to insecticides. Larval strains were assayed to dichlorodiphenyltrichloroethane (DDT), bendiocarb, temephos and permethrin, using the Centers for Disease Control and Prevention (CDC) time-mortality bioassay method. A Resistance Threshold (RT) was calculated for each insecticide in relation to the CAREC reference susceptible Ae. aegypti strain and larval strains with <80% mortality were considered to be resistant. Biochemical assays were performed to determine the activities of nonspecific esterases (α- and β-), PNPA-esterases, mixed function oxidases (MFO), glutathione-S-transferases (GST) and acetylcholinesterase (AChE) enzymes which are involved in insecticide resistance in mosquitoes. Enzyme profiles of each strain were compared with those of the CAREC reference susceptible strain by Kruskal-Wallis and Dunn's multiple comparison tests (p<0.05). The CAREC 99th percentile was calculated for each enzyme and the percentage of individuals with enzyme activities above that of the CAREC 99th percentile was calculated. Activities were classified as unaltered (<50%), incipiently altered (15-50%) or altered (>50%) for each strain. The established RTs for permethrin and bendiocarb were 30 and 75 min, respectively; and 120 min for DDT and temephos. All strains were resistant to DDT (1.00-40.25% mortality) and temephos (11.50-74.50% mortality) while six strains were resistant to bendiocarb (51.50-78.50% mortality) and five to permethrin (6.50-42.50% mortality). Biochemical assays revealed that the median activity levels for all enzymes varied significantly (p<0.05). The Curepe strain had incipiently altered levels of α-esterase while the other seven strains had altered activity with five of them registering 100%. The St Clair strain showed altered activity levels of β-esterase while three strains had incipiently altered levels. The majority of strains had altered activity of MFO enzymes but only the St Clair strain showed altered activity of GST. PNPA-esterases activity was unaltered in all strains and only the Haleland Park strain showed altered remaining AChE activity in the presence of propoxur. Elevated levels of enzymes (incipiently altered or altered), except in the case of PNPA-esterases, show that biochemical resistance may play an important role in the manifestation of insecticide resistance in Trinidadian populations of Ae. aegypti. It is therefore important for insecticide resistance surveillance to be ongoing as the detection of resistance before it spreads throughout an entire population makes it possible for early intervention.


Defibrotide reduces procoagulant activity and increases fibrinolytic properties of endothelial cells.
Defibrotide (DF), a polydeoxyribonucleotide with antithrombotic properties, has recently proven effective in patients with severe hepatic veno-occlusive disease (VOD), a life-threatening complication of high-dose chemo/radiotherapy regimens for stem cell transplantation. To understand the mechanism of its beneficial effect, we studied the impact of DF on the expression of tissue factor (TF) and fibrinolytic proteins (PAI-1 and t-PA) on endothelial cells. The in vitro response to DF of two types of human endothelial cells (ECs) of different origins, that is from macrovascular (HUVEC) and microvascular (HMEC-1 cell line) beds, was evaluated in the presence or absence of a proinflammatory stimulus (ie bacterial endotoxin, LPS). The results show that DF was able to significantly reduce the LPS-induced TF expression by HMEC-1, and less prominently by HUVEC. In addition, DF importantly influenced the fibrinolytic properties of both HMEC-1 and HUVEC. Specifically, it dose-dependently counteracted the LPS-induced increase in PAI-1 levels and decrease in t-PA activity expression. It also significantly incremented t-PA antigen in resting EC. Decreasing the procoagulant activity and increasing the fibrinolytic potential of EC favors an anticoagulant phenotype of the endothelium, which may protect from fibrin deposition and vascular occlusion.


Headache and seizure in a young woman postpartum.
A 19-year-old woman, 8 days postpartum, presented to the Emergency Department (ED) with a chief complaint of headache, decreased vision, and agitation. Past medical history was unremarkable. Physical examination was remarkable only for dry mucous membranes, decreased visual acuity, and trace pedal edema bilaterally. While in the ED, the patient experienced a generalized tonic-clonic seizure. Computed tomography (CT) scan of the head was suspicious for venous infarction. Magnetic resonance imaging (MRI) and MR venography revealed sagittal sinus and left transverse sinus thrombosis. The risk factors, clinical presentation, diagnostic evaluation, and management of cerebral venous thrombosis are reviewed.


Functional heterogeneity of human skin-resident memory T cells in health and disease.
The human skin is populated by a diverse pool of memory T cells, which can act rapidly in response to pathogens and cancer antigens. Tissue-resident memory T cells (TRM ) have been implicated in range of allergic, autoimmune and inflammatory skin diseases. Clonal expansion of cells with TRM properties is also known to contribute to cutaneous T-cell lymphoma. Here, we review the heterogeneous phenotypes, transcriptional programs, and effector functions of skin TRM . We summarize recent studies on TRM formation, longevity, plasticity, and retrograde migration and contextualize the findings to skin TRM and their role in maintaining skin homeostasis and altered functions in skin disease.


Cell cultured chondrocyte implantation and scaffold techniques for osteochondral talar lesions.
Cell cultured techniques have gained interest and popularity in osteochondral defects because, unlike bone marrow stimulation methods, where fibrocartilage fills the defect, they allow for the regeneration of "hyaline-like cartilage" with better stiffness, resilience, and wear characteristics. Osteochondral defects in the ankle are a rare but challenging problem to treat in young active patients. If left alone, they can cause pain and reduced function and risk progressive degenerative changes in the joint. Clinical results of cell cultured and scaffold technology in the ankle, although still limited by small studies and midterm follow-up, are certainly encouraging.


Chicken "erythroid" cells transformed by the Gag-Myb-Ets-encoding E26 leukemia virus are multipotent.
The E26 avian leukemia virus encodes a transcriptional activator-type oncoprotein consisting of Gag, Myb, and Ets domains, and transforms early erythroid cells as well as myeloblasts. Surprisingly, we have found that "early erythroid" transformants obtained in culture are multipotent, since they can be induced to differentiate into myeloblasts and eosinophils after superinfection with retroviruses containing kinase-type or ras oncogenes. In addition, TPA is an efficient inducer that generates predominantly eosinophils at low concentrations and myeloblasts at high concentrations. The determination process involves the complete extinction of erythroid/thrombocytic markers and the subsequent activation of myelomonocytic/eosinophilic properties, including the acquisition of specific growth factor requirements. "Erythroleukemic" cells from virus-infected animals were likewise found to be multipotent, making this a unique system to study the genesis of stem cell leukemias and the molecular basis of lineage commitment during hematopoiesis.


Temporal dynamics of base excision/single-strand break repair protein complex assembly/disassembly are modulated by the PARP/NAD+/SIRT6 axis.
Assembly and disassembly of DNA repair protein complexes at DNA damage sites are essential for maintaining genomic integrity. Investigating factors coordinating assembly of the base excision repair (BER) proteins DNA polymerase β (Polβ) and XRCC1 to DNA lesion sites identifies a role for Polβ in regulating XRCC1 disassembly from DNA repair complexes and, conversely, demonstrates Polβ's dependence on XRCC1 for complex assembly. LivePAR, a genetically encoded probe for live-cell imaging of poly(ADP-ribose) (PAR), reveals that Polβ and XRCC1 require PAR for repair-complex assembly, with PARP1 and PARP2 playing unique roles in complex dynamics. Further, BER complex assembly is modulated by attenuation/augmentation of NAD+ biosynthesis. Finally, SIRT6 does not modulate PARP1 or PARP2 activation but does regulate XRCC1 recruitment, leading to diminished Polβ abundance at sites of DNA damage. These findings highlight coordinated yet independent roles for PARP1, PARP2, and SIRT6 and their regulation by NAD+ bioavailability to facilitate BER.


De novo ceramides synthesis is not involved in skeletal muscle atrophy induced by short-term mechanical unloading.
Patients admitted to the intensive care unit commonly develop skeletal muscle weakness that can exacerbate illness and complicate their recovery. Beyond the primary disease or aging, weakness is promoted by a variety of prolonged hospitalization-associated conditions. These include altered nutritional status, pharmacologic side effects, physical inactivity, and prolonged bed rest. The two latter conditions (i.e. inactivity and bed rest) are the most ubiquitous, affecting all patients during a prolonged hospitalization. In both cases, skeletal muscle utilization is decreased with a concomitant reduction in fatty acid oxidation. Subsequent fatty acids accumulation converted to ceramides could be a cellular mechanism leading to muscle wasting. Indeed these sphingolipids act as second messengers in several of molecular signaling pathways involved in muscle atrophy. Consequently, the aim of this work is to determine the effects of immobilization on muscle ceramides accumulation, and identify the role of these ectopic lipids in molecular mechanisms involved in skeletal muscle atrophy. For this purpose, male Wistar rats were treated with an inhibitor of de novo synthesis of ceramides (i.e. myriocin) and subjected to hindlimb unloading for 7 days. We found that hindlimb unloading induced skeletal muscle atrophy, in part through proteolysis (i.e. decrease in AKT activation, increase in MuRF1 and polyubiquinated proteins content) and apoptosis activations (i.e. increase in Bax/Bcl-2 ratio and cleaved caspase-3). Myriocin treatment did not prevent skeletal muscle atrophy and concomitant induction of apoptosis and proteolysis. Data concerning muscle ceramides content are being analyzed. Together, these results suggest that de novo synthesis of ceramides is not involved in muscle atrophy induced by a short period of hindlimb unloading.


A Bayesian approach for application to method validation and measurement uncertainty.
The aim of this paper is to apply a new technique for the validation of quantitative analytical procedures based on Bayesian simulation and accuracy profile. Also, an original strategy for estimating measurement uncertainty by the same approach has been developed. The performance of our proposal was confirmed by application to analytical and bio-analytical methods. Compared to the classical strategy, the new approach has a more holistic character. It means that it is no longer necessary to know the various individual steps into which the analytical method can be broken down since this latter is taken as a whole. Furthermore, the Bayesian accuracy profile procedure allows to control the risk associated with the future use of the analytical method.


Evidence of sweepstakes reproductive success in a broadcast-spawning coral and its implications for coral metapopulation persistence.
Processes governing genetic diversity and adaptive potential in reef-building corals are of interest both for fundamental evolutionary biology and for reef conservation. Here, we investigated the possibility of "sweepstakes reproductive success" (SRS) in a broadcast spawning coral, Acropora hyacinthus, at Yap Island, Micronesia. SRS is an extreme yearly variation in the number of surviving offspring among parents. It is predicted to generate genetically differentiated, low-genetic-diversity recruit cohorts, containing close kin individuals. We have tested these predictions by comparing genetic composition of size classes (adults and juveniles) at several sites on the island of Yap. We did see the genome-wide dip in genetic diversity in juveniles compared to adults at two of the four sites; however, both adults and juveniles varied in genetic diversity across sites, and there was no detectable genetic structure among juveniles, which does not conform to the classical SRS scenario. Yet, we have identified a pair of juvenile siblings at the site where juveniles had the lowest genetic diversity compared to adults, an observation that is hard to explain without invoking SRS. While further support for SRS is needed to fully settle the issue, we show that incorporating SRS into the Indo-West Pacific coral metapopulation adaptation model had surprisingly little effect on mean rates of coral cover decline during warming. Still, SRS notably increases year-to-year variation in coral cover throughout the simulation.


Depression and Pseudodementia: Decoding the Intricate Bonds in an Italian Outpatient Setting.
In spite of the uncertainties of its diagnostic framework, pseudodementia may be conceptualized as a condition characterized by depressive symptoms and cognitive impairment in the absence of dementia. Given the controversies on this topic, the aim of the present study was to assess neurological and cognitive dysfunctions in a sample of elderly depressed subjects, and the eventual relationship between cognitive impairment and depressive symptoms. Fifty-seven elderly depressed outpatients of both sexes were included in the study. A series of rating scales were used to assess diagnoses, depressive and cognitive impairment. Comparisons for continuous variables were performed with the independent-sample Student's t-test. Comparisons for categorical variables were conducted by the χ2 test (or Fisher's exact test when appropriate). The correlations between between socio-demographic characteristics and clinical features, as well as between cognitive impairment and depressive symptoms were explored by Pearson's correlation coefficient or Spearman's rank correlation. Our data showed the presence of a mild-moderate depression and of a mild cognitive impairment that was only partially related to the severity of depression. These dysfunctions became more evident when analyzing behavioral responses, besides cognitive functions. A high educational qualification seemed to protect against cognitive decline, but not against depression. Single individuals were more prone to cognitive disturbance but were similar to married subjects in terms of the severity of depressive symptoms. Previous depressive episodes had no impact on the severity of depression or cognitive functioning. Although data are needed to draw firm conclusions, our findings strengthen the notion that pseudodementia represents a borderline condition between depression and cognitive decline that should be rapidly identified and adequately treated.


Morphologic evolution of Au nanocrystals grown in ionic liquid by plasma reduction.
Shape controlled Au nanocrystal growth in ionic liquid has been studied by glow discharge plasma reduction. In an 1-butyl-3-methylimidazolium tetrafluoroborate ([bmim][BF(4)]) solution with poly(vinyl pyrrolidone) (PVP) as surfactant, Au nanocrystals grow from faceted NPs to peanut-like particle aggregates, and eventually worm-like nanorods and nanowires as the growth time increases. This process is further found to be dependent on the concentration of employed surfactant. Based on systematic microscopic analysis and control experiments, a polymer-assisted oriented attachment mechanism is proposed to explain the observed morphologic evolution. Our work may have great implications for the employment of ionic liquids for shape controlled synthesis of nanocrystals.


DLP-based bioprinting of void-forming hydrogels for enhanced stem-cell-mediated bone regeneration.
The integration of 3D bioprinting and stem cells is of great promise in facilitating the reconstruction of cranial defects. However, the effectiveness of the scaffolds has been hampered by the limited cell behavior and functions. Herein, a therapeutic cell-laden hydrogel for bone regeneration is therefore developed through the design of a void-forming hydrogel. This hydrogel is prepared by digital light processing (DLP)-based bioprinting of the bone marrow stem cells (BMSCs) mixed with gelatin methacrylate (GelMA)/dextran emulsion. The 3D-bioprinted hydrogel can not only promote the proliferation, migration, and spreading of the encapsulated BMSCs, but also stimulate the YAP signal pathway, thus leading to the enhanced osteogenic differentiation of BMSCs. In addition, the in vivo therapeutic assessments reveal that the void-forming hydrogel shows great potential for BMSCs delivery and can significantly promote bone regeneration. These findings suggest that the unique 3D-bioprinted void-forming hydrogels are promising candidates for applications in bone regeneration.


Increased circulating level of interleukin-6 and CD8+ T cell exhaustion are associated with progression of COVID-19.
Coronavirus disease 2019 (COVID-19) is pandemic. It is critical to identify COVID-19 patients who are most likely to develop a severe disease. This study was designed to determine the clinical and epidemiological features of COVID-19 patients associated with the development of pneumonia and factors associated with disease progression. Seventy consecutive patients with etiologically confirmed COVID-19 admitted to PLA General Hospital in Beijing, China from December 27, 2019 to March 12, 2020 were enrolled in this study and followed-up to March 16, 2020. Differences in clinical and laboratory findings between COVID-19 patients with pneumonia and those without were determined by the χ2 test or the Fisher exact test (categorical variables) and independent group t test or Mann-Whitney U test (continuous variables). The Cox proportional hazard model and Generalized Estimating Equations were applied to evaluate factors that predicted the progression of COVID-19. The mean incubation was 8.67 (95% confidence interval, 6.78-10.56) days. Mean duration from the first test severe acute respiratory syndrome coronavirus 2-positive to conversion was 11.38 (9.86-12.90) days. Compared to pneumonia-free patients, pneumonia patients were 16.5 years older and had higher frequencies of having hypertension, fever, and cough and higher circulating levels of neutrophil proportion, interleukin-6, low count (< 190/µl) of CD8+ T cells, and neutrophil/lymphocyte ratio. Thirteen patients deteriorated during hospitalization. Cox regression analysis indicated that older age and higher serum levels of interleukin-6, C-reactive protein, procalcitonin, and lactate at admission significantly predicted the progression of COVID-19. During hospitalization, circulating counts of T lymphocytes, CD4+ T cells, and CD8+ T cells were lower, whereas neutrophil proportion, neutrophil/lymphocyte ratio, and the circulating levels of interleukin-6, C-reactive protein, and procalcitonin were higher, in pneumonia patients than in pneumonia-free patients. CD8+ lymphocyte count in pneumonia patients did not recover when discharged. Older age and higher levels of C-reactive protein, procalcitionin, interleukin-6, and lactate might predict COVID-19 progression. T lymphocyte, especially CD8+ cell-mediated immunity is critical in recovery of COVID-19. This study may help in predicting disease progression and designing immunotherapy for COVID-19.


Differences in Administration of Methotrexate and Impact on Outcome in Low-Risk Gestational Trophoblastic Neoplasia.
Methotrexate (MTX) is frequently used as first-line treatment for low-risk gestational trophoblastic neoplasia (GTN). Intravenous and intramuscular (im) routes of administration are the most common methods, although oral administration is used by some Scandinavian centers. The primary aim of this study was to assess the impact of form of administration (im/oral) on resistance to methotrexate (MTX-R) treatment in low-risk GTN. Secondary aims were time to hCG normalization, rates of toxicity-induced treatment switch, and rates of complete remission and recurrence. In total, 170 women treated at Karolinska University Hospital in Sweden and Aarhus University Hospital in Denmark between 1994 and 2018 were included, of whom 107 were given im and 63 oral MTX. MTX-R developed in 35% and 54% in the im and oral groups, respectively (p = 0.01). There was no difference in days to hCG normalization (42 vs. 41 days, p = 0.50) for MTX-sensitive women. Toxicity-induced treatment switch was only seen in the im group. Complete remission was obtained in 99.1% and 100% (p = 0.44), and recurrence rate within one year was 2.8% and 1.6% (p = 0.29). The form of administration of MTX had a significant impact on development of MTX-R and treatment-associated toxicity, but does not affect rates of complete remission, recurrence or survival.


Behavioral improvements in a valproic acid rat model of autism following vitamin D supplementation.
The aim was to identify the effects of early vitamin D supplementation on autism-like behaviors (ASD) induced by valproic acid (VPA, an anti-convulsant and a mood stabilizer) in rats. 10 male Wistar rat pups with prenatal exposure to saline were in control group, and 20 Pups with prenatal exposure to VPA were divided into ASD-N (0.9% saline treated) and ASD-D group (vitamin D 80,000 IU/kg treated) on postnatal day 12. Self-grooming, olfactory habituation/dishabituation, and social interaction tests were conducted to assess social interaction, communication, and repetitive behaviors. Serum 25-hydroxyvitamin D (25(OH)D3) was measured by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Results showed that compared with the control group, the ASD-N group exhibited increased self-grooming, and decreased pinning and serum 25(OH)D3. Furthermore, the repetitive behavior of the ASD-N group exhibited a negative linear relationship with serum 25(OH)D3 on PND 42. In conclusion, early vitamin D supplementation in infant rat with ASD induced by VPA significantly improved development and behavior of rats related with ASD.


Muscarinic receptor activation down-regulates the type I inositol 1,4,5-trisphosphate receptor by accelerating its degradation.
Stimulation of SH-SY5Y human neuroblastoma cells with carbachol, a muscarinic agonist, down-regulates the type I inositol 1,4,5-trisphosphate (InsP3) receptor by > 90% with maximal and half-maximal effects after approximately 6 h and approximately 1 h, respectively. Examination of the mechanistic basis of this down-regulation revealed that carbachol increased the rate of type I InsP3 receptor degradation (radiolabeled immunoprecipitable receptor was lost from cells with half-times of > 8 h and approximately 1 h in the absence and presence of carbachol, respectively) and that the concentration of type I InsP3 receptor mRNA, despite a transient decrease after 3 h, did not correlate with levels of the receptor. Only those muscarinic receptor subtypes coupled to stimulation of phosphoinositide hydrolysis were capable of causing type I InsP3 receptor down-regulation. Ca2+ mobilization was pivotal to the mechanisms of receptor down-regulation, since perturbation of Ca2+ homeostasis with either EGTA or thapsigargin blocked the ability of carbachol to accelerate receptor degradation. Studies with thapsigargin also revealed that both functional InsP3-sensitive Ca2+ stores and persistent elevation of InsP3 concentration were required for down-regulation to occur. In conclusion, phosphoinositidase C-linked muscarinic receptors down-regulate the type I InsP3 receptor by accelerating its degradation. It appears that this process is initiated by persistent discharge of intracellular Ca2+ stores via the channels formed by tetramerically complexed type I InsP3 receptors.


Environmental monitoring of toxic metals in roadside soil and dust in Ulsan, South Korea: pollution evaluation and distribution characteristics.
This study investigated toxic metal distribution in roadside soil and dust in the metropolitan city of Ulsan, South Korea, and the factors affecting distribution, using Korean waste-leaching tests, determination of total concentrations, sequential extraction, and statistical analysis. Composite grab samples were collected from high-traffic roads (7 sites), low-traffic roads (2 sites), and an uncontaminated control area (2 sites) in Ulsan. The pH of roadside soil and dust was slightly alkaline. The concentrations of copper, lead, and zinc in soil as determined by Korean waste-leaching tests decreased as soil depth increased, while those of arsenic, nickel, and chromium increased. Leaching concentrations in dust were lower than in soil, with the exception of copper. Total concentrations decreased as soil depth increased, and total concentrations of metals in dust were higher than in soil. The sampling sites that exceeded the regulation levels of soil contamination in South Korea were 7 points in topsoil, 3 points in middle soil, and 9 points in dust. TCLP tests showed that the concentrations of arsenic, cadmium, and lead in topsoil and dust at Duwang and Myeongchon intersections were higher than regulatory levels. The maximum correlation coefficient among two metals in soil and dust was 0.987 (p < 0.01), for cadmium and lead. Concentrations of cadmium, copper, arsenic, lead, nickel, and mercury, mostly from tire and brake-pad abrasion, were highly correlated. The strong positive correlation between traffic volume and metals in dust suggests that vehicle emissions may be responsible for metal contamination of soil and dust. Pollution indices of topsoil at 4 sites and all dust at 7 high-traffic sites were higher than 1.0, which is consistent with an effect of vehicle traffic on metal contamination in soil and dust.


Serotonin release is modulated by presynaptic autoreceptors.
The existence of presynaptic autoreceptors controlling the release of 5-hydroxytryptamine (5HT) from serotonergic nerve endings was investigated utilizing superfused hypothalamic synaptosomes. Extracellular 5HT reduced the high K+-induced release of previously accumulated 3H-5HT. The central 5HT receptor blocker methiothepin counteracted the inhibitory effect of 5HT. Other 5HT antagonists (cyproheptadine, methysergide and mianserin) were inactive and may therefore act preferentially at the postsynaptic receptors.


Effect of vagotomy upon the neurohistochemical and ultrastructural integrity of the integrity of the inbuilt intrinsic nervous apparatus of the choledocho-duodenal junction.
The neurons of the choledocho-duodenal junction of the cat were shown to be neurohistochemically and morphologically independent of their extrinsic vagal connections. The effect of vagotomies upon the intrinsic nerve nets was also quite negligible.


[Incidence and risk factors of exposure keratopathy in intensive care units: About 91 patients].
Exposure keratopathy is a sight-threatening complication which can occur in patients admitted to intensive care units. This is a prospective study over a period of 5 months in the intensive care unit of the specialty hospital of the IBN SINA medical center, with a capacity of 8 beds. The purpose of our study was to determine the incidence of and risk factors for exposure keratopathy in intensive care settings. Forty-two percent of patients developed exposure keratopathy, 48% exhibited chemosis, and 40% showed lagophthalmos with the cornea visible in 30% of cases. The frequency of keratopathy was significantly higher in patients with chemosis and lagophthalmos (P<0.0001). Lagophthalmos was significantly related to chemosis (P<0.0001). Chemosis in the ventilated patient, also known as "ventilator eye," is a serious complication leading to the risk of keratopathy. The risk factors for keratopathy found in our series were chemosis (OR=25.28, 95% CI=[3.339-191.52] P-value=0.002), lagophthalmos (OR=40.8, 95% CI=[4.347-383.666] P-value=0.001) and length of stay in intensive care (OR=12.28, 95% CI=[1.476-102.230] P-value=0.020). Several methods might be used and adapted to each case for prevention of exposure keratopathy, and we found that raising nursing staff awareness is of paramount importance.


The relationship between muscle power, functional performance, accelerometer-based measurement of physical activity and patient-reported outcomes in patients with hip osteoarthritis: A cross-sectional study.
Patients with unilateral hip osteoarthritis appear to have between-leg differences in leg extension power (LEP). The Nottingham Leg Extensor Power Rig provides reliable and valid results but requires sensitive equipment. It would be relevant to identify measures closely associated with this test. (i) To investigate if LEP is lower in the affected leg compared to the non-affected leg. Furthermore, to investigate the associations between LEP and the measures: (ii) Functional performance, (iii) accelerometer-based measurement of physical activity and (iv) patient-reported outcome measures (PROM). Cross-sectional study including 60 patients (30 men, 30 women) with hip osteoarthritis scheduled for hip replacement. The counter movement jump and 10-m sprint tests were used to determine functional performance, accelerometer-sensors were used to determine physical activity and the Copenhagen Hip and Groin Outcome Score (HAGOS) was used to determine PROM. (i) LEP in the affected leg corresponded to 79% [95% CI 74%; 85%] of the non-affected leg, (ii) LEP was positively associated with functional performance tests (β 0.63 to 0.78, p < 0.05), (iii) positively associated although non-significantly with physical activity (β 0.16 to 0.23, p > 0.05) and (iv) positively associated with the six HAGOS subscales (β 0.25 to 0.54, p < 0.05). Functional performance tests may be used as feasible, inexpensive and fast ways to assess LEP in clinical settings. These results may suggest that interventions aimed at improving LEP can improve functional performance and PROM, but not physical activity. Future research is needed to confirm the causality of these cross-sectional findings. Danish Data Protection Agency (1-16-02-640-14), ClinicalTrials.gov (NTC02301182) and approved by the Danish Biomedical Research Ethics Committee (1-10-72- 343-14) prior to data collection.


Clinical and laboratory diagnosis of bleeding disorders.
The diagnosis of hemostatic abnormalities requires a detailed clinical and laboratory evaluation of the patient. The clinical assessment includes a detailed history and a thorough physical examination. The patient's history may provide clues as to the time of onset of the bleeding tendency, the clinical severity of the abnormality, and the possible contributing role of other disease processes or of drugs. The nature of the bleeding symptoms may provide clues as to the nature of the hemostatic defect. Hemostatic screening tests are invaluable in helping to differentiate between platelet, vascular, coagulation, and fibrinolytic abnormalities. Specific tests, including specific factor assays, platelet aggregometry, and antiplatelet antibody assays are usually required to characterize the exact nature and severity of hemostatic defects.


Design and validation of a questionnaire to evaluate the usability of computerized critical care information systems.
The implementation of computerized critical care information systems (CCIS) can improve the quality of clinical care and staff satisfaction, but also holds risks of disrupting the workflow with consecutive negative impacts. The usability of CCIS is one of the key factors determining their benefits and weaknesses. However, no tailored instrument exists to measure the usability of such systems. Therefore, the aim of this study was to design and validate a questionnaire that measures the usability of CCIS. Following a mixed-method design approach, we developed a questionnaire comprising two evaluation models to assess the usability of CCIS: (1) the task-specific model rates the usability individually for several tasks which CCIS could support and which we derived by analyzing work processes in the ICU; (2) the characteristic-specific model rates the different aspects of the usability, as defined by the international standard "ergonomics of human-system interaction". We tested validity and reliability of the digital version of the questionnaire in a sample population. In the sample population of 535 participants both usability evaluation models showed a strong correlation with the overall rating of the system (multiple correlation coefficients ≥0.80) as well as a very high internal consistency (Cronbach's alpha ≥0.93). The novel questionnaire is a valid and reliable instrument to measure the usability of CCIS and can be used to study the influence of the usability on their implementation benefits and weaknesses.


Eight novel mutations in the ABCD1 gene and clinical characteristics of 25 Chinese patients with X-linked adrenoleukodystrophy.
X-linked adrenoleukodystrophy (X-ALD) is a fatal neurodegenerative disease caused by mutations in the adenosine triphosphate-binding cassette D1 (ABCD1) gene. This study aimed to retrospectively investigate the clinical characteristics of 25 patients with X-ALD including members of large pedigrees, to analyze ABCD1 gene mutations, the effect of gene novel variants on ALD protein (ALDP) structure and function, and to expand gene mutation spectrum of Chinese patients. Twenty-five male patients diagnosed with X-ALD were enrolled in this study. The clinical characteristics of the patients were retrospectively summarized by reviewing medical records or telephone consultation. ABCD1 gene mutations were analyzed. The pathogenicity of novel missense variants was analyzed using cobalt constraint-based multiple protein alignment tool, polymorphism phenotyping, sorting intolerant from tolerant, Align-Grantham variation and Grantham deviation, and Swiss-Program Database Viewer 4.04 software, respectively. Childhood cerebral form ALD (CCALD) is the most common phenotype (64%) in the 25 patients with X-ALD. The progressive deterioration of neurological and cognitive functions is the main clinical feature. The demyelination of the brain white matter and elevated plasma very long chain fatty acids (VLCFAs) were found in all patients. Different phenotypes were also presented within family members of the patients. Twenty-two different mutations including 8 novel mutations in the ABCD1 gene were identified in the 25 patients. Of the mutations, 63.6% were missense mutations and 34.8% located in exon 1. The amino acid residues of three novel missense mutations in eight species were highly conserved, and were predicted to be "probably" damaging to ALDP function. The other five novel mutations were splice, nonsense, deletion or duplication mutations. CCALD is the most common phenotype (64%) in our patients with X-ALD. Eight novel mutations in the ABCD1 gene identified are disease-causing mutations. Brain magnetic resonance imaging and plasma VLCFA determination should be performed for the patients who present with progressive deterioration of neurological development.


The spectrum of small bowel melanoma.
Twenty-one patients with documented secondary small bowel involvement by malignant melanoma are reviewed. Roentgen mainfestations are discussed and illustrated. A vigorous diagnostic approach emphasizing enteroclysis is described. Clinical awareness and aggressive radiologic investigation are providing these patients with earlier surgical and adjuvant therapy, which appears to be leading to increased length of survival.


TGF-β3 enhances cell-to-cell communication in chondrocytes via the ALK5/p-Smad3 axis.
Gap junctional intercellular communication (GJIC) is indispensable for the maintenance of physiological balance in articular cartilage. Transforming growth factor-β3 (TGF-β3), an important growth factor of TGF-β superfamily, is well recognized to play a unique regulatory role in cartilage development and diseases. However, the role of TGF-β3 in GJIC in adult chondrocytes remains elusive. This work aims to investigate the effect of TGF-β3 on gap-junction mediated intercellular communication in chondrocytes. We first showed that TGF-β3 could enhance the synaptic connections between chondrocytes by scanning electron microscopy (SEM) and promote the cell-to-cell communication in living chondrocytes by scrape loading/dye transfer assay. We then confirmed that TGF-β3 enhanced cell-to-cell communication via up-regulation of connexin 43 (Cx43). We next found that TGF-β3-enhanced GJIC required the participation of TGF-beta type I receptor ALK5 and depended on the activation of p-Smad3 signalling. Finally, through inhibitor experiments of SB525334 and SIS3, we demonstrated that TGF-β3-induced functional GJIC in chondrocytes via the axis of ALK5/p-Smad3 signalling. Taking together, these results demonstrate a strong correlation between TGF-β3 and GJIC in chondrocytes, which provides a new perspective on the importance of TGF-β3 on cartilage physiology and pathobiology.


Impurities in Bose-Einstein Condensates: From Polaron to Soliton.
We propose that impurities in a Bose-Einstein condensate which is coupled to a transversely laser-pumped multimode cavity form an experimentally accessible and analytically tractable model system for the study of impurities solvated in correlated liquids and the breakdown of linear-response theory [corrected]. As the strength of the coupling constant between the impurity and the Bose-Einstein condensate is increased, which is possible through Feshbach resonance methods, the impurity passes from a large to a small polaron state, and then to an impurity-soliton state. This last transition marks the breakdown of linear-response theory.


Analysis of predictors of response to ruxolitinib in patients with myelofibrosis in the phase 3b expanded-access JUMP study.
Data from the large, prospective, multinational, phase 3b JUMP study were analyzed to identify factors predictive of spleen and symptom responses in myelofibrosis patients receiving ruxolitinib. Factors associated with higher spleen response rates included International Prognostic Scoring System (IPSS) low/intermediate-1 risk vs intermediate-2/high risk (43.1% vs 30.6%; adjusted OR [aOR] 0.65 [95% CI 0.44-0.95]), ruxolitinib as first- vs second- or later-line therapy (40.2% vs 31.5%; aOR 0.53 [95% CI 0.38-0.75]), and a ruxolitinib total daily dose at Week 12 of >20 mg/day vs ≤20 mg/day (41.3% vs 30.4%; aOR 0.47 [95% CI 0.33-0.68]). No association was seen between baseline characteristics or total daily dose at Week 12 and symptom response. Ruxolitinib led to higher spleen response rates in patients with lower IPSS risk, and when used earlier in treatment. Higher doses of ruxolitinib were associated with higher spleen response rates, but not with symptom improvement.Trial registrationINC424 for patients with primary myelofibrosis, post polycythemia myelofibrosis or post-essential thrombocythemia myelofibrosis (JUMP).2010-024473-39; NCT01493414Date of registration: 16 December 2011https://www.clinicaltrialsregister.eu/ctr-search/search?query=2010-024473-39https://clinicaltrials.gov/ct2/show/NCT01493414.


Mast cells in meningiomas and brain inflammation.
Research focus in neuro-oncology has shifted in the last decades towards the exploration of tumor infiltration by a variety of immune cells and their products. T cells, macrophages, B cells, and mast cells (MCs) have been identified. A systematic review of the literature was conducted by searching PubMed, EMBASE, Google Scholar, and Turning Research into Practice (TRIP) for the presence of MCs in meningiomas using the terms meningioma, inflammation and mast cells. MCs have been detected in various tumors of the central nervous system (CNS), such as gliomas, including glioblastoma multiforme, hemangioblastomas, and meningiomas as well as metastatic brain tumors. MCs were present in as many as 90 % of all high-grade meningiomas mainly found in the perivascular areas of the tumor. A correlation between peritumoral edema and MCs was found. Accumulation of MCs in meningiomas could contribute to the aggressiveness of tumors and to brain inflammation that may be involved in the pathogenesis of additional disorders.


COVID-19: What should health professionals know? Assessment of Knowledge, attitude, and practice of community pharmacists in a developing country.
Healthcare professionals are the primary frontiers of the fight against every public health threat. These had made them vulnerable to various infectious agents, with many reports of morbidity and mortality also being evident. The current study aimed to assess the knowledge, attitude, and practice toward COVID-19 among community pharmacists in Gondar town, Ethiopia. A cross-sectional study was conducted. A structured, self-administered questionnaire was used for data collection, and the collected data were coded, validated, and analyzed using SPSS version 26. Quantitative data were summarized using means and standard deviations, while frequencies and proportions were used to summarize categorical data. A chi-square test was used to assess an association between the different variables, and a P-value of less than 0.05 was considered to declare statistical significance at a 95% confidence interval. A total of 46 community pharmacy professionals had participated in the study. The main sources of information about COVID-19 were news media 29 (63%), followed by social media 24 (52.2%). The mean knowledge score was 8.15 (standard deviation: 1.86), and among the total study participants, 29 (63%) were found to have good knowledge about COVID-19. The mean attitude score was 31.52 (standard deviation: 4.288). Only 30.4% of the study participants were found to have a good practice toward the prevention of COVID-19. Chi-square results showed that having a good knowledge of COVID-19 was significantly associated with practice (P = 0.035). Significant gaps were identified concerning community pharmacy professional's knowledge of COVID-19 and their practice toward prevention of the disease. Strategies should be implemented to equip community pharmacy professionals with all necessary information about COVID-19 through continuous training programs and other virtual platforms.


Taping versus electrical stimulation after botulinum toxin type A injection for wrist and finger spasticity. A case-control study.
To compare results from two approaches used in conjunction with botulinum toxin type A administration in rehabilitation: the application of a taping system and the electrical stimulation of the injected muscles and splinting. Case-control study. Two tertiary care rehabilitation hospitals in Italy. Sixty-five adult subjects affected by spasticity of the wrist and finger flexors. After injection with botulinum toxin type A, the group at hospital A (n=33) was treated with adhesive taping for six days and those at hospital B (n=32) with electrical stimulation and splinting for six days. Spastic hypertonia at the injected muscles was assessed before treatment, one week and one month post injection. Modified Ashworth Scale. In group A, the mean Modified Ashworth Scale reduction was 2.76 +/- 0.94 for wrist flexors and 2.45 +/- 0.92 for finger flexors; in group B the mean Modified Ashworth Scale reduction was 2.18 +/- 1.11 for wrist flexors and 2.1 +/- 0.98 for finger flexors. The observed difference between the two groups was statistically relevant (p < 0.05). Patients treated with adhesive taping and botulinum toxin type A achieved a greater reduction in spastic hypertonia as measured with Modified Ashworth Scale, with less time dedicated for the treatment.


A paclitaxel analogue with a 2(3-->20)abeotaxane skeleton: synthesis and biological evaluation.
A paclitaxel analogue having an unusual tricyclic [126.96.36.199] hexadecane skeleton was synthesized from deaminoacyltaxine A, a 2(3-->20) abeotaxane isolated in considerable amounts from the leaves of Taxus baccata L. In preliminary studies, this compound showed a much lower cytotoxicity than paclitaxel.


Enkephalin contributes to the locomotor stimulating effects of 3,4-methylenedioxy-N-methylamphetamine.
3,4-methylenedioxy-N-methylamphetamine (MDMA, 'Ecstasy') is a potent inhibitor of serotonin uptake, which induces both an increase in locomotion and a decrease in exploratory activity in rodents. Serotonin 5-HT1B receptors, located on the terminals of striatal efferent neurons, have been suggested to mediate these motor effects of MDMA. Striatal neurons projecting to the globus pallidus contain met-enkephalin, whilst those projecting to the substantia nigra contain substance P. We therefore analysed the levels of both peptides using radioimmunocytochemistry after MDMA administration (10 mg/kg, 3 h) in wild-type and 5-HT1B receptor knockout mice. Our results demonstrate that MDMA induces a decrease in pallidal met-enkephalin immunolabelling in wild-type, but not in 5-HT1B receptor knockout mice. Similar results were obtained following treatment with the 5-HT1A/1B agonist RU24969 (5 mg/kg, 3 h), suggesting that activation of 5-HT1B receptors leads to a reduction in met-enkephalin levels in the globus pallidus. In contrast, MDMA had no effect on the nigral substance P levels. We have previously shown that both MDMA and RU24969 fail to stimulate locomotor activity in 5-HT1B receptor knockout mice. Our present data indicate that the opioid antagonist naloxone suppressed the locomotor effects of MDMA. This study is the first to demonstrate that Enk contributes to MDMA-induced increases in locomotor activity. Such an effect may be related to the 5-HT control of pallidal met-enkephalin levels via the 5-HT1B receptors.


Anti-Programmed Cell Death (PD)-1 Immunotherapy for Malignant Tumor: A Systematic Review and Meta-Analysis.
This systematic review and meta-analysis evaluated anti-programmed cell death (PD)-1 immunotherapy (nivolumab or pembrolizumab) for overall efficacy, safety, and effective dose relative to standard chemotherapy or other conventional drugs in the treatment of malignant tumors. We searched the following databases, PubMed, Medline, Embase, Cochrane, Wangfang Data, Weipu, and China National Knowledge Infrastructure, and the reference lists of the selected articles for randomized controlled trials (RCTs) of anti-PD-1 therapies in humans. The outcome measures were overall survival, treatment response, and adverse events. Only four randomized controlled trials met our inclusion criteria. Three of these evaluated responses to nivolumab, whereas one tested pembrolizumab. The result of our analysis suggested that nivolumab may improve the overall response rate in treating melanoma relative to chemotherapy and has few associated adverse events. Similarly, in metastatic melanoma patients, nivolumab had a significant advantage over dacarbazine in terms of 1-year survival, progression-free survival, and objective response rate. Regarding dose levels of nivolumab for patients with metastatic renal cell carcinoma, the outcomes in response to 2 and 10 mg/kg were similar, but both had significant advantages over 0.3 mg/kg. In addition, pembrolizumab showed similar outcomes in response to 2- and 10-mg/kg treatment. Anti-PD-1 immunotherapy appears to be safe and effective for patients with melanoma or metastatic renal cell carcinoma. Our meta-analysis is limited, but additional clinical trials are warranted to verify this preliminary evidence of positive outcomes and before anti-PD-1 therapy can be recommended for routine clinical use.


Effect of multifactorial therapeutic approach on axonal regeneration and cell viability in an in-vitro model of spinal-derived neural injury.
The highly debilitated nature of spinal cord injuries (SCI) creates an inhibitory repair environment that limits the recovery rate and therefore single interventional treatment has been resulted in incomplete recovery. A multifactorial approach that combines several therapeutic approaches may address diverse aspects of SCI pathology and enhance the recovery rate over single therapy. Accordingly, in this study, we aimed to investigate the effect of combined olfactory ensheathing cells (OECs) (to transport trophic factor, mediate immunomodulation, provide a suitable environment for cell survival), G-CSF (to establish a favorable environment for cell survival) and lipopolysaccharide (LPS) (to boost the protective activity of OEC) therapy on the cell viability after a scratch injury caused by a cataract knife on cells in an in-vitro model of spinal-derived neural injury. In this study, we used mixed neuronal-glial cultures, which are widely used for an in vitro study of neuronal damage. Scratch insult was made on cells using a cataract knife. The cells were divided into 8 groups (two control groups with and without olfactory ensheathing cells (OECs) treatment, injury group, three injury groups with single therapy by using super low dose of LPS (SLD-LPS) (100 pg/ml), OEC group, and G-CSF (100 ng/ml) group, and two injury groups with combined therapy (OEC with SLD-LPS and with all three treatments)). We found a significant decrease in the survival rate of injured cells (p < 0.001) 24 h after scratching insult. Our results indicated morphological alterations in cells in the acute phase (1, 2 and 6 h) after injury, with significant increased gap size at 6 h after induction of injury. Our combined therapy, significantly prevented cell death and decreased the size of the gap over time. We found that combined therapy promoted cell survival following spinal injury by providing a neuroprotective environment for cells. Therefore, our findings provide new insight into the combined therapy, which can be considered for promising preclinical therapeutic strategy for SCI toward clinical trials.


Untreated head and neck cancer: Natural history and associated factors.
This study aims to provide modern estimates of survival for untreated head and neck cancer (HNC) and to identify patient- and tumor-specific factors associated with not receiving definitive treatment. Adults with mucosal HNC were identified from the Surveillance, Epidemiology, and End Results (SEER) database. Median overall survival of untreated patients was calculated and stratified by site and stage. Cause-specific survival and factors associated with no treatment were investigated with advanced statistics. The study cohort included 6477 patients who received no treatment. Overall, untreated patients had a median survival of 12 months, with significant variability by site and stage. Multivariable analysis identified advanced age, black race, unmarried status, and lack of private insurance as associated with no treatment. Survival of HNC without treatment is dismal, yet substantial variation exists by tumor site and stage. Higher rates of no treatment among the poor and racial minorities are potentially driven by disparities in care availability and provision.


[Ultrastructure of the oocyte nucleus and cytoplasm in the mollusk, Littorina saxatilis. I. The structure of the meiotic chromosomes and of the products of their activity in the period of great oocyte growth].
At the beginning of the early diplotene, the chromosomes are getting despiralized and are seen to be made of a mass of fibrous material and chromocentres. During previtellogenesis the chromosomes are made of fibrilles 4--20 nm in diameter. At the beginning of vitellogenesis nucleolus-like bodies are seen to appear on bivalents made of granules (25--30 nm) and fine fibrilles. At the same time, in association with chromosomes a structure of irregular form was observed made of granules 25--30 nm in diameter and a string 20 nm in diameter. This structure is analogous to granular loops of the lampbrush chromosomes of the amphibia. During the large growth of the oocyte perichromatin and interchromatin granules are seen in the nuclei. By the end of the large growth numerous protein spheres, formed by fibrilles 12--13 nm in diameter are seen on the chromosomes. It is concluded that chromosomes of the L. saxatilis oocytes function as the lampbrush chromosomes of the amphibia and other animals having the solitary type oogenesis.


Early clinical outcomes using proton radiation for children with central nervous system atypical teratoid rhabdoid tumors.
Atypical teratoid/rhabdoid tumor (AT/RT) is an uncommon and aggressive tumor that often affects infants. Irradiation improves survival but has traditionally been avoided in patients under the age of 3 due to the increasing risk of neurocognitive side effects. We report the first cohort of AT/RT patients treated with proton therapy. All patients with AT/RT treated at Massachusetts General Hospital (MGH) Frances H. Burr Proton Beam Therapy Benter between July 2004 and November 2011 were included in this study. All patients were treated with 3-dimensional conformal proton therapy (3D-CPT). Ten consecutive patients of a median 2.3 years of age and with a median follow-up of 27.3 months (range, 11.3-99.4 months) were identified. Two patients suffered distant relapse; 1 patient was successfully treated with involved field irradiation and chemotherapy, while the second patient died of disease. At last follow-up, 9 patients were alive without evidence of disease. Proton radiation demonstrated increasing sparing of the cerebrum, temporal lobe, cochlea, and hypothalamus. Initial clinical outcomes with proton therapy are favorable. The advantages of proton therapy are particularly suited to the treatment of AT/RT, a tumor that often requires irradiation treatment at an age when avoiding irradiation to healthy tissues is most desirable.


Construction of circRNA-Associated ceRNA Network Reveals Novel Biomarkers for Esophageal Cancer.
Esophageal cancer (ESCC) is reported to be the eighth most common malignant tumors worldwide with high mortality. However, the functions of majority circRNAs in ESCC requires to be further explored. This study identified differently expressed circRNAs in 3 paired ESCC using RNA-sequencing method. The interactions among circRNAs, miRNAs, and mRNAs were predicted using bioinformatics analysis. In this study, using RNA-sequencing method and integrated bioinformatics analysis, 418 overexpressed circRNAs and 637 reduced circRNAs in ESCC sample were identified. Based on the mechanism that circRNAs could play as ceRNAs to modulate targets expression, circRNA-miRNA and circRNA-miRNA-mRNA networks were constructed in this study. Based on the network analysis, 7 circRNAs, including circ_0002255, circ_0000530, circ_0001904, circ_0001005, circ_0000513, circ_0000075, and circ_0001121, were identified as key circRNAs in ESCC. We found that circ_0002255 was related to the regulation of substrate adhesion-dependent cell spreading. circ_0001121 was involved in regulating nucleocytoplasmic transport. circ_0000513 played a key role in regulating Adherens junction, B cell receptor signaling pathway. Meanwhile, we observed circ_0000075 was involved in regulating zinc II ion transport, transition metal ion homeostasis, and angiogenesis. We thought this study could provide novel biomarkers for the prognosis of ESCC.


Sequence analysis of the rat Brca1 homolog and its promoter region.
Since the identification of the human breast and ovarian cancer gene, BRCA1, a large spectrum of germline mutations has been characterized that predispose women to developing these diseases. We have determined the complete coding sequence for the rat BRCA1 homolog and compared it with those of the mouse, dog, and human to help identify the important functional domains of the BRCA1 protein. The overall rat Brcal amino acid identity compared with the predicted mouse, dog, and human gene products is 81%, 69%, and 58%, respectively. In spite of this low overall homology, the amino terminal RING finger domain and one of two nuclear localization signals are highly conserved among these species. In addition, two BRCT domains at the carboxy terminus and a highly acidic region are relatively well conserved. We have also identified several putative regulatory elements through comparison of the bidirectional BRCA1 promoter regions among the rat, mouse, and human genes. These include motifs for CCAAT and G/C boxes, as well as potential SP1, CREB, and NFkB transcription factor binding sites. Finally, analysis of splice variants from rat mammary gland, ovary, testis, spleen, and liver tissues revealed that, while alternative transcripts are detectable, full-length transcripts are the predominant steady-state form.


Association between BRCA mutations and endometrial carcinoma: a systematic review with meta-analysis.
To first investigate on the association between BRCA mutations and endometrial carcinoma. To first evaluate the contribution of tamoxifen use and risk-reducing bilateral salping-oophenrectomy (BSO) on endometrial carcinoma in BRCA carriers. A systematic search of electronic databases including the PubMed and EMBASE was conducted to identify publications exploring the association between BRCA mutations and endometrial carcinoma. Finally, single rate meta-analysis and diagnostic meta-analysis were performed. 11 retrospective studies and 3 prospective studies were included in the meta-analysis, single rate meta-analysis was performed on retrospective studies and prospective studies respectively. We got that incidence of BRCA mutations in patients with endometrial carcinoma is about 0.035, the incidence of endometrial carcinoma in BRCA carriers is about 0.004. Diagnostic meta-analysis performed on prospective studies found that tamoxifen increased incidence of endometrial carcinoma in BRCA carriers. The incidence of BRCA mutations in patients with endometrial carcinoma is about 0.035 according to present studies, the incidence of endometrial carcinoma in BRCA carriers is about 0.004. Tamoxifen use is a certain risk factor for subsequent endometrial carcinoma, while history of breast cancer or risk-reducing BSO is not associated with incidence of follow-up endometrial carcinoma. The necessity and rationality of prophylactic hysterectomy for BRCA carriers remained to be discussed.