Polyphenol-Rich Extracts from Toona sinensis Bark and Fruit Ameliorate Free Fatty Acid-Induced Lipogenesis through AMPK and LC3 Pathways.
Non-alcoholic fatty liver disease (NAFLD) is a chronic liver disease found worldwide. The present study aimed to evaluate the mechanisms of inhibiting lipid accumulation in free fatty acid (FFA)-treated HepG2 cells caused by bark and fruit extracts of Toona sinensis (TSB and TSF). FFA induced lipid and triglyceride (TG) accumulation, which was attenuated by TSB and TSF. TSB and/or TSF promoted phosphorylation of AMP-activated protein kinase (AMPK) and acetyl-coA carboxylase and peroxisome proliferator-activated receptor alpha upregulation. Furthermore, TSB and TSF suppressed FFA-induced liver X receptor, sterol regulatory element-binding transcription protein 1, fatty acid synthase, and stearoyl-CoA desaturase 1 protein expression. Moreover, TSB and/or TSF induced phosphorylation of Unc-51 like autophagy-activating kinase and microtubule-associated protein 1A/1B-light chain 3 expressions. Therefore, TSB and TSF relieve lipid accumulation by attenuating lipogenic protein expression, activating the AMPK pathway, and upregulating the autophagic flux to enhance lipid metabolism. Moreover, TSB and TSF reduced TG contents, implying the therapeutic use of TSB and TSF in NAFLD.


Rates of homicide and homicide associated with severe mental illness in NSW between 1993 and 2016.
To describe the characteristics of offenders found not guilty on the grounds of mental illness (NGMI) in New South Wales and rates of NGMI and other homicide verdicts. Demographic, legal and clinical data after referral to the NSW Mental Health Review Tribunal following an NGMI verdict for homicide matched with results from the National Homicide Monitoring Program. Between 1993 and 2016, a total of 2159 homicide offenders were dealt with by the NSW courts, including 169 (7.8%) who were found NGMI. Over this period, the rate of non-NGMI homicide convictions fell from 1.83 per 100,000 per annum to 0.65 per 100,000 per annum (Kendall's tau = -0.79, p ⩽ 0.001) while the rate of NGMI homicide fluctuated, with an average annual rate of about 0.1 per 100,000 per annum (Kendall's tau = 0.17, p = 0.23). There was no association between the annual rates of NGMI and non-NGMI homicides (Pearson r = -0.3, p = 0.16) but falling rate of non-NGMI homicide meant that the proportion of NGMI offences doubled from 5.5% in the first 12 years to 11% in the second 12 years. Nearly all (88.7%) of those found NGMI had a schizophrenia-related psychosis. However, there were high rates of psychiatric comorbidity including substance use disorder (60.7%) and a history of a prior head injury (41.1%). Most (83.4%) of the NGMI offenders had previous contact with mental health services, but only half of these had received treatment with antipsychotic medication. The fall in conviction for homicide offences in the last 24 years has not been matched by a reduction in NGMI homicide verdicts. More assertive treatment of emerging psychosis and comorbid substance use disorders, and improved continuity of care of chronic psychosis might prevent some homicides.


Comparative Effectiveness of Surgical Treatments for Pediatric Hydrocephalus.
Pediatric hydrocephalus represents a high health care burden in the United States (US). Surgery is the mainstay of treatment. To perform a comparative effectiveness analysis for endoscopic third ventriculostomy (ETV) and cerebrospinal fluid shunt placement in pediatric hydrocephalus patients in the US using a large administrative claims database through the application of propensity scores matching. The MarketScan® database (Truven Health Analytics, Atlanta, Georgia) 2003 to 2011 was used. Patients 19 yr or younger at first occurrence of ETV or shunt during the study period were included. The study outcome, surgery failure, was defined as further surgical treatment for hydrocephalus subsequent to initial ETV or shunt procedure. Age, etiology of hydrocephalus, and history of shunt were used to create matched samples for the ETV and shunt cohorts. Kaplan-Meier survival curves, stratified log-rank test, and Cox proportional-hazard models were used to analyze samples. There were 3231 eligible cases (478 ETV; 2753 shunt). Propensity scores matching produced 455 balanced pairs. For matched samples, 326 of 455 (72%) pairs were concordant, while 129 pairs were discordant in surgery outcomes within 3 mo. Among discordant pairs, ETV patients were more likely to experience surgery failure compared to patients receiving shunt (relative risk = 1.4, P value = .011). Furthermore, patients' age < 1 yr had lower ETV success rates than those with shunt (P value = .009). No similar pattern was found in patients' age ≥ 1 yr. There was no significant effect on time to failure between patients undergoing ETV and shunt, except in infants' age <1 yr.


An unusual case of a tibial metastasis as the clinical presentation of bronchogenic adenocarcinoma.
A patient with initial complaints of leg pain and difficulty walking was found to have a large right tibial metastatic tumor and poorly differentiated adenocarcinoma of the lung. Findings from total-body bone scintigraphy include a large area of increased uptake in the proximal half of the right tibia with a photon-deficient area medially, and focal areas of uptake in a right rib, in the femoral neck and the left ileum. An irregular area of increased uptake in the left lung mass was shown by thoracic bone SPECT. This is an unusual case of a tibial metastasis as the first clinical presentation of bronchogenic adenocarcinoma.


Robust, self-healing hydrogels synthesised from catechol rich polymers.
Coordinative interactions between polymer-bound catechols and metal ions are the basis for numerous bio-inspired soft materials. Here, we demonstrate rapid access to catechol rich polymers through reductive amination (RA) strategy. We employed chitosan to exemplify the utility of this protocol. Controlled grafting of catechol pendants (from as low as 18 mol% to as high as 80 mol%) onto chitosan was readily achieved in aqueous medium under ambient conditions by RA protocol. Because of the high density of catechol units grafted onto chitosan, we could accomplish the gelation of water even in acidic medium in the presence of transition metal ions, or on addition of chemical oxidant such as NaIO4. Increasing the mol% of catechol in polymer decreased the amounts of Fe(iii) or NaIO4 required to yield gels. UV-vis and Raman studies indicated the presence of mono-complex between Fe(iii) and catechol in the gels formed through coordinative crosslinking. Highly ductile hydrogels exhibiting excellent load bearing ability were obtained under these conditions. Electrostatic repulsions between the cationic polymer chains presumably prevented the gel to collapse upon the application of load. These gels were also completely self-healing due to the reversible nature of their coordinative interactions. Gels formed at higher pH are brittle and less resilient compared to those formed at lower pH. Ductility and self-healing ability of coordinative cross-linked gels are superior to those formed by oxidative crosslinking. We conclude that RA strategy offers rapid and easy access to catechol-rich systems, and retention of basic amine functionalities allows the preparation of robust bio inspired soft materials.


A Flexible Self-Powered Noncontact Sensor with an Ultrawide Sensing Range for Human-Machine Interactions in Harsh Environments.
Noncontact human-machine interactions (HMIs) provide a hygienic and intelligent approach to communicate between humans and machines. However, current noncontact HMIs are generally hampered by the interaction distance, and they lack the adaptability to environmental interference such as high humidity conditions. Here, we explore a self-powered electret-based noncontact sensor (ENS) with moisture-resisting ability and ultrawide sensing range exceeding 2.5 m. A megascopic air-bubble structure is designed to enhance charge-storage stability and charge-recovery ability of the ENS based on the heterocharge-synergy effect in electrets. Besides, multilayer electret films are introduced to strengthen the electric field by utilizing the electrostatic field superposition effect. Thanks to the above improved performances of the ENS, we demonstrate various noncontact HMI applications in harsh environments, including noncontact appliances, a moving trajectory and accidental fall tracking system, and a real-time machine learning-assisted gesture recognition system with accuracy as high as 99.21%. This research expands the way for noncontact sensor design and may further broaden applications in noncontact HMIs.


[Vascular bed of a lymph node and lymphodynamics after leg lengthening by the G. A. Ilizarov method].
In the experiments performed on dogs, the right shin has been elongated, lymph movement has been studied in the extremity, as well as in the arteries of the popliteal and medial iliac lymph nodes. The lymph movement rate immediately after fracture decreases 6 times, and during the elongation process it gradually restores with normalization on the 14th day of distraction. The integrity of the arterial bed in the lymph node is not disturbed during the elongation process of the shin. The intraorganic bed of the regional lymph nodes reacts to the shin elongation by increasing diameter of the arteries and the number of the functioning vessels. The changes in the arterial bed demonstrate an acceleration of the blood stream in the lymph nodes under the elongation of the shin. After osteogenesis normalization in the lymph stream takes place.


Investigation of orthologous pathogen recognition gene-rich regions in solanaceous species.
Pathogen receptor proteins such as receptor-like protein (RLP), receptor-like kinase (RLK), and nucleotide-binding leucine-rich repeat (NLR) play a leading role in plant immunity activation. The genome architecture of such genes has been extensively investigated in several plant species. However, we still know little about their elaborate reorganization that arose during the plant speciation process. Using recently released pepper and eggplant genome sequences, we were able to identify 1097 pathogen recognition genes (PRGs) in the cultivated pepper Zunla-1 and 775 in the eggplant line Nakate-Shinkuro. The retrieved genes were analysed for their tendency to cluster, using different methods to infer the means of grouping. Orthologous relationships among clustering loci were found, and interesting reshuffling within given loci was observed for each analysed species. The information obtained was integrated into a comparative map to highlight the evolutionary dynamics in which the PRG loci were involved. Diversification of 14 selected PRG-rich regions was also explored using a DNA target-enrichment approach. A large number of gene variants were found as well as rearrangements of sequences encoding single protein domain and changes in chromosome gene order among species. Gene duplication and transposition activity have clearly influenced plant genome R-gene architecture and diversification. Our findings contribute to addressing several biological questions concerning the parallel evolution that occurred between genomes of the family Solanaceae. Moreover, the integration of different methods proved a powerful approach to reconstruct the evolutionary history in plant families and to transfer important biology findings among plant genomes.


Flow in experimental berry aneurysms: method and model.
This study addresses two basic questions: What are the flow dynamics in aneurysms? Can these flows be modified to enhance retention of adhesive? Using Pyrex glass bifurcations, fluid flow was studied in a variety of aneurysms placed at varying positions around the bifurcations. Indicators injected into the slipstreams were recorded and studied both by stop-frame high-speed movie analysis and with 35 mm slides. Even at low-flow rates, a central slipstream strikes the apex of bifurcations, and may be partly responsible for the initial production of berry aneurysms. A low-pressure zone occurs at the lateral angle of bifurcations, probably explaining the formation of endovascular cushions. Flow into symmetrically placed narrow neck aneurysms did not occur. Indicator entered the aneurysm in a turbulent fashion only when there was orifice (lip) asymmetry. Both lip asymmetry and rapid flow favor intraaneurysmal turbulent flow.


A Study on the Macroscopic Morphometry of the Lesion Area on Diabetic Ulcers in Humans Treated with Photodynamic Therapy Using Two Methods of Measurement.
Diabetes Mellitus is a chronic disease that can lead to lower-limb ulceration. The photodynamic therapy (PDT) is based on light interaction with a photosensitizer capable to promote bacterial death and acceleration of wound healing. This study aims to accomplish a macroscopic morphometry evaluation on diabetic ulcer area in humans that were under PDT. A clinical study was conducted by convenience sample, experimental, controlled, and blinded, composed of 12 patients of both sexes with diabetic ulcers in lower limbs that were divided into two groups, control (n = 6) and PDT (n = 6). They were all treated with collagenase/chloramphenicol during the experimental period, in which six of them had received PDT with Methylene Blue dye (0.01%) associated with laser therapy (660 nm, 30 mW, 8 sec, 6 J/cm2, beam area of 0.04 mm2), three times per week, totaling 10 sessions. Ulcer areas were analyzed by their nominal area and photographed for analysis through the ImageJ® software. Then, the Ulcer Healing Index and the wound area reduction were calculated for both groups. Data from the UHI and wound area reduction parameters show that there is a statistically significant difference (p < 0.05) between the control group and PDT group, with the PDT group showing a greater reduction of the diabetic ulcer area than the control group. In the ImageJ software area analysis, both patient groups presented significant differences (p < 0.05) between pre and post-treatment in relation to lesion reduction area, the PDT being better. Results show that PDT accelerates the closure of ulcer wounds and to evaluate the wound area, different measurement methods can be used to follow-up the tissue repair process.


Addressing risk management difficulties in children with food allergies.
Risk is a concept inherent in every medical procedure. It can be defined as the probability of an adverse event in a defined population over a specified period of time. In the frame of food allergy management, it might be related to a diagnostic procedure, a treatment, or the consumption of foods. The risk of an adverse event can also be augmented by individual factors. This rostrum article discusses various aspects faced by children with food allergies in the light of risk, and their practical implications. Identifying personal risks for severe reaction, such as unstable asthma, and correcting them whenever possible also contribute to a reduction of the risk inherent to food allergy. Among the facets discussed, oral food challenges (OFC) are the most common diagnostic procedures implying an inherent risk. The risk of OFCs can be minimized by correct indication and timing of the test, a safe setting, as well as by ensuring that the patient is otherwise well without potential stressor potentially increasing the risk of a more severe reaction. Oral immunotherapy (OIT) has been studied as a potential treatment for increasing the threshold dose for reaction, and thus reducing the risk of accidental reaction. Nevertheless, the procedure is not devoid of risk as the patients may and do often react during the course of the procedure. Ingestion of trace amounts in processed foods, mainly in community settings such as restaurants, schools, or day care, represents a potential risk of reactions, although for a minority of patients. Precautionary allergen labeling (PAL) is a widespread strategy to reduce the potential risk of reactions due to traces. However, PAL is currently inefficient due to inconsistent labeling, also not indicating a clear maximum amount possibly present in the manufactured food. Finally, cost-effectiveness needs to be considered in risk management, as many risk reduction procedures are clearly not cost-effective.


Abnormalities in perineuronal nets and behavior in mice lacking CSGalNAcT1, a key enzyme in chondroitin sulfate synthesis.
Chondroitin sulfate (CS) is an important glycosaminoglycan and is mainly found in the extracellular matrix as CS proteoglycans. In the brain, CS proteoglycans are highly concentrated in perineuronal nets (PNNs), which surround synapses and modulate their functions. To investigate the importance of CS, we produced and precisely examined mice that were deficient in the CS synthesizing enzyme, CSGalNAcT1 (T1KO). Biochemical analysis of T1KO revealed that loss of this enzyme reduced the amount of CS by approximately 50% in various brain regions. The amount of CS in PNNs was also diminished in T1KO compared to wild-type mice, although the amount of a major CS proteoglycan core protein, aggrecan, was not changed. In T1KO, we observed abnormalities in several behavioral tests, including the open-field test, acoustic startle response, and social preference. These results suggest that T1 is important for plasticity, probably due to regulation of CS-dependent PNNs, and that T1KO is a good model for investigation of PNNs.


Household-based prevalence of helminths and parasitic protozoa in rural KwaZulu-Natal, South Africa, assessed from faecal vault sampling.
This study was undertaken to examine the family-based prevalence of environmentally persistent parasites in two rural communities of KwaZulu-Natal, South Africa. Samples were collected from 120 urine-diversion family toilets and screened for selected protozoa and helminths with immunomagnetic separation and the ammonium bicarbonate (AMBIC) protocol respectively. The parasites found were Ascaris lumbricoides (59%), Giardia intestinalis (54%), Trichuris trichiura (48%), Cryptosporidium spp. (21%) and Taenia spp. (18%). Only 14% of the household toilets were negative for these pathogens. The occurrence of A. lumbricoides and T. trichiura was lower (P<0.001) in the area with better hygiene behaviour, whereas G. intestinalis was more common (P<0.05) in families with at least one child aged five years or less and in families with more than four persons. Quantification of the parasites per gram was done for each sample and this provided realistic risk assessment data for the reuse of material from urine-diversion toilets. The high occurrence of parasites found in the two communities, in spite of sanitation and hygiene interventions in the areas, suggests an endemicity that will not be reduced without de-worming campaigns. Finally, the study showed that sampling directly from the deposited faecal material may be useful for parasitic prevalence estimations.


Data-driven statistical modeling of the emergent behavior of biohybrid microrobots.
Multi-agent biohybrid microrobotic systems, owing to their small size and distributed nature, offer powerful solutions to challenges in biomedicine, bioremediation, and biosensing. Synthetic biology enables programmed emergent behaviors in the biotic component of biohybrid machines, expounding vast potential benefits for building biohybrid swarms with sophisticated control schemes. The design of synthetic genetic circuits tailored toward specific performance characteristics is an iterative process that relies on experimental characterization of spatially homogeneous engineered cell suspensions. However, biohybrid systems often distribute heterogeneously in complex environments, which will alter circuit performance. Thus, there is a critically unmet need for simple predictive models that describe emergent behaviors of biohybrid systems to inform synthetic gene circuit design. Here, we report a data-driven statistical model for computationally efficient recapitulation of the motility dynamics of two types of Escherichia coli bacteria-based biohybrid swarms-NanoBEADS and BacteriaBots. The statistical model was coupled with a computational model of cooperative gene expression, known as quorum sensing (QS). We determined differences in timescales for programmed emergent behavior in BacteriaBots and NanoBEADS swarms, using bacteria as a comparative baseline. We show that agent localization and genetic circuit sensitivity strongly influence the timeframe and the robustness of the emergent behavior in both systems. Finally, we use our model to design a QS-based decentralized control scheme wherein agents make independent decisions based on their interaction with other agents and the local environment. We show that synergistic integration of synthetic biology and predictive modeling is requisite for the efficient development of biohybrid systems with robust emergent behaviors.


Short Review of Liposteroid: A Novel Targeted Glucocorticoid Preparation for Treatment of Autoimmune and Inflammatory Diseases.
This paper briefly reviews the safety and efficacy of liposteroid in different inflammatory and non-inflammatory diseases. Corticosteroids (CS) are the first-line therapy in many inflammatory and autoimmune disorders. Although highly efficacious, long-term use of CS is limited due to the occurrence of significant side effects. Liposteroid, which is a liposomal formulation of dexamethasone palmitate, possess more potent anti-inflammatory and immunosuppressive properties compared to dexamethasone sodium phosphate. These two formulations have markedly different lipid solubility, resulting in different pharmacokinetic and pharmacodynamic properties. Liposteroid has been used with success in patients with rheumatoid arthritis, macrophage activation syndrome, and idiopathic pulmonary hemosiderosis. In addition, liposteroid has been used in some non-inflammatory diseases. Moreover, we conceive that liposteroid may have a beneficial effect in patients, who are critically ill due to COVID-19, and suffer from the macrophage activation syndrome.


Differential requirements of the fused kinase for hedgehog signalling in the Drosophila embryo.
The two signalling proteins, Wingless and Hedgehog, play fundamental roles in patterning cells within each metamere of the Drosophila embryo. Within the ventral ectoderm, Hedgehog signals both to the anterior and posterior directions: anterior flanking cells express the wingless and patched Hedgehog target genes whereas posterior flanking cells express only patched. Furthermore, Hedgehog acts as a morphogen to pattern the dorsal cuticle, on the posterior side of cells where it is produced. Thus responsive embryonic cells appear to react according to their position relative to the Hedgehog source. The molecular basis of these differences is still largely unknown. In this paper we show that one component of the Hedgehog pathway, the Fused kinase accumulates preferentially in cells that could respond to Hedgehog but that Fused concentration is not a limiting step in the Hedgehog signalling. We present direct evidence that Fused is required autonomously in anterior cells neighbouring Hedgehog in order to maintain patched and wingless expression while Wingless is in turn maintaining engrailed and hedgehog expression. By expressing different components of the Hedgehog pathway only in anterior, wingless-expressing cells we could show that the Hedgehog signalling components Smoothened and Cubitus interruptus are required in cells posterior to Hedgehog domain to maintain patched expression whereas Fused is not necessary in these cells. This result suggests that Hedgehog responsive ventral cells in embryos can be divided into two distinct types depending on their requirement for Fused activity. In addition, we show that the morphogen Hedgehog can pattern the dorsal cuticle independently of Fused. In order to account for these differences in Fused requirements, we propose the existence of position-specific modulators of the Hedgehog response.


Cell wall peroxidases in the liverwort Dumortiera hirsuta are responsible for extracellular superoxide production, and can display tyrosinase activity.
In our earlier work, we showed that the liverwort Dumortiera hirsuta produces an extracellular oxidative burst of superoxide radicals during rehydration following desiccation stress. The oxidative burst is a common early response of organisms to biotic and abiotic stresses, with suggested roles in signal transduction, formation of protective substances such as suberin, melanin and lignin and defense against pathogens. To discover which enzymes are responsible for the extracellular superoxide production, we isolated apoplastic fractions from D. hirsuta, surveyed for the presence of potential redox enzymes, and performed non-denaturing polyacrylamide gel electrophoresis activity stains. Various isoforms of peroxidase (EC 18.104.22.168) and tyrosinase (o-diphenolase) (EC 220.127.116.11) were present at significant levels in the apoplast. In-gel activity staining revealed that some peroxidases isoforms could produce superoxide, while tryosinases could readily metabolize 3,4-dihydroxy phenyl l-alanine (l-dopa) into melanins. Interestingly, some peroxidase isoforms could oxidize the native tyrosinase substrate l-dopa at significant levels, even in the absence of hydrogen peroxide, while others could do so only in the presence of hydrogen peroxide. In D. hirsuta, peroxidases may play an important role in melanin formation. Possible functions for these diverse oxidases in liverwort biology are discussed.


Genetic association tests: a method for the joint analysis of family and case-control data.
With the trend in molecular epidemiology towards both genome-wide association studies and complex modelling, the need for large sample sizes to detect small effects and to allow for the estimation of many parameters within a model continues to increase. Unfortunately, most methods of association analysis have been restricted to either a family-based or a case-control design, resulting in the lack of synthesis of data from multiple studies. Transmission disequilibrium-type methods for detecting linkage disequilibrium from family data were developed as an effective way of preventing the detection of association due to population stratification. Because these methods condition on parental genotype, however, they have precluded the joint analysis of family and case-control data, although methods for case-control data may not protect against population stratification and do not allow for familial correlations. We present here an extension of a family-based association analysis method for continuous traits that will simultaneously test for, and if necessary control for, population stratification. We further extend this method to analyse binary traits (and therefore family and case-control data together) and accurately to estimate genetic effects in the population, even when using an ascertained family sample. Finally, we present the power of this binary extension for both family-only and joint family and case-control data, and demonstrate the accuracy of the association parameter and variance components in an ascertained family sample.


MR imaging of entrapment neuropathies of the lower extremity. Part 2. The knee, leg, ankle, and foot.
Entrapment neuropathies of the knee, leg, ankle, and foot are often underdiagnosed, as the results of clinical examination and electrophysiologic evaluation are not always reliable. The causes of most entrapment neuropathies in the lower extremity may be divided into two major categories: (a) mechanical causes, which occur at fibrous or fibro-osseous tunnels, and (b) dynamic causes related to nerve injury during specific limb positioning. Magnetic resonance (MR) imaging, including high-resolution MR neurography, allows detailed evaluation of the course and morphology of peripheral nerves, as well as accurate delineation of surrounding soft-tissue and osseous structures that may contribute to nerve entrapment. Familiarity with the normal MR imaging anatomy of the nerves in the knee, leg, ankle, and foot is essential for accurate assessment of the presence of peripheral entrapment syndromes. Common entrapment neuropathies in the knee, leg, ankle, and foot include those of the common peroneal nerve, deep peroneal nerve, superficial peroneal nerve, tibial nerve and its branches, and sural nerve.


Comparative analysis of molecular fingerprints in prediction of drug combination effects.
Application of machine and deep learning methods in drug discovery and cancer research has gained a considerable amount of attention in the past years. As the field grows, it becomes crucial to systematically evaluate the performance of novel computational solutions in relation to established techniques. To this end, we compare rule-based and data-driven molecular representations in prediction of drug combination sensitivity and drug synergy scores using standardized results of 14 high-throughput screening studies, comprising 64 200 unique combinations of 4153 molecules tested in 112 cancer cell lines. We evaluate the clustering performance of molecular representations and quantify their similarity by adapting the Centered Kernel Alignment metric. Our work demonstrates that to identify an optimal molecular representation type, it is necessary to supplement quantitative benchmark results with qualitative considerations, such as model interpretability and robustness, which may vary between and throughout preclinical drug development projects.


Epigenetic regulation of miR-34a expression in alcoholic liver injury.
Epigenetic changes are associated with the regulation of transcription of key cell regulatory genes [micro RNAs (miRNAs)] during different types of liver injury. This study evaluated the role of methylation-associated miRNA, miR-34a, in alcoholic liver diseases. We identified that ethanol feeding for 4 weeks significantly up-regulated 0.8% of known miRNA compared with controls, including miR-34a. Treatment of normal human hepatocytes (N-Heps) and cholangiocytes [human intrahepatic biliary epithelial cells (HiBECs)] with ethanol and lipopolysaccharide induced a significant increase of miR-34a expression. Overexpression of miR-34a decreased ethanol-induced apoptosis in both N-Heps and HiBECs. In support of the concept that the 5'-promoter region of miR-34a was noted to be embedded within a CpG island, the expression level of miR-34a was significantly increased after demethylation treatment in N-Heps and HiBECs. By methylation-specific PCR, we confirmed that miR-34a activation is associated with ethanol-linked hypomethylation of the miR-34a promoter. A combination of bioinformatics, dual-luciferase reporter assay, mass spectrometry, and Western blot analysis revealed that caspase-2 and sirtuin 1 are the direct targets of miR-34a. Furthermore, modulation of miR-34a also altered expression of matrix metalloproteases 1 and 2, the mediators involved in hepatic remodeling during alcoholic liver fibrosis. These findings provide the basis for an exciting field in which the epigenomic microRNAs of hepatic cells may be manipulated with potential therapeutic benefits in human alcoholic liver diseases.


Review of varicella zoster seroepidemiology in India and Southeast Asia.
Varicella zoster virus (VZV) predominantly affects children in temperate countries, with near-universal seroconversion occurring by late childhood. However, in tropical regions, VZV infection is common in adolescents and adults. This review identifies age-related VZV seroprevalence patterns in a number of Asian countries which indicate that seroconversion in tropical countries occurs at a later age than in temperate countries. Seasonal and regional variations in acute disease within some Asian countries suggest that temperate climates might favour transmission of the varicella virus, with incidence peaking during cooler months and in cooler, more temperate regions. VZV infection is often more severe in adults than in children, suggesting that tropical countries may be at risk of greater morbidity and mortality as a result of later-age seroconversion. Susceptibility of pregnant women and their infants, and of people infected with HIV/AIDS is also cause for concern. Vaccination may be beneficial in reducing the impact of VZV in Asian populations.


A smartphone aptasensor for fipronil detection in honey samples.
In this study, an electrochemical smartphone-based aptasensor for the determination of fipronil was developed by modifying a screen-printed carbon electrode (SPCE). Fipronil is a broad-spectrum insecticide that has been widely used in various applications such as agriculture, veterinary, and household pest control. Recently, its use has raised concerns over the potential impact on the environment and human health. The absence of effective methods for this purpose poses a significant obstacle. To tackle this problem, we have developed a cutting-edge aptamer-based portable sensor capable of rapidly and conveniently detecting fipronil in situ. Considering that the detection of small molecules, such as fipronil, can be a challenging task, a competitive replacement assay was set up based on the aptamer's preference for the free form of fipronil over the immobilized one on the electrode. The analytical performance provided by the sensor on standard solutions of a known fipronil content made it possible to estimate a limit of detection (LOD) equal to 1.07 μg kg-1 and a limit of quantification (LOQ) of 3.21 μg kg-1. Selectivity tests were conducted using atrazine as a possible interferent. The use and performance of the developed portable aptasensor was assessed on honey samples, which were simultaneously analyzed using an HPLC-MS method. This aptasensor could be an affordable and effective tool for accurately quantifying fipronil not only in honey samples but also in other food products.


Controlled Osteogenic Differentiation of Mouse Mesenchymal Stem Cells by Tetracycline-Controlled Transcriptional Activation of Amelogenin.
Regenerative dental therapies for bone tissues rely on efficient targeting of endogenous and transplanted mesenchymal stem cells (MSCs) to guide bone formation. Amelogenin is the primary component of Emdogain, which is used to regenerate periodontal defects; however, the mechanisms underlying the therapeutic effects on alveolar bone remain unclear. The tetracycline (Tet)-dependent transcriptional regulatory system is a good candidate to investigate distinct roles of genes of interest during stem cell differentiation. Here, we investigated amelogenin-dependent regulation of osteogenesis in MSCs by establishing a Tet-controlled transcriptional activation system. Clonal mouse bone marrow-derived MSCs were lentivirally transduced with the Tet repressor (TetR) expression vector followed by drug selection to obtain MSCs constitutively expressing TetR (MSCs-TetR). Expression vectors that contained the Tet operator and amelogenin-coding (Amelx) cDNA fragments were constructed using the Gateway system and lentivirally introduced into MSCs-TetR to generate a Tet regulation system in MSCs (MSCs-TetR/Amelx). MSCs-TetR/Amelx significantly overexpressed the Amelx gene and protein in the presence of the tetracycline derivative doxycycline. Concomitant expression of osterix, bone sialoprotein (BSP), osteopontin, and osteocalcin was modulated by addition or removal of doxycycline under osteogenic guidance. During osteogenic induction, MSCs-TetR/Amelx treated with doxycycline showed significantly increased gene expression of osterix, type I collagen, BSP, and osteocalcin in addition to increased alkaline phosphatase activity and mineralized nodule formation. Enhanced extracellular matrix calcification was observed when forced Amelx expression commenced at the early stage but not at the intermediate or late stages of osteogenesis. These results suggest that a Tet-controlled Amelx gene regulation system for mouse MSCs was successfully established, in which transcriptional activation of Amelx was associated with enhanced osteogenic differentiation, especially in the early stage of biomineralization.


Determination of lifetime injury mortality risk in Canada in 2002 by drinking amount per occasion and number of occasions.
Injury is the leading cause of alcohol-attributable mortality in Canada. Risk is determined by amount consumed per occasion and accumulates across drinking episodes. The authors estimated alcohol-attributable injury mortality in Canada for 2002 by combining the absolute risk of injury unrelated to alcohol with relative risks that were specific to gender and consumption per occasion, while taking into account lifetime number of drinking occasions. The absolute risk increased as number of drinking occasions and number of drinks per occasion increased. The absolute risk remained relatively low at fewer than 2 drinking occasions per month, regardless of number of drinks. Absolute risk levels reached 1 in 1,000 at 5 or more drinks once per month for men and at 5-7 drinks once per month for women. The probability of mortality was 1 in 100 for all levels of consumption above 3 drinks 3 times per week for men and above 5 drinks 3 times per week for women. No safe level of consumption is recommended based on these results, although risk is much lower for consuming 3 standard drinks or less fewer than 3 times per week. Absolute risk reflects long-term effects of drinking patterns and is important for risk-communication and alcohol-control policy.


A Prospective Study on the Effect of Corticosteroid Injection Dosage for Hand Disorders in Non-Insulin Dependent Diabetics.
Local administration of corticosteroids is an effective yet potentially dangerous intervention in the treatment of hand disorders in diabetics. Prolonged exposure to hyperglycemia contributes to non-enzymatic glycosylation of various organ systems, which may cause detrimental health effects such as blindness, renal failure, and peripheral neuropathy, contributing to the high cost of health care. The purpose of this study is to determine the effects of corticosteroid dosage on serum glucose levels when used to treat common hand disorders in diabetic patients. Twenty-one patients with non-insulin-dependent diabetes mellitus treated with a corticosteroid injection were prospectively enrolled. Either triamcinolone 10 mg (T-10 group, N = 11) or 40 mg (T-40 group, N = 10) was administered with a local anesthetic. Fasting morning serum glucose, QuickDASH scores, and visual analog scale (VAS) pain scores were recorded prior to injection. Post-prandial serum glucose was recorded the evening of the injection, and the fasting serum glucose was recorded each morning. Clinical outcomes were recorded at 6 weeks and again at an average of 26 months. Patients in both cohorts, on average, had improvements in their Quick- DASH and VAS scores after the injection without significant variation. There was a significant elevation in serum glucose in both groups. T-10 had an average glucose increase of 53 mg/dL (41%), which returned to baseline at 21 hours. T-40 had a maximum glucose increase of 50 mg/dL (40%), which returned to baseline in 58 hours. The difference in time to return to baseline was statistically significance. Both T-10 and T-40 are effective in relieving painful symptoms and improving patient functionality after injection. A lower dosage triamcinolone is associated with a quicker return of serum glucose to baseline and may be a safer alternative to higher dosages when considering prolonged hyperglycemia and its known detrimental effects of non-enzymatic glycosylation on various organ systems.


Level I of evidence does not support manual lymphatic drainage for total knee arthroplasty: a meta-analysis.
It is unclear whether manual lymphatic drainage (MLD) following primary total knee arthroplasty (TKA) is effective in reducing pain and swelling and improving knee function. The present study investigated the efficacy of MLD after TKA. The outcomes of interest are the range of motion (ROM), pain (visual analogue scale, VAS), and circumference of the lower leg. This meta-analysis was conducted according to the 2020 PRISMA statement. In November 2023, the following databases were accessed: PubMed, Web of Science, Google Scholar, and Embase, with no time constraint. Only level I evidence studies, according to the Oxford Centre of Evidence-Based Medicine, were considered. All the randomised controlled trials (RCTs) comparing patients who have received MLD versus a group of patients who did not undergo MLD following primary TKA were accessed. Data from four RCTs (197 TKAs) were retrieved. 67% (132 of 197 patients) were women. The mean length of follow-up was 7.0 ± 5.8 weeks. The mean age of the patients was 69.6 ± 2.7 years, and the mean BMI was 28.7 ± 0.9 kg/m2. At baseline, between-group comparability was evidenced in the male:female ratio, mean age, mean BMI, knee flexion, and VAS. No difference was found in flexion (P = 0.7) and VAS (P = 0.3). No difference was found in the circumference of the thigh (P = 0.8), knee (P = 0.4), calf (P = 0.4), and ankle (P = 0.3). The current level I of evidence does not support the use of MLD in primary TKA.


Regulation of alternative splicing of CD45 by antagonistic effects of SR protein splicing factors.
CD45 is a transmembrane glycoprotein possessing tyrosine phosphatase activity, which is involved in cell signaling. CD45 is expressed on the surface of most leukocytes and can be alternatively spliced by the inclusion or skipping of three variable exons (4, 5, and 6 or A, B, and C) to produce up to eight isoforms. In T cells, the splicing pattern of CD45 isoforms changes after activation; naive cells express high m.w. isoforms of CD45 which predominantly express exon A (CD45RA), whereas activated cells lose expression of exon A to form low m.w. isoforms of CD45 including CD45RO. Little is known about the specific factors controlling the switch in CD45 splicing which occurs on activation. In this study, we examined the influence of the SR family of splicing factors, which, like CD45, are expressed in tissue-specific patterns and have been shown to modulate the alternative splicing of a variety of transcripts. We show that specific SR proteins have antagonistic effects on CD45 splicing, leading either to exon inclusion or skipping. Furthermore, we were able to demonstrate specific changes in the SR protein expression pattern during T cell activation.


Speleotherapy for asthma.
Speleotherapy, the use of subterranean environments, is a therapeutic measure in the treatment of chronic obstructive airways diseases. It is virtually unknown in the UK or the US, but has considerable widespread use in some Central and Eastern European countries. To review evidence for the efficacy of speleotherapy in the treatment of asthma. We searched electronic databases (Medline, Embase, Cochrane Airways group database), contacted speleotherapy centres and experts in the field, hand searched proceedings, and checked bibliographies of articles obtained to identify possible relevant publications. We included controlled clinical trials (i.e., both randomized and those not reporting the method of allocation) that compared clinical effects of speleotherapy with another intervention or no intervention in patients with chronic asthma. Information concerning patients, interventions, results, and methodology were extracted in standardized manner by two independent reviewers and summarized descriptively. Three trials including a total of 124 asthmatic children met the inclusion criteria, but only one trial had reasonable methodological quality. Two trials reported that speleotherapy had a beneficial short-term effect on lung function. Other outcomes could not be assessed in a reliable manner. The available evidence does not permit a reliable conclusion as to whether speleo-therapeutic interventions are effective for the treatment of chronic asthma. Randomized controlled trials with long-term follow-up are necessary.


Dietary restriction and the pursuit of effective mimetics.
Dietary restriction (DR) has been shown to extend both median and maximum lifespan in a range of animals, although recent findings suggest that these effects are not universally enjoyed across all animals. In particular, the lifespan effect following DR in mice is highly strain-specific and there is little current evidence that DR induces a positive effect on all-cause mortality in non-human primates. However, the positive effects of DR on health appear to be highly conserved across the vast majority of species, including human subjects. Despite these effects on health, it is highly unlikely that DR will become a realistic or popular life choice for most human subjects given the level of restraint required. Consequently significant research is focusing on identifying compounds that will bestow the benefits of DR without the obligation to adhere to stringent reductions in daily food intake. Several such compounds, including rapamycin, metformin and resveratrol, have been identified as potential DR mimetics. Although these compounds show significant promise, there is a need to properly understand the mechanisms through which these drugs act. This review will discuss the importance in understanding the role that genetic background and heterogeneity play in mediating the lifespan and healthspan effects of DR. It will also provide an overview of the most promising current DR mimetics and their effects on healthy lifespan.


Pathologic heterogeneity persists in early active multiple sclerosis lesions.
Multiple sclerosis (MS) lesions demonstrate immunopathological heterogeneity in patterns of demyelination. Previous cross-sectional studies reported immunopatterns of demyelination were identical among multiple active demyelinating lesions from the same individual, but differed between individuals, leading to the hypothesis of intraindividual pathological homogeneity and interindividual heterogeneity. Other groups suggested a time-dependent heterogeneity of lesions. The objective of our present study was to analyze tissue samples collected longitudinally to determine whether patterns of demyelination persist over time within a given patient. Archival tissue samples derived from patients with pathologically confirmed central nervous system inflammatory demyelinating disease who had undergone either diagnostic serial biopsy or biopsy followed by autopsy were analyzed immunohistochemically. The inclusion criteria consisted of the presence of early active demyelinating lesions--required for immunopattern classification--obtained from the same patient at 2 or more time points. Among 1,321 surgical biopsies consistent with MS, 22 cases met the study inclusion criteria. Twenty-one patients (95%) showed a persistence of immunopathological patterns in tissue sampled from different time points. This persistence was demonstrated for all major patterns of demyelination. A single patient showed features suggestive of both pattern II and pattern III on biopsy, but only pattern II among all active lesions examined at autopsy. These findings continue to support the concept of patient-dependent immunopathological heterogeneity in early MS and suggest that the mechanisms and targets of tissue injury may differ among patient subgroups. These observations have potentially significant implications for individualized therapeutic approaches.


Neuroprotection in Glaucoma: Animal Models and Clinical Trials.
Glaucoma is a progressive neurodegenerative disease that frequently results in irreversible blindness. Glaucoma causes death of retinal ganglion cells (RGCs) and their axons in the optic nerve, resulting in visual field deficits and eventual loss of visual acuity. Glaucoma is a complex optic neuropathy, and a successful strategy for its treatment requires not only better management of known risk factors such as elevated intraocular pressure and the development of improved tools for detecting RGC injury but also treatments that address this injury (i.e., neuroprotection). Experimental models of glaucoma provide insight into the cellular and molecular mechanisms of glaucomatous optic neuropathy and aid the development of neuroprotective therapies.


Serum macrophage inhibitory cytokine 1 in rheumatoid arthritis: a potential marker of erosive joint destruction.
The transforming growth factor beta superfamily member macrophage inhibitory cytokine 1 (MIC-1) is expressed upon macrophage activation, regulated by the p53 pathway, and linked to clinical events in atherosclerosis and cancer. Since rheumatoid arthritis (RA) shares similar etiopathologic mechanisms with the above diseases, we sought to determine the clinical utility of determining MIC-1 serum levels and MIC-1 genotype in the management of RA. Ninety-one RA patients were recruited. Serum was collected from 83 of these patients and synovial biopsy samples were collected from the remaining 8 patients. Of the 83 patients from whom serum was collected, 61 were treated on an outpatient basis (defined as having nonsevere disease), and 22 patients went on to undergo hemopoietic stem cell transplantation (HSCT) (defined as having severe disease). Serum levels of MIC-1 were higher in RA patients and reflected disease severity independently of classic disease markers. MIC-1 was detected in rheumatoid synovial specimens, and allelic variation of MIC-1 was associated with earlier erosive disease and severe treatment-resistant chronic RA. Additionally, algorithms including serum and/or allelic variation in MIC-1 predicted response to HSCT, the presence of severe disease, and joint erosions. Determination of serum levels of MIC-1 and MIC-1 genotype may be clinically useful in the management of RA as well as in selection of patients for HSCT, since they predict disease course and response to therapy. The data indicate a potential role for MIC-1 in RA pathogenesis. These results warrant larger prospective studies to fully delineate and confirm a role for MIC-1 genotyping and serum estimation in patient selection for HSCT and in the management of RA.


Night shift work at specific age ranges and chronic disease risk factors.
We examined the association of night shift work history and age when night shift work was performed with cancer and cardiovascular disease risk factors among 54 724 women in the Nurses' Health Study (NHS) II. We calculated age-adjusted and socioeconomic status-adjusted means and percentages for cancer and cardiovascular risk factors in 2009 across categories of night shift work history. We used multivariable-adjusted logistic regression to estimate odds ratios (ORs) and 95% CIs for key risk factors among 54 724 participants (72% ever shift workers). We further examined these associations by age (20-25, 26-35, 36-45 and 46+ years) at which shift work was performed. Ever night shift workers had increased odds of obesity (body mass index ≥30 kg/m(2); OR=1.37, 95% CI 1.31 to 1.43); higher caffeine intake (≥131 mg/day; OR=1.16, 95% CI 1.12 to 1.22) and total calorie intake (≥1715 kcal/day; OR=1.09, 95% CI 1.04 to 1.13); current smoking (OR=1.30, 95% CI 1.19 to 1.42); and shorter sleep durations (≤7 h of sleep/day; OR=1.19, 95% CI 1.15 to 1.24) compared to never night shift workers. These estimates varied depending on age at which night work was performed, with a suggestion that night shift work before age 25 was associated with fewer risk factors compared to night shift work at older ages. Our results indicate that night shift work may contribute to an adverse chronic disease risk profile, and that risk factors may vary depending on the age at which night shift work was performed.


Blood metal levels and metal-influenced immune functions of harbour seals in captivity.
Immunological blood parameters and the effects of environmental pollutants on the immune system are important to assess the health status of seals. Animals living permanently in seal centres are useful for development and validation of diagnostic tools for free-ranging animals. In this study, parameters of cellular immunity as well as metal concentrations in blood and metal influence on cell proliferation of seven seals from a seal centre were investigated repeatedly using multi-element analysis and a lymphocyte proliferation assay. The metal concentrations, except for tin and chromium, were in general comparable to those of free-ranging animals of the North Sea. The unstimulated and mitogen-stimulated lymphocyte proliferation showed strong intra- and inter-individual variability, which reflected variability in activation of the immune status. Furthermore, both immunosuppressive and stimulative influences of metals on lymphocytes were found. Summarising, the methods used in this investigation provided useful information on these animals, and their application to free-ranging animals can be recommended.


Aphid-borne Viral Spread Is Enhanced by Virus-induced Accumulation of Plant Reactive Oxygen Species.
Most known plant viruses are spread from plant to plant by insect vectors. There is strong evidence that nonpersistently transmitted viruses manipulate the release of plant volatiles to attract insect vectors, thereby promoting virus spread. The mechanisms whereby aphid settling and feeding is altered on plants infected with these viruses, however, are unclear. Here we employed loss-of-function mutations in cucumber mosaic virus (CMV) and one of its host plants, tobacco (Nicotiana tabacum), to elucidate such mechanisms. We show that, relative to a CMVΔ2b strain with a deletion of the viral suppressor of RNAi 2b protein in CMV, plants infected with wild-type CMV produce higher concentrations of the reactive oxygen species (ROS) H2O2 in plant tissues. Aphids on wild-type CMV-infected plants engage in shorter probes, less phloem feeding, and exhibit other changes, as detected by electrical penetration graphing technology, relative to CMVΔ2b-infected plants. Therefore, the frequency of virus acquisition and the virus load per aphid were greater on CMV-infected plants than on CMVΔ2b-infected plants. Aphids also moved away from initial feeding sites more frequently on wild-type CMV infected versus CMVΔ2b-infected plants. The role of H2O2 in eliciting these effects on aphids was corroborated using healthy plants infused with H2O2 Finally, H2O2 levels were not elevated, and aphid behavior was unchanged, on CMV-infected RbohD-silenced tobacco plants, which are deficient in the induction of ROS production. These results suggest that CMV uses its viral suppressor of RNAi protein to increase plant ROS levels, thereby enhancing its acquisition and transmission by vector insects.


Long-term effects of unintended pregnancy on antiretroviral therapy outcomes among South African women living with HIV.
Unintended pregnancies are common among women living with HIV, but there are no data on their long-term impact on treatment outcomes. In a cohort of women initiating antiretroviral therapy (ART) during pregnancy, we examined the association between the intendedness of the current pregnancy, measured antenatally, and elevated viral load up to 5 years postpartum. Prospective study with enrolment at entry into antenatal care and follow-up at study visits separate from routine care. At enrolment women completed the London Measure of Unplanned Pregnancy. Mixed effects models examined the impact of the intendedness of the pregnancy (planned versus each of unplanned or ambivalent, respectively) on viral load 50 or more copies/ml across postpartum study visits. Overall, 459 women were followed for a median of 43 months postpartum, contributing 2535 viral load measures (median per woman: 6). Ambivalent and unplanned pregnancy were commonly reported (20 and 60%, respectively), and the proportion of women with elevated viral load increased over time (16% at 6 weeks to 43% by 36-60 months postpartum). Compared with those reporting a planned pregnancy, elevated viral load was more common among women reporting an unplanned pregnancy (odds ratio: 2.87; 95% confidence interval: 1.46-5.64), with a trend towards a higher odds among those reporting ambivalence (odds ratio: 2.19; 95% confidence interval: 0.97-4.82); associations persisted after adjustment for a wide range of demographic, clinical and psychosocial factors. These novel data suggest that unplanned pregnancy may be a prevalent and persistent predictor of poor ART outcomes among women initiating ART during pregnancy.


Anticarcinogenic properties of a daidzein-rich fraction isolated from soybean.
In a previous study, we demonstrated that the methanol extract of soybean powder contains an active component(s) that promotes the differentiation of HL-60 cells. Partial purification of the extract, using solvent fractionation and silica gel chromatography, produced an active fraction rich in daidzein. The daidzein-rich fraction (DRF) was evaluated for its cancer preventive potential by assessing its cytotoxic activity and effect on the expression of the transforming growth factor beta (TGF-beta) family of cytokines and their receptors. DRF appeared to exert cytotoxic activity via an apoptotic pathway as evaluated by a DNA fragmentation assay and caspase-3 induction. DRF also increased the expression of TGF-beta2, but had little effect on the expression of other members of the TGF-beta family of cytokines and their receptors, or on the expression of the vascular endothelial growth factor gene. In conclusion, the DRF isolated from the methanol extract of soybean may have the potential to prevent tumorigenesis and, therefore, deserves to undergo further evaluation of its active component(s) and in vivo evaluation for anticarcinogenic efficacy.


p300 and p300/cAMP-response element-binding protein-associated factor acetylate the androgen receptor at sites governing hormone-dependent transactivation.
The androgen receptor (AR) is a sequence-specific DNA-binding protein that plays a key role in prostate cancer cellular proliferation by dihydrotestosterone and the induction of secondary sexual characteristics. In this study we demonstrate that the AR can be modified by acetylation in vitro and in vivo. p300 and p300/cAMP-response element-binding protein acetylated the AR at a highly conserved lysine-rich motif carboxyl-terminal to the zinc finger DNA-binding domain. [(14)C]acetate-labeling experiments demonstrated that AR acetylation by p300 in cultured cells requires the same residues identified in vitro. Point mutation of the AR acetylation site (K632A/K633A) abrogated dihydrotestosterone-dependent transactivation of the AR in cultured cells. Mutation of the p300 CH3 region or the p300/cAMP-response element-binding protein histone acetylase domain reduced ligand-dependent AR function. The identification of the AR as a direct target of histone acetyltransferase co-activators has important implications for targeting inhibitors of AR function.


Do metformin and phenformin potentiate differently B-cell response to high glucose? An in vitro study on isolated rat pancreas.
The study investigated the effects of metformin and phenformin, at "therapeutic" concentrations, on the pancreatic A-, B- and D- cell response to glucose using the isolated perfused rat pancreas model. Changes in the rate of pancreatic lactate output after these biguanides were also evaluated. Metformin--at 1.5 micrograms/ml--and phenformin--at 100 ng/ml--were separately infused both at 160 mg/dl and 300 mg/dl glucose levels. Neither metformin nor phenformin affected glucagon or somatostatin secretion during these two metabolic stimuli with glucose, nor did they significantly influence insulin response to the lower glucose stimulus. Both metformin and phenformin enhanced insulin response to 300 mg/dl glucose infusion and increased the second phase of the B-cell secretory profile but only phenformin significantly enhanced the pancreatic lactate output rate during the 300 mg/dl glucose infusion. Infusion with dichloroacetate (a stimulator of the mitochondrial pyruvate oxidation) or with verapamil (a calcium antagonist) alone did not modify the insulin response to high glucose concentrations. During metformin infusion dichloroacetate neither modified metformin's effects on B-cell response to high glucose nor did it affect the pancreatic lactate output rate. On the other hand dichloroacetate opposed phenformin's effects on the B-cell response to high glucose and reversed the rise in the pancreatic lactate output rate. Verapamil inhibited the effect of metformin on the B-cell response to high glucose but failed to affect phenformin's influence on high-glucose induced insulin release. These data suggest both metformin and phenformin potentiate--at least in rats--the late phase of insulin secretory response to high glucose. However metformin seems to influence pancreatic B-cell activity mainly by facilitating the trans-membrane calcium ion influx responsible for the second phase of insulin release. Phenformin's influence seems indirect since it increases pancreatic lactate production which mediates the enhanced B-cell response to glucose.


Molecular properties of transmissible R factors of Haemophilus influenzae determing tetracycline resistance.
The tetracycline-resistant Haemophilus influenzae strains LU121 and FR16017, recently isolated in West Germany, each harbour a plasmid; that of the former (pLU12U) has a mol. wt of 31.5 X 10(6) and that of the latter (pFR16017) has a mol. wt of 33 X 10(6). Conjugation and DNA-DNA hybridization studies have shown that both plasmids are self-transmissible and carry tetracycline-resistance genes. The purified plasmid DNA of H. influenzae strain LU121 transformed a sensitive Escherichia coli strain to tetracycline resistance. The two R factors are closely related to the H. influenzae plasmid specifying ampicillin resistance (pKRE5367). Electron microscope DNA heteroduplex analysis indicated that pLU121 and pFR15017 probably carry the tetracycline-resistance transposon TnD and that pKRE5367 probably carries the ampicillin-resistance transposon TnA. There is more than one integration site for the insertion which probably represents TnD in pFR15017. All three plasmids have a similar plasmid core and could have a common evolutionary origin.


Management of complex CHD at the National Cardiothoracic Center of Excellence, University of Nigeria Teaching Hospital, Enugu: the role of foreign cardiac missions in 3.5 years.
CHD is defined as structural defect(s) in the heart and proximal blood vessels present at birth. The National Cardiothoracic Center of Excellence, University of Nigeria Teaching Hospital (UNTH), Enugu, through the aid of visiting Cardiac Missions has managed a significant number of patients within the last 3.5 years. Aim/Objective The objective of this study was to review surgical options and outcome of complex CHD among patients attending UNTH, Enugu, Enugu. Materials and Method During the period of 3.5 years (March, 2013 to June, 2016), a total of 20 cases of complex CHD were managed by cardiac missions that visited UNTH, Enugu. Their case notes and operating register were retrieved, reviewed, and analysed using SPSS version 19 (Chicago). There were eight females and 12 males, with a ratio of 2:5. The age range was from 5 months to 34 years with a mean of 1.7. Among all, five patients died giving a mortality rate of 25%. The operative procedures ranged from palliative shunts to complete repair. The outcome was relatively good. Complex CHD are present in our environment. Their surgical management in our centre is being made possible by periodic visits of foreign cardiac missions.


Improved parent-reported mobility and achievement of individual goals on activity and participation level after functional power-training in young children with cerebral palsy: a double-baseline controlled trial.
In children with cerebral palsy (CP), strength training programs to improve walking capacity and participation in activities of daily living are commonly used in clinical practice, despite lacking evidence of its effectiveness. It has been suggested that strength training with high movement velocity could be more effective than traditional resistance training to improve functional abilities such as walking. In a recently published study, we have demonstrated the positive effects of functional high-velocity resistance (power) training on muscle strength and walking capacity in young children with CP. Whether this type of training is also effective in achieving individual predefined goals in daily activities and self-reported mobility limitations, has not yet been described however. To evaluate the effect of functional power-training on parent-reported mobility and achievement of individual goals on activity and participation level in young children with CP. A double-baseline design was used to compare a 14-week period usual care with a 14-week period of functional power-training (3 times a week) and a follow-up period of 14 weeks. A rehabilitation center, two special needs schools for children with physical disabilities, and a university medical center outpatient clinic. Twenty-two children with spastic CP (13 bilateral, GMFCS level I [N.=10] and level II [N.=12], mean age 7.5 years [SD 1.8, range 4-10 years]) and their parents participated. Outcome measures were goal attainment scaling (GAS) of individual daily activity related treatment goals, mobility performance as measured using the Functional Mobility Scale (FMS-5 m, 50 m and 500 m), and the parent-reported Mobility Questionnaire (MobQues). After power-training, 86% of children achieved or exceeded their goal, compared with 14% in the usual care period (P<.001). The probability of improvement by one point or more on the FMS-500 meter after functional power-training was 10 times higher, compared with the usual care period (Relative Risk=10.0 with 95% CI 1.4 - 71.3). No changes were found in the FMS-5m and FMS-50m categories. Improvement on the MobQues was significantly greater after power-training compared with usual care (7.9% (95% CI 2.7 - 13.0, P=.005)). The improvement in performance in the activities defined in the treatment goals continued during the follow-up period. The results indicated that functional power-training is an effective training to achieve personalized treatment goals for activities in daily life and parent-reported mobility performance in young children with cerebral palsy.


Bacterial Endosymbionts Are Common Among, but not Necessarily Within, Insect Species.
Bacterial endosymbionts, particularly Wolbachia (Rickettsiales: Rickettsiaceae), Rickettsia (Rickettsiales: Rickettsiaceae), and Cardinium (Bacteroidales: Bacteroidaceae), are commonly found in several arthropod groups, including insects. Most estimates of the global infection rate of Wolbachia (52% [95% credible intervals: 44-60]) show that these bacteria infect more than half of all insect species. Other endosymbionts, such as Rickettsia (24% [confidence intervals [CIs] 20-42]) and Cardinium (13% [CIs 13-55]), infect a smaller but still substantial proportion of insect species. In spite of these observations, it is unclear what proportion of individuals within those species are infected. Here, we used published databases to estimate the proportion of individuals that are infected with either Wolbachia, Rickettsia, or Cardinium. We found that the majority (69%) of Wolbachia-infected species have less than half of their individuals infected with Wolbachia, indicating that although the bacterium may be common among species, it is not common within species. The same was true for Rickettsia (81%) and Cardinium (87%). This discrepancy was consistent across orders, in which less than 10% of individuals were typically infected, even though more than 50% of species within orders were infected. For example, according to our model, nearly 50% of beetle (Coleoptera) species are infected with Wolbachia (i.e., contain at least one individual that has tested positive for Wolbachia), but less than 5% of all individuals are infected. These results add to the growing knowledge base about endosymbionts in insects and should guide future sampling efforts and investigations on the role that these bacteria play in populations.


Increased morphological diversity of microglia in the activated hypothalamic supraoptic nucleus.
Microglia are the immune cells of the CNS. In the normal adult mammalian brain, the majority of these cells is quiescent and exhibits a ramified morphology. Microglia are perhaps best known for their swift transformation to an activated ameboid morphology in response to pathological insults. Here we have observed the responsiveness of these cells to events surrounding the normal activation of neurosecretory neurons in the hypothalamic supraoptic nucleus (SON), a well studied model of structural plasticity in the CNS. Neurons in the SON were activated by substituting 2% saline for drinking water. Brain sections were collected from four experimental groups [controls (C), 2 d-dehydrated (2D), 7 d-dehydrated (D7), and 7 d-dehydrated/21 d-rehydrated animals (R21)] and stained with Isolectin-B4-HRP to visualize microglial cells. Based on morphological criteria, we quantified ramified, hypertrophied, and ameboid microglia using unbiased stereological techniques. Statistical analyses showed significant increases in the number of hypertrophied microglia in the D2 and D7 groups. Moreover, there was a significant increase in the number of ameboid microglia in the D7 group. No changes were seen across conditions in the number of ramified cells, nor did we observe any significant phenotypic changes in a control area of the cingulate gyrus. Hence, increased morphological diversity of microglia was found specifically in the SON and was reversible with the cessation of stimulation. These results indicate that phenotypic plasticity of microglia may be a feature of the normal structural remodeling that accompanies neuronal activation in addition to the activation that accompanies brain pathology.


Chemical Profile and Antioxidant Properties of Extracts and Essential Oils from Citrus × limon (L.) Burm. cv. Femminello Comune.
Citrus × limon cv. Femminello Comune (Rutaceae) from Rocca Imperiale (Italy), one of the six Protected Geographical Indication (PGI) Italian lemon crops, has been recently received renewed interest. In this work, fresh and dried peels and leaves were extracted by hydrodistillation, supercritical fluid extraction (SFE), and Soxhlet apparatus. Chemical profile was assessed by gas chromatography (GC) and gas chromatography/mass spectrometry (GC/MS). Except for leaves extracts obtained by Soxhlet apparatus, the monoterpene hydrocarbons fraction dominated. Limonene, γ-terpinene, and β-pinene were the main identified compounds. The antioxidant activity was investigated using different in vitro assays namely 2,2-diphenyl-1-picrylhydrazyl (DPPH), ABTS, ferric reducing ability power (FRAP), and β-carotene bleaching test. In DPPH test, the essential oil obtained by hydrodistillation of fresh peel exhibited the highest activity (IC50 of 1.17 mg/ml). Leaves extracted by SFE showed a good activity in both DPPH and β-carotene bleaching test with IC50 values of 2.20 and 6.66 mg/ml, respectively. Monoterpene hydrocarbons fraction exhibited a positive Pearson's correlation coefficient with all antioxidant assays. Leaves, often considered waste material, should be considered from a different point because they represent a matrix of indisputable interest.


In vitro activity of ampicillin and ceftriaxone against ampicillin-susceptible Enterococcus faecium.
To assess activity of the combination of ceftriaxone and ampicillin against clinical isolates of ampicillin-susceptible Enterococcus faecium. Ampicillin-susceptible E. faecium (n = 29) and Enterococcus faecalis (n = 10) collected from locations in the USA and France were used for this analysis. Susceptibility testing was performed by gradient diffusion strip (GDS) and broth microdilution (BMD). Synergy with the combination of ceftriaxone and ampicillin was assessed in all isolates using GDS crossing and double disc diffusion methods. Selected isolates (nine E. faecium and three E. faecalis) were assessed for synergy in time-kill studies using ampicillin alone and in combination with ceftriaxone. In isolates of E. faecium, the median (range) ampicillin MIC by BMD was 0.5 (0.25-4) mg/L and by GDS it was 2 (1-8) mg/L. In E. faecalis, the median (range) ampicillin MIC by BMD was 0.5 (0.5-1) mg/L and by GDS it was 2 (0.75-3) mg/L. A total of 24/29 (82.8%) isolates of E. faecium displayed synergy by GDS and 22/29 (75.9%) by double disc diffusion. Seven of 10 (70%) isolates of E. faecalis displayed synergy by GDS and 4/10 (40%) by double disc diffusion. Time-kill studies found synergy in 3/9 (33.3%) E. faecium and 3/3 (100%) E. faecalis. In contrast to the demonstrated synergy in time-kill models of ceftriaxone and ampicillin for E. faecalis, this combination does not appear to provide uniform synergy in E. faecium. Antagonism was not observed. Clinical correlation is necessary and caution should be used when considering ampicillin and ceftriaxone for the treatment of infections caused by ampicillin-susceptible E. faecium.


Distinct metabolism of linoleic and linolenic acids in liver and adipose tissues of finishing Normande cull cows.
Feeding strategies based on the addition of plant lipids rich in polyunsaturated fatty acids (PUFAs) in diets of bovines during the finishing period are common to enhance the nutritional value of meat. However, following rumen biohydrogenations, these FAs could still be metabolised in various tissues/organs involved in the FA metabolism such as the liver and adipose tissues (ATs), thus affecting their subsequent deposition in muscles. In this context, the objective of this study was to characterise the various metabolic pathways of linoleic acid (LA) and α-linolenic acid (ALA) in the liver and ATs (subcutaneous (SC) and inter-muscular (IM)) of Normande cull cows fed a diet supplemented (LR) or not (C) with extruded linseeds and rapeseeds, using the ex vivo incubated tissue slice method. Hepatic uptake of both FAs was higher with the LR than with the C diet (P = 0.02). For the two diets, ALA uptake was higher than that of LA (+46%, P = 0.04). ALA was much more degraded by β-oxidation (>50% of ALA present in cells) than LA (∼27%) with both diets (P = 0.015). Whatever the diet, ALA was not converted into longer and/or more unsaturated FA, whereas about 14% of LA was converted into 20:4n-6. The intensity of the esterification pathway was higher (+70%, P = 0.004) with the LR than with the C diet, for both FAs. Hepatic secretion of ALA as part of the very-low-density lipoprotein particles was lower than that of LA (-58% and -23% for C and LR diets respectively, P = 0.02). In SC and IM ATs, dietary lipid supplementation did not alter metabolic pathways of LA and ALA. They were efficiently taken up by ATs (>68% of FA present in the medium), with uptake being higher for IM than for SC AT (+12%, P = 0.01). Moreover, LA uptake by ATs was higher than ALA uptake (+10.7%, P = 0.027). Both FAs were mainly esterified (>97% of FA present in adipocytes) into neutral lipids (>85% of esterified FA). Around 9.5% of LA was converted into 20:4n-6, whereas only around 1.3% of ALA was converted into 20:5n-3. We concluded that, in our experimental conditions, liver was highly active in ALA catabolism limiting its subsequent deposition in muscles. However, bovine liver and ATs were inefficient at converting ALA into long-chain n-3 PUFA, but actively converted LA into 20:4n-6.


Superior longitudinal fasciculus and cognitive dysfunction in adolescents born preterm and at term.
To understand the relationship between cognition and white-matter structure in adolescents born preterm without obvious brain injury. Thirty-two adolescents from a longitudinal study of child development were selected according to risk of developmental disorders at birth (born at term: eight males, five females; median age 16y 1mo, interquartile range 10mo; low risk preterm: four males, five females, median age 16y, range 4mo; high risk preterm: three males, seven females, median age 16y 2mo, range 1y 2mo) and reading ability (good: three males, eight females, median age 16y, range 7mo; average: six males, three females, median age 16y 10mo, range 1y; poor: six males, six females, median age 16y, range 6mo). Preterm birth was defined as a gestational age of 36 weeks or less and a birthweight of 1600g or less. All participants had normal clinical neuroimaging findings. We examined fractional anisotropy, radial diffusivity, and volume of three major white-matter fasciculi. The relationship between structural measures and birth risk, hemisphere, and cognitive ability (attention, lexical and sublexical decoding, auditory phonological awareness, and processing speed) were analysed using mixed-model regression. Left-hemisphere superior longitudinal fasciculus (SLF) fractional anisotropy and radial diffusivity were linked to reading-related skills (fractional anisotropy vs letter-word identification, r((30))=-0.37, p<0.05; fractional anisotropy vs phoneme reversal, r((30))=-0.34, p=0.05; radial diffusivity vs letter-word identification, r((30))=0.31, p<0.10; radial diffusivity vs phoneme reversal, r((30))=0.40, p<0.05), whereas right-hemisphere SLF fractional anisotropy was related to attention skills (fractional anisotropy vs inattentiveness, r((30))=-0.38, p<0.05). SLF volume decreased as these skills declined for adolescents born preterm (volume vs phoneme reversal, r((17))=0.58, p<0.01; volume vs inattentiveness, r((17))=-0.69, p<0.01), but not for those born at term. The relationship between cognitive skills and SLF volume suggests that in adolescents born preterm, cryptic white-matter injury may exist, possibly related to oligodendrocyte or axonal loss, despite normal clinical neuroimaging.


Team assessment of behaviour: a high stakes assessment with potential for poor implementation and impaired validity.
Team assessment of behaviour (TAB) is the multi-source feedback assessment of professional behaviours that all UK foundation doctors must engage in twice during their two-year programme. TAB can identify the few underperforming trainees and provide feedback to consolidate the good practice of most. For optimum validity, TAB must be undertaken by a range of assessors, as specified in the national UK Foundation Programme curriculum. This study reports an audit of invalid TAB submissions over a three-year cycle in the West Midlands' Foundation Programme. In 2010, large numbers of TABs were invalid, owing to an incorrect selection or number of assessors. Introduction of validity checking before sign-off greatly improved the numbers of valid assessments in 2011. This was partially sustained in 2012. Assurance of assessment validity is important to ensure delivery of appropriate constructive feedback and to allow early detection and remediation of signs of poor professional behaviours in foundation doctors.


Nicotine replacement therapy for smoking cessation.
The aim of nicotine replacement therapy (NRT) is to replace nicotine from cigarettes. This reduces withdrawal symptoms associated with smoking cessation thus helping resist the urge to smoke cigarettes. The aims of this review were to determine the effectiveness of the different forms of nicotine replacement therapy (chewing gum, transdermal patches, nasal spray, inhalers and tablets) in achieving abstinence from cigarettes; to determine whether the effect is influenced by the clinical setting in which the smoker is recruited and treated, the dosage and form of the NRT used, or the intensity of additional advice and support offered to the smoker; to determine whether combinations of NRT are more effective than one type alone; and to determine its effectiveness compared to other pharmacotherapies. We searched the Cochrane Tobacco Addiction Group trials register in April 2000. Randomized trials in which NRT was compared to placebo or no treatment, or where different doses of NRT were compared. We excluded trials which did not report cessation rates, and those with follow-up of less than six months. We extracted data in duplicate on the type of subjects, the dose and duration and form of nicotine therapy, the outcome measures, method of randomisation, and completeness of follow-up. The main outcome measure was abstinence from smoking after at least six months of follow-up. We used the most rigorous definition of abstinence for each trial, and biochemically validated rates if available. Where appropriate, we performed meta-analysis using a fixed effects model. We identified one hundred trials; of the 88 with a non NRT control group, 48 trials were of nicotine gum, 30 of transdermal nicotine patch, four of intranasal nicotine spray, four of inhaled nicotine and two of nicotine sublingual tablet. Four trials compared combinations of two forms of nicotine therapy with one form alone. The odds ratio for abstinence with NRT compared to control was 1.71 (95% confidence interval 1.60 to 1.82), The odds ratios for the different forms of NRT were 1.63 for gum, 1.73 for patches, 2.27 for nasal spray, 2.08 for inhaled nicotine and 1.73 for nicotine sublingual tablet. These odds were largely independent of the intensity of additional support provided or the setting in which the NRT was offered. Eight weeks of patch therapy was as effective as longer courses and there was no evidence that tapered therapy was better than abrupt withdrawal. Wearing the patch only during waking hours (16 hours/day) was as effective as wearing it for 24 hours/day. The odds ratio for abstinence in the trials which directly compared 4 mg versus 2 mg gum in highly dependent smokers found a significant benefit in favour of 4 mg gum (odds ratio 2.67, 95% confidence interval 1.69 to 4.22). There is no strong evidence that combinations of forms of NRT are more effective. Only one study directly compared NRT to an antidepressant (bupropion). In this study, bupropion was significantly more effective than nicotine patch or placebo. The combination of bupropion and nicotine patch was significantly more effective than nicotine patch alone. There was also a suggestion of greater of efficacy for bupropion and nicotine patch compared to bupropion alone, but the difference was not statistically significant. All of the commercially available forms of NRT (nicotine gum, transdermal patch, the nicotine nasal spray, nicotine inhaler and nicotine sublingual tablets) are effective as part of a strategy to promote smoking cessation. They increase quit rates approximately 1.5 to 2 fold regardless of setting. The effectiveness of NRT appears to be largely independent of the intensity of additional support provided to the smoker. Since all the trials of NRT reported so far have included at least some form of brief advice to the smoker, this represents the minimum which should be offered in order to ensure its effectiveness. Provision of more intense levels of support, although beneficial in facilitating the likelihood of quitting, is not essential to the success of NRT. There is promising evidence that bupropion may be more effective than NRT (either alone or in combination). However, its most appropriate place in the therapeutic armamentarium requires further study and consideration.


Isoforms of nitric oxide synthase in the optic nerves of rat eyes with chronic moderately elevated intraocular pressure.
To investigate the hypothesis that nitric oxide (NO) in the optic nerve heads of rats with chronic moderately elevated intraocular pressure (IOP) contributes to neurotoxicity of the retinal ganglion cells, the presence of the three isoforms of nitric oxide synthase (NOS) was determined in the tissue. Unilateral chronic moderately elevated IOP was produced in rats by cautery of three episcleral vessels. Histologic sections of optic nerves from eyes with normal IOP and with chronic moderately elevated IOP were studied by immunohistochemistry and by immunoblot analysis. Polyclonal antibodies to NOS-1, NOS-2, NOS-3, and glial fibrillary acidic protein (GFAP) were localized with immunoperoxidase. In the optic nerve of rat eyes with normal IOP, NOS-1 was constitutively present in astrocytes, pericytes and nerve terminals in the walls of the central artery. NOS-2 was not present in eyes with normal IOP. In these eyes, NOS-3 was constitutively present in the vascular endothelia of large and small vessels. Rat eyes treated with three-vessel cautery had sustained elevated IOP (1.6 fold) for at least 3 months. In these eyes, no obvious changes in NOS-1 or NOS-3 were noted. However, at time points as early as 4 days of chronic moderately elevated IOP, NOS-2 appeared in astrocytes in the optic nerve heads of these eyes and persisted for up to 3 months. Immunoblot analysis did not detect differences in NOS isoforms. The cellular distributions of constitutive NOS isoforms in the rat optic nerve suggest physiological roles for NO in this tissue. NOS-1 in astrocytes may produce NO as a mediator between neighboring cells. NO, produced by NOS-1 in pericytes and nitrergic nerve terminals and by NOS-3 in vascular endothelia, is probably a physiological vasodilator in this tissue. In eyes with chronic moderately elevated IOP, NOS-2 is apparently induced in astrocytes. The excessive NO production that is associated with this isoform may contribute to the neurotoxicity of the retinal ganglion cells in eyes with chronic moderately elevated IOP.


In search of tail-anchored protein machinery in plants: reevaluating the role of arsenite transporters.
Although the mechanisms underlying selective targeting of tail-anchored (TA) membrane proteins are well established in mammalian and yeast cells, little is known about their role in mediating intracellular membrane trafficking in plant cells. However, a recent study suggested that, in green algae, arsenite transporters located in the cytosol (ArsA1 and ArsA2) control the insertion of TA proteins into the membrane-bound organelles. In the present work, we overproduced and purified these hydrophilic proteins to near homogeneity. The analysis of their catalytic properties clearly demonstrates that C. reinhardtii ArsA proteins exhibit oxyanion-independent ATPase activity, as neither arsenite nor antimonite showed strong effects. Co-expression of ArsA proteins with TA-transmembrane regions showed not only that the former interact with the latter, but that ArsA1 does not share the same ligand specificity as ArsA2. Together with a structural model and molecular dynamics simulations, we propose that C. reinhadtii ArsA proteins are not arsenite transporters, but a TA-protein targeting factor. Further, we propose that ArsA targeting specificity is achieved at the ligand level, with ArsA1 mainly carrying TA-proteins to the chloroplast, while ArsA2 to the endoplasmic reticulum.


[Manifestations of local fibrosis in the clinical picture of carcinoids of digestive organs].
A complex of clinical, functional, and morphological studies was made in 20 patients aged 39 to 78 years who had histologically verified gastrointestinal carcinoids in the pre- and postoperative periods, by employing laboratory tests and instrumental techniques. Removed tumors were morphologically studied. Sixteen patients were operated on. Among them, 10 and 6 patients underwent planned and emergency operations, respectively. On referral for emergency laparotomy, the preoperative diagnoses were acute ileus in 2 cases, acute pancreatitis in 1, and acute appendicitis in 3. The local paracrine effect of serotonin on the intestinal wall leads to spasm and fibrosis, which manifests itself as the clinical picture of obstruction of a hollow organ or acute appendicitis. Cordlike or concentrated deformity as a kink is a characteristic sign of small intestinal carcinoid. Formation of fibrosis occurs not only in the pathways of evacuation of excess serotonin from hepatic metastases into the right ventricle of the heart (Hedinger's syndrome), but by the paracrine pathway in the immediate vicinity of a carcinoid tumor, in whatever organ the tumor is located.


PTEN, A Target of Microrna-374b, Contributes to the Radiosensitivity of Canine Oral Melanoma Cells.
Canine oral malignant melanoma (CoMM) is often treated by radiation therapy in veterinary medicine. However, not all cases are successfully managed by this treatment. For improved efficacy of radiation therapy, biomarkers predicting the radiosensitivity of melanoma cells need to be explored. Here, we, first, developed the radioresistant CoMM cell line, KMeC/R. We found that the expression level of phosphatase and tensin homolog (PTEN) of KMeC/R cells was significantly downregulated compared with KMeC cells. Overexpression of PTEN successfully restored the radiosensitivity of KMeC/R cells, and silencing of PTEN significantly increased the radioresistance of the CoMM cells tested. Next, we focused on microRNAs (miRNAs) to explore the mechanisms of downregulation of PTEN in KMeC/R cells. miR-374b was upregulated in KMeC/R cells compared with that in KMeC cells and in the irradiated CoMM cells tested. Furthermore, miR-374b directly targeted PTEN based on the luciferase activity assay. Moreover, the extrinsic miR-374b significantly increased the radioresistance of KMeC cells. In addition, the expression level of PTEN was significantly downregulated and that of miR-374b tended to be upregulated in recurrent CoMM tissues after radiation therapy compared with the pre-treatment tissues. Thus, the current study suggested that the miR-374b/PTEN signaling pathway possibly plays an important role in CoMM radiosensitivity.


Clonal variation of cadmium response in human tumor cell lines.
Subpopulations of human tumor-derived cell lines A101D, A204, and A549 were screened for Cd2+ cytotoxic response. Three of six A549, two of seven A101D, and four of seven A204 subpopulations were found to differ significantly from the parental line. A variant subpopulation of A101D (T3) was shown by flow cytometry to be comprised of cells having two distinct DNA histograms. One histogram, type 1, resembles that of normal human fibroblasts. The other, type 2, represents cells with one-third more DNA. Early passage T3 clonal populations were comprised primarily of type 1 cells. With passage, type 1 cells decreased relative to type 2 so that by passage 47 the culture was predominantly type 2. Correspondingly, the A101D T3 subpopulation became more Cd2+ sensitive with time in culture. Subclones having only type 1 DNA histograms were found to be Cd2+ resistant relative to subclones with type 2 histograms, and treatment of A101D T3 cultures having approximately equal amounts of type 1 and 2 cells with 2 microM Cd2+ resulted in the selection of type 1 cells. The enhanced Cd2+ resistance phenotype shown by A101D T3 type 1 cells correlated with reduced Cd2+ uptake and is not attributable to enhanced metallothionein synthesis.


U.s. State and federal laws targeting distracted driving.
Distracted driving has burgeoned with the proliferation of cell phones, global positioning systems and other in-vehicle and personal electronic devices. Annually more than 3,300 people are killed and an additional 400,000 are injured in the United States in distracted driving crashes. The United States (U.S.) federal and state governments have responded to this public health problem with policies and laws; however, a more comprehensive and more effective approach is still needed. Some restrictions on the use of electronic devices while driving by federal employees and some voluntary guidelines and recommendations have been issued. Public opinion polls show support for addressing the issue of distracted driving with state laws. The majority of states have laws banning text messaging while driving and prohibiting the use of an electronic device by teenage or novice drivers. Some states prohibit all drivers from using a hand-held cellphone. Currently no state has a total ban on the use of personal electronic devices while driving. Successful past traffic safety campaigns changing driver behavior have demonstrated the necessity to adopt a "three Es" approach of Enactment of a law, Education of the public about the law, and rigorous Enforcement of the law. Experience reveals that this approach, along with future federal regulation of in-vehicle electronic devices and the employment of technology to limit the use of electronic devices while driving, is needed to alter personal behavior in order to reduce distractions and keep drivers focused on the driving task.


Perioperative Corneal Abrasions After Nonocular Surgery: A Systematic Review.
To perform a systematic review of the international literature evaluating the risk factors, preventative steps, and treatments for perioperative corneal injuries for nonocular surgery. PubMed, Embase, and Evidence-Based Medicine Reviews databases were searched on April 13, 2018. Two hundred four articles were identified with 16 meeting the inclusion criteria. All studies were evaluated for quality and level of evidence. Two types of studies were included. The first were primary epidemiological studies that looked at the rates of perioperative corneal injuries after nonocular surgery and the second were trials that either studied preventative steps or treatments. A statistical analysis was completed to reveal trends in perioperative corneal abrasions. Rates ranged from 0.01% to 59% with a cumulative rate of 0.64% (95% confidence interval 0.36%-1.35%). Primary risk factors were identified as longer procedures, general anesthesia, and advanced age. The most commonly associated ocular injuries were found to include chemical injury, conjunctivitis, blurred vision, and conjunctival congestion. Treatment strategies for corneal abrasion in the literature recommended erythromycin ointment and ample ocular lubrication for the fastest recovery. Education interventions alone, as studied in 2 of the 16 articles, demonstrated a significant decrease in the rate of corneal abrasions. Standardized ocular protection, reporting, and education initiatives were found to maximally decrease rates of perioperative corneal abrasions after nonocular surgery. However, no gold standard currently exists for intraoperative ocular protection. More research needs to be conducted on specific prevention strategies and content of educational initiatives in hopes of standard development across facilities nationwide.


Enhancing effect of pretreatment with topical niacin in the treatment of rosacea-associated erythema by 585-nm pulsed dye laser in Koreans: a randomized, prospective, split-face trial.
Rosacea is a chronic dermatosis that is usually confined to the face. A pulsed dye laser (PDL) system has been proven to be effective in treating rosacea-associated erythema and telangiectasias. Niacin is a cutaneous vasodilator that can increase the chromophore through increased blood flow. We hypothesized that increased blood flow by pretreatment with topical niacin could enhance the effect of PDL in the treatment of rosacea. Eighteen Korean patients with rosacea were recruited. Three sessions of 585-nm PDL using a subpurpuragenic dose with and without pretreatment with niacin cream were performed on randomly assigned half-faces at 3-week intervals. Erythema was assessed objectively by a polarization colour imaging system, and evaluations were also made by three blinded dermatologists. Patient satisfaction was evaluated using a 10-point visual analogue scale. Fifteen patients completed this study. All patients showed an improvement in erythema after three sessions of PDL treatment both with and without niacin pretreatment (P = 0·023 and P = 0·009, respectively). There was no significant difference in the improvement of objective erythema between the two sides. However, based on physician assessment the overall clinical improvement on the niacin side was significantly higher (P = 0·005), and patient satisfaction was also higher on the niacin-pretreated side (P = 0·007). There were no remarkable side-effects, with the exception of transient erythema and oedema. Pretreatment with topical niacin safely enhanced the effect of 585-nm PDL treatment of rosacea-associated erythema in Koreans. Application of niacin can be helpful in overcoming the relatively lower effect of subpurpuragenic PDL in dark-skinned Asians.


Antimicrobial Peptide immunity protects human nasal and auricular cartilage against infection.
Despite being impervious to surveillance by the adaptive immune system because of its lack of vascularity, infection of the nasal and auricular cartilage after surgery such as rhinoplasty or otoplasty is rare. Why is this so? Our goal was to determine whether the expression of antimicrobial peptides provides a previously unrecognized nonepithelial layer of innate immune defense within the nasal and auricular cartilage. Seven samples of nasal septum cartilage and 2 biopsies from auricular cartilage grafts were harvested during rhinoplasty and otoplasty procedures. Ten cadaveric samples of auricular and 9 samples of nasal cartilage were also obtained. Immunohistochemical staining was directed against the human beta-defensin antimicrobial peptides (hBD) 1, 2, and 3. A semiquantitative analysis was performed to measure immunoreactivity. All 3 human beta-defensins were detected along the perichondral line and within the cartilage matrix in the nasal and auricular samples. Areas with positive immunohistochemical staining were also detected within chondrocyte cytoplasm. We provide the first evidence of antimicrobial peptide expression (hBD-1, -2 and -3) within the perichondrium and cartilage matrix layers of the nasal and auricular cartilage. This previously unrecognized innate immune function of perichondrocytes and chondrocytes may explain the resistance of the nasal and auricular cartilage to infection after surgical procedures despite the absence of a vascular system.