Red and blue light differentially impact retrograde signalling and photoprotection in rice.
Chloroplast-to-nucleus retrograde signalling (RS) is known to impact plant growth and development. In Arabidopsis, we and others have shown that RS affects seedling establishment by inhibiting deetiolation. In the presence of lincomycin, a chloroplast protein synthesis inhibitor that triggers RS, Arabidopsis light-grown seedlings display partial skotomorphogenesis with undeveloped plastids and closed cotyledons. By contrast, RS in monocotyledonous has been much less studied. Here, we show that emerging rice seedlings exposed to lincomycin do not accumulate chlorophyll but otherwise remain remarkably unaffected. However, by using high red (R) and blue (B) monochromatic lights in combination with lincomycin, we have uncovered a RS inhibition of length and a reduction in the B light-induced declination of the second leaf. Furthermore, we present data showing that seedlings grown in high B and R light display different non-photochemical quenching capacity. Our findings support the view that excess B and R light impact seedling photomorphogenesis differently to photoprotect and optimize the response to high-light stress. This article is part of the theme issue 'Retrograde signalling from endosymbiotic organelles'.


Analysis of a fractional-order couple model with acceleration in feelings.
A fractional-order nonlinear dynamical model of couple has been introduced. Upper bounds are obtained for a fractional-order nonlinear dynamical model. Also different from other models, a model with the order 2 α is discussed. We are expecting an acceleration in feelings; that is why we increase the order of the derivative between 1 < 2α ≤ 2. Stability analysis of the fractional-order nonlinear dynamical model of involving two persons is studied using the fractional Routh-Hurwitz criteria. By using stability analysis on fractional-order system, we obtain sufficient condition on the parameters for the locally asymptotic stability of equilibrium points. Finally, numerical simulations are presented to verify the obtained results.


Toxicities of influenza vaccine: peripheral leukocytic response to live and inactivated influenza viruses in mice.
Intraperitoneal or intravenous inoculation of live or inactivated influenza virus induced two characteristic responses of the peripheral leukocytes in mice, an early appearing leukopenic response and late appearing lymphopenia. The former response usually developed and subsided within several hours, though the change in leukocyte population was fairly complicated depending upon the activity of the inoculated material, while the latter began several hours after inoculation and reached its minimum level in 10 to 20 hr. The agent responsible for the former may be virus pyrogen, while the latter seems to be caused by some substance(s) other than that. The early appearing leukopenic response was similar to that due to bacterial endotoxin in respect to the characteristic pattern of the change in peripheral leukocyte population, though it was relatively easy to distinguish one from the other by the length of the latent period and by the heat stability of the causative agent. Live or inactivated influenza virus causing the early appearing leukopenic response was found also to have the mouse body weight-decreasing toxicity. The significance of these findings in the laboratory control test of influenza vaccine for untoward reactions often observed in human inoculated with some inactivated influenza vaccines was discussed. The possible roles of the two agents, virus pyrogen and endotoxin, in the febrile response were mentioned.


Annual distribution of births and deaths outcomes at Harare Maternity Hospital, Zimbabwe.
To assess socio-demographic and reproductive/obstetric risk factors for stillbirth, preterm births and low birth weight (LBW). To explore the probability of death at birth by antenatal care attendance, and by delivery with a Caesarean section. Using information available in obstetric records, we conducted a retrospective analysis of data, covering a twelve month period. Harare Maternity Hospital, Zimbabwe All deliveries (17,174 births) occurring from October 1997 to September 1998 Stillbirth, preterm births and LBW. The annual stillbirth rate was 57 per 1000, preterm birth 215 per 1000 and LBW was 243 per 1000 total births. Women delivering stillbirths, preterm births and LBW infants were less likely to attend antenatal care (adjusted relative risk [RR] = 2.56 95% confidence intervals [CI]: 2.18 to 3.00, RR= 3.02 95% CI: 2,77 to 3.29, and RR = 2.38; 95% CI: 1.58-3.60), or to be deliveredby Caesarean section (RR= 0.61; 95% CI: 0.49 to 0.76, RR = 0.68; 95% CI: 0.61 to 0.76 and RR = 0.82; 95% CI: 0.75 to 0.90), but more likely to deliver a breech (RR = 3.17; 95% CI: 2.62 to 3.85, RR = 4.16; 95% CI: 3.72 to 4.64 and RR = 5.34; 95% CI: 4.88 to 5.85), respectively. Stillbirths were more likely to be a preterm birth (RR = 6.53; 95% CI: 5.65 to 7.79) or a LBW infant (RR= 6.42; 95% CI: 5.49 to 7.52). The annual frequency of poor birth outcomes at Harare Maternity Hospital, which is a referral hospital, also catering for a proportion of patients coming from distant health centers and from rural areas, is high. This high prevalence of birth outcomes is associated with lack of prenatal care, and breech delivery. Infants born too early or small have an increased risk of mortality. Early prenatal care, improved obstetric care, accessibility to care, use of emergency care services, and early access to Caesarean section, could save life, and could assist in reducing the frequency of poor birth outcomes in this population.


Associations between the gut microbiome and metabolic, inflammatory, and appetitive effects of sleeve gastrectomy.
The complex and multifactorial etiology of obesity creates challenges for its effective long-term management. Increasingly, the gut microbiome is reported to play a key role in the maintenance of host health and wellbeing, with its dysregulation associated with chronic diseases such as obesity. The gut microbiome is hypothesized to contribute to obesity development and pathogenesis via several pathways involving food digestion, energy harvest and storage, production of metabolites influencing satiety, maintenance of gut barrier integrity, and bile acid metabolism. Moreover, the gut microbiome likely contributes to the metabolic, inflammatory, and satiety benefits and sustained weight-loss effects following bariatric procedures such as sleeve gastrectomy. While the field of gut microbiome research in relation to obesity and sleeve gastrectomy outcomes is largely in its infancy, the gut microbiome nonetheless holds great potential for understanding some of the mechanisms behind sleeve gastrectomy outcomes as well as for optimizing post-surgery benefits. This review will explore the current literature within the field as well as discuss the current limitations, including the small sample size, variability in methodological approaches, and lack of associative data, which need to be addressed in future studies.


A reappraisal of the clinical efficacy of nebulized flunisolide in pediatric asthma: the Italian experience.
Flunisolide (FLU) is a synthetic corticosteroid with potent topical anti-inflammatory activity. Its oral bioavailability is poor (6.7%). After gastrointestinal and lung absorption, the drug undergoes rapid and extensive first-pass metabolism by the liver to an inactive 6beta-hydroxylated metabolite. Plasma half-life is estimated to be 3.9 to 4.6 hours. FLU has a low volume of distribution at steady state and a short terminal half-life after inhalation (96 L and 1.6 hour, respectively). FLU, like budesonide, has a short pulmonary residence time and it is hypothesized that it may undergo esterification in the cell due to the presence of a free hydroxyl group at C21. Nebulization may offer important advantages over other inhalation methods. Nebulizers allow drug delivery in very young children through passive inhalation, depending less on patient coordination and cooperation. Comparative studies indicate that FLU is nebulized to a better advantage than beclomethasone dipropionate and budesonide. This is attributed to its elevated water solubility. The aim of this article is to outline the factors that influence drug nebulization and the pharmacokinetics-pharmacodynamics of FLU compared to other inhaled corticosteroids. In addition, we report a series of clinical data regarding the efficacy of nebulized FLU with focus on the Italian experience. Overall, the physicochemical characteristics and pharmacokinetic profile of FLU favor its use for nebulization. Clinical data indicate that nebulized FLU is effective in asthma treatment in infants and children. Side effects were not reported at the commonly used doses.


The impact of visuospatial perception on distance judgment and depth perception in an Augmented Reality environment in patients after stroke: an exploratory study.
Augmented Reality (AR)-based interventions are applied in neurorehabilitation with increasing frequency. Depth perception is required for the intended interaction within AR environments. Until now, however, it is unclear whether patients after stroke with impaired visuospatial perception (VSP) are able to perceive depth in the AR environment. Different aspects of VSP (stereovision and spatial localization/visuoconstruction) were assessed in 20 patients after stroke (mean age: 64 ± 14 years) and 20 healthy subjects (HS, mean age: 28 ± 8 years) using clinical tests. The group of HS was recruited to assess the validity of the developed AR tasks in testing stereovision. To measure perception of holographic objects, three distance judgment tasks and one three-dimensionality task were designed. The effect of impaired stereovision on performance in each AR task was analyzed. AR task performance was modeled by aspects of VSP using separate regression analyses for HS and for patients. In HS, stereovision had a significant effect on the performance in all AR distance judgment tasks (p = 0.021, p = 0.002, p = 0.046) and in the three-dimensionality task (p = 0.003). Individual quality of stereovision significantly predicted the accuracy in each distance judgment task and was highly related to the ability to perceive holograms as three-dimensional (p = 0.001). In stroke-survivors, impaired stereovision had a specific deterioration effect on only one distance judgment task (p = 0.042), whereas the three-dimensionality task was unaffected (p = 0.317). Regression analyses confirmed a lacking impact of patients' quality of stereovision on AR task performance, while spatial localization/visuoconstruction significantly prognosticated the accuracy in distance estimation of geometric objects in two AR tasks. Impairments in VSP reduce the ability to estimate distance and to perceive three-dimensionality in an AR environment. While stereovision is key for task performance in HS, spatial localization/visuoconstruction is predominant in patients. Since impairments in VSP are present after stroke, these findings might be crucial when AR is applied for neurorehabilitative treatment. In order to maximize the therapy outcome, the design of AR games should be adapted to patients' impaired VSP. Trial registration: The trial was not registered, as it was an observational study.


De Novo Generation and Characterization of New Zika Virus Isolate Using Sequence Data from a Microcephaly Case.
Zika virus (ZIKV) has recently emerged and is the etiological agent of congenital Zika syndrome (CZS), a spectrum of congenital abnormalities arising from neural tissue infections in utero. Herein, we describe the de novo generation of a new ZIKV isolate, ZIKVNatal, using a modified circular polymerase extension reaction protocol and sequence data obtained from a ZIKV-infected fetus with microcephaly. ZIKVNatal thus has no laboratory passage history and is unequivocally associated with CZS. ZIKVNatal could be used to establish a fetal brain infection model in IFNAR-/- mice (including intrauterine growth restriction) without causing symptomatic infections in dams. ZIKVNatal was also able to be transmitted by Aedes aegypti mosquitoes. ZIKVNatal thus retains key aspects of circulating pathogenic ZIKVs and illustrates a novel methodology for obtaining an authentic functional viral isolate by using data from deep sequencing of infected tissues. IMPORTANCE The major complications of an ongoing Zika virus outbreak in the Americas and Asia are congenital defects caused by the virus's ability to cross the placenta and infect the fetal brain. The ability to generate molecular tools to analyze viral isolates from the current outbreak is essential for furthering our understanding of how these viruses cause congenital defects. The majority of existing viral isolates and infectious cDNA clones generated from them have undergone various numbers of passages in cell culture and/or suckling mice, which is likely to result in the accumulation of adaptive mutations that may affect viral properties. The approach described herein allows rapid generation of new, fully functional Zika virus isolates directly from deep sequencing data from virus-infected tissues without the need for prior virus passaging and for the generation and propagation of full-length cDNA clones. The approach should be applicable to other medically important flaviviruses and perhaps other positive-strand RNA viruses.


The transformation of nascent disc shaped high density lipoproteins into mature spherical high density lipoproteins: conformational analysis of apolipoprotein A-I in model intermediate particles.
The conformation of apolipoprotein A-I (apo A-I) was studied in reassembled lipoprotein particles containing various proportions of apo A-I, phospholipids and cholesteryl esters or triglycerides, using both theoretical and experimental approaches. The proportions of the components were chosen to permit the formation of swollen discs mimicking the intermediate particles that must result from the lecithin:cholesterol acyl transferase (LCAT)-mediated transformation of nascent discoidal high-density lipoprotein (HDL) into mature spherical HDL. Three groups of particles were defined by the number of helices per apo A-I molecule that could fit the edge of the swollen discs. Groups I, II and III had six, seven and eight helices per apo A-I, respectively. The conformation of apo A-I was studied by quenching of tryptophan fluorescence in the presence of KI. In all groups the tryptophan (Trp) residues belonging to the helices located on the edge of the particles were very sparingly hydrated. In contrast, the degree of hydration of Trp residues in external position was different in each group. The structural organization of phospholipids was assessed by measurements of the fluorescence anisotropy of 1-(4-trimethylammonium phenyl)-6-phenyl-1,3,5-hexatriene (TMA-DPH) probe. In group I, which contained the less swollen particles, a segment of apo A-I of about 46 amino acids contributed to the area of the faces of the particles, resulting in a highly condensed and ordered phospholipid matrix. In groups II and III, the external segment of apo A-I did not interact with phospholipids.


Mutations of acetylcholinesterase1 contribute to prothiofos-resistance in Plutella xylostella (L.).
Insensitive acetylcholinesterase (AChE) is involved in the resistance of organophosphorous and carbamate insecticides. We cloned a novel full-length AChE cDNA encoding ace1 gene from adult heads of the diamondback moth (DBM, Plutella xylostella). The ace1 gene encoding 679 amino acids has conserved motifs including catalytic triad, choline-binding site and acyl pocket. Northern blot analysis revealed that the ace1 gene was expressed much higher than the ace2 in all examined body parts. The biochemical properties of expressed AChEs showed substrate specificity for acetylthiocholine iodide and inhibitor specificity for BW284C51 and eserine. Three mutations of AChE1 (D229G, A298S, and G324A) were identified in the prothiofos-resistant strain, two of which (A298S and G324A) were expected to be involved in the prothiofos-resistance through three-dimensional modeling. In vitro functional expression of AChEs in Sf9 cells revealed that only resistant AChE1 is less inhibited with paraoxon, suggesting that resistant AChE1 is responsible for prothiofos-resistance.


Characterization of rabphilin phosphorylation using phospho-specific antibodies.
Rab3A is a GTP-binding protein of synaptic vesicles that regulates neurotransmitter release and cycles on and off synaptic vesicles as a function of exocytosis. Rab3A presumably functions via GTP-dependent interactions with effectors. Two putative rab3A effectors have been described in neurons, rabphilin which is a soluble protein that moves onto and off synaptic vesicles in concert with rab3A, and RIM which is an active zone protein that only binds to rab3A on docked vesicles. Rabphilin is an abundant, evolutionarily conserved protein whose function has remained enigmatic since a knockout of rabphilin does not display the functional deficiencies observed in the rab3A knockout. However, previous studies have shown that rabphilin is phosphorylated by protein kinase A and CaM Kinase II, suggesting that it may have a regulatory role. In the present study, we have examined the site and regulation of rabphilin phosphorylation in living nerve terminals using phospho-specific antibodies raised against phospho-serine234 of rabphilin. With these antibodies, we demonstrate that rabphilin is physiologically phosphorylated on serine234, and that soluble rabphilin which is not bound to rab3A on synaptic vesicles is the primary target. However, different from synapsins which are induced to dissociate from synaptic vesicles by PKA phosphorylation, phosphorylation of rabphilin is not instrumental for dissociating rabphilin from synaptic vesicles. Our data support the notion that dissociated rabphilin is a synaptic phosphoprotein in vivo that may play a role in the regulation of nerve terminal protein-protein interactions.


[Vesicular intracellular transport in the digestive organs. Membrane vesicle--the universal mechanism of the functional transport].
On the basis of long-term research of the morpho-functional characteristics of the cells of the stomach, small intestine and gallbladder the mechanism and function of membrane vesicles in the implementation of the main functions of these organs sets out in this article: the secretion of hydrochloric acid by parietal cells, the absorption of nutrients in the small intestine and the fluid at a concentration of bile epitheliocytes of gallbladder. Proofs of the intracellular formation of hydrochloric acid in tubulovesicles of the parietal cells and turnover of its secretory membranes in the process of secretory cycle, that has ensured the re-use and explained the extraordinary life of these unique cells are presented. The credible mechanism of HCl output oppression by H(+)-K(+)-ATPase activity blockers has set out on this basis. The article provides detailed endocytosis mechanism of the ions and nutrients absorption by enterocytes. The mechanism of participation of the apical contractile complex of brush border of epithelial cells in the initiation of endocytosis and cytoplasmic microtubules in transport of membrane vesicles in the cytoplasm was analyzed. Based on our research and numerous of the world scientific proceedings the conclusion was done about the existence of two energy dependent types of transport in the absorptive epithelium of the digestive--transmembrane (ionic and nutritive) homeostatic type which is realized by the ATP-system of the basal plasmalemma, and vesicular (endocytosis) type which is impltmented by apical contractile complex of brush border and cytoplasmic microtubules. Both types of transport are interrelated and are under constant cellular control. This observation is relevant to the majority of cells, including those involved in the secretion of various substances: hydrochloric acid by parietal cells, enzymes by main cells of the gastric glands and exocrinocytes of the pancreas, hormone by endocrine cells of the APUD system and, finally, mediators by nerve cells. To clarify the problem of cell transport two types: homeostatic and functional transport should be distinguished. The first type is carried out mainly in relation to ions on basolateral membrane with the help of its ATPase systems and provides intracellular homeostasis and nutritive co- transportation, the second (functional) type defines the basic specific function by means of membrane vesicles.


Impact of a 6-wk olive oil supplementation in healthy adults on urinary proteomic biomarkers of coronary artery disease, chronic kidney disease, and diabetes (types 1 and 2): a randomized, parallel, controlled, double-blind study.
Olive oil (OO) consumption is associated with cardiovascular disease prevention because of both its oleic acid and phenolic contents. The capacity of OO phenolics to protect against low-density lipoprotein (LDL) oxidation is the basis for a health claim by the European Food Safety Authority. Proteomic biomarkers enable an early, presymptomatic diagnosis of disease, which makes them important and effective, but understudied, tools for primary prevention. We evaluated the impact of supplementation with OO, either low or high in phenolics, on urinary proteomic biomarkers of coronary artery disease (CAD), chronic kidney disease (CKD), and diabetes. Self-reported healthy participants (n = 69) were randomly allocated (stratified block random assignment) according to age and body mass index to supplementation with a daily 20-mL dose of OO either low or high in phenolics (18 compared with 286 mg caffeic acid equivalents per kg, respectively) for 6 wk. Urinary proteomic biomarkers were measured at baseline and 3 and 6 wk alongside blood lipids, the antioxidant capacity, and glycation markers. The consumption of both OOs improved the proteomic CAD score at endpoint compared with baseline (mean improvement: -0.3 for low-phenolic OO and -0.2 for high-phenolic OO; P < 0.01) but not CKD or diabetes proteomic biomarkers. However, there was no difference between groups for changes in proteomic biomarkers or any secondary outcomes including plasma triacylglycerols, oxidized LDL, and LDL cholesterol. In comparison with low-phenolic OO, supplementation for 6 wk with high-phenolic OO does not lead to an improvement in cardiovascular health markers in a healthy cohort.


Oral rehydration solutions for burn management in the field and underdeveloped regions: a review.
Burn injuries are the fourth most common type of trauma worldwide, and the appropriate care of burn injuries in resource-limited settings such as the battlefield, underdeveloped nations, or in mass casualtiesremains a significant challenge. Rehydration constitutes the primary treatment of the systemic effects of burns and is a major factor in patient recovery. The standard of care for the replenishment of fluid and electrolyte losses in burn injury remains intravenous fluid therapy, but oral rehydration solution therapy (ORST) demonstrates beneficial utility in saving the lives of burn patients when they are applied in the acute phase of burn injuries, especially when intravenous rehydration is unavailable or inaccessible. Advantages of ORST as compared to intravenous therapy include availability, ease of administration in the field, low risk of infections and complications, low cost, and no requirement for accessory or specialized equipment. These benefits position ORST very attractively for the provision of interim first aid until definitive medical assistance arrives. Extensive and comprehensive investigation may be warranted to elucidate, account for and quantify individual burn patient biochemical variables toward the potential realization of such an "omniuse" oral rehydration solution for the benefit of burn injuries worldwide.


Usage of a percutaneous closure device to repair an iatrogenic subclavian artery injury.
Inadvertent subclavian artery cannulation during attempted subclavian central venous access is more likely to occur during rushed trauma resuscitations when anatomic landmarks are used for placement. Traditional supraclavicular and infraclavicular approaches for direct repair of the resultant arteriotomy are painful, morbid procedures that should be replaced with more minimally invasive techniques. This case report describes the usage of a percutaneous suture-mediated device (Perclose Proglide, Abbott Laboratories) to repair an iatrogenic subclavian artery arteriotomy. Two patients had their injuries successfully repaired using a percutaneous closure device. The use of a percutaneous closure device to repair iatrogenic subclavian artery injuries is a safe and effective method of repair that precludes a more invasive exposure and repair.


Kinin antagonists: design and activities.
Bradykinin antagonists effective in a great variety of kinin-responsive tissues have been developed and used to study the roles of kinins in various physiological and pathological phenomena. Structural changes required to yield antagonists and to confer high potency, tissue selectivity, and resistance to enzymatic degradation have been explored in several hundred analogs. Certain of these offer promise for the development of drugs effective against pain, inflammation, and asthma.


Analytical Limit Distributions from Random Power-Law Interactions.
Nature is full of power-law interactions, e.g., gravity, electrostatics, and hydrodynamics. When sources of such fields are randomly distributed in space, the superposed interaction, which is what we observe, is naively expected to follow a Gauss or Lévy distribution. Here, we present an analytic expression for the actual distributions that converge to novel limits that are in between these already-known limit distributions, depending on physical parameters, such as the concentration of field sources and the size of the probe used to measure the interactions. By comparing with numerical simulations, the origin of non-Gauss and non-Lévy distributions are theoretically articulated.


Anticarcinogenic Effects of Dried Citrus Peel in Colon Carcinogenesis Due to Inhibition of Oxidative Stress.
Colorectal cancer is one of the leading causes of death worldwide. Reactive oxygen species produce oxidative stress and contribute to colorectal carcinogenesis. Because dietary citrus has been shown to reduce oxidative stress, we investigated the effects of citrus peel extract at dilutions of 1/200-1/500 on the activity of oxidative-stress-related transcription factors, including AP-1, NF-κB, NRF2, p53, and STAT3, in human colon cancer cell line HCT116 cells using a luciferase reporter gene assay. NRF2 transcriptional activities were 1.8- to 2.0-fold higher than the untreated control value. In addition, NF-κB, p53, and STAT3 transcriptional activities were 12-26% lower than the untreated control value. Administration of dried citrus peel in the diet of F344 rats at a dose of 1,000 ppm prevented the formation of azoxymethane-induced precancerous aberrant crypt foci (ACF) in the colon. The total number of ACF in rats fed with dried citrus peel was reduced to 75% of the control value. Moreover, the levels of oxidative-stress-related markers, reactive carbonyl species, in the serum of F344 rats were significantly reduced following the administration of dried citrus peel. These data suggest that citrus peel possesses an ability to suppress cellular oxidative stress through induction of NRF2, thereby preventing azoxymethane-induced colon carcinogenesis.


MosqTent: An individual portable protective double-chamber mosquito trap for anthropophilic mosquitoes.
Here, we describe the development of the MosqTent, an innovative double-chamber mosquito trap in which a human being attracts mosquitoes while is protected from being bitten within the inner chamber of the trap, while mosquitoes are lured to enter an outer chamber where they are trapped. The MosqTent previously collected an average of 3,000 anophelines/man-hour compared to 240 anophelines/man-hour for the human landing catch (HLC), thereby providing high numbers of human host-seeking mosquitoes while protecting the collector from mosquito bites. The MosqTent performed well by collecting a high number of specimens of Anopheles marajoara, a local vector and anthropophilic mosquito species present in high density, but not so well in collecting An. darlingi, an anthropophilic mosquito species considered the main vector in Brazil but is present in low-density conditions in the area. The HLC showed a higher efficiency in collecting An. darlingi in these low-density conditions. The MosqTent is light (<1 kg), portable (comes as a bag with two handles), flexible (can be used with other attractants), adaptable (can be deployed in a variety of environmental settings and weather conditions), and it can be used in the intra-, peri-, and in the extradomicile. Also, the MosqTent collected similar portions of parous females and anthropophilic mosquito species and collects specimens suitable for downstream analysis. Further developments may include testing for other fabric colors, different mesh sizes and dimensions for other hematophagous insects and conditions, additional chemical mosquito attractants, and even the replacement of the human attractant in favor of other attractants. MosqTent modifications that would allow the trap to be applied as a vector control tool with killing action could also be explored.


A Cell-Free Biosensor for Assessment of Hyperhomocysteinemia.
Hyperhomocysteinemia─a condition characterized by elevated levels of homocysteine in the blood─is associated with multiple health conditions including folate deficiency and birth defects, but there are no convenient, low-cost methods to measure homocysteine in plasma. A cell-free biosensor that harnesses the native homocysteine sensing machinery of Escherichia coli bacteria could satisfy the need for a detection platform with these characteristics. Here, we describe our efforts to engineer a cell-free biosensor for point-of-care, low-cost assessment of homocysteine status. This biosensor can detect physiologically relevant concentrations of homocysteine in plasma with a colorimetric output visible to the naked eye in under 1.5 h, making it a fast, convenient tool for point-of-use diagnosis and monitoring of hyperhomocysteinemia and related health conditions.


Fluorescence microlymphography.
Microneedles, 0.2 mm o.d., were connected to a microsyringe and mounted on a micromanipulator. Under microscopic control, 0.01 ml of a 25% solution of FITC-labeled dextran-40 or dextran-150 were injected into the subepidermis at the big toe near the nailfold or in the medial ankle region. Fluorescence intravital microscopy revealed a network of lymphatic microvessels. The comparison with recent anatomic studies reveals that the reticular network visualized by FITC-dextran corresponds to the network in the stratum papillare. In 20 healthy subjects lymphatic capillaries were detected in a restricted area on the lateral aspect of the big toe. In 10 patients with primary lymphedema, the dye expanded to almost the entire dorsal skin surface of the big toe. In two cases, enlarged and tortuous microvessels of pathologic shape were observed. Fluorescence microlymphography is a simple and nearly atraumatic approach for depicting the intravital anatomy of human skin lymphatic capillaries.


Auditory cortex interneuron development requires cadherins operating hair-cell mechanoelectrical transduction.
Many genetic forms of congenital deafness affect the sound reception antenna of cochlear sensory cells, the hair bundle. The resulting sensory deprivation jeopardizes auditory cortex (AC) maturation. Early prosthetic intervention should revive this process. Nevertheless, this view assumes that no intrinsic AC deficits coexist with the cochlear ones, a possibility as yet unexplored. We show here that many GABAergic interneurons, from their generation in the medial ganglionic eminence up to their settlement in the AC, express two cadherin-related (cdhr) proteins, cdhr23 and cdhr15, that form the hair bundle tip links gating the mechanoelectrical transduction channels. Mutant mice lacking either protein showed a major decrease in the number of parvalbumin interneurons specifically in the AC, and displayed audiogenic reflex seizures. Cdhr15- and Cdhr23-expressing interneuron precursors in Cdhr23-/- and Cdhr15-/- mouse embryos, respectively, failed to enter the embryonic cortex and were scattered throughout the subpallium, consistent with the cell polarity abnormalities we observed in vitro. In the absence of adhesion G protein-coupled receptor V1 (adgrv1), another hair bundle link protein, the entry of Cdhr23- and Cdhr15-expressing interneuron precursors into the embryonic cortex was also impaired. Our results demonstrate that a population of newborn interneurons is endowed with specific cdhr proteins necessary for these cells to reach the developing AC. We suggest that an "early adhesion code" targets populations of interneuron precursors to restricted neocortical regions belonging to the same functional area. These findings open up new perspectives for auditory rehabilitation and cortical therapies in patients.


EFFECT OF AIR VELOCITY ON INHALATION DOSES DUE TO RADON AND THORON PROGENY IN A TEST CHAMBER.
Inhalation doses due to radon and thoron are predominantly due to the inhalation of progeny of Radon and Thoron. The progeny/decay-products of radon and thoron are particulates unlike their parent gas and exhibit different physical properties like attachment to the aerosols and deposition on different surfaces. All these properties in turn depend on the environmental conditions such as air velocity, aerosol concentration, attachment rate, etc. The role of air velocity on deposition on surfaces decides the progeny particles left in the air for inhalation. Therefore, in the present work, we have studied the effect of air velocity on the inhalation dose due to radon and thoron progeny at the centre of a 0.5-m3 calibration chamber as well as on all surfaces. Hence, the studies were carried out at different air velocities, and inhalation doses were measured using deposition-based direct radon and thoron progeny sensors.


Mycotic enteritis in a chameleon and a brief review of phycomycoses of animals.
Rectal prolapse in an adult chameleon was surgically replaced. The animal was given tetracycline and dextrose orally, but became comatose 7 days postoperatively and was euthanatized. Necropsy revealed intussusception of the terminal portion of the colon. Phycomycotic hyphae accompanied by necrosis and a mixed leukocytic infiltrate were found in the area of intussusception.


The effect of crop rotation on pesticide leaching in a regional pesticide risk assessment.
New modeling approaches that include the use of GIS are under development in order to allow a more realistic assessment of environmental contamination by pesticides. This paper reports a regional GIS-based risk assessment using a software tool able to simulate complex and real crop rotations at the regional scale. A single pesticide leaching assessment has been done. The mean annual pesticide concentration in leachate has been analyzed using both stochastic and deterministic approaches. The outputs of these simulations were mapped over the sampling locations of the regional pesticide monitoring program, demonstrating that GIS-based risk assessment can be used to establish new monitoring programs. A multiple pesticide leaching assessment for analyzing the risk related to pest control strategies in six different maize-based rotations has been carried out. Additive toxic units approach has been used. Crop rotation allows to mediate the risk related to pesticide use because forces the use of different compounds with different fate and toxicology properties.


Multifocal nerve lesions and LZTR1 germline mutations in segmental schwannomatosis.
Schwannomatosis is a genetic disorder characterized by the occurrence of multiple peripheral schwannomas. Segmental schwannomatosis is diagnosed when schwannomas are restricted to 1 extremity and is thought to be caused by genetic mosaicism. We studied 5 patients with segmental schwannomatosis through microstructural magnetic resonance neurography and mutation analysis of NF2, SMARCB1, and LZTR1. In 4 of 5 patients, subtle fascicular nerve lesions were detected in clinically unaffected extremities. Two patients exhibited LZTR1 germline mutations. This appears contrary to a simple concept of genetic mosaicism and suggests more complex and heterogeneous mechanisms underlying the phenotype of segmental schwannomatosis than previously thought. Ann Neurol 2016;80:625-628.


Effect of water treatment residuals on soil phosphorus, copper and aluminium availability and toxicity.
Water treatment residuals (WTRs) are produced by the treatment of potable water with coagulating agents. Beneficial recycling in agriculture is hampered by the fact that WTRs contain potentially toxic contaminants (e.g. copper and aluminium) and they bind phosphorus strongly. These issues were investigated using a plant bioassay (Lactuca sativa), chemical extractions and an isotopic dilution technique. Two WTRs were applied to an acidic and a neutral pH soil at six rates. Reductions in plant growth in amended soils were due to WTR-induced P deficiency, rather than Al or Cu toxicity. The release of potentially toxic Al from WTRs was found to be mitigated by their alkaline nature and pH buffering capacity. However, acidification of WTRs was shown to release more soluble Al than soil naturally high in Al. Copper availability was relatively low in all treatments. However, the lability of WTR-Cu increased when the WTR was applied to the soil.


Locomotor activation induced by infusion of endorphins into the ventral tegmental area: evidence for opiate-dopamine interactions.
beta-Endorphin in nanomole quantities produced a stimulation of locomotor activity when infused into the region of the dopamine cell bodies of the ventral tegmental area (VTA) in rats. alpha-, gamma-, and des-Tyr-gamma-endorphin produced similar effects, but the D-alanine analogues of alpha and gamma-endorphin produced a larger and longer-lasting activation, presumably reflecting their resistance to degradation. This locomotor activation was reversible by pretreatment with naloxone and by destruction of the terminal projections of the mesocorticolimbic dopamine system originating in the VTA. These results demonstrate that locally infused endorphin can interact with the opioid receptors in the VTA, and they suggest a means by which endorphins activate limbic excitability.


1.36 million years of Mediterranean forest refugium dynamics in response to glacial-interglacial cycle strength.
The sediment record from Lake Ohrid (Southwestern Balkans) represents the longest continuous lake archive in Europe, extending back to 1.36 Ma. We reconstruct the vegetation history based on pollen analysis of the DEEP core to reveal changes in vegetation cover and forest diversity during glacial-interglacial (G-IG) cycles and early basin development. The earliest lake phase saw a significantly different composition rich in relict tree taxa and few herbs. Subsequent establishment of a permanent steppic herb association around 1.2 Ma implies a threshold response to changes in moisture availability and temperature and gradual adjustment of the basin morphology. A change in the character of G-IG cycles during the Early-Middle Pleistocene Transition is reflected in the record by reorganization of the vegetation from obliquity- to eccentricity-paced cycles. Based on a quantitative analysis of tree taxa richness, the first large-scale decline in tree diversity occurred around 0.94 Ma. Subsequent variations in tree richness were largely driven by the amplitude and duration of G-IG cycles. Significant tree richness declines occurred in periods with abundant dry herb associations, pointing to aridity affecting tree population survival. Assessment of long-term legacy effects between global climate and regional vegetation change reveals a significant influence of cool interglacial conditions on subsequent glacial vegetation composition and diversity. This effect is contrary to observations at high latitudes, where glacial intensity is known to control subsequent interglacial vegetation, and the evidence demonstrates that the Lake Ohrid catchment functioned as a refugium for both thermophilous and temperate tree species.


Is ulcer recurrence after simple closure of perforated duodenal ulcer predictable?
To study whether factors such as age, duration of pre-perforation symptoms, size of perforation and operative evidence of chronicity could predict recurrence of ulcer after simple closure of perforated duodenal ulcers, thus enabling patients at high risk of recurrence to be subjected to definitive surgery instead of simple closure of perforation. Retrospective. One hundred and fifty-two patients who had undergone simple closure of duodenal ulcer perforation in the last 10 years were included in the study. A postgraduate research and teaching institution. Patients were subjected to a personal interview and Visick grading of symptoms 1-10 years after simple closure of duodenal ulcer perforation. Ninety of them underwent esophagogastroduodenoscopic evaluation. Symptomatic ulcer recurrence rate was 23.1% by Visick grading and 42.2% on endoscopy. Ulcer symptom recurrence rate had no correlation with patient's age, duration of pre-perforation symptoms, or size of perforation, but correlated significantly with operative evidence of chronicity (p < 0.001). However, 7 of 35 symptomatic recurrences would have been missed by the use of this criterion alone. Endoscopic evidence of recurrence also correlated only with operative evidence of chronicity. False positives and false negatives with Visick grading showed that this symptomatic evaluation was unsatisfactory even for selecting patients for further endoscopic workup. Though not infallible, evidence of chronicity of ulcer at laparotomy may be a useful predictor of recurrence of ulcer after simple closure of perforated duodenal ulcer.


Serum cathepsin B to cystatin C ratio as a potential marker for the diagnosis of cholangiocarcinoma.
Cholangiocarcinoma (CCA) is a cancer of the bile duct epithelial cells. The highest incidence rate of CCA with a poor prognosis and poor response to chemotherapy is found in Southeast Asian countries, especially in northeastern Thailand and Lao PDR. Cathepsin B is a lysosomal cysteine protease which is regulated by cysteine proteinase inhibitors such as cystatin C. Elevation of cathepsin B levels in biological fluid has been observed in patients with inflammatory diseases and many cancers. We aimed to investigate the serum cathepsin B and cystatin C levels of CCA patients to evaluate the feasibility of using cathepsin B and cystatin C as markers for the diagnosis of CCA. Fifty-six sera from CCA patients, 17 with benign biliary diseases (BBD) and 13 from controls were collected and the cathepsin B and cystatin C levels were determined. In addition, cathepsin B expression was investigated immunohistochemically for 9 matched-pairs of cancerous and adjacent tissues of CCA patients. Serum cathepsin B, but not cystatin C, was significantly higher in CCA and BBD patient groups compared to that in the control group. Consistently, all cancerous tissues strongly expressed cathepsin B while adjacent tissues were negative in 7 out of 9 cases. In contrast, serum cystatin C levels were comparable between CCA and control groups, although serum cystatin C levels in the BBD group was higher than that in the control or CCA groups. When the serum cathepsin B to cystatin C ratio was calculated, that of the CCA group was significantly higher than that of the control group, and, although statistically not significant, the ratio of CCA group showed a trend to be higher than that of the BBD group. Thus, the cathepsin B to cystatin C ratio might be used as an alternative marker for aiding diagnosis of CCA.


Liposomes or traditional adjuvants: induction of bactericidal activity by the macrophage infectivity potentiator protein (Mip) of Neisseria meningitidis.
Currently, one of the main approaches to achieve a vaccine for serogroup B Neisseria meningitidis is based on outer membrane proteins with low antigenic variability among strains. Since these proteins tend to be minor components of the outer membrane, recombinant production is required to obtain them in sufficient amounts for evaluation and development of vaccines. In this study, we analysed the ability of recombinant macrophage infectivity potentiator (rMip) protein to induce protective bactericidal activity in mice. The rMip protein was cloned from N. meningitidis strain H44/76 and was used to immunise mice, and the sera obtained were tested against the homologous and several heterologous N. meningitidis strains. The sera were obtained using the rMip alone, with adjuvant Al(OH)3 , or after inclusion into liposomes. Bactericidal activity was variable depending on the strain, although high titres were seen against strains H44/76 and NmP27. Liposomes enhanced fourfold the reactivity against the homologous strain. The results presented suggest that the rMip protein should be considered a promising candidate for the improvement of future protein-based vaccines.


Efficacy of quantitative analysis of Epstein-Barr virus-infected peripheral blood lymphocytes by in situ hybridization of EBER1 after living-related liver transplantation: a case report.
We describe a 1-year-old female who underwent living-related liver transplantation for biliary atresia and developed Epstein-Barr virus (EBV)-related posttransplant lymphoproliferative disorder. This disorder was resolved after withdrawal of immunosuppression therapy and administration of a high dose of acyclovir. To quantify the extent of EBV activation and EBV load in peripheral blood, we measured the levels of EBV-infected peripheral lymphocytes by in situ hybridization (ISH) of EBV-encoded small mRNA 1 (EBER1). The decline in the number of EBER1-positive lymphocytes (from 362/50,000 mononuclear cells to 0/50,000) after treatment was in accord with the patient's clinical improvement. This finding showed that quantitative analysis of EBV-infected peripheral lymphocytes by ISH of EBER1 is very useful for monitoring the EBV load and response to treatment of patients with EBV-related disorders. Furthermore, ISH may become an important tool for the early diagnosis and prevention of life-threatening posttransplant lymphoproliferative disorder in posttransplant patients.


Objective Measures of Physical Distancing in the Hospital During the COVID-19 Pandemic.
During the COVID-19 pandemic, hospitals published physical-distancing guidance and created dedicated respiratory isolation units (RIUs) for patients with COVID-19. The degree to which such distancing occurred between clinicians and patients is unknown. In this study, heat sensors from an existing hospital hand-hygiene monitoring system objectively tracked room entries as a proxy for physical distancing in both RIUs and general medicine units before and during the pandemic. The RIUs saw a 60.6% reduction in entries per room per day (from 85.7 to 33.8). General medicine units that cared for patients under investigation for COVID-19 and other patients experienced a 14.7% reduction in entries per room per day (from 76.9 to 65.1). While gradual extinction was observed in both units as COVID-19 cases declined, the RIUs had a higher degree of physical distancing. Although the optimal level of physical distancing is unknown, sustaining physical distancing in the hospital may require re-education and real-time monitoring.


[Percutaneous nephrolithotripsy in patients after different methods of urinary diversion].
The study presents the results of percutaneous nephrolithotripsy (PNL) in patients who underwent different methods of urinary diversion. The study included the results of PNL in 16 patients (study group) with kidney stones who were subjected to different methods of urinary diversion. Earlier, 2 (12.5%) patients underwent bilateral ureterosigmoidostomy for bladder exstrophy, 12 (75%) experienced radical cystectomy with orthotopic ileocystoplasty and 2 (12.5%) - heterotopic cystoplasty. The control group comprised 30 patients with large kidney stones and normal function of the lower urinary tract. In the main group, the mean stone size was 2.5 (2.2-3.6) cm in the control group - 2.6 (2.4-3.8) cm. The puncture of the kidney cavity system and nephrostomy dilatation were performed under ultrasound and fluoroscopic guidance, respectively. We compared such factors as the surgery duration, the number of intra- and postoperative complications, blood loss and length of hospital stay. There were no hemorrhagic complications requiring blood transfusion. Exacerbation of chronic pyelonephritis occurred in 4 (25%) patients of the study group and in 3 (10%) of the control group (p<0.05). The two groups did not differ in the effectiveness of PNL (87.5 and 90.0% in the study and control groups, respectively), the blood loss volume and the length of hospital stay. PNL enables high effectiveness in removing large kidney stones with minimal complications in patients undergoing different methods of urinary diversion.


Understanding prostate-specific antigen dynamics in monitoring metastatic castration-resistant prostate cancer: implications for clinical practice.
Availability of novel hormonal therapies as well as docetaxel and cabazitaxel treatment for metastatic castration-resistant prostate cancer (CRPC) has changed the outlook for this group of patients with improvements in progression-free survival and overall survival. Physicians often diagnose the progression of prostate cancer using serum prostate-specific antigen (PSA). However, serum PSA is not always correlated with the clinical status in CRPC. To evaluate the PSA dynamics with greater precision, understanding of the control of PSA and of the mechanisms of development of CRPC is needed. Moreover, it is necessary to use new hormonal therapies with an appropriate timing to optimally improve the prognosis and the QOL of the patients. In the present review, we ascertain the PSA dynamics and the mechanisms of the development of CRPC to assist in optimal utilization of the new treatments for mCRPC.


Alcohol and drug problems in the schools: results of a national survey of school administrators.
A mail survey of public high-school administrators in the United States was conducted to gather information about the nature and extent of school problems with student alcohol and drug use. Of the 728 schools selected for the national metropolitan probability sample, 543 (75%) returned a completed questionnaire. The results indicated that from 1980 to 1985, about one in six students attended schools that reported a serious problem with student alcohol use. In contrast, the proportion of students attending schools with a serious drug problem decreased from about one in four in 1980-81 to about one in seven in 1984-85. The most common explanation provided for a decreasing student alcohol or drug problem was changes in the school's discipline policy or increased enforcement of the existing policy. Few respondents attributed a reduction in student alcohol or drug problems to prevention or treatment programs. Limitations of the survey data are discussed as well as directions for future research.


Peritoneal Dialysis Fluid and Some of Its Components Potentiate Fibrocyte Differentiation.
Long-term peritoneal dialysis (PD) often results in the development of peritoneal fibrosis. In many other fibrosing diseases, monocytes enter the fibrotic lesion and differentiate into fibroblast-like cells called fibrocytes. We find that peritoneal tissue from short-term PD patients contains few fibrocytes, while fibrocytes are readily observed in the peritoneal membrane of long-term PD patients. The PD fluid Dianeal (Baxter Healthcare Corporation, Deerfield, IL, USA) contains dextrose, a number of electrolytes including sodium chloride, and sodium lactate. We find that PD fluid potentiates human fibrocyte differentiation in vitro and implicates sodium lactate in this potentiation. The plasma protein serum amyloid P (SAP) inhibits fibrocyte differentiation. Peritoneal dialysis fluid and sodium chloride decrease the ability of human SAP to inhibit human fibrocyte differentiation in vitro Together, these results suggest that PD fluid contributes to the development of peritoneal fibrosis by potentiating fibrocyte differentiation.


Impact Of Socio-Economic Status On Determinants Of Medical Career.
Research on determinants of Medical profession has established some constant factors with universal impact and appeal. However, the dramatic changes in the dynamics of medical profession and medical education over the past years have necessitated a revisiting of the established conclusions. The knowledge of impact of economic status on these determinants would pave way to greater interest and research on the subject, especially in Hazara Division where it is practically nonexistent. This exploratory research followed deductive-inductive approach and was conducted upon 550 respondents from four public and private colleges in Hazara Division. Extensive study of related literature yielded 20 broad determinants of medical career. Pre-research pilot testing verified the reliability of a self-designed instrument. The data collection method adopted was a single shot survey. Personal data of respondents was collected during administration of the instrument. The sample was extracted through systematic random sampling technique giving due weight to proportional representation of each college as well as to the gender distribution. The results revealed a significant difference in the impact of reassuring social perceptions and social status and prestige of medical profession between upper middle class and lower middle class. The results also showed that the students of upper middle class faced significantly greater domestic and familial pressure to enter the medical profession than those from lower middle class. The findings showed that the upper middle class is under greater influence of societal forces and wider impressions about the medical profession. The students hailing from upper middle class enter the Medical career motivated by urge for social status and prestige and are most compelled by domestic and familial pressures to enter medicine.


Functional analysis of spontaneous movements in preterm infants.
Spontaneous movements of premature infants between 25 and 34 weeks conceptional age were observed for 1 hr on two or three occasions. Subjects had low-risk prognoses and were clinically stable at the time of testing. Behavioral acts were scored using a 0/1 time sampling technique in 60 continuous, 1-min time blocks. Temporal associations between individual movements were found using chi-square analyses. Some associated behaviors contained combinations consistent with neonatal action patterns, for example, single and bilateral leg kicking, head turning, and mouthing. Features of state organization were also evident in that general motor activity (GM), which has been used as a marker of active sleep (AS) in neonates, was found to cluster temporally with startle, facial, and head movements but not eye movements. Behavioral quiescence (> or = 5 s) was dissociated from AS-related behaviors (GM, facial, head, and eye movements). Combinations of state-segregated behaviors were more likely to exhibit co-occurrence within 1-min intervals in infants 30 weeks conceptional age and older.


A 51-year-old man with intramedullary spinal cord abscess having a patent foramen ovale.
The authors report a case of a 51-year-old man with intramedullary spinal cord abscess (ISCA) having a patent foramen ovale (PFO). He developed fever and tetraplegia after a recent dental treatment. MRI showed ISCA with longitudinal swelling from the upper cervical to the lumbar spinal cord. Cerebrospinal fluid (CSF) analysis indicated bacterial meningitis, and the culture of CSF revealed Streptococcus viridans. Transoesophageal echocardiography revealed the existence of a PFO. We suspected another possibility other than systemic bacteraemia, that paradoxical bacteric embolisation through PFO after the dental treatment caused ISCA. While several reports of brain abscess with PFO are available, this is the first report of ISCA with PFO.


RNF216 regulates meiosis and PKA stability in the testes.
Spermatogenesis is a highly sophisticated process that comprises of mitosis, meiosis, and spermiogenesis. RNF216 (ring finger protein 216), an E3 ubiquitin ligase, has been reported to be essential for spermatogenesis and male fertility in mice. However, the stages affected by Rnf216 deficiency and its underlying molecular pathological mechanisms are still unknown. In this study, we generated Rnf216-deficient mice (Rnf216-/- ) using CRISPR-Cas9 technology. Knockout of Rnf216 led to infertility in male but not female mice. Rnf216 knockout affected the prophase of meiosis I, as no genotypic difference was observed until 12 dpp (days postpartum). Rnf216-/- spermatocytes were incompletely arrested at the zygotene stage and underwent apoptosis at approximately the pachytene stage. The proportion of zygotene spermatocytes was significantly increased, whereas the proportion of pachytene spermatocytes was significantly decreased in Rnf216-/- testes. Nevertheless, there was no significantly genotypic difference in the number of diplotene spermatocytes. We further revealed that the PKA catalytic subunit β (PRKACB) was significantly increased, which subsequently resulted in elevated PKA activity in testes from adult as well as 9 dpp Rnf216-/- mice. RNF216 interacts with PRKACB and promotes its degradation through the ubiquitin-lysosome pathway. Collectively, our results revealed an important role for RNF216 in regulation of meiosis and PKA stability in the testes.


Relevant antibody subsets against MOG recognize conformational epitopes exclusively exposed in solid-phase ELISA.
A pathogenic role for circulating anti-myelin antibodies is difficult to establish in multiple sclerosis (MS). Here, we unravel a broad heterogeneity within the anti-myelin oligodendrocyte glycoprotein (MOG) antibodies in humans and non-human primates, and demonstrate that detection of important epitopes of MOG within the pathogenic repertoire is exclusively dependent on presentation on a solid-phase MOG conformer. Results of ELISA and those of a liquid-phase assay were compared using a MOG protein with identical sequence but different conformations. We tested sera from 50 human subjects, plasma of Callithrix jacchus marmosets known to contain antibodies reactive to either conformational or linearized MOG, and monoclonal, conformation-dependent anti-MOG antibodies. We have found no antibody reactivity against the soluble MOG conformer in human serum, and show that this lack of detection is not due to technical artifacts. Rather, dominant epitopes of MOG are not displayed in soluble phase, as shown by a complete lack of binding of conformation-dependent mAb. In MP4-immune marmosets that exhibit demyelinating pathology due to spreading of antibody determinants to myelin-embedded MOG, only ELISA can detect pathogenic circulating anti-MOG antibodies. Thus, the accurate detection of important subsets of pathogenic anti-MOG antibodies requires methods in which MOG is displayed similarly to its natural conformation in myelin.


[A New Teaching Model for Practicing Ultrasound-guided Regional Anesthesia Techniques].
We developed a simulator using "slime" composed of polyvinyl alcohol (PVA) and borax to evaluate this new ultrasound-guided nerve block training model. Seventeen subjects used the training model in the present study. They had no previous experience in performing ultrasound-guided nerve block. A plastic case measuring 25 x 18 x 12 cm was filled with 8 cm of slime. Three pieces of gauze were placed between the slime layers at 2 cm intervals. An in-plane approach was used to visualize the needle for the nerve block, and the amount of time required to stop the needle on the second gauze was measured 5 times for each subject. Significant differences were observed between the times for the first experiment and those for the third experiment to the fifth experiment In the fourth and fifth experiments, all subjects visualized the nerve block needle clearly above the target layer and were able to stop the needle at the target layer. The present simulation using our proposed ultrasound-guided nerve block training model was useful in terms of the amount of time required to perform the procedure and as well as in terms of its safety.


In vitro tumor cytotoxic activities of extracts from three Liriodendron plants.
The extracts prepared from Liriodendron tulipifera Linn., L. chinense (Hemsl.) Sarg., and their hybrid L. chinense x L. tulipifera, were investigated for their cytotoxic abilities in vitro against five human cancer cell lines: MDA-MB-231 and MCF-7 breast cancer cells, SGC-7901 gastric cancer cells, HuH-7 hepatocarcinoma cells, and HCT-15 colon carcinoma cells, and then measured their phenols and alkaloids contents. Of these plant extracts, some of them, especially the lower polar extracts from barks, exhibited potent cytotoxic effects on five tested tumor cell lines.


Injectable Hydrogels as Unique Platforms for Local Chemotherapeutics-Based Combination Antitumor Therapy.
Different strategies of chemotherapeutics-based combination cancer therapy have presented enhanced antitumor efficiency and are widely used in clinical cancer treatments. However, several drawbacks of the systems for systemic administration, including low drug accumulation at tumor sites and significant systemic side effects limit their efficacy and application in the clinic. Local drug co-delivery systems based on injectable hydrogels may have considerable advantages, such as a facile drug-delivery procedure, targeted delivery of antitumor agents to tumor sites in a sustained manner, and markedly reduced systemic toxicities. Thus, in recent years, these systems have received increasing attention and consequently various injectable hydrogels have been tested as platforms for local chemotherapeutics-based combination antitumor therapy. In this review, the focus is on recent advances in injectable hydrogel-based drug co-delivery systems for local combination antitumor therapy, including multiple chemotherapeutics combination therapy, chemo-immunotherapy, chemo-radiotherapy, and hyperthermia-chemotherapy. Moreover, the rationale and preparation of local co-delivery systems are summarized and discussed.


Chlorophyll-Based Organic-Inorganic Heterojunction Solar Cells.
Solid-state chlorophyll solar cells (CSCs) employing a carboxylated chlorophyll derivative, methyl trans-32 -carboxypyropheophorbide a, as a light-harvesting dye sensitizer chlorophyll (DSC) deposited on mesoporous TiO2 , on which four zinc hydroxylated chlorophyll derivatives were spin-coated for hole transporter chlorophylls (HTCs), are described. Key parameters, including the effective carrier mobility of the HTC films, as determined by the space charge-limited current method, and the frontier molecular orbitals of these DSCs and HTCs, as estimated from cyclic voltammetry and electronic absorption spectra, suggest that both charge separation and carrier transport are favorable. The power conversion efficiencies (PCEs) of the present CSCs with fluorine-doped tin oxide (FTO)/TiO2 /DSC/HTCs/Ag were determined to follow the order of HTC-1>HTC-2>HTC-3>HTC-4, which coincided perfectly with the order of their hole mobilities. The maximum PCE achieved was 0.86 % with HTC-1. The photovoltaic devices studied herein with two types of chlorophyll derivatives as dye sensitizers and hole transporters provide a unique solution for the utilization of solar energy with a view to truly realizing "green energy".


Josephson Diode Effect Induced by Valley Polarization in Twisted Bilayer Graphene.
Recently, the Josephson diode effect (JDE), in which the superconducting critical current magnitudes differ when the currents flow in opposite directions, has attracted great interest. In particular, it was demonstrated that gate-defined Josephson junctions based on magic-angle twisted bilayer graphene showed a strong nonreciprocal effect when the weak-link region is gated to a correlated insulating state at half filling (two holes per moiré cell). However, the mechanism behind such a phenomenon is not yet understood. In this Letter, we show that the interaction-driven valley polarization, together with the trigonal warping of the Fermi surface, induce the JDE. The valley polarization, which lifts the degeneracy of the states in the two valleys, induces a relative phase difference between the first and the second harmonics of the supercurrent and results in the JDE. We further show that the nontrivial current phase relation, which is responsible for the JDE, also generates the asymmetric Shapiro steps.


Lucy Maude Montgomery and Anne of Green Gables: An Early Description of Attention-Deficit/Hyperactivity Disorder.
Attention-deficit/hyperactivity disorder (ADHD) was added to the Diagnostic and Statistical Manual of Mental Disorders, third edition, revised in 1987. Similar disorders had appeared earlier, and many consider the first description of ADHD to be a lecture in 1902 about children with an "abnormal defect in moral control" but normal intelligence. This definition of ADHD is more alarming than the current one. Anne Shirley, the protagonist of the novel Anne of Green Gables (written by Lucy Maude Montgomery and published in 1908), shares the hyperactive and inattentive qualities that fit the current definition of ADHD. She also lacks the menacing characteristics of the 1902 description. This indicates that ADHD, by its modern definition, was probably present in the early 1900s. Furthermore, the character of Anne Shirley shares many biographical similarities with her author, suggesting that Montgomery herself may have had ADHD. Thus, looking at literature from the past not only provides insight into the timeline of ADHD, but also into the thought process of an individual with ADHD. By viewing literary classics through a medical lens, we may gain insight into other diseases as well. [Pediatr Ann. 2017; 46(7):e270-e272.].


In vivo Ca2+ imaging of astrocytic microdomains reveals a critical role of the amyloid precursor protein for mitochondria.
The investigation of amyloid precursor protein (APP) has been mainly confined to its neuronal functions, whereas very little is known about its physiological role in astrocytes. Astrocytes exhibit a particular morphology with slender extensions protruding from somata and primary branches. Along these fine extensions, spontaneous calcium transients occur in spatially restricted microdomains. Within these microdomains mitochondria are responsible for local energy supply and Ca2+ buffering. Using two-photon in vivo Ca2+ imaging, we report a significant decrease in the density of active microdomains, frequency of spontaneous Ca2+ transients and slower Ca2+ kinetics in mice lacking APP. Mechanistically, these changes could be potentially linked to mitochondrial malfunction as our in vivo and in vitro data revealed severe, APP-dependent structural mitochondrial fragmentation in astrocytes. Functionally, such mitochondria exhibited prolonged kinetics and morphology dependent signal size of ATP-induced Ca2+ transients. Our results highlight a prominent role of APP in the modulation of Ca2+ activity in astrocytic microdomains whose precise functioning is crucial for the reinforcement and modulation of synaptic function. This study provides novel insights in APP physiological functions which are important for the understanding of the effects of drugs validated in Alzheimer's disease treatment that affect the function of APP.


A Cyclopeptide and a Tetrahydroisoquinoline Alkaloid from Ophiorrhiza nutans.
A new cyclopeptide, ophiorrhisine A (1), a new tetrahydroisoquinoline alkaloid, 7',10-dide-O-methylcephaeline (2), two known β-carboline alkaloids, and four known tetrahydroisoquinoline alkaloids were isolated from Ophiorrhiza nutans (Rubiaceae). Compound 1 is a tetrapeptide possessing a 14-membered paracyclophane ring and a novel N,N,N-trimethyltyrosine residue in the side chain. The stereochemistry at the aryl-alkyl ether bond was different from that of other known 14-membered paracyclophanes. The structure of 2 was established by spectroscopic analysis and semisynthesis.


Effects of two freezing methods and two cryopreservation media on post-thaw quality of stallion spermatozoa.
Glycerol-based extenders are widely utilized for freezing equine semen, but media combining methylformamide may better preserve sperm motility and mitochondrial function. Semen is cryopreserved utilizing either a Styrofoam box filled with liquid nitrogen or an automatic freezer. The objective of this experiment was to compare the post-thaw characteristics of the same ejaculates cryopreserved in a Styrofoam box or in an automatic freezer, utilizing a glycerol-based extender (Gent) and an extender that combines methylformamide and glycerol (BotuCrio® ). For that, one ejaculate from 30 stallions collected in two different centres was used. For data analysis, a mixed linear model with laboratory, medium and freezing method and respective interactions as fixed effects was used. Stallion was taken into account as a random effect. There was no influence (p > .05) of laboratory, while stallion effect was marked. Semen frozen in BotuCrio® in the automatic freezer had higher (p < .001) VCL than semen cryopreserved in Gent using the Styrofoam box. VCL was also higher (p = .068) for semen frozen in BotuCrio® in the Styrofoam box than for semen cryopreserved in Gent using the same method. The difference between percentage of sperm with intact plasma membrane frozen in Gent using the Styrofoam box (44.43% ± 2.44%) compared to spermatozoa cryopreserved in BotuCrio® using the same method (40.78% ± 2.42%) approached significance (p = .0507). The percentage of sperm with intact acrosome membrane was higher (p < .05) in semen frozen in BotuCrio® (79.08% ± 1.79%) than semen frozen in Gent (75.15% ± 1.80%). A higher (p = .0125) percentage (32.24% ± 2.18%) of semen extended in Gent and cryopreserved in the Styrofoam box had high mitochondrial membrane potential than semen frozen in BotuCrio® using the same method (26.02% ± 2.15%). Fertility studies are warranted to assess whether differences found have any effect on the fertility of inseminated mares.


Owning a Dog as a Determinant of Physical Activity and Metabolic Control in Patients With Type 1 and Type 2 Diabetes Mellitus.
Physical activity is recommended for patients with type 1 (T1D) and type 2 diabetes (T2D). We wanted to assess whether owning a dog influences duration or intensity of physical activity and metabolic control of diabetes mellitus. 143 patients with T1D (age 50±16 y.; BMI 25.7±4.5 kg/m2, HbA1c 8.6±1.6%) and 303 with T2D (age 63±11 y., BMI 33.7±7.3 kg/m2, HbA1c 9.0±1.6%, 232 [76.6%] insulin-treated, 89±61 IU/d), respectively, participated. A standardized questionnaire assessed diabetes history and treatment, details regarding time spent (per week) and intensity (MET, metabolic equivalent of task) of physical activity ("walking the dog" and other activities), anthropometric (BMI) and laboratory measures. 31.5% of T1D 23.1% of T2D patients were dog owners. Dog owners with T1D and T2D diabetes spent 19.0±3.3 and 19.8±2.6 MET.h per week walking the dog, which represented 61.3±5.7 and 62.9±4.9% of their total physical activity. Participants not owning a dog compensated by performing significantly more other activities. Taken together, total physical activity was similar in dog owners with T1D (p=0.80), but higher in dog owners with T2D (30.1±2.8 vs. 18.6±1.4 MET.h per week in those not owning a dog; p=0.0001). Body-mass-index or HbA1c were not significantly different in either patients with T1D or T2D either owning a dog or not. Owning a dog motivates to a significant amount of physical activity, but this was fully compensated for by other forms of physical activity in (younger) patients with T1D. Even the higher physical activity in dog owners with T2D did not result in improved glycaemic or body weight control.


Minimally invasive: minimally reimbursed? An examination of six laparoscopic surgical procedures.
It is generally believed that minimally invasive surgery (MIS) results in less postoperative pain, fewer complications, and shorter recovery periods compared with open procedures. Yet despite these benefits, the level of reimbursement assigned to the surgeon by United States health-care payers is often lower than that for open procedures. Furthermore, the cost of performing a MIS may be higher vs an open procedure because specialized equipment, increased surgical time, or both may be required. In this report, we examine the issue by comparing reimbursements for MIS with open procedures, summarizing the medical literature on MIS vs open surgical procedures, and offering recommendations for payers who establish reimbursement policies. The review is focused on six MIS procedures where outcomes data exist: laparoscopic cholecystectomy (lap chole), laparoscopic colectomy (LC), laparoscopic fundoplication (LF), laparoscopic hysterectomy (LH), laparoscopic ventral hernia repair (LVHR), and laparoscopic appendectomy (LA). Outcomes summarized were length of hospital stay (LOS), operating room time, operating room costs, complications, and return to work or normal activities. The level of scientific evidence was assigned to each study using predetermined criteria. A total of 112 articles were reviewed: 14 for lap chole, 26 for LC, 7 for LF, 19 for LH, 9 for LVHR, and 37 for LA. The data demonstrate that these procedures result in reduced hospital stay, reduced hospital costs, and faster return to work or normal activities. Yet, the operating room time and costs are frequently higher for MIS. These findings suggest that as both the outcomes value and level of operating room resources are greater, MIS warrants reimbursement that meets or exceeds that of open procedures.


Decreased salivary lactoferrin levels are specific to Alzheimer's disease.
Evidences of infectious pathogens in Alzheimer's disease (AD) brains may suggest a deteriorated innate immune system in AD pathophysiology. We previously demonstrated reduced salivary lactoferrin (Lf) levels, one of the major antimicrobial proteins, in AD patients. To assess the clinical utility of salivary Lf for AD diagnosis, we examine the relationship between salivary Lf and cerebral amyloid-β (Aβ) load using amyloid-Positron-Emission Tomography (PET) neuroimaging, in two different cross-sectional cohorts including patients with different neurodegenerative disorders. The diagnostic performance of salivary Lf in the cohort 1 had an area under the curve [AUC] of 0•95 (0•911-0•992) for the differentiation of the prodromal AD/AD group positive for amyloid-PET (PET+) versus healthy group, and 0•97 (0•924-1) versus the frontotemporal dementia (FTD) group. In the cohort 2, salivary Lf had also an excellent diagnostic performance in the health control group versus prodromal AD comparison: AUC 0•93 (0•876-0•989). Salivary Lf detected prodromal AD and AD dementia distinguishing them from FTD with over 87% sensitivity and 91% specificity. Salivary Lf seems to have a very good diagnostic performance to detect AD. Our findings support the possible utility of salivary Lf as a new non-invasive and cost-effective AD biomarker. Instituto de Salud Carlos III (FIS15/00780, FIS18/00118), FEDER, Comunidad de Madrid (S2017/BMD-3700; NEUROMETAB-CM), and CIBERNED (PI2016/01) to E.C.; Spanish Ministry of Economy and Competitiveness (SAF2017-85310-R) to J.L.C., and (PSI2017-85311-P) to M.A.; International Centre on ageing CENIE-POCTEP (0348_CIE_6_E) to M.A.; Instituto de Salud Carlos III (PIE16/00021, PI17/01799), to H.B.


α,α-disubstituted β-amino amides eliminate Staphylococcus aureus biofilms by membrane disruption and biomass removal.
Bacterial biofilms account for up to 80% of all infections and complicate successful therapies due to their intrinsic tolerance to antibiotics. Biofilms also cause serious problems in the industrial sectors, for instance due to the deterioration of metals or microbial contamination of products. Efforts are put in finding novel strategies in both avoiding and fighting biofilms. Biofilm control is achieved by killing and/or removing biofilm or preventing transition to the biofilm lifestyle. Previous research reported on the anti-biofilm potency of α,α-disubstituted β-amino amides A1, A2 and A3, which are small antimicrobial peptidomimetics with a molecular weight below 500 Da. In the current study it was investigated if these derivatives cause a fast disintegration of biofilm bacteria and removal of Staphylococcus aureus biofilms. One hour incubation of biofilms with all three derivatives resulted in reduced metabolic activity and membrane permeabilization in S. aureus (ATCC 25923) biofilms. Bactericidal properties of these derivatives were attributed to a direct effect on membranes of biofilm bacteria. The green fluorescence protein expressing Staphylococcus aureus strain AH2547 was cultivated in a CDC biofilm reactor and utilized for disinfectant efficacy testing of A3, following the single tube method (American Society for Testing and Materials designation number E2871). A3 at a concentration of 90 μM acted as fast as 100 μM chlorhexidine and was equally effective. Confocal laser scanning microscopy studies showed that chlorhexidine treatment lead to fluorescence fading indicating membrane permeabilization but did not cause biomass removal. In contrast, A3 treatment caused a simultaneous biofilm fluorescence loss and biomass removal. These dual anti-biofilm properties make α,α-disubstituted β-amino amides promising scaffolds in finding new control strategies against recalcitrant biofilms.


From leukocyte recruitment to resolution of inflammation: the cardinal role of integrins.
Integrins constitute a large group of adhesion receptors that are formed as heterodimers of α and β subunits. Their presence and activation status on the surface of leukocytes modulate a broad spectrum of processes in inflammation and immunity. This mini review critically outlines research advances with regard to the function of leukocyte integrins in regulating and integrating the onset and resolution of acute inflammation. Specifically, we summarize and discuss relevant, current literature that supports the multifunctional role of integrins and their partners. The latter include molecules that physically associate with integrins or regulate their activity in the context of the following: 1) leukocyte recruitment to an inflamed tissue, 2) recognition and phagocytosis of apoptotic neutrophils (efferocytosis), and 3) egress of efferocytic macrophages from the inflamed site to lymphoid tissues. The understanding of the fine-tuning mechanisms of the aforementioned processes by integrins and their functional partners may enable the design of therapeutic tools to counteract destructive inflammation and promote more efficient resolution of inflammation.


The phylogenetically invariant ACAGAGA and AGC sequences of U6 small nuclear RNA are more tolerant of mutation in human cells than in Saccharomyces cerevisiae.
U6 small nuclear RNA (snRNA) is the most highly conserved of the five spliceosomal snRNAs that participate in nuclear mRNA splicing. The proposal that U6 snRNA plays a key catalytic role in splicing [D. Brow and C. Guthrie, Nature (London) 337:14-15, 1989] is supported by the phylogenetic conservation of U6, the sensitivity of U6 to mutation, cross-linking of U6 to the vicinity of the 5' splice site, and genetic evidence for extensive base pairing between U2 and U6 snRNAs. We chose to mutate the phylogenetically invariant 41-ACAGAGA-47 and 53-AGC-55 sequences of human U6 because certain point mutations within the homologous regions of Saccharomyces cerevisiae U6 selectively block the first or second step of mRNA splicing. We found that both sequences are more tolerant to mutation in human cells (assayed by transient expression in vivo) than in S. cerevisiae (assayed by effects on growth or in vitro splicing). These differences may reflect different rate-limiting steps in the particular assays used or differential reliance on redundant RNA-RNA or RNA-protein interactions. The ability of mutations in U6 nucleotides A-45 and A-53 to selectively block step 2 of splicing in S. cerevisiae had previously been construed as evidence that these residues might participate directly in the second chemical step of splicing; an indirect, structural role seems more likely because the equivalent mutations have no obvious phenotype in the human transient expression assay.


Statistics and medical experimentation.
Starting from a review of four major experimental areas in medicine (Polio, Coronary Surgery, Diabetes, Breast Cancer), some of the special statistical problems arising in the design and analysis of clinical experiments are reviewed, and the limitations of current formulations are emphasized. Particular attention is given to the ethical dilemma and an alternative formulation of the inference/decision problem is proposed.


Electron collisions with trifluorides: BF3 and PF3 molecules.
Absolute total cross sections (TCSs) for electron scattering from boron trifluoride (BF(3)) and phosphorus trifluoride (PF(3)) molecules have been measured using a linear transmission method. The electron energy ranges from 0.6 to 370 eV for BF(3) and from 0.5 to 370 eV for PF(3). The TCS energy dependence for BF(3) exhibits two very pronounced enhancements: resonantlike narrow feature located near 3.6 eV with the maximum value of 19.2 x 10(-20) m(2), and intermediate energy very broad enhancement with two humps, one centered around 21 eV (18.8 x 10(-20) m(2) in the maximum) and the other near 45 eV (19.5 x 10(-20) m(2)). For PF(3) the TCS has quite different low-energy dependence: at 0.5 eV it has a high value of 70 x 10(-20) m(2) and decreases steeply towards higher energies. Beyond the minimum near 5.5 eV, the TCS reveals two distinct humps: the resonant one centered near 11 eV with the peak value of 32.9 x 10(-20) m(2) and the second one much broader around 35 eV (27.9 x 10(-20) m(2)). The present TCSs for trifluorides are compared to each other as well as to previous TCS data for selected perfluorides and to results for their perhydrided counterparts. The differences and similarities in the shape and magnitude of TCSs are pointed out.


Echocardiographic monitoring for clozapine cardiac toxicity: lessons from real-world experience.
We aimed to identify the baseline prevalence of cardiac dysfunction in patients commencing clozapine, assess adherence with echocardiographic monitoring recommendations, and evaluate the utility and cost of echocardiographic monitoring for the development of clozapine-associated myocarditis and cardiomyopathy. A retrospective longitudinal cohort study was undertaken of 159 consecutive patients from a major tertiary centre commencing clozapine in the period January 2002 to July 2009. Some 73% of patients had a baseline study, and 11% had a six-month follow-up study. Nine patients had abnormal left ventricular function at baseline. Myocarditis was identified in three patients, with all cases occurring within the first month of treatment and suspected on clinical grounds before an echocardiogram was performed. One case of possible cardiomyopathy was identified. The cost of echocardiographic screening in the first year of treatment was estimated at $AUD 209,356 per case of cardiomyopathy detected. The prevalence of cardiac dysfunction in patients commencing clozapine is high, and there are challenges in adhering with the recommended protocol for monitoring. Routine echocardiography is not useful in the detection of clozapine-associated myocarditis. Although cardiomyopathy may be identified, it is rare and associated with significant cost. Recommendations for routine echocardiographic monitoring should be re-examined.


Peek through the smoke: a report of moyamoya disease in a 32-year-old female patient presenting with ischaemic stroke.
Moyamoya disease is a vasculopathy causing chronic progressive stenosis and occlusion of the large arteries of the circle of Willis that could lead to brain ischaemia. The condition may also present with haemorrhagic strokes. This is a case report of moyamoya disease in a 32-year-old woman presenting with ischaemic stroke. The report describes her inpatient stay and investigations and findings.The report reviews the main aspects of moyamoya disease definition, epidemiology, clinical features, diagnosis, classification and treatment. This case is interesting because her first presentation occurred after 3 months of her second delivery. Whether the different physiological stresses of pregnancy, child birth and puerperium have had some effect in accelerating the pathogenesis of her moyamoya disease remains unknown. 1.


Barium sulfate aspiration: Is early bronchioalveolar lavage a life-saving procedure?
Aspiration of barium sulfate is a well-known complication, occurring accidentally during contrast-enhanced examinations of the upper gastrointestinal system. Massive aspiration of contrast material causes mechanical obstruction that increases alveolar dead space leading to altered ventilation/perfusion (V/Q) ratio with secondary respiratory failure and death. The potential treatment strategies include early recognition of predisposing factors, pretreatment with antireflux medications, such as domperidone, correct choice of contrast media, and use of retroesophageal suction catheter during barium swallow study. We report a case of barium swallow aspiration, which was successfully managed by early institution of bronchioalveolar lavage (BAL) and positive pressure mechanical ventilation. Till date, BAL has not been recommended for the same, with studies showing it can worsen the clinical scenario.


Rotational Inerfia of Continents: A Proposed Link between Polar Wandering and Plate Tectonics.
A mechanism is proposed whereby displacement between continents and the earth's pole of rotation (polar wandering) gives rise to latitudinal transport of continental plates (continental drift) because of their relatively greater rotational inertia. When extended to short-term polar wobble, the hypothesis predicts an energy change nearly equivalent to the seismic energy rate.


Computational Study of the Isomerization Reactions of Borirane.
Borirane is isoelectronic to the cyclopropyl cation, but is stable toward electrocyclic ring opening to 2-bora-propa-1,3-diyl A, the boron analogue of the allyl cation. A computational investigation using density functional theory (B3LYP) in combination with highly correlated electronic structure theory methods of the coupled-cluster [CCSD(T)] and multireference configuration interaction (MRCI) type in conjunction with basis sets of up to quadruple-ζ quality reveal that formation of A is endothermic by roughly 15 kcal mol-1 and that A collapses almost without barrier (0.2 kcal mol-1) to borirane. The vinylborane isomer B is more stable than borirane and its formation is associated with a barrier of 36-38 kcal mol-1. Methyl methylideneborane E, only slightly less stable than B, can only be accessed by pathways involving barriers of at least 60 kcal mol-1.


T-cells "à la CAR-T(e)" - Genetically engineering T-cell response against cancer.
The last decade will be remembered as the dawn of the immunotherapy era during which we have witnessed the approval by regulatory agencies of genetically engineered CAR T-cells and of checkpoint inhibitors for cancer treatment. Understandably, T-lymphocytes represent the essential player in these approaches. These cells can mediate impressive tumor regression in terminally-ill cancer patients. Moreover, they are amenable to genetic engineering to improve their function and specificity. In the present review, we will give an overview of the most recent developments in the field of T-cell genetic engineering including TCR-gene transfer and CAR T-cells strategies. We will also elaborate on the development of other types of genetic modifications to enhance their anti-tumor immune response such as the use of co-stimulatory chimeric receptors (CCRs) and unconventional CARs built on non-antibody molecules. Finally, we will discuss recent advances in genome editing and synthetic biology applied to T-cell engineering and comment on the next challenges ahead.


Co-application of selenite and phosphate reduces arsenite uptake in hydroponically grown rice seedlings: toxicity and defence mechanism.
The study empirically evaluates the abatement of As(III) uptake in rice seedlings (7d), in presence of Se and phosphate (P) under hydroponic condition. Positive correlation between As(III) translocation to the shoots of As(III) and P treated seedlings, shows P dependent As(III) translocation in rice. Whereas, presence of both P (5 and 10μgml(-1)) and (0.75μgml(-1)) of Se significantly reduces the As(III) uptake in rice seedlings. Application of Se alone also reduces As(III) uptake both in shoots and roots significantly, however, the seedlings suffers from lipid peroxidation. Among all the studied treatments, lower rates of P (5μgml(-1)) and Se (0.75μgml(-1)) when co-applied, significantly reduced As(III) translocation to the shoots without inflicting much toxicity in the seedlings which is manifested as significant increase in biomass with lower thio-barbituric reactive substances (TBARS). Also, significantly lower TBARS in seedlings receiving As(4)+Se(0.75) and higher TBARS in As(4)+Se(1.5), demonstrates that Se applied at lower rates (0.75μgml(-1)), lowers As induced toxicity. Higher SOD, APX and guaiacol peroxidase (POD) activities in As(4)+P(5)+Se(0.75) compared to that of As(4)+P(5) and As(4)+Se(0.75), supports that lower rate of P and Se provides tolerance towards As induced stress. The nitrogen metabolism in As(4)+P+Se treated seedlings is affected adversely at higher rates of Se and P application. Overall study concluded that application of lower rates of P (5μgml(-1)) and Se (0.75μgml(-1)) provides maximum amelioration of As(III) toxicity in rice seedlings.


An Augmented Reality-Based Approach for Surgical Telementoring in Austere Environments.
Telementoring can improve treatment of combat trauma injuries by connecting remote experienced surgeons with local less-experienced surgeons in an austere environment. Current surgical telementoring systems force the local surgeon to regularly shift focus away from the operating field to receive expert guidance, which can lead to surgery delays or even errors. The System for Telementoring with Augmented Reality (STAR) integrates expert-created annotations directly into the local surgeon's field of view. The local surgeon views the operating field by looking at a tablet display suspended between the patient and the surgeon that captures video of the surgical field. The remote surgeon remotely adds graphical annotations to the video. The annotations are sent back and displayed to the local surgeon while being automatically anchored to the operating field elements they describe. A technical evaluation demonstrates that STAR robustly anchors annotations despite tablet repositioning and occlusions. In a user study, participants used either STAR or a conventional telementoring system to precisely mark locations on a surgical simulator under a remote surgeon's guidance. Participants who used STAR completed the task with fewer focus shifts and with greater accuracy. The STAR reduces the local surgeon's need to shift attention during surgery, allowing him or her to continuously work while looking "through" the tablet screen.


Single Live Cell Imaging of Multidrug Resistance Using Silver Ultrasmall Nanoparticles as Biosensing Probes in Triple-Negative Breast Cancer Cells.
Silver ultrasmall nanoparticles (Ag UNPs) (size < 5 nm) were used as biosensing probes to analyze the efflux kinetics contributing to multidrug resistance (MDR) in single live triple-negative breast cancer (TNBC) cells by using dark-field optical microscopy to follow their size-dependent localized surface plasmon resonance. TNBC cells lack expression of estrogen (ER-), progesterone (PR-), and human epidermal growth factor 2 (HER2-) receptors and are more likely to acquire resistance to anticancer drugs due to their ability to transport harmful substances outside the cell. The TNBC cells displayed greater nuclear and cytoplasmic efflux, resulting in less toxicity of Ag UNPs in a concentration-independent manner. In contrast, more Ag UNPs and an increase in cytotoxic effects were observed in the receptor-positive breast cancer cells that have receptors for ER+, PR+, and HER2+ and are known to better respond to anticancer therapies. Ag UNPs accumulated in receptor-positive breast cancer cells in a time-and concentration-dependent mode and caused decreased cellular growth, whereas the TNBC cells due to the efflux were able to continue to grow. The TNBC cells demonstrated a marked increase in survival due to their ability to have MDR determined by efflux of Ag UNPs outside the nucleus and the cytoplasm of the cells. Further evaluation of the nuclear efflux kinetics of TNBC cells with Ag UNPs as biosensing probes is critical to gain a better understanding of MDR and potential for enhancement of cancer drug delivery.


Decision making and physician prescribing characteristics: a pilot study of Japanese physicians.
The factors that affect physicians' prescribing remain unclear. Although previous reports suggest that prescription decisions are associated with various clinical situation, most of these studies analyzed simulated patient models rather than actual clinical practice. Here, we retrospectively analyzed actual cases of statin prescription for hyperlipidemia at Tohoku University Hospital between Apr 1, 2004 and Mar 31, 2008. Twelve physicians (6 cardiologists, 3 nephrologist, and 3 diabetologist) made decisions on whether to prescribe statins to 187 patients in 788 visits. As expected, cardiologists started prescribing statins at significantly lower serum total cholesterol levels than other specialists (221.7mg/dL vs. 244.7mg/dL, P<0.05). Interestingly, the total cholesterol levels that triggered prescribing differed significantly among cardiologists (p<0.05). These results suggested that prescription decisions differed not only among specialties but also among individuals.


A study of platelet fibronectin immunofluorescence in peripheral vascular disease and diabetes mellitus.
Platelets hyperaggregability and hypersecretion fibronectin (Fn) are known to occur in peripheral vascular disease (PVD) and diabetes mellitus (DM) with microangiopathy. To determine whether an increase in platelet membrane bound Fn constitutes to hyperaggregability of platelets, washed platelets from normal subjects and from patients with peripheral vascular disease and patients with diabetes mellitus were examined for the presence of fibronectin (Fn) by means of fluorescein linked antibody to Fn. Platelets from peripheral vascular disease and diabetes mellitus patients tended to aggregate during preparation and apparently exhibited greater fluorescence in platelet "smears" than was observed in smears from controls. In contrast, when washed platelet "smears" were prepared from platelet preparations containing iloprost, an analogue of prostacyclin, platelet aggregates did not form and the 'excess' of fluorescence disappeared from all the three groups. When platelets were stained for Fn fluorescence in suspensions, no fluorescence was observed on the surface of platelets from peripheral vascular disease and diabetes mellitus patients or controls. On stimulation with thrombin washed platelet suspension showed fluorescence for Fn. Platelet activation leads to Fn appearing on platelet surface but this effect cannot be quantified by optical fluorescence microscopy.


A quality improvement project to reduce falls and improve medication management.
This paper describes the implementation of a medication management model within a medical-center based home health agency. The model was integrated into the agency's quality improvement falls prevention program and was selected in part because it directly addressed two medication-related accreditation standards for home health care agencies. During a five-month period, a staff pharmacist conducted medication reviews for 228 HHA patients who met the program's inclusion criteria. Thirty-three percent of these patients required some type of follow-up to resolve potential medication-related problems. By far, falls were the most common reason for referral, with 71 patients, or 30% of all participating patients, referred to the pharmacist due to a recent fall. From a quality improvement standpoint, the program met and even exceeded expectations in that it enabled staff to identify a serious threat to patient safety-medication-related problems, especially falls--and gave them the tools to resolve these potential problems.


Stationary phase of cultured mammalian cells (L5178Y).
The stationary phase of the mammalian cells L5178Y in culture can be divided into two stages: (a) an early phase characterized by the decline of mitotic index, followed by a stabilization of the cell number, and (b) a late stage, occurring several hours after the flattening of the growth curve, during which dead or dying cells appear in the cultures. The estimates of rates of cell progress showed that the rates from G(1) to S and from G(2) to M were affected in the early stationary phase. The main cause of cessation of increase in cell number in the early stationary phase is resulted from the decline in mitotic index, which is caused by prolongation of the G(2) period. The importance of the G(2) stage in regulating the cell growth is discussed in relation to other known situations in the literature.